-DOCSTART-	O

Number	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
lymphocytes	NN	O	B-cell_type
and	NN	O	O
their	NN	O	O
sensitivity	NN	O	O
to	NN	O	O
hormone	NN	O	O
action	NN	O	O
.	NN	O	O

The	NN	O	O
study	NN	O	O
demonstrated	NN	O	O
a	NN	O	O
decreased	NN	O	O
level	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
(	NN	O	O
GR	NN	O	B-protein
)	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
from	NN	O	O
hypercholesterolemic	NN	O	O
subjects	NN	O	O
,	NN	O	O
and	NN	O	O
an	NN	O	O
elevated	NN	O	O
level	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
acute	NN	O	O
myocardial	NN	O	O
infarction	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
lymphocytes	NN	O	B-cell_type
with	NN	O	O
a	NN	O	O
high	NN	O	O
GR	NN	O	B-protein
number	NN	O	O
,	NN	O	O
dexamethasone	NN	O	O
inhibited	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
-thymidine	NN	O	O
and	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
-acetate	NN	O	O
incorporation	NN	O	O
into	NN	O	O
DNA	NN	O	O
and	NN	O	O
cholesterol	NN	O	O
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
in	NN	O	O
the	NN	O	O
same	NN	O	O
manner	NN	O	O
as	NN	O	O
in	NN	O	O
the	NN	O	O
control	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

On	NN	O	O
the	NN	O	O
other	NN	O	O
hand	NN	O	O
,	NN	O	O
a	NN	O	O
decreased	NN	O	O
GR	NN	O	B-protein
number	NN	O	O
resulted	NN	O	O
in	NN	O	O
a	NN	O	O
less	NN	O	O
efficient	NN	O	O
dexamethasone	NN	O	O
inhibition	NN	O	O
of	NN	O	O
the	NN	O	O
incorporation	NN	O	O
of	NN	O	O
labeled	NN	O	O
compounds	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
showed	NN	O	O
that	NN	O	O
the	NN	O	O
sensitivity	NN	O	O
of	NN	O	O
lymphocytes	NN	O	B-cell_type
to	NN	O	O
glucocorticoids	NN	O	O
changed	NN	O	O
only	NN	O	O
with	NN	O	O
a	NN	O	O
decrease	NN	O	O
of	NN	O	O
GR	NN	O	B-protein
level	NN	O	O
.	NN	O	O

-DOCSTART-	O

[	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
lymphocytes	NN	O	B-cell_type
and	NN	O	O
T-	NN	O	B-cell_type
and	NN	O	I-cell_type
B-lymphocyte	NN	O	I-cell_type
count	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
glomerulonephritis	NN	O	O
]	NN	O	O

Content	NN	O	O
of	NN	O	O
receptors	NN	O	O
to	NN	O	O
hormonal	NN	O	O
form	NN	O	O
of	NN	O	O
vitamin	NN	O	O
D3	NN	O	O
,	NN	O	O
1.25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
,	NN	O	O
constituted	NN	O	O
27.3	NN	O	O
fmole/mg	NN	O	O
of	NN	O	O
protein	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
of	NN	O	O
peripheric	NN	O	O
blood	NN	O	O
of	NN	O	O
children	NN	O	O
with	NN	O	O
glomerulonephritis	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
patients	NN	O	O
concentration	NN	O	O
of	NN	O	O
total	NN	O	O
and	NN	O	O
ionized	NN	O	O
form	NN	O	O
of	NN	O	O
Ca2+	NN	O	O
was	NN	O	O
decreased	NN	O	O
down	NN	O	O
to	NN	O	O
2.04	NN	O	O
mmole/L	NN	O	O
and	NN	O	O
1.09	NN	O	O
mmole/L	NN	O	O
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
while	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
parathormone	NN	O	O
(	NN	O	O
PTH	NN	O	O
)	NN	O	O
by	NN	O	O
36	NN	O	O
%	NN	O	O
and	NN	O	O
a	NN	O	O
distinct	NN	O	O
decrease	NN	O	O
in	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
D	NN	O	O
concentration	NN	O	O
(	NN	O	O
lower	NN	O	O
than	NN	O	O
1.25	NN	O	O
ng/ml	NN	O	O
)	NN	O	O
was	NN	O	O
found	NN	O	O
in	NN	O	O
blood	NN	O	O
;	NN	O	O
content	NN	O	O
of	NN	O	O
cAMP	NN	O	O
was	NN	O	O
also	NN	O	O
decreased	NN	O	O
in	NN	O	O
lymphocytes	NN	O	O
by	NN	O	O
33	NN	O	O
%	NN	O	O
.	NN	O	O

At	NN	O	O
the	NN	O	O
same	NN	O	O
time	NN	O	O
,	NN	O	O
total	NN	O	O
content	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
was	NN	O	O
decreased	NN	O	O
1.5-fold	NN	O	O
in	NN	O	O
peripheric	NN	O	O
blood	NN	O	O
.	NN	O	O

Treatment	NN	O	O
with	NN	O	O
I-hydroxyvitamin	NN	O	O
D3	NN	O	O
(	NN	O	O
1-1.5	NN	O	O
mg	NN	O	O
daily	NN	O	O
,	NN	O	O
within	NN	O	O
4	NN	O	O
weeks	NN	O	O
)	NN	O	O
led	NN	O	O
to	NN	O	O
normalization	NN	O	O
of	NN	O	O
total	NN	O	O
and	NN	O	O
ionized	NN	O	O
form	NN	O	O
of	NN	O	O
Ca2+	NN	O	O
and	NN	O	O
of	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
D	NN	O	O
,	NN	O	O
but	NN	O	O
did	NN	O	O
not	NN	O	O
affect	NN	O	O
the	NN	O	O
PTH	NN	O	O
content	NN	O	O
in	NN	O	O
blood	NN	O	O
.	NN	O	O

Concentration	NN	O	O
of	NN	O	O
the	NN	O	O
receptors	NN	O	B-protein
to	NN	O	O
1.25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
was	NN	O	O
elevated	NN	O	O
up	NN	O	O
to	NN	O	O
39.7	NN	O	O
fmole/mg	NN	O	O
after	NN	O	O
I	NN	O	O
week	NN	O	O
of	NN	O	O
the	NN	O	O
treatment	NN	O	O
,	NN	O	O
whereas	NN	O	O
it	NN	O	O
was	NN	O	O
decreased	NN	O	O
to	NN	O	O
the	NN	O	O
initial	NN	O	O
level	NN	O	O
24.8	NN	O	O
fmole/mg	NN	O	O
within	NN	O	O
4	NN	O	O
weeks	NN	O	O
;	NN	O	O
simultaneous	NN	O	O
alteration	NN	O	O
in	NN	O	O
the	NN	O	O
cAMP	NN	O	O
content	NN	O	O
was	NN	O	O
observed	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
.	NN	O	O

Treatment	NN	O	O
with	NN	O	O
1-	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
D3	NN	O	O
normalized	NN	O	O
also	NN	O	O
the	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
content	NN	O	O
in	NN	O	O
peripheric	NN	O	O
blood	NN	O	O
.	NN	O	O

The	NN	O	O
data	NN	O	O
obtained	NN	O	O
suggest	NN	O	O
that	NN	O	O
under	NN	O	O
conditions	NN	O	O
of	NN	O	O
glomerulonephritis	NN	O	O
only	NN	O	O
high	NN	O	O
content	NN	O	O
of	NN	O	O
receptors	NN	O	B-protein
to	NN	O	O
1.25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
enabled	NN	O	O
to	NN	O	O
perform	NN	O	O
the	NN	O	O
cell	NN	O	O
response	NN	O	O
to	NN	O	O
the	NN	O	O
hormone	NN	O	O
effect	NN	O	O
.	NN	O	O

-DOCSTART-	O

Tumor	NN	O	O
and	NN	O	O
serum	NN	O	O
beta-2-microglobulin	NN	O	O
expression	NN	O	O
in	NN	O	O
women	NN	O	O
with	NN	O	O
breast	NN	O	O
cancer	NN	O	O
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
whether	NN	O	O
the	NN	O	O
tumor	NN	O	B-cell_type
expression	NN	O	O
of	NN	O	O
beta-2-microglobulin	NN	O	B-protein
(	NN	O	O
beta	NN	O	B-protein
2-M	NN	O	I-protein
)	NN	O	O
could	NN	O	O
serve	NN	O	O
as	NN	O	O
a	NN	O	O
marker	NN	O	O
of	NN	O	O
tumor	NN	O	O
biologic	NN	O	O
behavior	NN	O	O
,	NN	O	O
the	NN	O	O
authors	NN	O	O
studied	NN	O	O
specimens	NN	O	O
of	NN	O	O
breast	NN	O	B-cell_type
carcinomas	NN	O	I-cell_type
from	NN	O	O
60	NN	O	O
consecutive	NN	O	O
female	NN	O	O
patients	NN	O	O
.	NN	O	O

Presence	NN	O	O
of	NN	O	O
beta	NN	O	B-protein
2-M	NN	O	I-protein
was	NN	O	O
analyzed	NN	O	O
by	NN	O	O
immunohistochemistry	NN	O	O
.	NN	O	O

No	NN	O	O
significant	NN	O	O
correlations	NN	O	O
were	NN	O	O
found	NN	O	O
between	NN	O	O
tumor	NN	O	B-protein
beta	NN	O	I-protein
2-M	NN	O	I-protein
expression	NN	O	O
and	NN	O	O
several	NN	O	O
histologic	NN	O	O
attributes	NN	O	O
such	NN	O	O
as	NN	O	O
type	NN	O	O
,	NN	O	O
histologic	NN	O	O
and	NN	O	O
nuclear	NN	O	O
grades	NN	O	O
,	NN	O	O
mitotic	NN	O	O
index	NN	O	O
,	NN	O	O
necrosis	NN	O	O
,	NN	O	O
vascular	NN	O	O
invasion	NN	O	O
,	NN	O	O
and	NN	O	O
lymphocytic	NN	O	O
infiltration	NN	O	O
.	NN	O	O

Likewise	NN	O	O
,	NN	O	O
beta	NN	O	B-protein
2-M	NN	O	I-protein
was	NN	O	O
not	NN	O	O
associated	NN	O	O
with	NN	O	O
markers	NN	O	O
of	NN	O	O
disease	NN	O	O
extension	NN	O	O
such	NN	O	O
as	NN	O	O
TNM	NN	O	O
,	NN	O	O
(	NN	O	O
UICC	NN	O	O
,	NN	O	O
classification	NN	O	O
of	NN	O	O
malignant	NN	O	B-cell_type
tumors	NN	O	I-cell_type
)	NN	O	O
staging	NN	O	O
and	NN	O	O
axillary	NN	O	O
lymph	NN	O	O
node	NN	O	O
involvement	NN	O	O
or	NN	O	O
with	NN	O	O
estrogen	NN	O	O
,	NN	O	O
progesterone	NN	O	O
,	NN	O	O
and	NN	O	O
glucocorticoid	NN	O	O
receptor	NN	O	O
levels	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
there	NN	O	O
was	NN	O	O
a	NN	O	O
significantly	NN	O	O
positive	NN	O	O
association	NN	O	O
between	NN	O	O
tumor	NN	O	B-protein
beta	NN	O	I-protein
2-M	NN	O	I-protein
expression	NN	O	O
and	NN	O	O
the	NN	O	O
degree	NN	O	O
of	NN	O	O
lymphocytic	NN	O	O
infiltration	NN	O	O
in	NN	O	O
the	NN	O	O
tumor	NN	O	O
tissue	NN	O	O
.	NN	O	O

Beta	NN	O	B-protein
2-M	NN	O	I-protein
serum	NN	O	O
levels	NN	O	O
were	NN	O	O
determined	NN	O	O
by	NN	O	O
an	NN	O	O
enzyme-linked	NN	O	O
immunosorbent	NN	O	O
assay	NN	O	O
in	NN	O	O
samples	NN	O	O
from	NN	O	O
22	NN	O	O
of	NN	O	O
the	NN	O	O
above	NN	O	O
women	NN	O	O
.	NN	O	O

Although	NN	O	O
some	NN	O	O
of	NN	O	O
the	NN	O	O
highest	NN	O	O
values	NN	O	O
had	NN	O	O
been	NN	O	O
obtained	NN	O	O
in	NN	O	O
women	NN	O	O
with	NN	O	O
larger	NN	O	O
(	NN	O	O
T4	NN	O	O
)	NN	O	O
primary	NN	O	O
tumors	NN	O	O
,	NN	O	O
the	NN	O	O
authors	NN	O	O
failed	NN	O	O
to	NN	O	O
detect	NN	O	O
any	NN	O	O
statistical	NN	O	O
relationship	NN	O	O
between	NN	O	O
beta	NN	O	B-protein
2-M	NN	O	I-protein
expression	NN	O	O
in	NN	O	O
the	NN	O	O
tumor	NN	O	O
with	NN	O	O
serum	NN	O	O
levels	NN	O	O
or	NN	O	O
between	NN	O	O
serum	NN	O	B-protein
beta	NN	O	I-protein
2-M	NN	O	I-protein
and	NN	O	O
the	NN	O	O
above	NN	O	O
histologic	NN	O	O
,	NN	O	O
laboratory	NN	O	O
,	NN	O	O
and	NN	O	O
clinical	NN	O	O
factors	NN	O	O
.	NN	O	O

-DOCSTART-	O

[	NN	O	O
Preliminary	NN	O	O
observation	NN	O	O
of	NN	O	O
level	NN	O	O
free-form	NN	O	B-protein
E	NN	O	I-protein
receptor	NN	O	I-protein
levels	NN	O	O
in	NN	O	O
serum	NN	O	O
of	NN	O	O
normal	NN	O	O
childbearing-aged	NN	O	O
and	NN	O	O
pregnant	NN	O	O
women	NN	O	O
]	NN	O	O

In	NN	O	O
137	NN	O	O
cases	NN	O	O
of	NN	O	O
childbearing-aged	NN	O	O
and	NN	O	O
pregnant	NN	O	O
women	NN	O	O
,	NN	O	O
free	NN	O	B-protein
form	NN	O	I-protein
E	NN	O	I-protein
receptor	NN	O	I-protein
levels	NN	O	O
(	NN	O	O
sE	NN	O	B-protein
)	NN	O	O
in	NN	O	O
serum	NN	O	O
were	NN	O	O
measured	NN	O	O
by	NN	O	O
ELISA	NN	O	O
.	NN	O	O

The	NN	O	O
level	NN	O	O
of	NN	O	O
sE	NN	O	B-protein
was	NN	O	O
significantly	NN	O	O
decreased	NN	O	O
during	NN	O	O
the	NN	O	O
first	NN	O	O
trimester	NN	O	O
,	NN	O	O
slightly	NN	O	O
higher	NN	O	O
in	NN	O	O
the	NN	O	O
second	NN	O	O
trimester	NN	O	O
,	NN	O	O
and	NN	O	O
recovered	NN	O	O
to	NN	O	O
normal	NN	O	O
in	NN	O	O
the	NN	O	O
third	NN	O	O
trimester	NN	O	O
.	NN	O	O

The	NN	O	O
level	NN	O	O
remained	NN	O	O
lower	NN	O	O
in	NN	O	O
29	NN	O	O
PIH	NN	O	O
women	NN	O	O
but	NN	O	O
appeared	NN	O	O
higher	NN	O	O
in	NN	O	O
overdue	NN	O	O
pregnancies	NN	O	O
as	NN	O	O
compared	NN	O	O
with	NN	O	O
the	NN	O	O
normal	NN	O	O
3rd	NN	O	O
trimester	NN	O	O
range	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
there	NN	O	O
is	NN	O	O
a	NN	O	O
relationship	NN	O	O
between	NN	O	O
a	NN	O	O
change	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
function	NN	O	O
and	NN	O	O
pregnancy	NN	O	O
.	NN	O	O

-DOCSTART-	O

Kappa	NN	O	B-protein
B-specific	NN	O	I-protein
DNA	NN	O	I-protein
binding	NN	O	I-protein
proteins	NN	O	I-protein
:	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
human	NN	O	B-DNA
interleukin-2	NN	O	I-DNA
gene	NN	O	I-DNA
expression	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
interleukin-2	NN	O	I-DNA
(	NN	O	I-DNA
IL-2	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
like	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
receptor	NN	O	I-DNA
alpha	NN	O	I-DNA
(	NN	O	I-DNA
IL-2R	NN	O	I-DNA
alpha	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
type	NN	O	O
1	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
(	NN	O	O
HIV-1	NN	O	O
)	NN	O	O
,	NN	O	O
is	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
modulated	NN	O	O
by	NN	O	O
a	NN	O	O
kappa	NN	O	B-protein
B-like	NN	O	I-protein
enhancer	NN	O	I-protein
element	NN	O	I-protein
.	NN	O	O

Mutation	NN	O	O
of	NN	O	O
a	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
core	NN	O	I-DNA
sequence	NN	O	I-DNA
identified	NN	O	O
in	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
(	NN	O	O
-206	NN	O	O
to	NN	O	O
-195	NN	O	O
)	NN	O	O
partially	NN	O	O
inhibits	NN	O	O
both	NN	O	O
mitogen-	NN	O	O
and	NN	O	O
HTLV-I	NN	O	O
Tax-mediated	NN	O	O
activation	NN	O	O
of	NN	O	O
this	NN	O	O
transcription	NN	O	B-protein
unit	NN	O	I-protein
and	NN	O	O
blocks	NN	O	O
the	NN	O	O
specific	NN	O	O
binding	NN	O	O
of	NN	O	O
two	NN	O	O
inducible	NN	O	B-protein
cellular	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

These	NN	O	O
kappa	NN	O	B-protein
B-specific	NN	O	I-protein
proteins	NN	O	I-protein
(	NN	O	O
80	NN	O	O
to	NN	O	O
90	NN	O	O
and	NN	O	O
50	NN	O	O
to	NN	O	O
55	NN	O	O
kilodaltons	NN	O	O
)	NN	O	O
similarly	NN	O	O
interact	NN	O	O
with	NN	O	O
the	NN	O	O
functional	NN	O	O
kappa	NN	O	B-protein
B	NN	O	I-protein
enhancer	NN	O	I-protein
present	NN	O	O
in	NN	O	O
the	NN	O	O
IL-2R	NN	O	B-DNA
alpha	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
these	NN	O	O
kappa	NN	O	B-protein
B-specific	NN	O	I-protein
proteins	NN	O	I-protein
have	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
coordinate	NN	O	O
regulation	NN	O	O
of	NN	O	O
this	NN	O	O
growth	NN	O	O
factor-growth	NN	O	O
factor	NN	O	O
receptor	NN	O	O
gene	NN	O	O
system	NN	O	O
that	NN	O	O
controls	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
proliferation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Novel	NN	O	O
region	NN	O	O
within	NN	O	O
the	NN	O	O
V	NN	O	B-DNA
kappa	NN	O	I-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
is	NN	O	O
responsible	NN	O	O
for	NN	O	O
tissue	NN	O	O
and	NN	O	O
stage-specific	NN	O	O
expression	NN	O	O
of	NN	O	O
immunoglobulin	NN	O	B-DNA
genes	NN	O	I-DNA
in	NN	O	O
human	NN	O	B-cell_type
lymphoid	NN	O	I-cell_type
neoplasms	NN	O	I-cell_type
.	NN	O	O

Immunoglobulin	NN	O	B-protein
gene-specific	NN	O	I-protein
transacting	NN	O	I-protein
factors	NN	O	I-protein
have	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
lymphoid	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
.	NN	O	O

The	NN	O	O
role	NN	O	O
of	NN	O	O
these	NN	O	O
factors	NN	O	O
in	NN	O	O
B-cell	NN	O	B-cell_type
differentiation	NN	O	O
and	NN	O	O
stage-specific	NN	O	O
expression	NN	O	O
of	NN	O	O
these	NN	O	O
genes	NN	O	O
is	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
not	NN	O	O
fully	NN	O	O
understood	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
used	NN	O	O
a	NN	O	O
model	NN	O	O
of	NN	O	O
human	NN	O	O
lymphoid	NN	O	O
neoplasia	NN	O	O
to	NN	O	O
address	NN	O	O
this	NN	O	O
question	NN	O	O
.	NN	O	O

Different	NN	O	O
fragments	NN	O	O
of	NN	O	O
unrearranged	NN	O	B-DNA
human	NN	O	I-DNA
variable	NN	O	I-DNA
region	NN	O	I-DNA
of	NN	O	O
immunoglobulin	NN	O	B-DNA
kappa	NN	O	I-DNA
gene	NN	O	I-DNA
(	NN	O	O
V	NN	O	B-DNA
kappa	NN	O	I-DNA
)	NN	O	O
were	NN	O	O
used	NN	O	O
for	NN	O	O
cell-free	NN	O	O
in	NN	O	O
vitro	NN	O	O
transcription	NN	O	O
and	NN	O	O
DNA	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
.	NN	O	O

Previously	NN	O	O
described	NN	O	O
enhancement	NN	O	O
of	NN	O	O
in	NN	O	O
vitro	NN	O	O
transcription	NN	O	O
that	NN	O	O
was	NN	O	O
only	NN	O	O
seen	NN	O	O
with	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
derived	NN	O	O
from	NN	O	O
B-cell	NN	O	B-cell_type
neoplasms	NN	O	I-cell_type
corresponding	NN	O	O
to	NN	O	O
the	NN	O	O
late	NN	O	O
stages	NN	O	O
of	NN	O	O
B-cell	NN	O	B-cell_type
differentiation	NN	O	O
was	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
actions	NN	O	O
of	NN	O	O
these	NN	O	O
factor	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
on	NN	O	O
the	NN	O	O
DNA	NN	O	O
region	NN	O	O
within	NN	O	O
the	NN	O	O
V	NN	O	B-DNA
kappa	NN	O	I-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
region	NN	O	O
is	NN	O	O
located	NN	O	O
within	NN	O	O
the	NN	O	O
920	NN	O	O
bp	NN	O	O
fragment	NN	O	O
located	NN	O	O
210	NN	O	O
bp	NN	O	O
upstream	NN	O	O
from	NN	O	O
the	NN	O	O
coding	NN	O	O
region	NN	O	O
and	NN	O	O
this	NN	O	O
fragment	NN	O	O
represents	NN	O	O
a	NN	O	O
possible	NN	O	O
novel	NN	O	O
DNA	NN	O	O
region	NN	O	O
,	NN	O	O
which	NN	O	O
plays	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
stage-	NN	O	O
and	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
.	NN	O	O

-DOCSTART-	O

[	NN	O	O
Determination	NN	O	O
of	NN	O	O
the	NN	O	O
sensitivity	NN	O	O
to	NN	O	O
glucocorticoids	NN	O	B-protein
in	NN	O	O
vitro	NN	O	O
]	NN	O	O

A	NN	O	O
modified	NN	O	O
method	NN	O	O
for	NN	O	O
the	NN	O	O
determination	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
lymphocytes	NN	O	I-cell_type
is	NN	O	O
suggested	NN	O	O
.	NN	O	O

The	NN	O	O
principal	NN	O	O
distinction	NN	O	O
of	NN	O	O
the	NN	O	O
method	NN	O	O
is	NN	O	O
standardization	NN	O	O
by	NN	O	O
the	NN	O	O
lymphocyte	NN	O	B-cell_type
count	NN	O	O
in	NN	O	O
a	NN	O	O
sample	NN	O	O
(	NN	O	O
1	NN	O	O
mln	NN	O	O
)	NN	O	O
and	NN	O	O
the	NN	O	O
labeled	NN	O	O
hormone	NN	O	B-protein
concentration	NN	O	O
.	NN	O	O

The	NN	O	O
modification	NN	O	O
saves	NN	O	O
time	NN	O	O
and	NN	O	O
money	NN	O	O
,	NN	O	O
limits	NN	O	O
the	NN	O	O
range	NN	O	O
of	NN	O	O
the	NN	O	O
data	NN	O	O
variations	NN	O	O
,	NN	O	O
and	NN	O	O
makes	NN	O	O
use	NN	O	O
of	NN	O	O
a	NN	O	O
lesser	NN	O	O
volume	NN	O	O
of	NN	O	O
blood	NN	O	O
.	NN	O	O

Examinations	NN	O	O
of	NN	O	O
70	NN	O	O
children	NN	O	O
aged	NN	O	O
4	NN	O	O
to	NN	O	O
15	NN	O	O
suffering	NN	O	O
from	NN	O	O
the	NN	O	O
nephrotic	NN	O	O
form	NN	O	O
of	NN	O	O
glomerulonephritis	NN	O	O
have	NN	O	O
made	NN	O	O
it	NN	O	O
possible	NN	O	O
to	NN	O	O
distinguish	NN	O	O
two	NN	O	O
groups	NN	O	O
of	NN	O	O
patients	NN	O	O
:	NN	O	O
with	NN	O	O
relatively	NN	O	O
high	NN	O	O
values	NN	O	O
of	NN	O	O
specific	NN	O	O
binding	NN	O	O
X	NN	O	O
=	NN	O	O
6820.1	NN	O	O
+/-	NN	O	O
530.0	NN	O	O
(	NN	O	O
n	NN	O	O
=	NN	O	O
30	NN	O	O
,	NN	O	O
p	NN	O	O
=	NN	O	O
0.95	NN	O	O
,	NN	O	O
t	NN	O	O
=	NN	O	O
2.04	NN	O	O
)	NN	O	O
,	NN	O	O
this	NN	O	O
corresponding	NN	O	O
to	NN	O	O
a	NN	O	O
clinical	NN	O	O
form	NN	O	O
of	NN	O	O
hormone	NN	O	B-protein
-sensitive	NN	O	O
glomerulonephritis	NN	O	O
,	NN	O	O
and	NN	O	O
with	NN	O	O
relatively	NN	O	O
low	NN	O	O
values	NN	O	O
of	NN	O	O
specific	NN	O	O
binding	NN	O	O
X	NN	O	O
=	NN	O	O
1815.2	NN	O	O
+/-	NN	O	O
302.8	NN	O	O
(	NN	O	O
n	NN	O	O
=	NN	O	O
40	NN	O	O
,	NN	O	O
p	NN	O	O
=	NN	O	O
0.95	NN	O	O
,	NN	O	O
t	NN	O	O
=	NN	O	O
1.96	NN	O	O
)	NN	O	O
,	NN	O	O
that	NN	O	O
corresponds	NN	O	O
to	NN	O	O
hormone	NN	O	B-protein
-resistant	NN	O	O
glomerulonephritis	NN	O	O
.	NN	O	O

Dynamic	NN	O	O
studies	NN	O	O
have	NN	O	O
not	NN	O	O
shown	NN	O	O
any	NN	O	O
statistically	NN	O	O
significant	NN	O	O
changes	NN	O	O
in	NN	O	O
the	NN	O	O
specific	NN	O	O
binding	NN	O	O
values	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
permit	NN	O	O
regarding	NN	O	O
the	NN	O	O
specific	NN	O	O
binding	NN	O	O
value	NN	O	O
as	NN	O	O
a	NN	O	O
prognostic	NN	O	O
criterion	NN	O	O
in	NN	O	O
the	NN	O	O
assessment	NN	O	O
of	NN	O	O
corticosteroid	NN	O	B-protein
therapy	NN	O	O
;	NN	O	O
this	NN	O	O
allows	NN	O	O
a	NN	O	O
wide	NN	O	O
employment	NN	O	O
of	NN	O	O
the	NN	O	O
described	NN	O	O
method	NN	O	O
in	NN	O	O
practical	NN	O	O
nephrology	NN	O	O
.	NN	O	O

-DOCSTART-	O

Octamer-binding	NN	O	B-protein
proteins	NN	O	I-protein
from	NN	O	O
B	NN	O	B-cell_line
or	NN	O	I-cell_line
HeLa	NN	O	I-cell_line
cells	NN	O	I-cell_line
stimulate	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
immunoglobulin	NN	O	B-DNA
heavy-chain	NN	O	I-DNA
promoter	NN	O	I-DNA
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

The	NN	O	O
B-cell	NN	O	B-cell_type
-type	NN	O	O
specificity	NN	O	O
of	NN	O	O
the	NN	O	O
immunoglobulin	NN	O	B-protein
(	NN	O	O
Ig	NN	O	B-protein
)	NN	O	O
heavy-chain	NN	O	O
and	NN	O	O
light-chain	NN	O	O
promoters	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
an	NN	O	O
octanucleotide	NN	O	B-DNA
(	NN	O	I-DNA
OCTA	NN	O	I-DNA
)	NN	O	I-DNA
element	NN	O	I-DNA
,	NN	O	O
ATGCAAAT	NN	O	O
,	NN	O	O
that	NN	O	O
is	NN	O	O
also	NN	O	O
a	NN	O	O
functional	NN	O	O
component	NN	O	O
of	NN	O	O
other	NN	O	O
RNA	NN	O	B-DNA
polymerase	NN	O	I-DNA
II	NN	O	I-DNA
promoters	NN	O	I-DNA
,	NN	O	O
such	NN	O	O
as	NN	O	O
snRNA	NN	O	O
and	NN	O	O
histone	NN	O	O
H2B	NN	O	O
promoters	NN	O	O
.	NN	O	O

Two	NN	O	O
nuclear	NN	O	B-protein
proteins	NN	O	I-protein
that	NN	O	O
bind	NN	O	O
specifically	NN	O	O
and	NN	O	O
with	NN	O	O
high	NN	O	O
affinity	NN	O	O
to	NN	O	O
the	NN	O	O
OCTA	NN	O	B-DNA
element	NN	O	I-DNA
have	NN	O	O
been	NN	O	O
identified	NN	O	O
.	NN	O	O

NF-A1	NN	O	B-protein
is	NN	O	O
present	NN	O	O
in	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
cell	NN	O	O
types	NN	O	O
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
NF-A2	NN	O	B-protein
is	NN	O	O
essentially	NN	O	O
confined	NN	O	O
to	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
leading	NN	O	O
to	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
NF-A2	NN	O	B-protein
activates	NN	O	O
cell-type-specific	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
Ig	NN	O	B-DNA
promoter	NN	O	I-DNA
and	NN	O	O
NF-A1	NN	O	B-protein
mediates	NN	O	O
the	NN	O	O
other	NN	O	O
responses	NN	O	O
of	NN	O	O
the	NN	O	O
OCTA	NN	O	B-DNA
element	NN	O	I-DNA
.	NN	O	O

Extracts	NN	O	O
of	NN	O	O
the	NN	O	O
B-cell	NN	O	B-cell_line
line	NN	O	I-cell_line
,	NN	O	O
BJA-B	NN	O	B-cell_line
,	NN	O	O
contain	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
NF-A2	NN	O	B-protein
and	NN	O	O
specifically	NN	O	O
transcribe	NN	O	O
Ig	NN	O	B-DNA
promoters	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
extracts	NN	O	O
from	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
transcribed	NN	O	O
the	NN	O	O
Ig	NN	O	B-DNA
promoter	NN	O	I-DNA
poorly	NN	O	O
.	NN	O	O

Surprisingly	NN	O	O
,	NN	O	O
addition	NN	O	O
of	NN	O	O
either	NN	O	O
affinity-enriched	NN	O	O
NF-A2	NN	O	O
or	NN	O	O
NF-A1	NN	O	O
to	NN	O	O
either	NN	O	O
a	NN	O	O
HeLa	NN	O	O
extract	NN	O	O
or	NN	O	O
a	NN	O	O
partially	NN	O	O
purified	NN	O	O
reaction	NN	O	O
system	NN	O	O
specifically	NN	O	O
stimulates	NN	O	O
the	NN	O	O
Ig	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
suggests	NN	O	O
that	NN	O	O
the	NN	O	O
constitutive	NN	O	O
OCTA-binding	NN	O	B-protein
factor	NN	O	I-protein
NF-A1	NN	O	I-protein
can	NN	O	O
activate	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
Ig	NN	O	B-DNA
promoter	NN	O	I-DNA
and	NN	O	O
that	NN	O	O
B-cell	NN	O	B-cell_type
-specific	NN	O	O
transcription	NN	O	O
of	NN	O	O
this	NN	O	O
promoter	NN	O	O
,	NN	O	O
at	NN	O	O
least	NN	O	O
in	NN	O	O
vitro	NN	O	O
,	NN	O	O
is	NN	O	O
partially	NN	O	O
due	NN	O	O
to	NN	O	O
a	NN	O	O
quantitative	NN	O	O
difference	NN	O	O
in	NN	O	O
the	NN	O	O
amount	NN	O	O
of	NN	O	O
OCTA-binding	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

Because	NN	O	O
NF-A1	NN	O	B-protein
can	NN	O	O
stimulate	NN	O	O
Ig	NN	O	B-protein
transcription	NN	O	O
,	NN	O	O
the	NN	O	O
inability	NN	O	O
of	NN	O	O
this	NN	O	O
factor	NN	O	O
to	NN	O	O
activate	NN	O	O
in	NN	O	O
vivo	NN	O	O
the	NN	O	O
Ig	NN	O	B-DNA
promoter	NN	O	I-DNA
to	NN	O	O
the	NN	O	O
same	NN	O	O
degree	NN	O	O
as	NN	O	O
the	NN	O	O
snRNA	NN	O	B-DNA
promoters	NN	O	I-DNA
probably	NN	O	O
reflects	NN	O	O
a	NN	O	O
difference	NN	O	O
in	NN	O	O
the	NN	O	O
context	NN	O	O
of	NN	O	O
the	NN	O	O
OCTA	NN	O	B-DNA
element	NN	O	I-DNA
in	NN	O	O
these	NN	O	O
two	NN	O	O
types	NN	O	O
of	NN	O	O
promoters	NN	O	B-DNA
.	NN	O	O

-DOCSTART-	O

Identification	NN	O	O
of	NN	O	O
a	NN	O	O
putative	NN	O	O
regulator	NN	O	O
of	NN	O	O
early	NN	O	B-DNA
T	NN	O	I-DNA
cell	NN	O	I-DNA
activation	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

Molecules	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
antigen	NN	O	O
receptor-dependent	NN	O	O
regulation	NN	O	O
of	NN	O	O
early	NN	O	B-DNA
T	NN	O	I-DNA
cell	NN	O	I-DNA
activation	NN	O	I-DNA
genes	NN	O	I-DNA
were	NN	O	O
investigated	NN	O	O
with	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
functional	NN	O	O
sequences	NN	O	O
of	NN	O	O
the	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
activation-specific	NN	O	I-protein
enhancer	NN	O	I-protein
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

One	NN	O	O
of	NN	O	O
these	NN	O	O
sequences	NN	O	O
forms	NN	O	O
a	NN	O	O
protein	NN	O	O
complex	NN	O	O
,	NN	O	O
NFAT-1	NN	O	B-protein
,	NN	O	O
specifically	NN	O	O
with	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
of	NN	O	O
activated	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

This	NN	O	O
complex	NN	O	O
appeared	NN	O	O
10	NN	O	O
to	NN	O	O
25	NN	O	O
minutes	NN	O	O
before	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

Studies	NN	O	O
with	NN	O	O
inhibitors	NN	O	O
of	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
indicated	NN	O	O
that	NN	O	O
the	NN	O	O
time	NN	O	O
of	NN	O	O
synthesis	NN	O	O
of	NN	O	O
the	NN	O	O
activator	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
corresponds	NN	O	O
to	NN	O	O
the	NN	O	O
time	NN	O	O
of	NN	O	O
appearance	NN	O	O
of	NN	O	O
NFAT-1	NN	O	B-protein
.	NN	O	O

NFAT-1	NN	O	B-protein
,	NN	O	O
or	NN	O	O
a	NN	O	O
very	NN	O	O
similar	NN	O	O
protein	NN	O	O
,	NN	O	O
bound	NN	O	O
functional	NN	O	O
sequences	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
type	NN	O	O
1	NN	O	O
;	NN	O	O
the	NN	O	O
LTR	NN	O	B-DNA
of	NN	O	O
this	NN	O	O
virus	NN	O	O
is	NN	O	O
known	NN	O	O
to	NN	O	O
be	NN	O	O
stimulated	NN	O	O
during	NN	O	O
early	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
activation	NN	O	O
.	NN	O	O

The	NN	O	O
binding	NN	O	B-DNA
site	NN	O	I-DNA
for	NN	O	O
this	NN	O	O
complex	NN	O	O
activated	NN	O	O
a	NN	O	O
linked	NN	O	O
promoter	NN	O	O
after	NN	O	O
transfection	NN	O	O
into	NN	O	O
antigen	NN	O	B-cell_type
receptor-activated	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
but	NN	O	O
not	NN	O	O
other	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

These	NN	O	O
characteristics	NN	O	O
suggest	NN	O	O
that	NN	O	O
NFAT-1	NN	O	B-protein
transmits	NN	O	O
signals	NN	O	O
initiated	NN	O	O
at	NN	O	O
the	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
antigen	NN	O	I-protein
receptor	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Characterization	NN	O	O
of	NN	O	O
thyroid	NN	O	B-protein
hormone	NN	O	I-protein
receptors	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_line
IM-9	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
.	NN	O	O

Although	NN	O	O
putatively	NN	O	O
identified	NN	O	O
more	NN	O	O
than	NN	O	O
10	NN	O	O
years	NN	O	O
ago	NN	O	O
,	NN	O	O
thyroid	NN	O	B-protein
hormone	NN	O	I-protein
receptors	NN	O	I-protein
in	NN	O	O
human	NN	O	O
tissues	NN	O	O
remain	NN	O	O
poorly	NN	O	O
characterized	NN	O	O
.	NN	O	O

As	NN	O	O
a	NN	O	O
first	NN	O	O
step	NN	O	O
towards	NN	O	O
understanding	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
thyroid	NN	O	O
hormone	NN	O	O
action	NN	O	O
in	NN	O	O
man	NN	O	O
we	NN	O	O
have	NN	O	O
characterized	NN	O	O
T3	NN	O	B-protein
binding	NN	O	I-protein
sites	NN	O	I-protein
in	NN	O	O
nuclei	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
lymphoblastoid	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
IM-9	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
whole	NN	O	O
cell	NN	O	O
experiments	NN	O	O
at	NN	O	O
37	NN	O	O
degrees	NN	O	O
C	NN	O	O
,	NN	O	O
nuclear	NN	O	O
binding	NN	O	O
of	NN	O	O
[	NN	O	B-protein
125I	NN	O	I-protein
]	NN	O	I-protein
T3	NN	O	I-protein
was	NN	O	O
saturable	NN	O	O
(	NN	O	O
Kd	NN	O	O
34	NN	O	O
+/-	NN	O	O
6	NN	O	O
pmol/l	NN	O	O
)	NN	O	O
and	NN	O	O
of	NN	O	O
finite	NN	O	O
capacity	NN	O	O
(	NN	O	O
approximately	NN	O	O
equal	NN	O	O
to	NN	O	O
350	NN	O	O
sites/cell	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
binding	NN	O	B-protein
sites	NN	O	I-protein
were	NN	O	O
extracted	NN	O	O
from	NN	O	O
a	NN	O	O
nuclear	NN	O	O
pellet	NN	O	O
by	NN	O	O
treatment	NN	O	O
with	NN	O	O
0.4	NN	O	O
mol/l	NN	O	O
KCl	NN	O	O
and	NN	O	O
sonication	NN	O	O
.	NN	O	O

Separation	NN	O	O
of	NN	O	O
bound	NN	O	O
from	NN	O	O
free	NN	O	O
[	NN	O	B-protein
125I	NN	O	I-protein
]	NN	O	I-protein
T3	NN	O	I-protein
in	NN	O	O
the	NN	O	O
extracts	NN	O	O
was	NN	O	O
achieved	NN	O	O
using	NN	O	O
the	NN	O	O
calcium	NN	O	O
phosphate	NN	O	O
matrix	NN	O	O
,	NN	O	O
hydroxyapatite	NN	O	O
at	NN	O	O
a	NN	O	O
concentration	NN	O	O
of	NN	O	O
0.3	NN	O	O
ml	NN	O	O
of	NN	O	O
a	NN	O	O
150	NN	O	O
g/l	NN	O	O
slurry	NN	O	O
.	NN	O	O

Rectilinear	NN	O	O
Scatchard	NN	O	O
plots	NN	O	O
were	NN	O	O
obtained	NN	O	O
only	NN	O	O
when	NN	O	O
the	NN	O	O
hydroxyapatite	NN	O	O
was	NN	O	O
washed	NN	O	O
with	NN	O	O
a	NN	O	O
buffer	NN	O	O
containing	NN	O	O
0.5	NN	O	O
%	NN	O	O
Triton	NN	O	O
X-100	NN	O	O
.	NN	O	O

Under	NN	O	O
these	NN	O	O
conditions	NN	O	O
T3	NN	O	B-protein
binding	NN	O	I-protein
sites	NN	O	I-protein
in	NN	O	O
the	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
were	NN	O	O
present	NN	O	O
at	NN	O	O
a	NN	O	O
concentration	NN	O	O
of	NN	O	O
22.4	NN	O	O
+/-	NN	O	O
8.6	NN	O	O
fmol/mg	NN	O	O
protein	NN	O	O
and	NN	O	O
showed	NN	O	O
an	NN	O	O
affinity	NN	O	O
of	NN	O	O
(	NN	O	O
Kd	NN	O	O
,	NN	O	O
room	NN	O	O
temperature	NN	O	O
)	NN	O	O
140	NN	O	O
+/-	NN	O	O
10	NN	O	O
pmol/l	NN	O	O
.	NN	O	O

The	NN	O	O
same	NN	O	O
assay	NN	O	O
system	NN	O	O
was	NN	O	O
used	NN	O	O
to	NN	O	O
determine	NN	O	O
the	NN	O	O
hierarchy	NN	O	O
of	NN	O	O
affinities	NN	O	O
for	NN	O	O
a	NN	O	O
range	NN	O	O
of	NN	O	O
natural	NN	O	O
and	NN	O	O
synthetic	NN	O	O
analogues	NN	O	O
.	NN	O	O

Calling	NN	O	O
T3	NN	O	O
100	NN	O	O
,	NN	O	O
the	NN	O	O
order	NN	O	O
of	NN	O	O
potencies	NN	O	O
observed	NN	O	O
was	NN	O	O
:	NN	O	O
Triac	NN	O	O
,	NN	O	O
500	NN	O	O
;	NN	O	O
3	NN	O	O
,	NN	O	O
5-diiodo-3'-isopropylthyronine	NN	O	O
,	NN	O	O
89	NN	O	O
;	NN	O	O
T4	NN	O	O
,	NN	O	O
32	NN	O	O
;	NN	O	O
3	NN	O	O
,	NN	O	O
5-dimethyl-3'isopropylthyronine	NN	O	O
2	NN	O	O
;	NN	O	O
3	NN	O	O
,	NN	O	O
5-T2	NN	O	O
,	NN	O	O
0.7	NN	O	O
,	NN	O	O
rT3	NN	O	O
,	NN	O	O
0.4	NN	O	O
;	NN	O	O
3'5'-T2	NN	O	O
,	NN	O	O
less	NN	O	O
than	NN	O	O
0.01	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
T3	NN	O	B-protein
binding	NN	O	I-protein
sites	NN	O	I-protein
present	NN	O	O
in	NN	O	O
human	NN	O	B-cell_line
IM-9	NN	O	I-cell_line
lymphocyte	NN	O	I-cell_line
nuclei	NN	O	O
and	NN	O	O
extracts	NN	O	O
thereof	NN	O	O
are	NN	O	O
thyroid	NN	O	B-protein
hormone	NN	O	I-protein
receptors	NN	O	I-protein
.	NN	O	O

These	NN	O	O
cells	NN	O	O
may	NN	O	O
be	NN	O	O
a	NN	O	O
useful	NN	O	O
tool	NN	O	O
to	NN	O	O
increase	NN	O	O
our	NN	O	O
understanding	NN	O	O
of	NN	O	O
human	NN	O	B-protein
T3	NN	O	I-protein
receptors	NN	O	I-protein

-DOCSTART-	O

Definition	NN	O	O
of	NN	O	O
T-cell	NN	O	B-protein
specific	NN	O	I-protein
DNA-binding	NN	O	I-protein
factors	NN	O	I-protein
that	NN	O	O
interact	NN	O	O
with	NN	O	O
a	NN	O	O
3'-silencer	NN	O	B-DNA
in	NN	O	O
the	NN	O	O
CD4+	NN	O	B-DNA
T-cell	NN	O	I-DNA
gene	NN	O	I-DNA
Rpt-1	NN	O	I-DNA
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
the	NN	O	O
region	NN	O	O
3	NN	O	O
'	NN	O	O
to	NN	O	O
the	NN	O	O
CD4+	NN	O	B-DNA
T-cell	NN	O	I-DNA
gene	NN	O	I-DNA
Rpt-1	NN	O	I-DNA
(	NN	O	O
encoding	NN	O	O
regulatory	NN	O	O
protein	NN	O	O
T-lymphocyte	NN	O	B-protein
1	NN	O	I-protein
)	NN	O	O
led	NN	O	O
to	NN	O	O
the	NN	O	O
definition	NN	O	O
of	NN	O	O
a	NN	O	O
silencer	NN	O	B-DNA
element	NN	O	I-DNA
that	NN	O	O
inhibits	NN	O	O
heterologous	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
certain	NN	O	O
CD4+	NN	O	B-cell_type
T-cell	NN	O	I-cell_type
lines	NN	O	I-cell_type
but	NN	O	O
not	NN	O	O
in	NN	O	O
B-cell	NN	O	B-cell_type
or	NN	O	O
non-lymphoid	NN	O	B-cell_type
cell	NN	O	I-cell_type
lines	NN	O	I-cell_type
.	NN	O	O

Functional	NN	O	O
silencer	NN	O	B-DNA
activity	NN	O	O
in	NN	O	O
vivo	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
a	NN	O	O
specific	NN	O	O
silencer-DNA-protein	NN	O	B-protein
complex	NN	O	I-protein
in	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
with	NN	O	O
T-cell	NN	O	B-cell_type
extracts	NN	O	O
.	NN	O	O

Formation	NN	O	O
of	NN	O	O
this	NN	O	O
complex	NN	O	O
was	NN	O	O
selectively	NN	O	O
inhibited	NN	O	O
by	NN	O	O
the	NN	O	O
region	NN	O	O
in	NN	O	O
HIV-1	NN	O	B-DNA
containing	NN	O	O
a	NN	O	O
silencer	NN	O	B-DNA
element	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
discuss	NN	O	O
the	NN	O	O
possibility	NN	O	O
that	NN	O	O
DNA-binding	NN	O	B-protein
factors	NN	O	I-protein
may	NN	O	O
coregulate	NN	O	O
HIV-1	NN	O	O
and	NN	O	O
Rpt-1	NN	O	O
gene	NN	O	O
expression	NN	O	O
through	NN	O	O
a	NN	O	O
common	NN	O	O
transcriptional	NN	O	B-DNA
silencer	NN	O	I-DNA
element	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Congenital	NN	O	O
immunodeficiencies	NN	O	O
associated	NN	O	O
with	NN	O	O
absence	NN	O	O
of	NN	O	O
HLA	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
antigens	NN	O	I-protein
on	NN	O	O
lymphocytes	NN	O	B-cell_type
result	NN	O	O
from	NN	O	O
distinct	NN	O	O
mutations	NN	O	O
in	NN	O	O
trans-acting	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

Coordinate	NN	O	O
regulation	NN	O	O
of	NN	O	O
HLA	NN	O	B-DNA
class	NN	O	I-DNA
II	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
during	NN	O	O
development	NN	O	O
and	NN	O	O
coinduction	NN	O	O
of	NN	O	O
class	NN	O	B-DNA
II	NN	O	I-DNA
genes	NN	O	I-DNA
by	NN	O	O
soluble	NN	O	B-protein
factors	NN	O	I-protein
suggests	NN	O	O
that	NN	O	O
common	NN	O	O
trans-acting	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	O
s	NN	O	O
)	NN	O	O
control	NN	O	O
expression	NN	O	O
of	NN	O	O
these	NN	O	O
genes	NN	O	O
.	NN	O	O

In	NN	O	O
B-lymphoblastoid	NN	O	B-cell_type
cell	NN	O	I-cell_type
lines	NN	O	I-cell_type
derived	NN	O	O
from	NN	O	O
two	NN	O	O
independent	NN	O	O
class	NN	O	O
II-deficient	NN	O	O
bare	NN	O	O
lymphocyte	NN	O	O
syndrome	NN	O	O
patients	NN	O	O
,	NN	O	O
we	NN	O	O
observed	NN	O	O
a	NN	O	O
drastic	NN	O	O
decrease	NN	O	O
in	NN	O	O
transcription	NN	O	O
rates	NN	O	O
of	NN	O	O
the	NN	O	O
class	NN	O	B-DNA
II	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

When	NN	O	O
these	NN	O	O
cell	NN	O	O
lines	NN	O	O
are	NN	O	O
fused	NN	O	O
,	NN	O	O
class	NN	O	B-DNA
II	NN	O	I-DNA
genes	NN	O	I-DNA
are	NN	O	O
reexpressed	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
immunodeficiencies	NN	O	O
in	NN	O	O
bare	NN	O	O
lymphocyte	NN	O	O
syndrome	NN	O	O
patients	NN	O	O
are	NN	O	O
the	NN	O	O
result	NN	O	O
of	NN	O	O
two	NN	O	O
distinct	NN	O	O
mutations	NN	O	O
.	NN	O	O

Further	NN	O	O
studies	NN	O	O
show	NN	O	O
that	NN	O	O
genes	NN	O	O
governing	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
class	NN	O	B-protein
II	NN	O	I-protein
antigens	NN	O	I-protein
fall	NN	O	O
into	NN	O	O
at	NN	O	O
least	NN	O	O
three	NN	O	O
complementation	NN	O	O
groups	NN	O	O
;	NN	O	O
two	NN	O	O
of	NN	O	O
these	NN	O	O
were	NN	O	O
previously	NN	O	O
unidentified	NN	O	O
in	NN	O	O
mutant	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
generated	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
we	NN	O	O
report	NN	O	O
the	NN	O	O
identification	NN	O	O
of	NN	O	O
two	NN	O	O
discrete	NN	O	O
complexes	NN	O	O
,	NN	O	O
NFX1.1	NN	O	B-protein
and	NN	O	O
NFX1.2	NN	O	B-protein
,	NN	O	O
that	NN	O	O
bind	NN	O	O
to	NN	O	O
the	NN	O	O
DRA	NN	O	B-DNA
X	NN	O	I-DNA
consensus	NN	O	I-DNA
element	NN	O	I-DNA
.	NN	O	O

Though	NN	O	O
the	NN	O	O
mutation	NN	O	O
in	NN	O	O
at	NN	O	O
least	NN	O	O
one	NN	O	O
mutant	NN	O	B-cell_line
line	NN	O	I-cell_line
generated	NN	O	O
in	NN	O	O
vitro	NN	O	O
(	NN	O	O
RJ2.2.5	NN	O	B-cell_line
)	NN	O	O
affects	NN	O	O
products	NN	O	O
functioning	NN	O	O
via	NN	O	O
interaction	NN	O	O
with	NN	O	O
the	NN	O	O
X	NN	O	B-DNA
box	NN	O	I-DNA
,	NN	O	O
clear	NN	O	O
alterations	NN	O	O
in	NN	O	O
either	NN	O	O
NFX1.1	NN	O	B-protein
or	NN	O	O
NFX1.2	NN	O	B-protein
are	NN	O	O
not	NN	O	O
found	NN	O	O
in	NN	O	O
any	NN	O	O
of	NN	O	O
the	NN	O	O
mutant	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

In	NN	O	O
vivo	NN	O	O
responsiveness	NN	O	O
to	NN	O	O
glucocorticoid	NN	O	O
correlated	NN	O	O
with	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
content	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
leukocytes	NN	O	I-cell_type
in	NN	O	O
normal	NN	O	O
humans	NN	O	O
.	NN	O	O

Dexamethasone	NN	O	O
loading	NN	O	O
tests	NN	O	O
(	NN	O	O
0.1	NN	O	O
mg	NN	O	O
dexamethasone/kg	NN	O	O
,	NN	O	O
iv	NN	O	O
)	NN	O	O
were	NN	O	O
performed	NN	O	O
in	NN	O	O
18	NN	O	O
normal	NN	O	O
males	NN	O	O
to	NN	O	O
evaluate	NN	O	O
the	NN	O	O
individual	NN	O	O
responsiveness	NN	O	O
to	NN	O	O
glucocorticoid	NN	O	O
.	NN	O	O

There	NN	O	O
were	NN	O	O
inter-individual	NN	O	O
differences	NN	O	O
in	NN	O	O
increase	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
polymorphonuclear	NN	O	I-cell_type
leukocyte	NN	O	I-cell_type
count	NN	O	O
,	NN	O	O
decrease	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
lymphocyte	NN	O	I-cell_type
count	NN	O	O
,	NN	O	O
and	NN	O	O
increase	NN	O	O
in	NN	O	O
plasma	NN	O	O
free	NN	O	O
fatty	NN	O	O
acids	NN	O	O
levels	NN	O	O
after	NN	O	O
dexamethasone	NN	O	O
injection	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
there	NN	O	O
was	NN	O	O
a	NN	O	O
significant	NN	O	O
correlation	NN	O	O
between	NN	O	O
the	NN	O	O
maximum	NN	O	O
increase	NN	O	O
in	NN	O	O
polymorphonuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
and	NN	O	O
the	NN	O	O
maximum	NN	O	O
decrease	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
(	NN	O	O
r	NN	O	O
=	NN	O	O
0.7514	NN	O	O
,	NN	O	O
p	NN	O	O
less	NN	O	O
than	NN	O	O
0.0003	NN	O	O
)	NN	O	O
.	NN	O	O

Simultaneous	NN	O	O
measurements	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
content	NN	O	O
by	NN	O	O
whole-cell	NN	O	O
assay	NN	O	O
revealed	NN	O	O
that	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
content	NN	O	O
in	NN	O	O
polymorphonuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
linearly	NN	O	O
correlated	NN	O	O
with	NN	O	O
that	NN	O	O
in	NN	O	O
the	NN	O	O
corresponding	NN	O	O
lymphocytes	NN	O	B-cell_type
(	NN	O	O
r	NN	O	O
=	NN	O	O
0.9482	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

There	NN	O	O
were	NN	O	O
also	NN	O	O
significant	NN	O	O
correlations	NN	O	O
between	NN	O	O
the	NN	O	O
maximum	NN	O	O
increase	NN	O	O
in	NN	O	O
polymorphonuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
and	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
content	NN	O	O
in	NN	O	O
polymorphonuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
(	NN	O	O
r	NN	O	O
=	NN	O	O
0.7239	NN	O	O
,	NN	O	O
p	NN	O	O
less	NN	O	O
than	NN	O	O
0.0007	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
between	NN	O	O
the	NN	O	O
maximum	NN	O	O
decrease	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
and	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
content	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
(	NN	O	O
r	NN	O	O
=	NN	O	O
0.7703	NN	O	O
,	NN	O	O
p	NN	O	O
less	NN	O	O
than	NN	O	O
0.0002	NN	O	O
)	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
individual	NN	O	O
differences	NN	O	O
are	NN	O	O
preserved	NN	O	O
both	NN	O	O
in	NN	O	O
glucocorticoid	NN	O	O
responsiveness	NN	O	O
and	NN	O	O
in	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
content	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
leukocytes	NN	O	I-cell_type
in	NN	O	O
normal	NN	O	O
humans	NN	O	O
.	NN	O	O

-DOCSTART-	O

Estradiol	NN	O	O
receptors	NN	O	O
in	NN	O	O
the	NN	O	O
cytosol	NN	O	O
of	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
hepatitis	NN	O	O
B	NN	O	O
virus	NN	O	O
carriers	NN	O	O
treated	NN	O	O
with	NN	O	O
interferon-alpha	NN	O	B-protein
.	NN	O	O

Estradiol	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
the	NN	O	O
cytosol	NN	O	O
of	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
interferon-alpha	NN	O	B-protein
(	NN	O	O
IFN-alpha	NN	O	B-protein
)	NN	O	O
on	NN	O	O
estradiol	NN	O	B-protein
receptors	NN	O	I-protein
were	NN	O	O
studied	NN	O	O
in	NN	O	O
asymptomatic	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
carriers	NN	O	O
,	NN	O	O
patients	NN	O	O
with	NN	O	O
chronic	NN	O	O
hepatitis	NN	O	O
B	NN	O	O
and	NN	O	O
normal	NN	O	O
controls	NN	O	O
.	NN	O	O

The	NN	O	O
level	NN	O	O
of	NN	O	O
estradiol	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
the	NN	O	O
cytosol	NN	O	O
of	NN	O	O
mononuclear	NN	O	B-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
significantly	NN	O	O
lower	NN	O	O
in	NN	O	O
asymptomatic	NN	O	O
HBV	NN	O	O
carriers	NN	O	O
and	NN	O	O
patients	NN	O	O
with	NN	O	O
chronic	NN	O	O
hepatitis	NN	O	O
B	NN	O	O
,	NN	O	O
compared	NN	O	O
to	NN	O	O
normal	NN	O	O
controls	NN	O	O
.	NN	O	O

This	NN	O	O
low	NN	O	O
level	NN	O	O
of	NN	O	O
cytosol	NN	O	O
estradiol	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
patients	NN	O	O
with	NN	O	O
chronic	NN	O	O
hepatitis	NN	O	O
B	NN	O	O
was	NN	O	O
increased	NN	O	O
by	NN	O	O
the	NN	O	O
administration	NN	O	O
of	NN	O	O
IFN-alpha	NN	O	B-protein
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
when	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
patients	NN	O	O
with	NN	O	O
chronic	NN	O	O
hepatitis	NN	O	O
B	NN	O	O
were	NN	O	O
incubated	NN	O	O
with	NN	O	O
IFN-alpha	NN	O	B-protein
in	NN	O	O
vitro	NN	O	O
,	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
cytosol	NN	O	O
estradiol	NN	O	B-protein
receptors	NN	O	I-protein
also	NN	O	O
increased	NN	O	O
by	NN	O	O
increasing	NN	O	O
the	NN	O	O
concentration	NN	O	O
of	NN	O	O
IFN-alpha	NN	O	B-protein
.	NN	O	O

We	NN	O	O
previously	NN	O	O
reported	NN	O	O
that	NN	O	O
the	NN	O	O
response	NN	O	O
of	NN	O	O
mononuclear	NN	O	B-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
estrogen	NN	O	O
is	NN	O	O
impaired	NN	O	O
in	NN	O	O
HBV	NN	O	O
carriers	NN	O	O
,	NN	O	O
and	NN	O	O
our	NN	O	O
present	NN	O	O
results	NN	O	O
suggested	NN	O	O
that	NN	O	O
this	NN	O	O
may	NN	O	O
be	NN	O	O
due	NN	O	O
to	NN	O	O
the	NN	O	O
low	NN	O	O
level	NN	O	O
of	NN	O	O
estradiol	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
the	NN	O	O
cytosol	NN	O	O
of	NN	O	O
mononuclear	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Association	NN	O	O
of	NN	O	O
increased	NN	O	O
lytic	NN	O	O
effector	NN	O	O
cell	NN	O	O
function	NN	O	O
with	NN	O	O
high	NN	O	O
estrogen	NN	O	B-protein
receptor	NN	O	I-protein
levels	NN	O	O
in	NN	O	O
tumor-bearing	NN	O	O
patients	NN	O	O
with	NN	O	O
breast	NN	O	O
cancer	NN	O	O
.	NN	O	O

Tumor-bearing	NN	O	O
patients	NN	O	O
with	NN	O	O
breast	NN	O	O
cancer	NN	O	O
were	NN	O	O
assayed	NN	O	O
for	NN	O	O
their	NN	O	O
natural	NN	O	B-cell_type
killer	NN	O	I-cell_type
(	NN	O	I-cell_type
NK	NN	O	I-cell_type
)	NN	O	I-cell_type
cell	NN	O	I-cell_type
activity	NN	O	O
and	NN	O	O
for	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
activated	NN	O	O
cytotoxic	NN	O	B-cell_type
T-cells	NN	O	I-cell_type
,	NN	O	O
as	NN	O	O
assessed	NN	O	O
by	NN	O	O
lectin-dependent	NN	O	O
cellular	NN	O	O
cytotoxicity	NN	O	O
(	NN	O	O
LDCC	NN	O	O
)	NN	O	O
.	NN	O	O

Tumor-bearing	NN	O	O
patients	NN	O	O
with	NN	O	O
breast	NN	O	O
cancer	NN	O	O
had	NN	O	O
a	NN	O	O
significant	NN	O	O
increase	NN	O	O
in	NN	O	O
NK	NN	O	B-cell_type
activity	NN	O	O
and	NN	O	O
in	NN	O	O
LDCC	NN	O	O
,	NN	O	O
as	NN	O	O
compared	NN	O	O
with	NN	O	O
healthy	NN	O	O
control	NN	O	O
individuals	NN	O	O
.	NN	O	O

Although	NN	O	O
the	NN	O	O
enhanced	NN	O	O
NK	NN	O	B-cell_type
cell	NN	O	I-cell_type
activity	NN	O	O
and	NN	O	O
LDCC	NN	O	O
were	NN	O	O
closely	NN	O	O
associated	NN	O	O
with	NN	O	O
high	NN	O	O
levels	NN	O	O
(	NN	O	O
greater	NN	O	O
than	NN	O	O
31	NN	O	O
fmol/mg	NN	O	O
)	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
content	NN	O	O
in	NN	O	O
the	NN	O	O
primary	NN	O	O
tumor	NN	O	O
,	NN	O	O
no	NN	O	O
other	NN	O	O
clinical	NN	O	O
or	NN	O	O
histologic	NN	O	O
correlation	NN	O	O
between	NN	O	O
the	NN	O	O
increase	NN	O	O
in	NN	O	O
either	NN	O	O
parameter	NN	O	O
of	NN	O	O
cytotoxic	NN	O	B-cell_type
effector	NN	O	I-cell_type
cell	NN	O	I-cell_type
function	NN	O	O
could	NN	O	O
be	NN	O	O
found	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
ER	NN	O	B-protein
levels	NN	O	O
greater	NN	O	O
than	NN	O	O
31	NN	O	O
fmol/mg	NN	O	O
might	NN	O	O
be	NN	O	O
associated	NN	O	O
with	NN	O	O
increased	NN	O	O
cytotoxic	NN	O	B-cell_type
effector	NN	O	I-cell_type
cell	NN	O	I-cell_type
function	NN	O	O
in	NN	O	O
tumor-bearing	NN	O	O
patients	NN	O	O
with	NN	O	O
breast	NN	O	O
cancer	NN	O	O
.	NN	O	O

-DOCSTART-	O

Properties	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
Epstein-Barr	NN	O	B-cell_line
virus-transformed	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
from	NN	O	O
patients	NN	O	O
with	NN	O	O
familial	NN	O	O
cortisol	NN	O	O
resistance	NN	O	O
.	NN	O	O

In	NN	O	O
a	NN	O	O
previous	NN	O	O
report	NN	O	O
of	NN	O	O
two	NN	O	O
patients	NN	O	O
with	NN	O	O
familial	NN	O	O
glucocorticoid	NN	O	O
resistance	NN	O	O
due	NN	O	O
to	NN	O	O
reduced	NN	O	O
numbers	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
(	NN	O	O
GR	NN	O	B-protein
)	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
shown	NN	O	O
decreased	NN	O	O
numbers	NN	O	O
of	NN	O	O
GR	NN	O	B-protein
in	NN	O	O
peripheral	NN	O	B-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
cultured	NN	O	B-cell_line
fibroblasts	NN	O	I-cell_line
but	NN	O	O
normal	NN	O	O
affinity	NN	O	O
of	NN	O	O
GR	NN	O	B-protein
in	NN	O	O
both	NN	O	O
patients	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
peripheral	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
from	NN	O	O
these	NN	O	O
patients	NN	O	O
,	NN	O	O
one	NN	O	O
patient	NN	O	O
's	NN	O	O
son	NN	O	O
and	NN	O	O
daughter	NN	O	O
,	NN	O	O
and	NN	O	O
normal	NN	O	O
subjects	NN	O	O
were	NN	O	O
transformed	NN	O	O
with	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
.	NN	O	O

Reduced	NN	O	O
numbers	NN	O	O
and	NN	O	O
normal	NN	O	O
affinity	NN	O	O
of	NN	O	O
GR	NN	O	B-protein
were	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
Epstein-Barr	NN	O	B-cell_line
virus-transformed	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
from	NN	O	O
both	NN	O	O
patients	NN	O	O
while	NN	O	O
the	NN	O	O
son	NN	O	O
and	NN	O	O
daughter	NN	O	O
had	NN	O	O
normal	NN	O	O
numbers	NN	O	O
and	NN	O	O
affinity	NN	O	O
of	NN	O	O
GR	NN	O	B-protein
.	NN	O	O

The	NN	O	O
thermal	NN	O	O
stability	NN	O	O
of	NN	O	O
GR	NN	O	B-protein
and	NN	O	O
thermal	NN	O	O
activation	NN	O	O
of	NN	O	O
cytosolic	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
both	NN	O	O
patients	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
normal	NN	O	O
.	NN	O	O

Although	NN	O	O
the	NN	O	O
percentages	NN	O	O
of	NN	O	O
nuclear	NN	O	O
bound	NN	O	O
GR	NN	O	B-protein
were	NN	O	O
similar	NN	O	O
in	NN	O	O
both	NN	O	O
patients	NN	O	O
and	NN	O	O
normal	NN	O	O
controls	NN	O	O
,	NN	O	O
the	NN	O	O
absolute	NN	O	O
amounts	NN	O	O
of	NN	O	O
nuclear	NN	O	O
bound	NN	O	O
GR	NN	O	B-protein
of	NN	O	O
the	NN	O	O
patients	NN	O	O
were	NN	O	O
about	NN	O	O
one-half	NN	O	O
that	NN	O	O
of	NN	O	O
normal	NN	O	O
controls	NN	O	O
.	NN	O	O

These	NN	O	O
abnormal	NN	O	O
properties	NN	O	O
of	NN	O	O
GR	NN	O	B-protein
(	NN	O	O
reduced	NN	O	O
numbers	NN	O	O
of	NN	O	O
GR	NN	O	B-protein
)	NN	O	O
were	NN	O	O
preserved	NN	O	O
in	NN	O	O
the	NN	O	O
transformed	NN	O	B-cell_line
cells	NN	O	I-cell_line
from	NN	O	O
the	NN	O	O
patients	NN	O	O
.	NN	O	O

-DOCSTART-	O

Octamer	NN	O	O
transcription	NN	O	O
factors	NN	O	O
1	NN	O	O
and	NN	O	O
2	NN	O	O
each	NN	O	O
bind	NN	O	O
to	NN	O	O
two	NN	O	O
different	NN	O	O
functional	NN	O	B-DNA
elements	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
immunoglobulin	NN	O	B-DNA
heavy-chain	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Immunoglobulin	NN	O	B-DNA
heavy-chain	NN	O	I-DNA
genes	NN	O	I-DNA
contain	NN	O	O
two	NN	O	O
conserved	NN	O	B-DNA
sequence	NN	O	I-DNA
elements	NN	O	I-DNA
5	NN	O	O
'	NN	O	O
to	NN	O	O
the	NN	O	O
site	NN	O	O
of	NN	O	O
transcription	NN	O	O
initiation	NN	O	O
:	NN	O	O
the	NN	O	O
octamer	NN	O	O
ATGCAAAT	NN	O	O
and	NN	O	O
the	NN	O	O
heptamer	NN	O	O
CTCATGA	NN	O	O
.	NN	O	O

Both	NN	O	O
of	NN	O	O
these	NN	O	O
elements	NN	O	O
are	NN	O	O
required	NN	O	O
for	NN	O	O
normal	NN	O	B-DNA
cell-specific	NN	O	I-DNA
promoter	NN	O	I-DNA
function	NN	O	O
.	NN	O	O

The	NN	O	O
present	NN	O	O
study	NN	O	O
demonstrates	NN	O	O
that	NN	O	O
both	NN	O	O
the	NN	O	O
ubiquitous	NN	O	B-protein
and	NN	O	I-protein
lymphoid-cell-specific	NN	O	I-protein
octamer	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
(	NN	O	O
OTF-1	NN	O	B-protein
and	NN	O	O
OTF-2	NN	O	B-protein
,	NN	O	O
respectively	NN	O	O
)	NN	O	O
interact	NN	O	O
specifically	NN	O	O
with	NN	O	O
each	NN	O	O
of	NN	O	O
the	NN	O	O
two	NN	O	O
conserved	NN	O	B-DNA
sequence	NN	O	I-DNA
elements	NN	O	I-DNA
,	NN	O	O
forming	NN	O	O
either	NN	O	O
homo-	NN	O	B-protein
or	NN	O	I-protein
heterodimeric	NN	O	I-protein
complexes	NN	O	I-protein
.	NN	O	O

This	NN	O	O
was	NN	O	O
surprising	NN	O	O
,	NN	O	O
since	NN	O	O
the	NN	O	O
heptamer	NN	O	O
and	NN	O	O
octamer	NN	O	O
sequence	NN	O	O
motifs	NN	O	O
bear	NN	O	O
no	NN	O	O
obvious	NN	O	O
similarity	NN	O	O
to	NN	O	O
each	NN	O	O
other	NN	O	O
.	NN	O	O

Binding	NN	O	O
of	NN	O	O
either	NN	O	O
factor	NN	O	O
to	NN	O	O
the	NN	O	O
octamer	NN	O	B-DNA
element	NN	O	I-DNA
occurred	NN	O	O
independently	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
OTF	NN	O	B-protein
interaction	NN	O	O
with	NN	O	O
the	NN	O	O
heptamer	NN	O	B-DNA
sequence	NN	O	I-DNA
appeared	NN	O	O
to	NN	O	O
require	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
an	NN	O	O
intact	NN	O	O
octamer	NN	O	O
motif	NN	O	O
and	NN	O	O
occurred	NN	O	O
with	NN	O	O
a	NN	O	O
spacing	NN	O	O
of	NN	O	O
either	NN	O	O
2	NN	O	B-DNA
or	NN	O	I-DNA
14	NN	O	I-DNA
base	NN	O	I-DNA
pairs	NN	O	I-DNA
between	NN	O	O
the	NN	O	O
two	NN	O	O
elements	NN	O	O
,	NN	O	O
suggesting	NN	O	O
coordinate	NN	O	O
binding	NN	O	O
resulting	NN	O	O
from	NN	O	O
protein-protein	NN	O	O
interactions	NN	O	O
.	NN	O	O

The	NN	O	O
degeneracy	NN	O	O
in	NN	O	O
sequences	NN	O	O
recognized	NN	O	O
by	NN	O	O
the	NN	O	O
OTFs	NN	O	B-protein
may	NN	O	O
be	NN	O	O
important	NN	O	O
in	NN	O	O
widening	NN	O	O
the	NN	O	O
range	NN	O	O
over	NN	O	O
which	NN	O	O
gene	NN	O	O
expression	NN	O	O
can	NN	O	O
be	NN	O	O
modulated	NN	O	O
and	NN	O	O
in	NN	O	O
establishing	NN	O	O
cell	NN	O	O
type	NN	O	O
specificity	NN	O	O
.	NN	O	O

-DOCSTART-	O

Identification	NN	O	O
of	NN	O	O
a	NN	O	O
novel	NN	O	O
lymphoid	NN	O	B-protein
specific	NN	O	I-protein
octamer	NN	O	I-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
OTF-2B	NN	O	B-protein
)	NN	O	O
by	NN	O	O
proteolytic	NN	O	O
clipping	NN	O	O
bandshift	NN	O	O
assay	NN	O	O
(	NN	O	O
PCBA	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
octamer	NN	O	O
sequence	NN	O	O
ATGCAAAT	NN	O	O
is	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
promoters	NN	O	O
of	NN	O	O
immunoglobulin	NN	O	B-DNA
(	NN	O	I-DNA
Ig	NN	O	I-DNA
)	NN	O	I-DNA
heavy	NN	O	I-DNA
and	NN	O	I-DNA
light	NN	O	I-DNA
chain	NN	O	I-DNA
genes	NN	O	I-DNA
and	NN	O	O
in	NN	O	O
the	NN	O	O
heavy	NN	O	B-DNA
chain	NN	O	I-DNA
enhancer	NN	O	I-DNA
and	NN	O	O
is	NN	O	O
a	NN	O	O
major	NN	O	O
determinant	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	O
type	NN	O	O
specific	NN	O	O
expression	NN	O	O
of	NN	O	O
Ig	NN	O	B-DNA
genes	NN	O	I-DNA
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

An	NN	O	O
apparent	NN	O	O
paradox	NN	O	O
is	NN	O	O
that	NN	O	O
the	NN	O	O
same	NN	O	O
sequence	NN	O	O
serves	NN	O	O
as	NN	O	O
an	NN	O	O
upstream	NN	O	B-DNA
promoter	NN	O	I-DNA
or	NN	O	I-DNA
enhancer	NN	O	I-DNA
element	NN	O	I-DNA
in	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
housekeeping	NN	O	B-DNA
genes	NN	O	I-DNA
such	NN	O	O
as	NN	O	O
the	NN	O	O
histone	NN	O	B-DNA
H2B	NN	O	I-DNA
and	NN	O	I-DNA
U	NN	O	I-DNA
snRNA	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
differential	NN	O	O
usage	NN	O	O
of	NN	O	O
this	NN	O	O
regulatory	NN	O	O
sequence	NN	O	O
motif	NN	O	O
is	NN	O	O
thought	NN	O	O
to	NN	O	O
be	NN	O	O
mediated	NN	O	O
by	NN	O	O
different	NN	O	O
species	NN	O	O
of	NN	O	O
octamer	NN	O	B-protein
binding	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

One	NN	O	O
species	NN	O	O
of	NN	O	O
100	NN	O	O
kd	NN	O	O
,	NN	O	O
designated	NN	O	O
OTF-1	NN	O	B-protein
,	NN	O	O
is	NN	O	O
present	NN	O	O
in	NN	O	O
all	NN	O	O
cell	NN	O	O
types	NN	O	O
and	NN	O	O
may	NN	O	O
exert	NN	O	O
its	NN	O	O
activating	NN	O	O
function	NN	O	O
only	NN	O	O
when	NN	O	O
it	NN	O	O
can	NN	O	O
interact	NN	O	O
with	NN	O	O
additional	NN	O	O
adjacent	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

The	NN	O	O
lymphoid	NN	O	B-protein
cell	NN	O	I-protein
specific	NN	O	I-protein
octamer	NN	O	I-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
of	NN	O	O
60	NN	O	O
kd	NN	O	O
(	NN	O	O
OTF-2A	NN	O	B-protein
)	NN	O	O
specifically	NN	O	O
stimulates	NN	O	O
Ig	NN	O	B-DNA
promoters	NN	O	I-DNA
which	NN	O	O
consist	NN	O	O
essentially	NN	O	O
of	NN	O	O
a	NN	O	O
TATA-box	NN	O	B-DNA
and	NN	O	O
an	NN	O	O
octamer	NN	O	B-DNA
sequence	NN	O	I-DNA
upstream	NN	O	O
of	NN	O	O
it	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
present	NN	O	O
evidence	NN	O	O
for	NN	O	O
yet	NN	O	O
another	NN	O	O
B	NN	O	B-protein
cell	NN	O	I-protein
specific	NN	O	I-protein
octamer	NN	O	I-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
of	NN	O	O
75	NN	O	O
kd	NN	O	O
(	NN	O	O
OTF-2B	NN	O	B-protein
)	NN	O	O
.	NN	O	O

From	NN	O	O
several	NN	O	O
findings	NN	O	O
,	NN	O	O
including	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
OTF-2B	NN	O	B-protein
(	NN	O	O
but	NN	O	O
not	NN	O	O
OTF-2A	NN	O	B-protein
)	NN	O	O
from	NN	O	O
a	NN	O	O
lymphocyte	NN	O	B-cell_line
line	NN	O	I-cell_line
that	NN	O	O
can	NN	O	O
not	NN	O	O
respond	NN	O	O
to	NN	O	O
the	NN	O	O
IgH	NN	O	B-DNA
enhancer	NN	O	I-DNA
,	NN	O	O
we	NN	O	O
propose	NN	O	O
a	NN	O	O
role	NN	O	O
of	NN	O	O
the	NN	O	O
novel	NN	O	B-protein
octamer	NN	O	I-protein
factor	NN	O	I-protein
in	NN	O	O
the	NN	O	O
long	NN	O	O
range	NN	O	O
activation	NN	O	O
by	NN	O	O
the	NN	O	O
IgH	NN	O	B-DNA
enhancer	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
have	NN	O	O
used	NN	O	O
the	NN	O	O
proteolytic	NN	O	O
clipping	NN	O	O
bandshift	NN	O	O
assay	NN	O	O
(	NN	O	O
PCBA	NN	O	O
)	NN	O	O
technique	NN	O	O
to	NN	O	O
distinguish	NN	O	O
the	NN	O	O
three	NN	O	O
different	NN	O	O
forms	NN	O	O
found	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

This	NN	O	O
analysis	NN	O	O
indicates	NN	O	O
that	NN	O	O
the	NN	O	O
75	NN	O	O
kd-species	NN	O	O
OTF-2B	NN	O	B-protein
is	NN	O	O
closely	NN	O	O
related	NN	O	O
to	NN	O	O
the	NN	O	O
60	NN	O	B-protein
kd	NN	O	I-protein
species	NN	O	I-protein
OTF-2A	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Inhibition	NN	O	O
of	NN	O	O
interleukin	NN	O	B-protein
2	NN	O	I-protein
-induced	NN	O	O
proliferation	NN	O	O
of	NN	O	O
cloned	NN	O	B-cell_line
murine	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
by	NN	O	O
glucocorticoids	NN	O	O
.	NN	O	O

Possible	NN	O	O
involvement	NN	O	O
of	NN	O	O
an	NN	O	O
inhibitory	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

The	NN	O	O
ability	NN	O	O
of	NN	O	O
glucocorticoids	NN	O	O
to	NN	O	O
inhibit	NN	O	O
interleukin	NN	O	B-protein
2	NN	O	I-protein
(	NN	O	O
IL	NN	O	B-protein
2	NN	O	I-protein
)	NN	O	O
-induced	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
proliferation	NN	O	O
in	NN	O	O
two	NN	O	O
cytotoxic	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
(	NN	O	I-cell_line
CTL	NN	O	I-cell_line
)	NN	O	I-cell_line
clones	NN	O	I-cell_line
has	NN	O	O
been	NN	O	O
studied	NN	O	O
.	NN	O	O

A	NN	O	O
complete	NN	O	O
inhibition	NN	O	O
of	NN	O	O
DNA	NN	O	O
synthesis	NN	O	O
by	NN	O	O
dexamethasone	NN	O	O
(	NN	O	O
Dx	NN	O	O
)	NN	O	O
could	NN	O	O
be	NN	O	O
observed	NN	O	O
when	NN	O	O
IL	NN	O	B-cell_line
2-depleted	NN	O	I-cell_line
cultures	NN	O	I-cell_line
of	NN	O	O
CTL	NN	O	B-cell_line
were	NN	O	O
either	NN	O	O
incubated	NN	O	O
for	NN	O	O
6	NN	O	O
h	NN	O	O
with	NN	O	O
the	NN	O	O
hormone	NN	O	B-protein
prior	NN	O	O
to	NN	O	O
the	NN	O	O
addition	NN	O	O
of	NN	O	O
IL	NN	O	B-protein
2	NN	O	I-protein
or	NN	O	O
treated	NN	O	O
simultaneously	NN	O	O
with	NN	O	O
Dx	NN	O	O
and	NN	O	O
a	NN	O	O
low	NN	O	O
concentration	NN	O	O
of	NN	O	O
IL	NN	O	B-protein
2	NN	O	I-protein
.	NN	O	O

No	NN	O	O
significant	NN	O	O
reduction	NN	O	O
in	NN	O	O
the	NN	O	O
number	NN	O	O
and	NN	O	O
affinity	NN	O	O
of	NN	O	O
IL	NN	O	B-protein
2	NN	O	I-protein
receptors	NN	O	I-protein
was	NN	O	O
seen	NN	O	O
after	NN	O	O
6	NN	O	O
h	NN	O	O
incubation	NN	O	O
with	NN	O	O
Dx	NN	O	O
.	NN	O	O

The	NN	O	O
order	NN	O	O
of	NN	O	O
potency	NN	O	O
observed	NN	O	O
with	NN	O	O
the	NN	O	O
different	NN	O	O
steroids	NN	O	O
indicated	NN	O	O
that	NN	O	O
this	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
was	NN	O	O
mediated	NN	O	O
through	NN	O	O
binding	NN	O	O
to	NN	O	O
a	NN	O	O
specific	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
.	NN	O	O

The	NN	O	O
action	NN	O	O
of	NN	O	O
these	NN	O	O
hormones	NN	O	B-protein
possibly	NN	O	O
involves	NN	O	O
the	NN	O	O
synthesis	NN	O	O
of	NN	O	O
an	NN	O	O
inhibitory	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	O
s	NN	O	O
)	NN	O	O
,	NN	O	O
since	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
cycloheximide	NN	O	O
during	NN	O	O
the	NN	O	O
incubation	NN	O	O
with	NN	O	O
Dx	NN	O	O
prevented	NN	O	O
the	NN	O	O
inhibition	NN	O	O
of	NN	O	O
DNA	NN	O	O
synthesis	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
supernatant	NN	O	O
from	NN	O	O
Dx-treated	NN	O	O
CTL	NN	O	B-cell_line
contained	NN	O	O
a	NN	O	O
nondialyzable	NN	O	O
factor	NN	O	O
which	NN	O	O
inhibited	NN	O	O
DNA	NN	O	O
synthesis	NN	O	O
and	NN	O	O
cell	NN	O	O
growth	NN	O	O
of	NN	O	O
CTL	NN	O	B-cell_line
clones	NN	O	I-cell_line
induced	NN	O	O
by	NN	O	O
IL	NN	O	B-protein
2	NN	O	I-protein
.	NN	O	O

Blocking	NN	O	O
of	NN	O	O
IL	NN	O	B-protein
2	NN	O	I-protein
synthesis	NN	O	O
and	NN	O	O
IL	NN	O	B-protein
2	NN	O	I-protein
receptor	NN	O	I-protein
formation	NN	O	O
have	NN	O	O
been	NN	O	O
proposed	NN	O	O
as	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
major	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
glucocorticoid-induced	NN	O	O
immunosuppression	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
these	NN	O	O
hormones	NN	O	B-protein
may	NN	O	O
also	NN	O	O
affect	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
proliferation	NN	O	O
by	NN	O	O
inhibiting	NN	O	O
IL	NN	O	B-protein
2	NN	O	I-protein
activity	NN	O	O
.	NN	O	O

-DOCSTART-	O

Identification	NN	O	O
and	NN	O	O
purification	NN	O	O
of	NN	O	O
a	NN	O	O
human	NN	O	B-protein
immunoglobulin-enhancer-binding	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
that	NN	O	O
activates	NN	O	O
transcription	NN	O	O
from	NN	O	O
a	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
promoter	NN	O	I-DNA
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

The	NN	O	O
enhancer-binding	NN	O	B-protein
factor	NN	O	I-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
found	NN	O	O
only	NN	O	O
in	NN	O	O
cells	NN	O	O
that	NN	O	O
transcribe	NN	O	O
immunoglobulin	NN	O	B-DNA
light	NN	O	I-DNA
chain	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
has	NN	O	O
been	NN	O	O
purified	NN	O	O
from	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
of	NN	O	O
Namalwa	NN	O	B-cell_line
cells	NN	O	I-cell_line
(	NN	O	O
human	NN	O	B-cell_line
Burkitt	NN	O	I-cell_line
lymphoma	NN	O	I-cell_line
cells	NN	O	I-cell_line
)	NN	O	O
by	NN	O	O
sequence-specific	NN	O	O
DNA	NN	O	O
affinity	NN	O	O
chromatography	NN	O	O
.	NN	O	O

The	NN	O	O
purified	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
has	NN	O	O
been	NN	O	O
identified	NN	O	O
as	NN	O	O
a	NN	O	O
51-kDa	NN	O	B-protein
polypeptide	NN	O	I-protein
by	NN	O	O
UV-crosslinking	NN	O	O
analysis	NN	O	O
.	NN	O	O

``	NN	O	O
Footprint	NN	O	O
''	NN	O	O
and	NN	O	O
methylation-interference	NN	O	O
analyses	NN	O	O
have	NN	O	O
shown	NN	O	O
that	NN	O	O
purified	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
has	NN	O	O
a	NN	O	O
binding	NN	O	O
activity	NN	O	O
specific	NN	O	O
for	NN	O	O
the	NN	O	O
kappa	NN	O	B-DNA
light	NN	O	I-DNA
chain	NN	O	I-DNA
enhancer	NN	O	I-DNA
sequence	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
purified	NN	O	O
factor	NN	O	O
activated	NN	O	O
in	NN	O	O
vitro	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
immunodeficiency	NN	O	I-DNA
virus	NN	O	I-DNA
type	NN	O	I-DNA
I	NN	O	I-DNA
promoter	NN	O	I-DNA
by	NN	O	O
binding	NN	O	O
to	NN	O	O
an	NN	O	O
upstream	NN	O	B-DNA
NF-kappa	NN	O	I-DNA
B-binding	NN	O	I-DNA
site	NN	O	I-DNA

-DOCSTART-	O

Lymphocyte	NN	O	B-cell_type
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
binding	NN	O	O
in	NN	O	O
depression	NN	O	O
:	NN	O	O
normal	NN	O	O
values	NN	O	O
following	NN	O	O
recovery	NN	O	O
.	NN	O	O

The	NN	O	O
number	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
sites	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
and	NN	O	O
plasma	NN	O	O
cortisol	NN	O	O
concentrations	NN	O	O
were	NN	O	O
measured	NN	O	O
in	NN	O	O
20	NN	O	O
patients	NN	O	O
who	NN	O	O
had	NN	O	O
recovered	NN	O	O
from	NN	O	O
major	NN	O	O
depressive	NN	O	O
disorder	NN	O	O
and	NN	O	O
20	NN	O	O
healthy	NN	O	O
control	NN	O	O
subjects	NN	O	O
.	NN	O	O

The	NN	O	O
number	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
sites	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
from	NN	O	O
the	NN	O	O
recovered	NN	O	O
depressed	NN	O	O
group	NN	O	O
was	NN	O	O
not	NN	O	O
significantly	NN	O	O
different	NN	O	O
from	NN	O	O
that	NN	O	O
of	NN	O	O
the	NN	O	O
control	NN	O	O
group	NN	O	O
.	NN	O	O

Although	NN	O	O
the	NN	O	O
mean	NN	O	O
plasma	NN	O	O
cortisol	NN	O	O
concentration	NN	O	O
in	NN	O	O
recovered	NN	O	O
depressives	NN	O	O
was	NN	O	O
higher	NN	O	O
than	NN	O	O
in	NN	O	O
control	NN	O	O
subjects	NN	O	O
,	NN	O	O
the	NN	O	O
difference	NN	O	O
only	NN	O	O
just	NN	O	O
reached	NN	O	O
significance	NN	O	O
.	NN	O	O

This	NN	O	O
study	NN	O	O
shows	NN	O	O
that	NN	O	O
the	NN	O	O
reduction	NN	O	O
in	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
numbers	NN	O	O
which	NN	O	O
occurs	NN	O	O
during	NN	O	O
acute	NN	O	O
depressive	NN	O	O
illness	NN	O	O
does	NN	O	O
not	NN	O	O
persist	NN	O	O
on	NN	O	O
recovery	NN	O	O
and	NN	O	O
is	NN	O	O
,	NN	O	O
therefore	NN	O	O
,	NN	O	O
state-dependent	NN	O	O
.	NN	O	O

-DOCSTART-	O

Identification	NN	O	O
and	NN	O	O
purification	NN	O	O
of	NN	O	O
a	NN	O	O
human	NN	O	B-protein
lymphoid-specific	NN	O	I-protein
octamer-binding	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
OTF-2	NN	O	B-protein
)	NN	O	O
that	NN	O	O
activates	NN	O	O
transcription	NN	O	O
of	NN	O	O
an	NN	O	O
immunoglobulin	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

The	NN	O	O
octamer	NN	O	O
sequence	NN	O	O
5'-ATGCAAAT	NN	O	O
,	NN	O	O
in	NN	O	O
either	NN	O	O
orientation	NN	O	O
,	NN	O	O
serves	NN	O	O
as	NN	O	O
an	NN	O	O
upstream	NN	O	B-DNA
element	NN	O	I-DNA
in	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
promoters	NN	O	B-DNA
and	NN	O	O
also	NN	O	O
occurs	NN	O	O
as	NN	O	O
a	NN	O	O
modular	NN	O	B-DNA
enhancer	NN	O	I-DNA
element	NN	O	I-DNA
.	NN	O	O

It	NN	O	O
is	NN	O	O
of	NN	O	O
particular	NN	O	O
interest	NN	O	O
in	NN	O	O
immunoglobulin	NN	O	B-DNA
genes	NN	O	I-DNA
since	NN	O	O
it	NN	O	O
is	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
upstream	NN	O	B-DNA
regions	NN	O	I-DNA
of	NN	O	O
all	NN	O	O
heavy	NN	O	B-DNA
and	NN	O	I-DNA
light	NN	O	I-DNA
chain	NN	O	I-DNA
promoters	NN	O	I-DNA
and	NN	O	O
in	NN	O	O
the	NN	O	O
heavy	NN	O	B-DNA
chain	NN	O	I-DNA
enhancer	NN	O	I-DNA
,	NN	O	O
both	NN	O	O
of	NN	O	O
which	NN	O	O
are	NN	O	O
known	NN	O	O
to	NN	O	O
be	NN	O	O
necessary	NN	O	O
for	NN	O	O
cell-specific	NN	O	O
expression	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
here	NN	O	O
the	NN	O	O
chromatographic	NN	O	O
separation	NN	O	O
of	NN	O	O
ubiquitous	NN	O	B-protein
and	NN	O	I-protein
B	NN	O	I-protein
cell-specific	NN	O	I-protein
octamer-binding	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

The	NN	O	O
B	NN	O	B-protein
cell	NN	O	I-protein
factor	NN	O	I-protein
was	NN	O	O
purified	NN	O	O
to	NN	O	O
homogeneity	NN	O	O
using	NN	O	O
affinity	NN	O	O
chromatography	NN	O	O
and	NN	O	O
consists	NN	O	O
of	NN	O	O
three	NN	O	O
peptides	NN	O	O
of	NN	O	O
62	NN	O	O
,	NN	O	O
61	NN	O	O
,	NN	O	O
and	NN	O	O
58.5	NN	O	O
+/-	NN	O	O
1.5	NN	O	O
kd	NN	O	O
.	NN	O	O

Each	NN	O	O
of	NN	O	O
the	NN	O	O
polypeptides	NN	O	O
was	NN	O	O
renatured	NN	O	O
after	NN	O	O
SDS-PAGE	NN	O	O
and	NN	O	O
shown	NN	O	O
to	NN	O	O
bind	NN	O	O
to	NN	O	O
the	NN	O	O
octamer	NN	O	O
sequence	NN	O	O
.	NN	O	O

The	NN	O	O
specific	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
pure	NN	O	O
B	NN	O	B-protein
cell-specific	NN	O	I-protein
factor	NN	O	I-protein
was	NN	O	O
indistinguishable	NN	O	O
from	NN	O	O
that	NN	O	O
of	NN	O	O
the	NN	O	O
affinity-purified	NN	O	B-protein
ubiquitous	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

This	NN	O	O
B	NN	O	B-protein
cell-specific	NN	O	I-protein
octamer-binding	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
in	NN	O	O
pure	NN	O	O
form	NN	O	O
,	NN	O	O
activated	NN	O	O
transcription	NN	O	O
from	NN	O	O
a	NN	O	O
kappa	NN	O	B-DNA
light	NN	O	I-DNA
chain	NN	O	I-DNA
promoter	NN	O	I-DNA
in	NN	O	O
vitro	NN	O	O
,	NN	O	O
thus	NN	O	O
demonstrating	NN	O	O
that	NN	O	O
it	NN	O	O
is	NN	O	O
indeed	NN	O	O
a	NN	O	O
B	NN	O	B-protein
cell-specific	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
for	NN	O	O
this	NN	O	O
gene	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
to	NN	O	O
the	NN	O	O
ubiquitous	NN	O	B-protein
and	NN	O	I-protein
B	NN	O	I-protein
cell-specific	NN	O	I-protein
octamer-binding	NN	O	I-protein
factors	NN	O	I-protein
,	NN	O	O
we	NN	O	O
identified	NN	O	O
several	NN	O	O
additional	NN	O	O
proteins	NN	O	O
,	NN	O	O
one	NN	O	O
of	NN	O	O
which	NN	O	O
is	NN	O	O
B	NN	O	B-cell_type
cell	NN	O	I-cell_type
-specific	NN	O	O
,	NN	O	O
that	NN	O	O
interact	NN	O	O
with	NN	O	O
the	NN	O	O
kappa	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Decreased	NN	O	O
deoxyribonucleic	NN	O	O
acid	NN	O	O
binding	NN	O	O
of	NN	O	O
glucocorticoid-receptor	NN	O	B-protein
complex	NN	O	I-protein
in	NN	O	O
cultured	NN	O	O
skin	NN	O	B-cell_line
fibroblasts	NN	O	I-cell_line
from	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	O
resistance	NN	O	O
syndrome	NN	O	O
.	NN	O	O

A	NN	O	O
patient	NN	O	O
with	NN	O	O
the	NN	O	O
syndrome	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	O
resistance	NN	O	O
was	NN	O	O
studied	NN	O	O
.	NN	O	O

A	NN	O	O
27-yr-old	NN	O	O
woman	NN	O	O
initially	NN	O	O
was	NN	O	O
diagnosed	NN	O	O
as	NN	O	O
having	NN	O	O
Cushing	NN	O	O
's	NN	O	O
disease	NN	O	O
,	NN	O	O
based	NN	O	O
on	NN	O	O
the	NN	O	O
findings	NN	O	O
of	NN	O	O
high	NN	O	O
plasma	NN	O	O
ACTH	NN	O	O
and	NN	O	O
serum	NN	O	O
cortisol	NN	O	O
levels	NN	O	O
,	NN	O	O
increased	NN	O	O
urinary	NN	O	O
cortisol	NN	O	O
secretion	NN	O	O
,	NN	O	O
resistance	NN	O	O
to	NN	O	O
adrenal	NN	O	O
suppression	NN	O	O
with	NN	O	O
dexamethasone	NN	O	O
,	NN	O	O
and	NN	O	O
bilateral	NN	O	O
adrenal	NN	O	O
hyperplasia	NN	O	O
by	NN	O	O
computed	NN	O	O
tomography	NN	O	O
and	NN	O	O
scintigraphy	NN	O	O
of	NN	O	O
the	NN	O	O
adrenal	NN	O	O
glands	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
she	NN	O	O
had	NN	O	O
no	NN	O	O
signs	NN	O	O
or	NN	O	O
symptoms	NN	O	O
of	NN	O	O
Cushing	NN	O	O
's	NN	O	O
syndrome	NN	O	O
.	NN	O	O

During	NN	O	O
a	NN	O	O
5-yr	NN	O	O
follow-up	NN	O	O
,	NN	O	O
no	NN	O	O
clinical	NN	O	O
abnormalities	NN	O	O
developed	NN	O	O
,	NN	O	O
although	NN	O	O
hypercortisolism	NN	O	O
persisted	NN	O	O
.	NN	O	O

End-organ	NN	O	O
resistance	NN	O	O
to	NN	O	O
cortisol	NN	O	O
was	NN	O	O
suspected	NN	O	O
.	NN	O	O

To	NN	O	O
explain	NN	O	O
the	NN	O	O
end-organ	NN	O	O
resistance	NN	O	O
to	NN	O	O
cortisol	NN	O	O
,	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
(	NN	O	O
GR	NN	O	B-protein
)	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
mononuclear	NN	O	I-cell_type
leukocytes	NN	O	I-cell_type
and	NN	O	O
cultured	NN	O	B-cell_line
skin	NN	O	I-cell_line
fibroblasts	NN	O	I-cell_line
from	NN	O	O
a	NN	O	O
forearm	NN	O	O
skin	NN	O	O
biopsy	NN	O	O
were	NN	O	O
characterized	NN	O	O
and	NN	O	O
compared	NN	O	O
with	NN	O	O
the	NN	O	O
results	NN	O	O
of	NN	O	O
similar	NN	O	O
studies	NN	O	O
in	NN	O	O
normal	NN	O	O
subjects	NN	O	O
.	NN	O	O

The	NN	O	O
patient	NN	O	O
's	NN	O	O
GR	NN	O	B-protein
in	NN	O	O
whole	NN	O	O
cell	NN	O	O
assays	NN	O	O
had	NN	O	O
an	NN	O	O
increased	NN	O	O
dissociation	NN	O	O
constant	NN	O	O
(	NN	O	O
Kd	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
cytosol	NN	O	O
of	NN	O	O
cultured	NN	O	B-cell_line
skin	NN	O	I-cell_line
fibroblasts	NN	O	I-cell_line
from	NN	O	O
the	NN	O	O
patient	NN	O	O
,	NN	O	O
there	NN	O	O
was	NN	O	O
also	NN	O	O
decreased	NN	O	O
binding	NN	O	O
capacity	NN	O	O
.	NN	O	O

The	NN	O	O
thermal	NN	O	O
stability	NN	O	O
and	NN	O	O
the	NN	O	O
sedimentation	NN	O	O
coefficient	NN	O	O
in	NN	O	O
a	NN	O	O
sucrose	NN	O	O
density	NN	O	O
gradient	NN	O	O
of	NN	O	O
the	NN	O	O
receptors	NN	O	O
in	NN	O	O
the	NN	O	O
cytosol	NN	O	O
of	NN	O	O
cultured	NN	O	B-cell_line
skin	NN	O	I-cell_line
fibroblasts	NN	O	I-cell_line
from	NN	O	O
the	NN	O	O
patient	NN	O	O
and	NN	O	O
normal	NN	O	O
subjects	NN	O	O
were	NN	O	O
similar	NN	O	O
.	NN	O	O

GR	NN	O	B-cell_line
complex	NN	O	I-cell_line
activation	NN	O	I-cell_line
,	NN	O	O
analyzed	NN	O	O
by	NN	O	O
DEAE-cellulose	NN	O	B-cell_line
chromatography	NN	O	I-cell_line
,	NN	O	O
was	NN	O	O
decreased	NN	O	O
in	NN	O	O
the	NN	O	O
patient	NN	O	O
.	NN	O	O

DNA	NN	O	B-DNA
binding	NN	O	O
of	NN	O	O
the	NN	O	O
GR	NN	O	B-protein
complex	NN	O	I-protein
after	NN	O	O
temperature-induced	NN	O	O
activation	NN	O	O
was	NN	O	O
lower	NN	O	O
in	NN	O	O
the	NN	O	O
patient	NN	O	O
than	NN	O	O
in	NN	O	O
normal	NN	O	O
subjects	NN	O	O
.	NN	O	O

Nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
GR	NN	O	B-protein
complexes	NN	O	I-protein
from	NN	O	O
the	NN	O	O
patient	NN	O	O
was	NN	O	O
also	NN	O	O
slightly	NN	O	O
decreased	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
patient	NN	O	O
's	NN	O	O
glucocorticoid	NN	O	O
resistance	NN	O	O
was	NN	O	O
due	NN	O	O
to	NN	O	O
a	NN	O	O
decrease	NN	O	O
in	NN	O	O
the	NN	O	O
affinity	NN	O	O
of	NN	O	O
the	NN	O	O
receptor	NN	O	O
for	NN	O	O
glucocorticoids	NN	O	O
and	NN	O	O
a	NN	O	O
decrease	NN	O	O
in	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
the	NN	O	O
GR	NN	O	B-protein
complex	NN	O	I-protein
to	NN	O	O
DNA	NN	O	B-DNA
.	NN	O	O

-DOCSTART-	O

Granulocyte-macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

Sensitive	NN	O	O
and	NN	O	O
receptor-mediated	NN	O	O
regulation	NN	O	O
by	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
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
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
most	NN	O	O
hormonally	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
modulates	NN	O	O
sensitively	NN	O	O
and	NN	O	O
specifically	NN	O	O
both	NN	O	O
the	NN	O	O
protein	NN	O	O
and	NN	O	O
messenger	NN	O	B-RNA
RNA	NN	O	I-RNA
accumulation	NN	O	O
of	NN	O	O
the	NN	O	O
multilineage	NN	O	B-protein
growth	NN	O	I-protein
factor	NN	O	I-protein
granulocyte-macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
GM-CSF	NN	O	B-protein
)	NN	O	O
.	NN	O	O

The	NN	O	O
regulation	NN	O	O
of	NN	O	O
GM-CSF	NN	O	B-protein
expression	NN	O	O
is	NN	O	O
seen	NN	O	O
in	NN	O	O
both	NN	O	O
normal	NN	O	B-cell_type
human	NN	O	I-cell_type
mitogen-activated	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
T	NN	O	B-cell_line
lymphocytes	NN	O	I-cell_line
from	NN	O	I-cell_line
a	NN	O	I-cell_line
line	NN	O	I-cell_line
(	NN	O	I-cell_line
S-LB1	NN	O	I-cell_line
)	NN	O	I-cell_line
transformed	NN	O	I-cell_line
with	NN	O	I-cell_line
human	NN	O	I-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
lymphotropic	NN	O	I-cell_line
virus	NN	O	I-cell_line
1	NN	O	I-cell_line
(	NN	O	O
HTLV-1	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
cells	NN	O	O
from	NN	O	O
a	NN	O	O
HTLV-1	NN	O	B-cell_line
transformed	NN	O	I-cell_line
T	NN	O	I-cell_line
lymphocyte	NN	O	I-cell_line
line	NN	O	I-cell_line
(	NN	O	O
Ab-VDR	NN	O	B-cell_line
)	NN	O	O
established	NN	O	O
from	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
vitamin	NN	O	O
D-resistant	NN	O	O
rickets	NN	O	O
type	NN	O	O
II	NN	O	O
with	NN	O	O
undetectable	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
cellular	NN	O	I-protein
receptors	NN	O	I-protein
are	NN	O	O
resistant	NN	O	O
to	NN	O	O
the	NN	O	O
action	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

Inhibition	NN	O	O
of	NN	O	O
GM-CSF	NN	O	B-protein
expression	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
can	NN	O	O
occur	NN	O	O
independently	NN	O	O
of	NN	O	O
interleukin	NN	O	B-protein
2	NN	O	I-protein
regulation	NN	O	O
and	NN	O	O
is	NN	O	O
probably	NN	O	O
mediated	NN	O	O
through	NN	O	O
cellular	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
receptors	NN	O	I-protein
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
may	NN	O	O
be	NN	O	O
important	NN	O	O
in	NN	O	O
the	NN	O	O
physiology	NN	O	O
of	NN	O	O
hematopoiesis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Altered	NN	O	O
interaction	NN	O	O
between	NN	O	O
triiodothyronine	NN	O	O
and	NN	O	O
its	NN	O	O
nuclear	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
absence	NN	O	O
of	NN	O	O
cortisol	NN	O	O
:	NN	O	O
a	NN	O	O
proposed	NN	O	O
mechanism	NN	O	O
for	NN	O	O
increased	NN	O	O
thyrotropin	NN	O	O
secretion	NN	O	O
in	NN	O	O
corticosteroid	NN	O	O
deficiency	NN	O	O
states	NN	O	O
.	NN	O	O

Thyroid	NN	O	O
hormones	NN	O	O
occasionally	NN	O	O
appear	NN	O	O
less	NN	O	O
effective	NN	O	O
when	NN	O	O
administered	NN	O	O
alone	NN	O	O
to	NN	O	O
patients	NN	O	O
with	NN	O	O
panhypopituitarism	NN	O	O
,	NN	O	O
and	NN	O	O
manifestations	NN	O	O
suggestive	NN	O	O
of	NN	O	O
hypothyroidism	NN	O	O
have	NN	O	O
been	NN	O	O
reported	NN	O	O
in	NN	O	O
patients	NN	O	O
suffering	NN	O	O
from	NN	O	O
untreated	NN	O	O
Addison	NN	O	O
's	NN	O	O
disease	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
latter	NN	O	O
condition	NN	O	O
,	NN	O	O
thyrotropin	NN	O	O
secretion	NN	O	O
is	NN	O	O
increased	NN	O	O
:	NN	O	O
this	NN	O	O
occurs	NN	O	O
already	NN	O	O
after	NN	O	O
as	NN	O	O
little	NN	O	O
as	NN	O	O
2	NN	O	O
days	NN	O	O
of	NN	O	O
temporary	NN	O	O
withdrawal	NN	O	O
of	NN	O	O
therapy	NN	O	O
with	NN	O	O
substitution	NN	O	O
doses	NN	O	O
of	NN	O	O
corticosteroids	NN	O	O
while	NN	O	O
circulating	NN	O	O
levels	NN	O	O
of	NN	O	O
thyroid	NN	O	O
hormones	NN	O	O
remain	NN	O	O
within	NN	O	O
normal	NN	O	O
limits	NN	O	O
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
a	NN	O	O
possible	NN	O	O
role	NN	O	O
of	NN	O	O
cortisol	NN	O	O
in	NN	O	O
interaction	NN	O	O
between	NN	O	O
triiodothyronine	NN	O	O
and	NN	O	O
its	NN	O	O
nuclear	NN	O	B-protein
receptors	NN	O	I-protein
was	NN	O	O
examined	NN	O	O
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
circulating	NN	O	O
lymphocytes	NN	O	O
obtained	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
primary	NN	O	O
or	NN	O	O
secondary	NN	O	O
adrenocortical	NN	O	O
failure	NN	O	O
.	NN	O	O

The	NN	O	O
affinity	NN	O	O
of	NN	O	O
these	NN	O	O
receptors	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
decreased	NN	O	O
,	NN	O	O
by	NN	O	O
more	NN	O	O
than	NN	O	O
50	NN	O	O
%	NN	O	O
on	NN	O	O
average	NN	O	O
,	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
cortisol	NN	O	O
treatments	NN	O	O
.	NN	O	O

This	NN	O	O
change	NN	O	O
was	NN	O	O
promptly	NN	O	O
corrected	NN	O	O
upon	NN	O	O
resumption	NN	O	O
of	NN	O	O
therapy	NN	O	O
.	NN	O	O

The	NN	O	O
number	NN	O	O
of	NN	O	O
binding	NN	O	O
sites	NN	O	O
was	NN	O	O
not	NN	O	O
significantly	NN	O	O
modified	NN	O	O
.	NN	O	O

The	NN	O	O
influence	NN	O	O
of	NN	O	O
cortisol	NN	O	O
on	NN	O	O
thyroid	NN	O	B-protein
hormone	NN	O	I-protein
receptors	NN	O	I-protein
discussed	NN	O	O
here	NN	O	O
might	NN	O	O
account	NN	O	O
for	NN	O	O
the	NN	O	O
clinical	NN	O	O
observations	NN	O	O
mentioned	NN	O	O
above	NN	O	O
.	NN	O	O

-DOCSTART-	O

Inhibition	NN	O	O
by	NN	O	O
cortisol	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
natural	NN	O	I-cell_type
killer	NN	O	I-cell_type
(	NN	O	I-cell_type
NK	NN	O	I-cell_type
)	NN	O	I-cell_type
cell	NN	O	I-cell_type
activity	NN	O	O
.	NN	O	O

The	NN	O	O
effects	NN	O	O
of	NN	O	O
cortisol	NN	O	O
on	NN	O	O
the	NN	O	O
natural	NN	O	B-cell_type
killer	NN	O	I-cell_type
(	NN	O	O
NK	NN	O	O
)	NN	O	O
activity	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
(	NN	O	I-cell_type
PBM	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
studied	NN	O	O
in	NN	O	O
vitro	NN	O	O
using	NN	O	O
a	NN	O	O
direct	NN	O	O
4-h	NN	O	O
51Cr-release	NN	O	O
assay	NN	O	O
and	NN	O	O
K	NN	O	B-cell_line
562	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
as	NN	O	O
a	NN	O	O
target	NN	O	O
.	NN	O	O

Preincubation	NN	O	O
for	NN	O	O
20	NN	O	O
h	NN	O	O
of	NN	O	O
PBM	NN	O	B-cell_type
cells	NN	O	I-cell_type
drawn	NN	O	O
from	NN	O	O
healthy	NN	O	O
donors	NN	O	O
with	NN	O	O
1	NN	O	O
X	NN	O	O
10	NN	O	O
(	NN	O	O
-8	NN	O	O
)	NN	O	O
to	NN	O	O
1	NN	O	O
X	NN	O	O
10	NN	O	O
(	NN	O	O
-5	NN	O	O
)	NN	O	O
M	NN	O	O
cortisol	NN	O	O
resulted	NN	O	O
in	NN	O	O
a	NN	O	O
significant	NN	O	O
decrease	NN	O	O
of	NN	O	O
NK	NN	O	B-cell_type
cell	NN	O	I-cell_type
activity	NN	O	O
.	NN	O	O

The	NN	O	O
magnitude	NN	O	O
of	NN	O	O
the	NN	O	O
suppression	NN	O	O
was	NN	O	O
directly	NN	O	O
related	NN	O	O
to	NN	O	O
the	NN	O	O
steroid	NN	O	O
concentration	NN	O	O
and	NN	O	O
inversely	NN	O	O
related	NN	O	O
to	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
effector	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Cortisol	NN	O	O
was	NN	O	O
able	NN	O	O
to	NN	O	O
minimize	NN	O	O
the	NN	O	O
enhancement	NN	O	O
of	NN	O	O
NK	NN	O	O
cytotoxicity	NN	O	O
obtainable	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
immune	NN	O	O
interferon	NN	O	O
(	NN	O	O
IFN-gamma	NN	O	O
)	NN	O	O
.	NN	O	O

A	NN	O	O
significantly	NN	O	O
higher	NN	O	O
suppression	NN	O	O
was	NN	O	O
achieved	NN	O	O
after	NN	O	O
sequential	NN	O	O
exposure	NN	O	O
of	NN	O	O
PBM	NN	O	B-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
cortisol	NN	O	O
and	NN	O	O
equimolar	NN	O	O
levels	NN	O	O
of	NN	O	O
prostaglandin	NN	O	O
E2	NN	O	O
(	NN	O	O
PgE2	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
concomitant	NN	O	O
incubation	NN	O	O
with	NN	O	O
theophylline	NN	O	O
and	NN	O	O
isobutyl-methylxanthine	NN	O	O
failed	NN	O	O
to	NN	O	O
enhance	NN	O	O
the	NN	O	O
cortisol-induced	NN	O	O
suppression	NN	O	O
,	NN	O	O
whereas	NN	O	O
PgE2-dependent	NN	O	O
inhibition	NN	O	O
significantly	NN	O	O
increased	NN	O	O
after	NN	O	O
exposure	NN	O	O
of	NN	O	O
PBM	NN	O	B-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
methyl-xanthines	NN	O	O
.	NN	O	O

The	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
of	NN	O	O
cortisol	NN	O	O
was	NN	O	O
partially	NN	O	O
or	NN	O	O
totally	NN	O	O
prevented	NN	O	O
by	NN	O	O
the	NN	O	O
concomitant	NN	O	O
incubation	NN	O	O
with	NN	O	O
equimolar	NN	O	O
amounts	NN	O	O
of	NN	O	O
11-deoxycortisol	NN	O	O
and	NN	O	O
RU	NN	O	O
486	NN	O	O
but	NN	O	O
not	NN	O	O
of	NN	O	O
progesterone	NN	O	O
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
NK	NN	O	B-cell_type
effectors	NN	O	I-cell_type
with	NN	O	O
a	NN	O	O
monoclonal	NN	O	B-protein
anti-human	NN	O	I-protein
corticosteroid-binding	NN	O	I-protein
globulin	NN	O	I-protein
(	NN	O	I-protein
CBG	NN	O	I-protein
)	NN	O	I-protein
antibody	NN	O	I-protein
produced	NN	O	O
an	NN	O	O
enhancement	NN	O	O
of	NN	O	O
the	NN	O	O
spontaneous	NN	O	O
NK	NN	O	B-cell_type
activity	NN	O	O
and	NN	O	O
a	NN	O	O
partial	NN	O	O
suppression	NN	O	O
of	NN	O	O
cortisol-mediated	NN	O	O
effects	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
endogenous	NN	O	O
glucocorticoids	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
NK	NN	O	O
cell-mediated	NN	O	O
cytotoxicity	NN	O	O
.	NN	O	O

Since	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
cortisol	NN	O	O
was	NN	O	O
additive	NN	O	O
to	NN	O	O
that	NN	O	O
of	NN	O	O
PgE2	NN	O	O
and	NN	O	O
was	NN	O	O
not	NN	O	O
changed	NN	O	O
by	NN	O	O
phosphodiesterase	NN	O	B-protein
inhibitors	NN	O	O
,	NN	O	O
it	NN	O	O
is	NN	O	O
conceivable	NN	O	O
that	NN	O	O
the	NN	O	O
hormone	NN	O	O
acts	NN	O	O
at	NN	O	O
a	NN	O	O
level	NN	O	O
different	NN	O	O
from	NN	O	O
the	NN	O	O
adenylate	NN	O	B-protein
cyclase	NN	O	I-protein
-	NN	O	O
phosphodiesterase	NN	O	B-protein
system	NN	O	O
.	NN	O	O

Data	NN	O	O
obtained	NN	O	O
with	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
antiglucocorticoids	NN	O	O
and	NN	O	O
the	NN	O	O
anti-CBG	NN	O	B-protein
antibody	NN	O	I-protein
are	NN	O	O
compatible	NN	O	O
with	NN	O	O
a	NN	O	O
role	NN	O	O
both	NN	O	O
of	NN	O	O
high-affinity	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
and	NN	O	O
of	NN	O	O
CBG	NN	O	B-protein
in	NN	O	O
mediating	NN	O	O
cortisol	NN	O	O
action	NN	O	O
on	NN	O	O
the	NN	O	O
human	NN	O	B-cell_type
NK	NN	O	I-cell_type
cell	NN	O	I-cell_type
activity	NN	O	O
.	NN	O	O

-DOCSTART-	O

Interaction	NN	O	O
of	NN	O	O
cell-type-specific	NN	O	B-protein
nuclear	NN	O	I-protein
proteins	NN	O	I-protein
with	NN	O	O
immunoglobulin	NN	O	B-DNA
VH	NN	O	I-DNA
promoter	NN	O	I-DNA
region	NN	O	I-DNA
sequences	NN	O	I-DNA
.	NN	O	O

All	NN	O	O
human	NN	O	B-DNA
and	NN	O	I-DNA
murine	NN	O	I-DNA
immunoglobulin	NN	O	I-DNA
heavy	NN	O	I-DNA
chain	NN	O	I-DNA
variable	NN	O	I-DNA
region	NN	O	I-DNA
(	NN	O	I-DNA
VH	NN	O	I-DNA
)	NN	O	I-DNA
genes	NN	O	I-DNA
contain	NN	O	O
the	NN	O	O
sequence	NN	O	O
ATGCAAAT	NN	O	O
approximately	NN	O	O
70	NN	O	O
nucleotides	NN	O	O
5	NN	O	O
'	NN	O	O
from	NN	O	O
the	NN	O	O
site	NN	O	O
of	NN	O	O
transcription	NN	O	O
initiation	NN	O	O
.	NN	O	O

This	NN	O	O
octanucleotide	NN	O	O
,	NN	O	O
in	NN	O	O
reverse	NN	O	O
orientation	NN	O	O
,	NN	O	O
is	NN	O	O
also	NN	O	O
found	NN	O	O
in	NN	O	O
all	NN	O	O
light	NN	O	B-DNA
chain	NN	O	I-DNA
variable	NN	O	I-DNA
region	NN	O	I-DNA
(	NN	O	I-DNA
VL	NN	O	I-DNA
)	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
in	NN	O	O
the	NN	O	O
immunoglobulin	NN	O	B-DNA
heavy	NN	O	I-DNA
chain	NN	O	I-DNA
transcriptional	NN	O	I-DNA
enhancer	NN	O	I-DNA
.	NN	O	O

Transfection	NN	O	O
studies	NN	O	O
have	NN	O	O
established	NN	O	O
that	NN	O	O
this	NN	O	O
octamer	NN	O	O
is	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
lymphoid-specific	NN	O	O
transcription	NN	O	O
of	NN	O	O
immunoglobulin	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

Octamer-containing	NN	O	B-DNA
fragments	NN	O	I-DNA
have	NN	O	O
been	NN	O	O
reported	NN	O	O
to	NN	O	O
bind	NN	O	O
a	NN	O	O
factor	NN	O	O
present	NN	O	O
in	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
of	NN	O	O
human	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
;	NN	O	O
however	NN	O	O
,	NN	O	O
identical	NN	O	O
binding	NN	O	O
activity	NN	O	O
was	NN	O	O
detected	NN	O	O
in	NN	O	O
both	NN	O	O
B	NN	O	B-cell_line
lymphoid	NN	O	I-cell_line
and	NN	O	I-cell_line
non-lymphoid	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Here	NN	O	O
we	NN	O	O
establish	NN	O	O
that	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
from	NN	O	O
distinct	NN	O	O
cell	NN	O	O
types	NN	O	O
differ	NN	O	O
in	NN	O	O
their	NN	O	O
ability	NN	O	O
to	NN	O	O
interact	NN	O	O
with	NN	O	O
octamer-containing	NN	O	B-DNA
fragments	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
have	NN	O	O
also	NN	O	O
detected	NN	O	O
a	NN	O	O
DNA-protein	NN	O	O
interaction	NN	O	O
that	NN	O	O
may	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
cell-type	NN	O	O
specificity	NN	O	O
of	NN	O	O
immunoglobulin	NN	O	B-protein
expression	NN	O	O
,	NN	O	O
and	NN	O	O
we	NN	O	O
have	NN	O	O
determined	NN	O	O
that	NN	O	O
a	NN	O	O
sequence	NN	O	O
upstream	NN	O	O
of	NN	O	O
the	NN	O	O
octamer	NN	O	O
participates	NN	O	O
in	NN	O	O
an	NN	O	O
interaction	NN	O	O
with	NN	O	O
a	NN	O	O
nuclear	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	O
s	NN	O	O
)	NN	O	O
.	NN	O	O

-DOCSTART-	O

Preferential	NN	O	O
transcription	NN	O	O
of	NN	O	O
HTLV-I	NN	O	B-DNA
LTR	NN	O	I-DNA
in	NN	O	O
cell-free	NN	O	O
extracts	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
producing	NN	O	O
HTLV-I	NN	O	B-protein
viral	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

The	NN	O	O
promoters	NN	O	B-DNA
of	NN	O	O
the	NN	O	O
adenovirus	NN	O	B-DNA
2	NN	O	I-DNA
major	NN	O	I-DNA
late	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
the	NN	O	O
mouse	NN	O	B-DNA
beta-globin	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
the	NN	O	O
mouse	NN	O	B-DNA
immunoglobulin	NN	O	I-DNA
VH	NN	O	I-DNA
gene	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
LTR	NN	O	B-DNA
of	NN	O	O
the	NN	O	O
human	NN	O	O
T-lymphotropic	NN	O	O
retrovirus	NN	O	O
type	NN	O	O
I	NN	O	O
were	NN	O	O
tested	NN	O	O
for	NN	O	O
their	NN	O	O
transcription	NN	O	O
activities	NN	O	O
in	NN	O	O
cell-free	NN	O	O
extracts	NN	O	O
of	NN	O	O
four	NN	O	O
cell	NN	O	O
lines	NN	O	O
;	NN	O	O
HeLa	NN	O	B-cell_line
,	NN	O	O
CESS	NN	O	B-cell_line
(	NN	O	O
Epstein-Barr	NN	O	B-cell_line
virus-transformed	NN	O	I-cell_line
human	NN	O	I-cell_line
B	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
)	NN	O	O
,	NN	O	O
MT-1	NN	O	B-cell_line
(	NN	O	O
HTLV-I-infected	NN	O	B-cell_line
human	NN	O	I-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
without	NN	O	O
viral	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
MT-2	NN	O	B-cell_line
(	NN	O	O
HTLV-I-infected	NN	O	B-cell_line
human	NN	O	I-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
producing	NN	O	O
viral	NN	O	B-protein
proteins	NN	O	I-protein
)	NN	O	O
.	NN	O	O

LTR	NN	O	B-DNA
was	NN	O	O
preferentially	NN	O	O
transcribed	NN	O	O
in	NN	O	O
the	NN	O	O
extracts	NN	O	O
of	NN	O	O
MT-2	NN	O	B-cell_line
although	NN	O	O
the	NN	O	O
other	NN	O	O
three	NN	O	O
genes	NN	O	O
were	NN	O	O
transcribed	NN	O	O
with	NN	O	O
relatively	NN	O	O
constant	NN	O	O
efficiencies	NN	O	O
in	NN	O	O
different	NN	O	O
extracts	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
agree	NN	O	O
well	NN	O	O
with	NN	O	O
the	NN	O	O
previous	NN	O	O
in	NN	O	O
vivo	NN	O	O
studies	NN	O	O
on	NN	O	O
the	NN	O	O
promoter	NN	O	O
activity	NN	O	O
of	NN	O	O
HTLV-I	NN	O	B-DNA
LTR	NN	O	I-DNA
.	NN	O	O

Mixing	NN	O	O
of	NN	O	O
HeLa	NN	O	O
and	NN	O	O
MT-2	NN	O	O
extracts	NN	O	O
revealed	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
a	NN	O	O
LTR-specific	NN	O	O
stimulating	NN	O	O
activity	NN	O	O
in	NN	O	O
MT-2	NN	O	O
extracts	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
that	NN	O	O
binds	NN	O	O
to	NN	O	O
a	NN	O	O
conserved	NN	O	O
sequence	NN	O	O
motif	NN	O	O
in	NN	O	O
transcriptional	NN	O	B-DNA
control	NN	O	I-DNA
elements	NN	O	I-DNA
of	NN	O	O
immunoglobulin	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

Trans-acting	NN	O	B-protein
factors	NN	O	I-protein
that	NN	O	O
mediate	NN	O	O
B-cell	NN	O	O
specific	NN	O	O
transcription	NN	O	O
of	NN	O	O
immunoglobulin	NN	O	B-DNA
genes	NN	O	I-DNA
have	NN	O	O
been	NN	O	O
postulated	NN	O	O
based	NN	O	O
on	NN	O	O
an	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
exogenously	NN	O	O
introduced	NN	O	O
immunoglobulin	NN	O	B-DNA
gene	NN	O	I-DNA
recombinants	NN	O	I-DNA
in	NN	O	O
lymphoid	NN	O	B-cell_type
and	NN	O	I-cell_type
non-lymphoid	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Two	NN	O	O
B-cell-specific	NN	O	O
,	NN	O	O
cis-acting	NN	O	B-DNA
transcriptional	NN	O	I-DNA
regulatory	NN	O	I-DNA
elements	NN	O	I-DNA
have	NN	O	O
been	NN	O	O
identified	NN	O	O
.	NN	O	O

One	NN	O	O
element	NN	O	O
is	NN	O	O
located	NN	O	O
in	NN	O	O
the	NN	O	O
intron	NN	O	O
between	NN	O	O
the	NN	O	O
variable	NN	O	O
(	NN	O	O
V	NN	O	O
)	NN	O	O
and	NN	O	O
constant	NN	O	O
(	NN	O	O
C	NN	O	O
)	NN	O	O
regions	NN	O	O
of	NN	O	O
both	NN	O	O
heavy	NN	O	B-DNA
and	NN	O	I-DNA
kappa	NN	O	I-DNA
light-chain	NN	O	I-DNA
genes	NN	O	I-DNA
and	NN	O	O
acts	NN	O	O
as	NN	O	O
a	NN	O	O
transcriptional	NN	O	O
enhancer	NN	O	O
.	NN	O	O

The	NN	O	O
second	NN	O	O
element	NN	O	B-DNA
is	NN	O	O
found	NN	O	O
upstream	NN	O	O
of	NN	O	O
both	NN	O	O
heavy	NN	O	B-DNA
and	NN	O	I-DNA
kappa	NN	O	I-DNA
light-chain	NN	O	I-DNA
gene	NN	O	I-DNA
promoters	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
element	NN	O	O
directs	NN	O	O
lymphoid-specific	NN	O	O
transcription	NN	O	O
even	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
viral	NN	O	B-DNA
enhancers	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
have	NN	O	O
sought	NN	O	O
nuclear	NN	O	O
factors	NN	O	O
that	NN	O	O
might	NN	O	O
bind	NN	O	O
specifically	NN	O	O
to	NN	O	O
these	NN	O	O
two	NN	O	O
regulatory	NN	O	B-DNA
elements	NN	O	I-DNA
by	NN	O	O
application	NN	O	O
of	NN	O	O
a	NN	O	O
modified	NN	O	O
gel	NN	O	O
electrophoresis	NN	O	O
DNA	NN	O	O
binding	NN	O	O
assay	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
here	NN	O	O
the	NN	O	O
identification	NN	O	O
of	NN	O	O
a	NN	O	O
human	NN	O	B-protein
B-cell	NN	O	I-protein
nuclear	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
IgNF-A	NN	O	B-protein
)	NN	O	O
that	NN	O	O
binds	NN	O	O
to	NN	O	O
DNA	NN	O	B-DNA
sequences	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
upstream	NN	O	B-DNA
regions	NN	O	I-DNA
of	NN	O	O
both	NN	O	O
the	NN	O	O
mouse	NN	O	B-DNA
heavy	NN	O	I-DNA
and	NN	O	I-DNA
kappa	NN	O	I-DNA
light-chain	NN	O	I-DNA
gene	NN	O	I-DNA
promoters	NN	O	I-DNA
and	NN	O	O
also	NN	O	O
to	NN	O	O
the	NN	O	O
mouse	NN	O	B-DNA
heavy-chain	NN	O	I-DNA
gene	NN	O	I-DNA
enhancer	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
sequence-specific	NN	O	O
binding	NN	O	O
is	NN	O	O
probably	NN	O	O
mediated	NN	O	O
by	NN	O	O
a	NN	O	O
highly	NN	O	O
conserved	NN	O	O
sequence	NN	O	O
motif	NN	O	O
,	NN	O	O
ATTTGCAT	NN	O	O
,	NN	O	O
present	NN	O	O
in	NN	O	O
all	NN	O	O
three	NN	O	O
transcriptional	NN	O	B-DNA
elements	NN	O	I-DNA
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
a	NN	O	O
factor	NN	O	O
showing	NN	O	O
similar	NN	O	O
binding	NN	O	O
specificity	NN	O	O
to	NN	O	O
IgNF-A	NN	O	B-DNA
is	NN	O	O
also	NN	O	O
present	NN	O	O
in	NN	O	O
human	NN	O	B-cell_line
HeLa	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
new	NN	O	O
world	NN	O	O
primates	NN	O	O
as	NN	O	O
animal	NN	O	O
models	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	O
resistance	NN	O	O
.	NN	O	O

Many	NN	O	O
New	NN	O	O
World	NN	O	O
primate	NN	O	O
species	NN	O	O
have	NN	O	O
greatly	NN	O	O
increased	NN	O	O
plasma	NN	O	O
cortisol	NN	O	O
concentrations	NN	O	O
,	NN	O	O
decreased	NN	O	O
plasma	NN	O	O
cortisol	NN	O	O
binding	NN	O	O
globulin	NN	O	O
capacity	NN	O	O
and	NN	O	O
affinity	NN	O	O
,	NN	O	O
marked	NN	O	O
resistance	NN	O	O
of	NN	O	O
the	NN	O	O
hypothalamic-pituitary-adrenal	NN	O	O
axis	NN	O	O
to	NN	O	O
suppression	NN	O	O
by	NN	O	O
dexamethasone	NN	O	O
,	NN	O	O
and	NN	O	O
no	NN	O	O
biological	NN	O	O
evidence	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	O
excess	NN	O	O
.	NN	O	O

These	NN	O	O
primates	NN	O	O
also	NN	O	O
have	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
circulating	NN	O	O
progesterone	NN	O	O
,	NN	O	O
estrogen	NN	O	O
,	NN	O	O
mineralocorticoid	NN	O	O
,	NN	O	O
androgen	NN	O	O
and	NN	O	O
vitamin	NN	O	O
D	NN	O	O
.	NN	O	O

The	NN	O	O
glucocorticoid	NN	O	O
target	NN	O	O
tissues	NN	O	O
that	NN	O	O
have	NN	O	O
been	NN	O	O
examined	NN	O	O
(	NN	O	O
circulating	NN	O	B-cell_type
mononuclear	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
cultured	NN	O	B-cell_line
skin	NN	O	I-cell_line
fibroblasts	NN	O	I-cell_line
)	NN	O	O
have	NN	O	O
normal	NN	O	O
concentrations	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
with	NN	O	O
decreased	NN	O	O
affinity	NN	O	O
for	NN	O	O
dexamethasone	NN	O	O
.	NN	O	O

Transformation	NN	O	O
of	NN	O	O
B-lymphocytes	NN	O	B-cell_type
with	NN	O	O
the	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
leads	NN	O	O
to	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
induction	NN	O	O
that	NN	O	O
is	NN	O	O
less	NN	O	O
than	NN	O	O
that	NN	O	O
observed	NN	O	O
with	NN	O	O
cells	NN	O	O
from	NN	O	O
Old	NN	O	O
World	NN	O	O
primates	NN	O	O
.	NN	O	O

The	NN	O	O
receptor	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
has	NN	O	O
a	NN	O	O
low	NN	O	O
affinity	NN	O	O
for	NN	O	O
dexamethasone	NN	O	O
.	NN	O	O

The	NN	O	O
low	NN	O	O
affinity	NN	O	O
leads	NN	O	O
to	NN	O	O
an	NN	O	O
increased	NN	O	O
loss	NN	O	O
of	NN	O	O
specific	NN	O	O
bound	NN	O	O
ligand	NN	O	O
during	NN	O	O
thermal	NN	O	O
activation	NN	O	O
.	NN	O	O

Meroreceptor	NN	O	O
generation	NN	O	O
is	NN	O	O
normal	NN	O	O
.	NN	O	O

The	NN	O	O
molecular	NN	O	O
weight	NN	O	O
of	NN	O	O
the	NN	O	O
receptor	NN	O	O
,	NN	O	O
determined	NN	O	O
by	NN	O	O
SDS-PAGE	NN	O	O
,	NN	O	O
is	NN	O	O
similar	NN	O	O
to	NN	O	O
that	NN	O	O
of	NN	O	O
Old	NN	O	O
World	NN	O	O
primates	NN	O	O
(	NN	O	O
approximately	NN	O	O
92	NN	O	O
,	NN	O	O
000	NN	O	O
)	NN	O	O
and	NN	O	O
the	NN	O	O
activation	NN	O	O
pattern	NN	O	O
per	NN	O	O
se	NN	O	O
,	NN	O	O
examined	NN	O	O
in	NN	O	O
vitro	NN	O	O
by	NN	O	O
heating	NN	O	O
cytosol	NN	O	O
and	NN	O	O
performing	NN	O	O
phosphocellulose	NN	O	O
chromatography	NN	O	O
,	NN	O	O
appears	NN	O	O
similar	NN	O	O
to	NN	O	O
that	NN	O	O
of	NN	O	O
human	NN	O	O
controls	NN	O	O
.	NN	O	O

The	NN	O	O
ratios	NN	O	O
of	NN	O	O
nuclear	NN	O	O
to	NN	O	O
cytosolic	NN	O	B-protein
hormone-receptor-complexes	NN	O	I-protein
and	NN	O	O
of	NN	O	O
cytosolic	NN	O	O
activated	NN	O	O
to	NN	O	O
unactivated	NN	O	B-protein
receptor	NN	O	I-protein
complexes	NN	O	I-protein
in	NN	O	O
intact	NN	O	O
cells	NN	O	O
are	NN	O	O
similar	NN	O	O
to	NN	O	O
Old	NN	O	O
World	NN	O	O
primates	NN	O	O
.	NN	O	O

Results	NN	O	O
from	NN	O	O
mixing	NN	O	O
studies	NN	O	O
do	NN	O	O
not	NN	O	O
support	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
a	NN	O	O
binding	NN	O	O
inhibitor	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
or	NN	O	O
a	NN	O	O
deficient	NN	O	O
cytosolic	NN	O	O
positive	NN	O	O
modifier	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
of	NN	O	O
binding	NN	O	O
underlies	NN	O	O
the	NN	O	O
findings	NN	O	O
in	NN	O	O
these	NN	O	O
primates	NN	O	O
.	NN	O	O

The	NN	O	O
New	NN	O	O
World	NN	O	O
primates	NN	O	O
,	NN	O	O
unlike	NN	O	O
men	NN	O	O
with	NN	O	O
the	NN	O	O
syndrome	NN	O	O
of	NN	O	O
primary	NN	O	O
cortisol	NN	O	O
resistance	NN	O	O
,	NN	O	O
have	NN	O	O
compensated	NN	O	O
for	NN	O	O
their	NN	O	O
condition	NN	O	O
with	NN	O	O
intra-adrenal	NN	O	O
and	NN	O	O
mineralocorticoid	NN	O	O
receptor	NN	O	O
adaptations	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
unlike	NN	O	O
Old	NN	O	O
World	NN	O	O
primates	NN	O	O
,	NN	O	O
cortisol	NN	O	O
in	NN	O	O
New	NN	O	O
World	NN	O	O
primates	NN	O	O
has	NN	O	O
only	NN	O	O
weak	NN	O	O
sodium-retaining	NN	O	O
potency	NN	O	O
because	NN	O	O
the	NN	O	O
aldosterone	NN	O	B-protein
receptor	NN	O	I-protein
has	NN	O	O
a	NN	O	O
low	NN	O	O
affinity	NN	O	O
for	NN	O	O
cortisol	NN	O	O
.	NN	O	O

The	NN	O	O
common	NN	O	O
element	NN	O	O
that	NN	O	O
would	NN	O	O
explain	NN	O	O
the	NN	O	O
apparent	NN	O	O
resistance	NN	O	O
to	NN	O	O
six	NN	O	O
steroid	NN	O	O
hormones	NN	O	O
in	NN	O	O
New	NN	O	O
World	NN	O	O
primates	NN	O	O

-DOCSTART-	O

Acetylation	NN	O	O
and	NN	O	O
modulation	NN	O	O
of	NN	O	O
erythroid	NN	O	B-protein
Kruppel-like	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
EKLF	NN	O	B-protein
)	NN	O	O
activity	NN	O	O
by	NN	O	O
interaction	NN	O	O
with	NN	O	O
histone	NN	O	B-protein
acetyltransferases	NN	O	I-protein
.	NN	O	O

Erythroid	NN	O	B-protein
Kruppel-like	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
EKLF	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
red	NN	O	B-protein
cell-specific	NN	O	I-protein
transcriptional	NN	O	I-protein
activator	NN	O	I-protein
that	NN	O	O
is	NN	O	O
crucial	NN	O	O
for	NN	O	O
consolidating	NN	O	O
the	NN	O	O
switch	NN	O	O
to	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
adult	NN	O	O
beta-globin	NN	O	O
expression	NN	O	O
during	NN	O	O
erythroid	NN	O	O
ontogeny	NN	O	O
.	NN	O	O

EKLF	NN	O	B-protein
is	NN	O	O
required	NN	O	O
for	NN	O	O
integrity	NN	O	O
of	NN	O	O
the	NN	O	O
chromatin	NN	O	O
structure	NN	O	O
at	NN	O	O
the	NN	O	O
beta-like	NN	O	B-DNA
globin	NN	O	I-DNA
locus	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
it	NN	O	O
interacts	NN	O	O
with	NN	O	O
a	NN	O	O
positive-acting	NN	O	B-protein
factor	NN	O	I-protein
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

We	NN	O	O
find	NN	O	O
that	NN	O	O
EKLF	NN	O	B-protein
is	NN	O	O
an	NN	O	O
acetylated	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
and	NN	O	O
that	NN	O	O
it	NN	O	O
interacts	NN	O	O
in	NN	O	O
vivo	NN	O	O
with	NN	O	O
CBP	NN	O	B-protein
,	NN	O	O
p300	NN	O	B-protein
,	NN	O	O
and	NN	O	O
P/CAF	NN	O	B-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
its	NN	O	O
interactions	NN	O	O
with	NN	O	O
these	NN	O	O
histone	NN	O	B-protein
acetyltransferases	NN	O	I-protein
are	NN	O	O
not	NN	O	O
equivalent	NN	O	O
,	NN	O	O
as	NN	O	O
CBP	NN	O	B-protein
and	NN	O	O
p300	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
P/CAF	NN	O	B-protein
,	NN	O	O
utilize	NN	O	O
EKLF	NN	O	B-protein
as	NN	O	O
a	NN	O	O
substrate	NN	O	O
for	NN	O	O
in	NN	O	O
vitro	NN	O	O
acetylation	NN	O	O
within	NN	O	O
its	NN	O	O
trans-activation	NN	O	O
region	NN	O	O
.	NN	O	O

The	NN	O	O
functional	NN	O	O
effects	NN	O	O
of	NN	O	O
these	NN	O	O
interactions	NN	O	O
are	NN	O	O
that	NN	O	O
CBP	NN	O	B-protein
and	NN	O	O
p300	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
P/CAF	NN	O	B-protein
,	NN	O	O
enhance	NN	O	O
EKLF	NN	O	B-protein
's	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
beta-globin	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
erythroid	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
results	NN	O	O
establish	NN	O	O
EKLF	NN	O	B-protein
as	NN	O	O
a	NN	O	O
tissue-specific	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
that	NN	O	O
undergoes	NN	O	O
post-translational	NN	O	O
acetylation	NN	O	O
and	NN	O	O
suggest	NN	O	O
a	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
EKLF	NN	O	B-protein
is	NN	O	O
able	NN	O	O
to	NN	O	O
alter	NN	O	O
chromatin	NN	O	O
structure	NN	O	O
and	NN	O	O
induce	NN	O	O
beta-globin	NN	O	O
expression	NN	O	O
within	NN	O	O
the	NN	O	O
beta-like	NN	O	B-DNA
globin	NN	O	I-DNA
cluster	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Recognition	NN	O	O
of	NN	O	O
herpes	NN	O	B-protein
simplex	NN	O	I-protein
virus	NN	O	I-protein
type	NN	O	I-protein
2	NN	O	I-protein
tegument	NN	O	I-protein
proteins	NN	O	I-protein
by	NN	O	O
CD4	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
infiltrating	NN	O	O
human	NN	O	O
genital	NN	O	O
herpes	NN	O	O
lesions	NN	O	O
.	NN	O	O

The	NN	O	O
local	NN	O	O
cellular	NN	O	O
immune	NN	O	O
response	NN	O	O
to	NN	O	O
herpes	NN	O	O
simplex	NN	O	O
virus	NN	O	O
(	NN	O	O
HSV	NN	O	O
)	NN	O	O
is	NN	O	O
important	NN	O	O
in	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
recurrent	NN	O	O
HSV	NN	O	O
infection	NN	O	O
.	NN	O	O

The	NN	O	O
antiviral	NN	O	O
functions	NN	O	O
of	NN	O	O
infiltrating	NN	O	B-cell_type
CD4-bearing	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
may	NN	O	O
include	NN	O	O
cytotoxicity	NN	O	O
,	NN	O	O
inhibition	NN	O	O
of	NN	O	O
viral	NN	O	O
growth	NN	O	O
,	NN	O	O
lymphokine	NN	O	O
secretion	NN	O	O
,	NN	O	O
and	NN	O	O
support	NN	O	O
of	NN	O	O
humoral	NN	O	O
and	NN	O	O
CD8	NN	O	O
responses	NN	O	O
.	NN	O	O

The	NN	O	O
antigens	NN	O	O
recognized	NN	O	O
by	NN	O	O
many	NN	O	O
HSV-specific	NN	O	B-cell_type
CD4	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
localizing	NN	O	O
to	NN	O	O
genital	NN	O	O
HSV-2	NN	O	O
lesions	NN	O	O
are	NN	O	O
unknown	NN	O	O
.	NN	O	O

T	NN	O	B-cell_type
cells	NN	O	I-cell_type
recognizing	NN	O	O
antigens	NN	O	O
encoded	NN	O	O
within	NN	O	O
map	NN	O	O
units	NN	O	O
0.67	NN	O	O
to	NN	O	O
0.73	NN	O	O
of	NN	O	O
HSV	NN	O	B-DNA
DNA	NN	O	I-DNA
are	NN	O	O
frequently	NN	O	O
recovered	NN	O	O
from	NN	O	O
herpetic	NN	O	O
lesions	NN	O	O
.	NN	O	O

Expression	NN	O	O
cloning	NN	O	O
with	NN	O	O
this	NN	O	O
region	NN	O	O
of	NN	O	O
DNA	NN	O	O
now	NN	O	O
shows	NN	O	O
that	NN	O	O
tegument	NN	O	B-protein
protein	NN	O	I-protein
VP22	NN	O	B-protein
and	NN	O	O
the	NN	O	O
viral	NN	O	B-protein
dUTPase	NN	O	I-protein
,	NN	O	O
encoded	NN	O	O
by	NN	O	O
genes	NN	O	B-DNA
UL49	NN	O	I-DNA
and	NN	O	I-DNA
UL50	NN	O	I-DNA
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
are	NN	O	O
T-cell	NN	O	B-protein
antigens	NN	O	I-protein
.	NN	O	O

Separate	NN	O	O
epitopes	NN	O	B-protein
in	NN	O	O
VP22	NN	O	B-protein
were	NN	O	O
defined	NN	O	O
for	NN	O	O
T-cell	NN	O	B-cell_line
clones	NN	O	I-cell_line
from	NN	O	O
each	NN	O	O
of	NN	O	O
three	NN	O	O
patients	NN	O	O
.	NN	O	O

Reactivity	NN	O	O
with	NN	O	O
the	NN	O	O
tegument	NN	O	B-protein
protein	NN	O	I-protein
encoded	NN	O	O
by	NN	O	O
UL21	NN	O	B-DNA
was	NN	O	O
identified	NN	O	O
for	NN	O	O
an	NN	O	O
additional	NN	O	O
patient	NN	O	O
.	NN	O	O

Three	NN	O	O
new	NN	O	O
epitopes	NN	O	O
were	NN	O	O
identified	NN	O	O
in	NN	O	O
VP16	NN	O	B-protein
,	NN	O	O
a	NN	O	O
tegument	NN	O	B-protein
protein	NN	O	I-protein
associated	NN	O	O
with	NN	O	O
VP22	NN	O	B-protein
.	NN	O	O

Some	NN	O	O
tegument-specific	NN	O	B-cell_line
CD4	NN	O	I-cell_line
T-cell	NN	O	I-cell_line
clones	NN	O	I-cell_line
exhibited	NN	O	O
cytotoxic	NN	O	O
activity	NN	O	O
against	NN	O	O
HSV-infected	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
herpes	NN	O	B-protein
simplex	NN	O	I-protein
tegument	NN	O	I-protein
proteins	NN	O	I-protein
are	NN	O	O
processed	NN	O	O
for	NN	O	O
antigen	NN	O	O
presentation	NN	O	O
in	NN	O	O
vivo	NN	O	O
and	NN	O	O
are	NN	O	O
possible	NN	O	O
candidate	NN	O	O
compounds	NN	O	O
for	NN	O	O
herpes	NN	O	O
simplex	NN	O	O
vaccines	NN	O	O
.	NN	O	O

-DOCSTART-	O

Fibrinogen	NN	O	B-protein
activates	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
in	NN	O	O
mononuclear	NN	O	B-cell_type
phagocytes	NN	O	I-cell_type
.	NN	O	O

Adhesion	NN	O	O
to	NN	O	O
extracellular	NN	O	O
matrices	NN	O	O
is	NN	O	O
known	NN	O	O
to	NN	O	O
modulate	NN	O	O
leukocyte	NN	O	O
activation	NN	O	O
,	NN	O	O
although	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
are	NN	O	O
not	NN	O	O
fully	NN	O	O
understood	NN	O	O
.	NN	O	O

Mononuclear	NN	O	B-cell_type
phagocytes	NN	O	I-cell_type
are	NN	O	O
exposed	NN	O	O
to	NN	O	O
fibrinous	NN	O	O
provisional	NN	O	O
matrix	NN	O	O
throughout	NN	O	O
migration	NN	O	O
into	NN	O	O
inflammatory	NN	O	O
foci	NN	O	O
,	NN	O	O
so	NN	O	O
this	NN	O	O
study	NN	O	O
was	NN	O	O
undertaken	NN	O	O
to	NN	O	O
determine	NN	O	O
whether	NN	O	O
fibrinogen	NN	O	O
triggers	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
differentiated	NN	O	O
with	NN	O	O
PMA	NN	O	O
in	NN	O	O
nonadherent	NN	O	B-cell_line
culture	NN	O	I-cell_line
were	NN	O	O
shown	NN	O	O
to	NN	O	O
express	NN	O	O
two	NN	O	O
fibrinogen-binding	NN	O	B-protein
integrins	NN	O	I-protein
,	NN	O	O
predominately	NN	O	O
CD11b/CD18	NN	O	B-protein
,	NN	O	O
and	NN	O	O
to	NN	O	O
a	NN	O	O
lesser	NN	O	O
extent	NN	O	O
,	NN	O	O
CD11c/CD18	NN	O	B-protein
.	NN	O	O

Cells	NN	O	O
stimulated	NN	O	O
with	NN	O	O
fibrinogen	NN	O	B-protein
(	NN	O	O
10-100	NN	O	O
microg/ml	NN	O	O
)	NN	O	O
/Mn2+	NN	O	O
(	NN	O	O
50	NN	O	O
microM	NN	O	O
)	NN	O	O
for	NN	O	O
2	NN	O	O
h	NN	O	O
were	NN	O	O
examined	NN	O	O
by	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
.	NN	O	O

NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
,	NN	O	O
minimal	NN	O	O
in	NN	O	O
unstimulated	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
was	NN	O	O
substantially	NN	O	O
up-regulated	NN	O	O
by	NN	O	O
fibrinogen	NN	O	B-protein
.	NN	O	O

Fibrinogen	NN	O	B-protein
also	NN	O	O
caused	NN	O	O
activation	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
SP1	NN	O	B-protein
or	NN	O	O
cAMP	NN	O	B-protein
response	NN	O	I-protein
element-binding	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	I-protein
CREB	NN	O	I-protein
)	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

Blocking	NN	O	O
mAbs	NN	O	B-protein
against	NN	O	O
CD18	NN	O	B-protein
and	NN	O	O
CD11b	NN	O	B-protein
abrogated	NN	O	O
fibrinogen	NN	O	B-protein
-induced	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

To	NN	O	O
determine	NN	O	O
the	NN	O	O
effects	NN	O	O
on	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
,	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
transfected	NN	O	O
with	NN	O	O
a	NN	O	O
plasmid	NN	O	O
containing	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
enhancer	NN	O	I-DNA
(	NN	O	O
bearing	NN	O	O
two	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
sites	NN	O	I-DNA
)	NN	O	O
coupled	NN	O	O
to	NN	O	O
a	NN	O	O
chloramphenicol	NN	O	B-DNA
acetyltransferase	NN	O	I-DNA
(	NN	O	I-DNA
CAT	NN	O	I-DNA
)	NN	O	I-DNA
reporter	NN	O	I-DNA
.	NN	O	O

Cells	NN	O	O
were	NN	O	O
subsequently	NN	O	O
stimulated	NN	O	O
with	NN	O	O
1	NN	O	O
)	NN	O	O
PMA	NN	O	O
for	NN	O	O
24	NN	O	O
h	NN	O	O
,	NN	O	O
inducing	NN	O	O
CAT	NN	O	O
activity	NN	O	O
by	NN	O	O
2.6-fold	NN	O	O
,	NN	O	O
2	NN	O	O
)	NN	O	O
fibrinogen	NN	O	B-protein
/Mn2+	NN	O	O
for	NN	O	O
2	NN	O	O
h	NN	O	O
,	NN	O	O
inducing	NN	O	O
CAT	NN	O	O
activity	NN	O	O
by	NN	O	O
3.2-fold	NN	O	O
,	NN	O	O
or	NN	O	O
3	NN	O	O
)	NN	O	O
costimulation	NN	O	O
with	NN	O	O
fibrinogen	NN	O	B-protein
and	NN	O	O
PMA	NN	O	O
,	NN	O	O
inducing	NN	O	O
5.7-fold	NN	O	O
the	NN	O	O
CAT	NN	O	O
activity	NN	O	O
induced	NN	O	O
by	NN	O	O
PMA	NN	O	O
alone	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
contact	NN	O	O
with	NN	O	O
fibrinogen-derived	NN	O	B-protein
proteins	NN	O	I-protein
may	NN	O	O
contribute	NN	O	O
to	NN	O	O
mononuclear	NN	O	O
phagocyte	NN	O	O
activation	NN	O	O
by	NN	O	O
signaling	NN	O	O
through	NN	O	O
CD11b/CD18	NN	O	B-protein
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
selective	NN	O	O
activation	NN	O	O
of	NN	O	O
transcriptional	NN	O	B-protein
regulatory	NN	O	I-protein
factors	NN	O	I-protein
,	NN	O	O
including	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
and	NN	O	O
B	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
derived	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
lupus	NN	O	O
express	NN	O	O
estrogen	NN	O	O
receptor	NN	O	O
transcripts	NN	O	O
similar	NN	O	O
to	NN	O	O
those	NN	O	O
of	NN	O	O
normal	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

OBJECTIVE	NN	O	O
:	NN	O	O
To	NN	O	O
identify	NN	O	O
and	NN	O	O
characterize	NN	O	O
estrogen	NN	O	B-RNA
receptor	NN	O	I-RNA
(	NN	O	I-RNA
ER	NN	O	I-RNA
)	NN	O	I-RNA
transcripts	NN	O	I-RNA
expressed	NN	O	O
in	NN	O	O
immune	NN	O	B-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
patients	NN	O	O
with	NN	O	O
systemic	NN	O	O
lupus	NN	O	O
erythematosus	NN	O	O
(	NN	O	O
SLE	NN	O	O
)	NN	O	O
and	NN	O	O
healthy	NN	O	O
donors	NN	O	O
.	NN	O	O

METHODS	NN	O	O
:	NN	O	O
Peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
and	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
prepared	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
SLE	NN	O	O
(	NN	O	O
n	NN	O	O
=	NN	O	O
6	NN	O	O
)	NN	O	O
and	NN	O	O
healthy	NN	O	O
donors	NN	O	O
(	NN	O	O
n	NN	O	O
=	NN	O	O
8	NN	O	O
)	NN	O	O
.	NN	O	O

T	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
separated	NN	O	O
into	NN	O	O
CD4	NN	O	B-cell_type
and	NN	O	O
CD8	NN	O	B-cell_type
.	NN	O	O

Some	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
stimulated	NN	O	O
with	NN	O	O
estradiol	NN	O	O
,	NN	O	O
PMA	NN	O	O
,	NN	O	O
and	NN	O	O
ionomycin	NN	O	O
.	NN	O	O

Epstein-Barr	NN	O	O
virus-transformed	NN	O	O
B	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
(	NN	O	O
n	NN	O	O
=	NN	O	O
7	NN	O	O
)	NN	O	O
and	NN	O	O
B	NN	O	B-cell_line
cell	NN	O	I-cell_line
hybridomas	NN	O	I-cell_line
(	NN	O	O
n	NN	O	O
=	NN	O	O
2	NN	O	O
)	NN	O	O
established	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
SLE	NN	O	O
and	NN	O	O
a	NN	O	O
healthy	NN	O	O
individual	NN	O	O
were	NN	O	O
used	NN	O	O
as	NN	O	O
a	NN	O	O
B	NN	O	O
cell	NN	O	O
source	NN	O	O
.	NN	O	O

These	NN	O	O
cells	NN	O	O
were	NN	O	O
examined	NN	O	O
for	NN	O	O
ER	NN	O	B-RNA
mRNA	NN	O	I-RNA
by	NN	O	O
reverse	NN	O	O
transcription	NN	O	O
nested	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
.	NN	O	O

Amplified	NN	O	O
cDNA	NN	O	B-DNA
were	NN	O	O
sequenced	NN	O	O
by	NN	O	O
standard	NN	O	O
methods	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
In	NN	O	O
all	NN	O	O
cells	NN	O	O
tested	NN	O	O
,	NN	O	O
ER	NN	O	B-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
expressed	NN	O	O
without	NN	O	O
prior	NN	O	O
in	NN	O	O
vitro	NN	O	O
stimulation	NN	O	O
.	NN	O	O

Partial	NN	O	O
sequences	NN	O	O
from	NN	O	O
exons	NN	O	B-DNA
1-8	NN	O	I-DNA
were	NN	O	O
nearly	NN	O	O
identical	NN	O	O
to	NN	O	O
the	NN	O	O
published	NN	O	O
sequence	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-RNA
ER	NN	O	I-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

There	NN	O	O
were	NN	O	O
no	NN	O	O
notable	NN	O	O
differences	NN	O	O
in	NN	O	O
the	NN	O	O
ER	NN	O	B-RNA
transcripts	NN	O	I-RNA
between	NN	O	O
patients	NN	O	O
and	NN	O	O
healthy	NN	O	O
controls	NN	O	O
.	NN	O	O

Variant	NN	O	O
receptor	NN	O	O
transcripts	NN	O	O
lacking	NN	O	O
exon	NN	O	B-DNA
5	NN	O	I-DNA
or	NN	O	O
exon	NN	O	B-DNA
7	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
encodes	NN	O	O
the	NN	O	O
hormone	NN	O	B-protein
binding	NN	O	I-protein
domain	NN	O	I-protein
,	NN	O	O
were	NN	O	O
identified	NN	O	O
in	NN	O	O
the	NN	O	O
majority	NN	O	O
of	NN	O	O
the	NN	O	O
cells	NN	O	O
.	NN	O	O

Precise	NN	O	O
deletion	NN	O	O
of	NN	O	O
the	NN	O	O
exons	NN	O	O
suggests	NN	O	O
that	NN	O	O
they	NN	O	O
are	NN	O	O
alternatively	NN	O	O
spliced	NN	O	O
transcripts	NN	O	O
.	NN	O	O

Whether	NN	O	O
the	NN	O	O
detected	NN	O	O
transcripts	NN	O	O
are	NN	O	O
translated	NN	O	O
into	NN	O	O
functional	NN	O	B-protein
receptor	NN	O	I-protein
proteins	NN	O	I-protein
remains	NN	O	O
to	NN	O	O
be	NN	O	O
determined	NN	O	O
.	NN	O	O

In	NN	O	O
vitro	NN	O	O
stimulation	NN	O	O
did	NN	O	O
not	NN	O	O
affect	NN	O	O
ER	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
.	NN	O	O

The	NN	O	O
presence	NN	O	O
of	NN	O	O
variants	NN	O	O
did	NN	O	O
not	NN	O	O
correlate	NN	O	O
with	NN	O	O
disease	NN	O	O
activity	NN	O	O
or	NN	O	O
medication	NN	O	O
.	NN	O	O

CONCLUSION	NN	O	O
:	NN	O	O
Monocytes	NN	O	B-cell_type
,	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
patients	NN	O	O
express	NN	O	O
transcripts	NN	O	O
of	NN	O	O
the	NN	O	O
normal	NN	O	O
wild	NN	O	B-protein
type	NN	O	I-protein
ER	NN	O	I-protein
and	NN	O	O
the	NN	O	O
hormone	NN	O	B-protein
binding	NN	O	I-protein
domain	NN	O	I-protein
variants	NN	O	I-protein
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

-DOCSTART-	O

DNA	NN	O	O
damaging	NN	O	O
agents	NN	O	O
induce	NN	O	O
expression	NN	O	O
of	NN	O	O
Fas	NN	O	B-protein
ligand	NN	O	I-protein
and	NN	O	O
subsequent	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
via	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
AP-1	NN	O	B-protein
.	NN	O	O

Apoptosis	NN	O	O
induced	NN	O	O
by	NN	O	O
DNA	NN	O	O
damage	NN	O	O
and	NN	O	O
other	NN	O	O
stresses	NN	O	O
can	NN	O	O
proceed	NN	O	O
via	NN	O	O
expression	NN	O	O
of	NN	O	O
Fas	NN	O	B-protein
ligand	NN	O	I-protein
(	NN	O	O
FasL	NN	O	B-protein
)	NN	O	O
and	NN	O	O
ligation	NN	O	O
of	NN	O	O
its	NN	O	O
receptor	NN	O	O
,	NN	O	O
Fas	NN	O	O
(	NN	O	O
CD95	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
that	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
two	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
and	NN	O	I-protein
AP-1	NN	O	I-protein
is	NN	O	O
crucially	NN	O	O
involved	NN	O	O
in	NN	O	O
FasL	NN	O	B-protein
expression	NN	O	O
induced	NN	O	O
by	NN	O	O
etoposide	NN	O	O
,	NN	O	O
teniposide	NN	O	O
,	NN	O	O
and	NN	O	O
UV	NN	O	O
irradiation	NN	O	O
.	NN	O	O

A	NN	O	O
nondegradable	NN	O	B-protein
mutant	NN	O	I-protein
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
blocked	NN	O	O
both	NN	O	O
FasL	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
apoptosis	NN	O	O
induced	NN	O	O
by	NN	O	O
DNA	NN	O	O
damage	NN	O	O
but	NN	O	O
not	NN	O	O
Fas	NN	O	O
ligation	NN	O	O
.	NN	O	O

These	NN	O	O
stimuli	NN	O	O
also	NN	O	O
induced	NN	O	O
the	NN	O	O
stress-activated	NN	O	O
kinase	NN	O	O
pathway	NN	O	O
(	NN	O	O
SAPK/JNK	NN	O	O
)	NN	O	O
,	NN	O	O
which	NN	O	O
was	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
maximal	NN	O	O
induction	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

A	NN	O	O
1.2	NN	O	O
kb	NN	O	O
FasL	NN	O	B-DNA
promoter	NN	O	I-DNA
responded	NN	O	O
to	NN	O	O
DNA	NN	O	O
damage	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
coexpression	NN	O	O
with	NN	O	O
p65	NN	O	B-protein
Rel	NN	O	I-protein
or	NN	O	O
Fos/Jun	NN	O	B-protein
.	NN	O	O

Mutations	NN	O	O
in	NN	O	O
the	NN	O	O
relevant	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
and	NN	O	I-DNA
AP-1	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
eliminated	NN	O	O
these	NN	O	O
responses	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
AP-1	NN	O	B-protein
contributes	NN	O	O
to	NN	O	O
stress-induced	NN	O	O
apoptosis	NN	O	O
via	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
FasL	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
small	NN	O	O
,	NN	O	O
nonpeptidyl	NN	O	O
mimic	NN	O	O
of	NN	O	O
granulocyte-colony-stimulating	NN	O	B-protein
factor	NN	O	I-protein
[	NN	O	O
see	NN	O	O
commetns	NN	O	O
]	NN	O	O

A	NN	O	O
nonpeptidyl	NN	O	O
small	NN	O	O
molecule	NN	O	O
SB	NN	O	O
247464	NN	O	O
,	NN	O	O
capable	NN	O	O
of	NN	O	O
activating	NN	O	O
granulocyte-colony-stimulating	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	O
G-CSF	NN	O	B-protein
)	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathways	NN	O	O
,	NN	O	O
was	NN	O	O
identified	NN	O	O
in	NN	O	O
a	NN	O	O
high-throughput	NN	O	O
assay	NN	O	O
in	NN	O	O
cultured	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Like	NN	O	O
G-CSF	NN	O	B-protein
,	NN	O	O
SB	NN	O	O
247464	NN	O	O
induced	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
multiple	NN	O	B-protein
signaling	NN	O	I-protein
proteins	NN	O	I-protein
and	NN	O	O
stimulated	NN	O	O
primary	NN	O	B-cell_type
murine	NN	O	I-cell_type
bone	NN	O	I-cell_type
marrow	NN	O	I-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
form	NN	O	O
granulocytic	NN	O	B-cell_type
colonies	NN	O	I-cell_type
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

It	NN	O	O
also	NN	O	O
elevated	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
neutrophil	NN	O	I-cell_type
counts	NN	O	O
in	NN	O	O
mice	NN	O	O
.	NN	O	O

The	NN	O	O
extracellular	NN	O	O
domain	NN	O	O
of	NN	O	O
the	NN	O	O
murine	NN	O	B-protein
G-CSF	NN	O	I-protein
receptor	NN	O	I-protein
was	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
SB	NN	O	O
247464	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
the	NN	O	O
compound	NN	O	O
acts	NN	O	O
by	NN	O	O
oligomerizing	NN	O	B-protein
receptor	NN	O	I-protein
chains	NN	O	I-protein
.	NN	O	O

The	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
a	NN	O	O
small	NN	O	O
molecule	NN	O	O
can	NN	O	O
activate	NN	O	O
a	NN	O	O
receptor	NN	O	O
that	NN	O	O
normally	NN	O	O
binds	NN	O	O
a	NN	O	O
relatively	NN	O	O
large	NN	O	O
protein	NN	O	O
ligand	NN	O	O
.	NN	O	O

-DOCSTART-	O

Minimal	NN	O	O
residual	NN	O	O
disease	NN	O	O
in	NN	O	O
acute	NN	O	O
myelogenous	NN	O	O
leukemia	NN	O	O
with	NN	O	O
PML/RAR	NN	O	B-protein
alpha	NN	O	I-protein
or	NN	O	O
AML1/ETO	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
phenotypic	NN	O	O
analysis	NN	O	O
of	NN	O	O
possible	NN	O	O
T	NN	O	B-cell_type
and	NN	O	I-cell_type
natural	NN	O	I-cell_type
killer	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
bone	NN	O	O
marrow	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
studied	NN	O	O
minimal	NN	O	O
residual	NN	O	O
disease	NN	O	O
(	NN	O	O
MRD	NN	O	O
)	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
acute	NN	O	O
myeloid	NN	O	O
leukemia	NN	O	O
(	NN	O	O
AML	NN	O	O
)	NN	O	O
who	NN	O	O
have	NN	O	O
PML/RAR	NN	O	B-protein
alpha	NN	O	I-protein
or	NN	O	O
AML1/ETO	NN	O	B-protein
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
phenotypic	NN	O	O
analysis	NN	O	O
of	NN	O	O
lymphocyte	NN	O	B-cell_type
subsets	NN	O	I-cell_type
involved	NN	O	O
in	NN	O	O
antitumor	NN	O	O
immunity	NN	O	O
.	NN	O	O

Eight	NN	O	O
patients	NN	O	O
in	NN	O	O
long-term	NN	O	O
(	NN	O	O
LT	NN	O	O
;	NN	O	O
3	NN	O	O
to	NN	O	O
15	NN	O	O
years	NN	O	O
)	NN	O	O
and	NN	O	O
15	NN	O	O
patients	NN	O	O
in	NN	O	O
short-term	NN	O	O
(	NN	O	O
ST	NN	O	O
;	NN	O	O
up	NN	O	O
to	NN	O	O
3	NN	O	O
years	NN	O	O
)	NN	O	O
remission	NN	O	O
were	NN	O	O
studied	NN	O	O
.	NN	O	O

Using	NN	O	O
the	NN	O	O
reverse	NN	O	O
transcription-polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
(	NN	O	O
RT	NN	O	O
)	NN	O	O
assay	NN	O	O
,	NN	O	O
the	NN	O	O
limit	NN	O	O
of	NN	O	O
detection	NN	O	O
was	NN	O	O
10	NN	O	O
(	NN	O	O
-5	NN	O	O
)	NN	O	O
to	NN	O	O
10	NN	O	O
(	NN	O	O
-6	NN	O	O
)	NN	O	O
for	NN	O	O
PML/RAR	NN	O	B-RNA
alpha	NN	O	I-RNA
transcript	NN	O	I-RNA
and	NN	O	O
10	NN	O	O
(	NN	O	O
-4	NN	O	O
)	NN	O	O
to	NN	O	O
10	NN	O	O
(	NN	O	O
-5	NN	O	O
)	NN	O	O
for	NN	O	O
the	NN	O	O
AML1/ETO	NN	O	B-RNA
transcript	NN	O	I-RNA
.	NN	O	O

Simultaneously	NN	O	O
,	NN	O	O
T	NN	O	B-cell_type
lymphocyte	NN	O	I-cell_type
subsets	NN	O	I-cell_type
and	NN	O	O
NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
the	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
(	NN	O	O
PB	NN	O	O
)	NN	O	O
and	NN	O	O
bone	NN	O	O
marrow	NN	O	O
(	NN	O	O
BM	NN	O	O
)	NN	O	O
were	NN	O	O
investigated	NN	O	O
by	NN	O	O
flow	NN	O	O
cytometric	NN	O	O
analysis	NN	O	O
.	NN	O	O

Four	NN	O	O
of	NN	O	O
the	NN	O	O
eight	NN	O	O
patients	NN	O	O
in	NN	O	O
LT	NN	O	O
and	NN	O	O
7	NN	O	O
of	NN	O	O
the	NN	O	O
15	NN	O	O
patients	NN	O	O
in	NN	O	O
ST	NN	O	O
remission	NN	O	O
were	NN	O	O
MRD-positive	NN	O	O
.	NN	O	O

Although	NN	O	O
all	NN	O	O
MRD-positive	NN	O	O
patients	NN	O	O
in	NN	O	O
LT	NN	O	O
remission	NN	O	O
are	NN	O	O
still	NN	O	O
until	NN	O	O
now	NN	O	O
event-free	NN	O	O
,	NN	O	O
3	NN	O	O
of	NN	O	O
the	NN	O	O
7	NN	O	O
MRD-positive	NN	O	O
(	NN	O	O
MRD+	NN	O	O
)	NN	O	O
patients	NN	O	O
in	NN	O	O
ST	NN	O	O
remission	NN	O	O
soon	NN	O	O
relapsed	NN	O	O
.	NN	O	O

The	NN	O	O
total	NN	O	O
populations	NN	O	O
of	NN	O	O
CD4	NN	O	B-protein
+	NN	O	O
,	NN	O	O
CD8	NN	O	B-protein
+	NN	O	O
and	NN	O	O
CD56	NN	O	B-protein
+	NN	O	O
[	NN	O	O
possible	NN	O	O
T-cell	NN	O	B-cell_type
and	NN	O	I-cell_type
natural	NN	O	I-cell_type
killer	NN	O	I-cell_type
(	NN	O	I-cell_type
T/NK	NN	O	I-cell_type
)	NN	O	I-cell_type
populations	NN	O	I-cell_type
]	NN	O	O
in	NN	O	O
the	NN	O	O
BM	NN	O	O
of	NN	O	O
ST	NN	O	O
patients	NN	O	O
and	NN	O	O
MRD+/LT	NN	O	O
patients	NN	O	O
were	NN	O	O
significantly	NN	O	O
(	NN	O	O
p	NN	O	O
<	NN	O	O
.01	NN	O	O
)	NN	O	O
low	NN	O	O
.	NN	O	O

The	NN	O	O
CD8+	NN	O	B-cell_type
CD28+	NN	O	I-cell_type
population	NN	O	I-cell_type
showed	NN	O	O
the	NN	O	O
same	NN	O	O
tendency	NN	O	O
(	NN	O	O
p	NN	O	O
<	NN	O	O
.01-.02	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
T/NK	NN	O	B-cell_type
subsets	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
BM	NN	O	O
of	NN	O	O
MRD-negative	NN	O	O
(	NN	O	O
MRD-	NN	O	O
)	NN	O	O
LT	NN	O	O
(	NN	O	O
MRD-/LT	NN	O	O
)	NN	O	O
patients	NN	O	O
showed	NN	O	O
similar	NN	O	O
numbers	NN	O	O
of	NN	O	O
cells	NN	O	O
as	NN	O	O
normal	NN	O	O
volunteers	NN	O	O
.	NN	O	O

Basically	NN	O	O
,	NN	O	O
the	NN	O	O
total	NN	O	O
percentage	NN	O	O
of	NN	O	O
the	NN	O	O
CD4+	NN	O	B-cell_type
,	NN	O	I-cell_type
CD8+	NN	O	I-cell_type
and	NN	O	I-cell_type
CD56+	NN	O	I-cell_type
cell	NN	O	I-cell_type
populations	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
BM	NN	O	O
was	NN	O	O
increased	NN	O	O
and	NN	O	O
in	NN	O	O
the	NN	O	O
following	NN	O	O
order	NN	O	O
:	NN	O	O
MRD-/LT	NN	O	O
patients	NN	O	O
,	NN	O	O
normal	NN	O	O
volunteers	NN	O	O
,	NN	O	O
MRD+/LT	NN	O	O
patients	NN	O	O
and	NN	O	O
MRD+	NN	O	O
or	NN	O	O
-/ST	NN	O	O
patients	NN	O	O
.	NN	O	O

The	NN	O	O
percentages	NN	O	O
of	NN	O	O
the	NN	O	O
T/NK-cell	NN	O	B-cell_type
subsets	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
PB	NN	O	O
were	NN	O	O
not	NN	O	O
significantly	NN	O	O
different	NN	O	O
among	NN	O	O
these	NN	O	O
groups	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
difference	NN	O	O
of	NN	O	O
the	NN	O	O
possible	NN	O	O
T/NK-cell	NN	O	B-cell_type
phenotype	NN	O	O
in	NN	O	O
the	NN	O	O
BM	NN	O	O
may	NN	O	O
strongly	NN	O	O
influence	NN	O	O
clinical	NN	O	O
and	NN	O	O
molecular	NN	O	O
remission	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
still	NN	O	O
remain	NN	O	O
to	NN	O	O
be	NN	O	O
confirmed	NN	O	O
by	NN	O	O
further	NN	O	O
studies	NN	O	O
of	NN	O	O
the	NN	O	O
functional	NN	O	O
anti-tumor	NN	O	O
immunity	NN	O	O
of	NN	O	O
T/NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
AML	NN	O	B-cell_type
in	NN	O	O
remission	NN	O	O
.	NN	O	O

-DOCSTART-	O

Mycobacterium	NN	O	O
tuberculosis	NN	O	O
mannose-capped	NN	O	O
lipoarabinomannan	NN	O	O
can	NN	O	O
induce	NN	O	O
NF-kappaB	NN	O	B-protein
-dependent	NN	O	O
activation	NN	O	O
of	NN	O	O
human	NN	O	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

Tuberculosis	NN	O	O
has	NN	O	O
emerged	NN	O	O
as	NN	O	O
an	NN	O	O
epidemic	NN	O	O
,	NN	O	O
extended	NN	O	O
by	NN	O	O
the	NN	O	O
large	NN	O	O
number	NN	O	O
of	NN	O	O
individuals	NN	O	O
infected	NN	O	O
with	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
(	NN	O	O
HIV-1	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
major	NN	O	O
goal	NN	O	O
of	NN	O	O
this	NN	O	O
study	NN	O	O
was	NN	O	O
to	NN	O	O
determine	NN	O	O
whether	NN	O	O
the	NN	O	O
mycobacterial	NN	O	O
cell	NN	O	O
wall	NN	O	O
component	NN	O	O
mannose-capped	NN	O	O
lipoarabinomannan	NN	O	O
(	NN	O	O
ManLAM	NN	O	O
)	NN	O	O
of	NN	O	O
Mycobacterium	NN	O	O
tuberculosis	NN	O	O
(	NN	O	O
M.	NN	O	O
tuberculosis	NN	O	O
)	NN	O	O
could	NN	O	O
activate	NN	O	O
transcription	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
an	NN	O	O
in	NN	O	O
vitro	NN	O	O
cell	NN	O	O
culture	NN	O	O
system	NN	O	O
.	NN	O	O

These	NN	O	O
experiments	NN	O	O
are	NN	O	O
of	NN	O	O
prime	NN	O	O
importance	NN	O	O
considering	NN	O	O
that	NN	O	O
CD4	NN	O	B-protein
-expressing	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
represent	NN	O	O
the	NN	O	O
major	NN	O	O
virus	NN	O	O
reservoir	NN	O	O
in	NN	O	O
the	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
of	NN	O	O
infected	NN	O	O
individuals	NN	O	O
.	NN	O	O

Using	NN	O	O
the	NN	O	O
1G5	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
harbouring	NN	O	O
the	NN	O	O
luciferase	NN	O	O
reporter	NN	O	O
gene	NN	O	O
under	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
LTR	NN	O	I-DNA
,	NN	O	O
it	NN	O	O
was	NN	O	O
first	NN	O	O
found	NN	O	O
that	NN	O	O
culture	NN	O	O
protein	NN	O	O
filtrates	NN	O	O
(	NN	O	O
CFP	NN	O	O
)	NN	O	O
from	NN	O	O
M.	NN	O	O
tuberculosis	NN	O	O
or	NN	O	O
purified	NN	O	O
ManLAM	NN	O	O
could	NN	O	O
activate	NN	O	O
HIV-1	NN	O	B-DNA
LTR-dependent	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
unlike	NN	O	O
similarly	NN	O	O
prepared	NN	O	O
CFP	NN	O	O
extracts	NN	O	O
devoid	NN	O	O
of	NN	O	O
ManLAM	NN	O	O
.	NN	O	O

The	NN	O	O
implication	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	I-protein
s	NN	O	I-protein
)	NN	O	I-protein
,	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
A	NN	O	I-protein
and/or	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
was	NN	O	O
highlighted	NN	O	O
by	NN	O	O
the	NN	O	O
abrogation	NN	O	O
of	NN	O	O
the	NN	O	O
ManLAM-mediated	NN	O	O
activation	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
LTR-driven	NN	O	O
gene	NN	O	O
expression	NN	O	O
using	NN	O	O
herbimycin	NN	O	O
A	NN	O	O
and	NN	O	O
H7	NN	O	O
.	NN	O	O

It	NN	O	O
was	NN	O	O
also	NN	O	O
determined	NN	O	O
,	NN	O	O
using	NN	O	O
electrophoresis	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
,	NN	O	O
that	NN	O	O
M.	NN	O	O
tuberculosis	NN	O	O
ManLAM	NN	O	O
led	NN	O	O
to	NN	O	O
the	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappaB	NN	O	I-protein
.	NN	O	O

M.	NN	O	O
tuberculosis	NN	O	O
ManLAM	NN	O	O
resulted	NN	O	O
in	NN	O	O
clear	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
luciferase	NN	O	B-DNA
gene	NN	O	I-DNA
placed	NN	O	O
under	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
the	NN	O	O
wild-type	NN	O	B-DNA
,	NN	O	I-DNA
but	NN	O	I-DNA
not	NN	O	I-DNA
the	NN	O	I-DNA
kappaB-mutated	NN	O	I-DNA
,	NN	O	I-DNA
HIV-1	NN	O	I-DNA
LTR	NN	O	I-DNA
region	NN	O	I-DNA
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
the	NN	O	O
ManLAM-mediated	NN	O	O
activation	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
LTR	NN	O	O
transcription	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
independent	NN	O	O
of	NN	O	O
the	NN	O	O
autocrine	NN	O	O
or	NN	O	O
paracrine	NN	O	O
action	NN	O	O
of	NN	O	O
endogenous	NN	O	B-protein
TNF-alpha	NN	O	I-protein
.	NN	O	O

The	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
M.	NN	O	O
tuberculosis	NN	O	O
can	NN	O	O
upregulate	NN	O	O
HIV-1	NN	O	O
expression	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
could	NN	O	O
thus	NN	O	O
have	NN	O	O
the	NN	O	O
potential	NN	O	O
to	NN	O	O
influence	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
infection	NN	O	O
.	NN	O	O

-DOCSTART-	O

Human	NN	O	B-DNA
immunodeficiency	NN	O	I-DNA
virus	NN	O	I-DNA
type	NN	O	I-DNA
1	NN	O	I-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
quasispecies	NN	O	O
differ	NN	O	O
in	NN	O	O
basal	NN	O	O
transcription	NN	O	O
and	NN	O	O
nuclear	NN	O	O
factor	NN	O	O
recruitment	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
glial	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
generation	NN	O	O
of	NN	O	O
genomic	NN	O	O
diversity	NN	O	O
during	NN	O	O
the	NN	O	O
course	NN	O	O
of	NN	O	O
infection	NN	O	O
has	NN	O	O
the	NN	O	O
potential	NN	O	O
to	NN	O	O
affect	NN	O	O
all	NN	O	O
aspects	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
,	NN	O	O
including	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
proviral	NN	O	O
genome	NN	O	O
.	NN	O	O

To	NN	O	O
gain	NN	O	O
a	NN	O	O
better	NN	O	O
understanding	NN	O	O
of	NN	O	O
the	NN	O	O
impact	NN	O	O
of	NN	O	O
long	NN	O	B-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
(	NN	O	O
LTR	NN	O	B-DNA
)	NN	O	O
sequence	NN	O	O
diversity	NN	O	O
on	NN	O	O
LTR-directed	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
cells	NN	O	O
of	NN	O	O
the	NN	O	O
central	NN	O	O
nervous	NN	O	O
system	NN	O	O
(	NN	O	O
CNS	NN	O	O
)	NN	O	O
and	NN	O	O
immune	NN	O	O
system	NN	O	O
,	NN	O	O
we	NN	O	O
amplified	NN	O	O
and	NN	O	O
cloned	NN	O	O
LTRs	NN	O	B-DNA
from	NN	O	O
proviral	NN	O	O
DNA	NN	O	O
in	NN	O	O
HIV-1-infected	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
.	NN	O	O

Sequence	NN	O	O
analysis	NN	O	O
of	NN	O	O
nineteen	NN	O	O
LTRs	NN	O	B-DNA
cloned	NN	O	O
from	NN	O	O
2	NN	O	O
adult	NN	O	O
and	NN	O	O
3	NN	O	O
pediatric	NN	O	O
patients	NN	O	O
revealed	NN	O	O
an	NN	O	O
average	NN	O	O
of	NN	O	O
33	NN	O	O
nucleotide	NN	O	O
changes	NN	O	O
(	NN	O	O
with	NN	O	O
respect	NN	O	O
to	NN	O	O
the	NN	O	O
sequence	NN	O	O
of	NN	O	O
the	NN	O	O
LAI	NN	O	B-DNA
LTR	NN	O	I-DNA
)	NN	O	O
within	NN	O	O
the	NN	O	O
455-bp	NN	O	B-DNA
U3	NN	O	I-DNA
region	NN	O	I-DNA
.	NN	O	O

Transient	NN	O	O
expression	NN	O	O
analyses	NN	O	O
in	NN	O	O
cells	NN	O	O
of	NN	O	O
neuroglial	NN	O	O
and	NN	O	O
lymphocytic	NN	O	O
origin	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
some	NN	O	O
of	NN	O	O
these	NN	O	O
LTRs	NN	O	B-DNA
had	NN	O	O
activities	NN	O	O
which	NN	O	O
varied	NN	O	O
significantly	NN	O	O
from	NN	O	O
the	NN	O	O
LAI	NN	O	B-DNA
LTR	NN	O	I-DNA
in	NN	O	O
U-373	NN	O	B-cell_line
MG	NN	O	I-cell_line
cells	NN	O	I-cell_line
(	NN	O	O
an	NN	O	O
astrocytoma	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
)	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
(	NN	O	O
a	NN	O	O
CD4-positive	NN	O	B-cell_line
lymphocyte	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
)	NN	O	O
.	NN	O	O

While	NN	O	O
LTRs	NN	O	B-DNA
which	NN	O	O
demonstrated	NN	O	O
the	NN	O	O
highest	NN	O	O
activities	NN	O	O
in	NN	O	O
U-373	NN	O	B-cell_line
MG	NN	O	I-cell_line
cells	NN	O	I-cell_line
also	NN	O	O
yielded	NN	O	O
high	NN	O	O
activities	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
the	NN	O	O
LTRs	NN	O	B-DNA
were	NN	O	O
generally	NN	O	O
more	NN	O	O
active	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
when	NN	O	O
compared	NN	O	O
to	NN	O	O
the	NN	O	O
LAI	NN	O	B-DNA
LTR	NN	O	I-DNA
.	NN	O	O

Differences	NN	O	O
in	NN	O	O
LTR	NN	O	B-DNA
sequence	NN	O	I-DNA
also	NN	O	O
resulted	NN	O	O
in	NN	O	O
differences	NN	O	O
in	NN	O	O
transcription	NN	O	O
factor	NN	O	O
recruitment	NN	O	O
to	NN	O	O
cis-acting	NN	O	B-DNA
sites	NN	O	I-DNA
within	NN	O	O
the	NN	O	O
U3	NN	O	B-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
LTR	NN	O	B-DNA
,	NN	O	O
as	NN	O	O
demonstrated	NN	O	O
by	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
.	NN	O	O

In	NN	O	O
particular	NN	O	O
,	NN	O	O
naturally	NN	O	O
occurring	NN	O	O
sequence	NN	O	O
variation	NN	O	O
impacted	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
binding	NN	O	O
to	NN	O	O
an	NN	O	O
activating	NN	O	B-protein
transcription	NN	O	I-protein
factor	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
(	NN	O	O
ATF	NN	O	B-protein
/	NN	O	O
CREB	NN	O	B-protein
)	NN	O	O
binding	NN	O	O
site	NN	O	O
(	NN	O	O
located	NN	O	O
between	NN	O	O
the	NN	O	O
LEF-1	NN	O	B-DNA
and	NN	O	I-DNA
distal	NN	O	I-DNA
NF-kappaB	NN	O	I-DNA
transcription	NN	O	I-DNA
factor	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
)	NN	O	O
that	NN	O	O
we	NN	O	O
identified	NN	O	O
in	NN	O	O
previous	NN	O	O
studies	NN	O	O
of	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
LTR	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
LTR	NN	O	B-DNA
sequence	NN	O	I-DNA
changes	NN	O	O
can	NN	O	O
significantly	NN	O	O
affect	NN	O	O
basal	NN	O	O
LTR	NN	O	O
function	NN	O	O
and	NN	O	O
transcription	NN	O	O
factor	NN	O	O
recruitment	NN	O	O
,	NN	O	O
which	NN	O	O
may	NN	O	O
,	NN	O	O
in	NN	O	O
turn	NN	O	O
,	NN	O	O
alter	NN	O	O
the	NN	O	O
course	NN	O	O
of	NN	O	O
viral	NN	O	O
replication	NN	O	O
in	NN	O	O
cells	NN	O	O
of	NN	O	O
CNS	NN	O	O
and	NN	O	O
immune	NN	O	O
system	NN	O	O
origin	NN	O	O
.	NN	O	O

-DOCSTART-	O

HMG	NN	O	B-protein
box	NN	O	I-protein
containing	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
in	NN	O	O
lymphocyte	NN	O	B-cell_type
differentiation	NN	O	O
.	NN	O	O

The	NN	O	O
identification	NN	O	O
of	NN	O	O
the	NN	O	O
mammalian	NN	O	B-DNA
sex-determining	NN	O	I-DNA
gene	NN	O	I-DNA
Sry	NN	O	I-DNA
has	NN	O	O
led	NN	O	O
to	NN	O	O
the	NN	O	O
discovery	NN	O	O
of	NN	O	O
a	NN	O	O
large	NN	O	O
family	NN	O	O
of	NN	O	O
related	NN	O	O
(	NN	O	O
'	NN	O	O
HMG	NN	O	B-protein
box	NN	O	I-protein
'	NN	O	O
)	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
that	NN	O	O
control	NN	O	O
developmental	NN	O	O
events	NN	O	O
in	NN	O	O
yeast	NN	O	O
,	NN	O	O
C.	NN	O	O
elegans	NN	O	O
,	NN	O	O
Drosophila	NN	O	O
and	NN	O	O
vertebrates	NN	O	O
.	NN	O	O

In	NN	O	O
lymphocyte	NN	O	B-cell_type
differentiation	NN	O	O
,	NN	O	O
several	NN	O	O
HMG	NN	O	B-protein
box	NN	O	I-protein
proteins	NN	O	I-protein
play	NN	O	O
a	NN	O	O
decisive	NN	O	O
role	NN	O	O
.	NN	O	O

Sox-4	NN	O	B-protein
is	NN	O	O
important	NN	O	O
for	NN	O	O
very	NN	O	O
early	NN	O	O
B-cell	NN	O	O
differentiation	NN	O	O
,	NN	O	O
while	NN	O	O
TCF-1	NN	O	B-protein
/	NN	O	O
LEF-1	NN	O	B-protein
play	NN	O	O
a	NN	O	O
crucial	NN	O	O
role	NN	O	O
in	NN	O	O
early	NN	O	O
thymocyte	NN	O	O
development	NN	O	O
.	NN	O	O

TCF/LEF	NN	O	B-protein
proteins	NN	O	I-protein
have	NN	O	O
recently	NN	O	O
been	NN	O	O
found	NN	O	O
to	NN	O	O
constitute	NN	O	O
a	NN	O	O
downstream	NN	O	O
component	NN	O	O
of	NN	O	O
the	NN	O	O
Wingless/Wnt	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathway	NN	O	O
.	NN	O	O

In	NN	O	O
flies	NN	O	O
,	NN	O	O
this	NN	O	O
pathway	NN	O	O
controls	NN	O	O
segment	NN	O	O
polarity	NN	O	O
;	NN	O	O
in	NN	O	O
Xenopus	NN	O	O
it	NN	O	O
controls	NN	O	O
the	NN	O	O
definition	NN	O	O
of	NN	O	O
the	NN	O	O
body	NN	O	O
axis	NN	O	O
.	NN	O	O

Deregulation	NN	O	O
of	NN	O	O
the	NN	O	O
pathway	NN	O	O
occurs	NN	O	O
in	NN	O	O
several	NN	O	O
human	NN	O	B-cell_type
tumors	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
insights	NN	O	O
in	NN	O	O
the	NN	O	O
molecular	NN	O	O
events	NN	O	O
that	NN	O	O
are	NN	O	O
involved	NN	O	O
in	NN	O	O
TCF/LEF	NN	O	B-protein
function	NN	O	O
in	NN	O	O
these	NN	O	O
organisms	NN	O	O
may	NN	O	O
eventually	NN	O	O
lead	NN	O	O
to	NN	O	O
the	NN	O	O
understanding	NN	O	O
of	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
these	NN	O	O
HMG	NN	O	B-protein
box	NN	O	I-protein
proteins	NN	O	I-protein
in	NN	O	O
lymphoid	NN	O	O
development	NN	O	O

-DOCSTART-	O

Transcriptional	NN	O	O
regulation	NN	O	O
by	NN	O	O
C/EBP	NN	O	B-protein
alpha	NN	O	I-protein
and	NN	O	I-protein
-beta	NN	O	I-protein
in	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
for	NN	O	O
the	NN	O	O
MRP14	NN	O	B-protein
myeloid	NN	O	I-protein
calcium	NN	O	I-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

Transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
for	NN	O	O
the	NN	O	O
myeloid	NN	O	B-protein
calcium	NN	O	I-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
MRP14	NN	O	B-protein
,	NN	O	O
was	NN	O	O
investigated	NN	O	O
in	NN	O	O
human	NN	O	B-cell_line
monocytic	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
MRP14	NN	O	B-DNA
gene	NN	O	I-DNA
was	NN	O	O
not	NN	O	O
expressed	NN	O	O
in	NN	O	O
monoblastic	NN	O	B-cell_line
ML-1	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
promonocytic	NN	O	B-cell_line
U-937	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
or	NN	O	O
promyelocytic	NN	O	B-cell_line
HL-60	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

On	NN	O	O
the	NN	O	O
other	NN	O	O
hand	NN	O	O
,	NN	O	O
the	NN	O	O
gene	NN	O	O
was	NN	O	O
expressed	NN	O	O
in	NN	O	O
monocytic	NN	O	B-cell_line
THP-1	NN	O	I-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
in	NN	O	O
the	NN	O	O
HL-60	NN	O	B-cell_line
cells	NN	O	I-cell_line
treated	NN	O	O
with	NN	O	O
1	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
.	NN	O	O

The	NN	O	O
level	NN	O	O
of	NN	O	O
MRP14	NN	O	B-protein
in	NN	O	O
VD3-treated	NN	O	O
HL-60	NN	O	B-cell_line
cells	NN	O	I-cell_line
was	NN	O	O
two-fold	NN	O	O
higher	NN	O	O
than	NN	O	O
that	NN	O	O
in	NN	O	O
THP-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Among	NN	O	O
several	NN	O	O
known	NN	O	O
transcription	NN	O	B-DNA
factor	NN	O	I-DNA
binding	NN	O	I-DNA
motifs	NN	O	I-DNA
,	NN	O	O
nuclear	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	I-protein
s	NN	O	I-protein
)	NN	O	I-protein
of	NN	O	O
VD3-treated	NN	O	B-cell_line
HL-60	NN	O	I-cell_line
cells	NN	O	I-cell_line
and	NN	O	I-cell_line
THP-1	NN	O	I-cell_line
cells	NN	O	I-cell_line
bound	NN	O	O
to	NN	O	O
the	NN	O	O
CCAAT/enhancer	NN	O	B-DNA
binding	NN	O	I-DNA
protein	NN	O	I-DNA
(	NN	O	I-DNA
C/EBP	NN	O	I-DNA
)	NN	O	I-DNA
-binding	NN	O	I-DNA
motif	NN	O	I-DNA
that	NN	O	O
was	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
the	NN	O	O
MRP14	NN	O	B-DNA
gene	NN	O	I-DNA
(	NN	O	O
-81	NN	O	O
)	NN	O	O
,	NN	O	O
as	NN	O	O
evidenced	NN	O	O
by	NN	O	O
the	NN	O	O
competitive	NN	O	O
gel	NN	O	O
mobility-shift	NN	O	O
assay	NN	O	O
.	NN	O	O

An	NN	O	O
antibody	NN	O	O
for	NN	O	O
C/EBP	NN	O	B-protein
alpha	NN	O	I-protein
super-shifted	NN	O	O
the	NN	O	O
nucleoprotein	NN	O	B-protein
complex	NN	O	I-protein
in	NN	O	O
THP-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
but	NN	O	O
not	NN	O	O
in	NN	O	O
the	NN	O	O
VD3-treated	NN	O	B-cell_line
HL-60	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
whereas	NN	O	O
an	NN	O	O
antibody	NN	O	O
for	NN	O	O
C/EBP	NN	O	B-protein
beta	NN	O	I-protein
blocked	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
the	NN	O	O
complex	NN	O	O
with	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	O
the	NN	O	O
HL-60	NN	O	B-cell_line
cells	NN	O	I-cell_line
but	NN	O	O
not	NN	O	O
with	NN	O	O
that	NN	O	O
of	NN	O	O
THP-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

An	NN	O	O
anti-C/EBP	NN	O	B-protein
delta	NN	O	I-protein
antibody	NN	O	I-protein
had	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
complex	NN	O	O
in	NN	O	O
either	NN	O	O
cell	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
it	NN	O	O
was	NN	O	O
concluded	NN	O	O
that	NN	O	O
C/EBP	NN	O	B-protein
alpha	NN	O	I-protein
and	NN	O	I-protein
-beta	NN	O	I-protein
were	NN	O	O
able	NN	O	O
to	NN	O	O
bind	NN	O	O
to	NN	O	O
the	NN	O	O
C/EBP	NN	O	B-protein
motif	NN	O	I-protein
,	NN	O	O
and	NN	O	O
that	NN	O	O
C/EBP	NN	O	B-protein
alpha	NN	O	I-protein
bound	NN	O	O
to	NN	O	O
the	NN	O	O
motif	NN	O	O
in	NN	O	O
THP-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
C/EBP	NN	O	B-protein
beta	NN	O	I-protein
bound	NN	O	O
to	NN	O	O
that	NN	O	O
in	NN	O	O
the	NN	O	O
VD3-treated	NN	O	B-cell_line
HL-60	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
to	NN	O	O
examine	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
C/EBP	NN	O	B-protein
motif	NN	O	I-protein
,	NN	O	O
we	NN	O	O
transfected	NN	O	O
several	NN	O	O
constructed	NN	O	O
luciferase	NN	O	B-DNA
reporter	NN	O	I-DNA
DNAs	NN	O	I-DNA
into	NN	O	O
HL-60	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
THP-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
luciferase	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
C/EBP	NN	O	B-protein
motif	NN	O	I-protein
in	NN	O	O
HL-60	NN	O	B-cell_line
cells	NN	O	I-cell_line
was	NN	O	O
increased	NN	O	O
by	NN	O	O
VD3	NN	O	O
treatment	NN	O	O
.	NN	O	O

The	NN	O	O
C/EBP	NN	O	B-protein
motif	NN	O	I-protein
in	NN	O	O
the	NN	O	O
MRP14	NN	O	B-DNA
gene	NN	O	I-DNA
was	NN	O	O
confirmed	NN	O	O
to	NN	O	O
function	NN	O	O
as	NN	O	O
a	NN	O	O
regulatory	NN	O	O
region	NN	O	O
in	NN	O	O
VD3-treated	NN	O	B-cell_line
HL-60	NN	O	I-cell_line
cells	NN	O	I-cell_line
and	NN	O	I-cell_line
THP-1	NN	O	I-cell_line
cells	NN	O	I-cell_line
by	NN	O	O
the	NN	O	O
assay	NN	O	O
.	NN	O	O

Since	NN	O	O
C/EBP	NN	O	B-protein
beta	NN	O	I-protein
was	NN	O	O
also	NN	O	O
detected	NN	O	O
in	NN	O	O
VD3-untreated	NN	O	B-cell_line
HL-60	NN	O	I-cell_line
cells	NN	O	I-cell_line
by	NN	O	O
immunoblotting	NN	O	O
,	NN	O	O
VD3	NN	O	O
activated	NN	O	O
C/EBP	NN	O	B-protein
beta	NN	O	I-protein
to	NN	O	O
bind	NN	O	O
to	NN	O	O
the	NN	O	O
motif	NN	O	O
,	NN	O	O
probably	NN	O	O
through	NN	O	O
post-translational	NN	O	O
modification	NN	O	O
.	NN	O	O

-DOCSTART-	O

Patients	NN	O	O
with	NN	O	O
high-risk	NN	O	O
myelodysplastic	NN	O	O
syndrome	NN	O	O
can	NN	O	O
have	NN	O	O
polyclonal	NN	O	O
or	NN	O	O
clonal	NN	O	O
haemopoiesis	NN	O	O
in	NN	O	O
complete	NN	O	O
haematological	NN	O	O
remission	NN	O	O
.	NN	O	O

The	NN	O	O
clonality	NN	O	O
of	NN	O	O
mature	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood-derived	NN	O	I-cell_type
myeloid	NN	O	I-cell_type
and	NN	O	I-cell_type
lymphoid	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
bone	NN	O	B-cell_type
marrow	NN	O	I-cell_type
haemopoietic	NN	O	I-cell_type
progenitors	NN	O	I-cell_type
from	NN	O	O
18	NN	O	O
females	NN	O	O
with	NN	O	O
myelodysplasia	NN	O	O
(	NN	O	O
MDS	NN	O	O
)	NN	O	O
(	NN	O	O
five	NN	O	O
refractory	NN	O	O
anaemia	NN	O	O
,	NN	O	O
RA	NN	O	O
;	NN	O	O
one	NN	O	O
RA	NN	O	O
with	NN	O	O
ringed	NN	O	B-cell_type
sideroblasts	NN	O	I-cell_type
,	NN	O	O
RARS	NN	O	O
;	NN	O	O
three	NN	O	O
chronic	NN	O	O
myelomonocytic	NN	O	O
leukaemia	NN	O	O
,	NN	O	O
CMML	NN	O	O
;	NN	O	O
four	NN	O	O
RA	NN	O	O
with	NN	O	O
excess	NN	O	O
of	NN	O	O
blasts	NN	O	O
,	NN	O	O
RAEB	NN	O	O
;	NN	O	O
five	NN	O	O
RAEB	NN	O	O
in	NN	O	O
transformation	NN	O	O
,	NN	O	O
RAEB-t	NN	O	O
)	NN	O	O
was	NN	O	O
studied	NN	O	O
by	NN	O	O
X-chromosome	NN	O	B-DNA
inactivation	NN	O	O
analysis	NN	O	O
.	NN	O	O

Using	NN	O	O
the	NN	O	O
human	NN	O	O
androgen-receptor	NN	O	O
(	NN	O	O
HUMARA	NN	O	O
)	NN	O	O
assay	NN	O	O
,	NN	O	O
we	NN	O	O
analysed	NN	O	O
the	NN	O	O
clonal	NN	O	O
patterns	NN	O	O
of	NN	O	O
highly	NN	O	B-cell_type
purified	NN	O	I-cell_type
immature	NN	O	I-cell_type
CD34+	NN	O	I-cell_type
38-	NN	O	I-cell_type
and	NN	O	I-cell_type
committed	NN	O	I-cell_type
CD34+	NN	O	I-cell_type
38+	NN	O	I-cell_type
marrow-derived	NN	O	I-cell_type
progenitors	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
CD16+	NN	O	B-cell_type
14-	NN	O	I-cell_type
granulocytes	NN	O	I-cell_type
,	NN	O	O
CD14+	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
,	NN	O	O
CD3+	NN	O	B-cell_type
T	NN	O	I-cell_type
and	NN	O	I-cell_type
CD19+	NN	O	I-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
from	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
.	NN	O	O

In	NN	O	O
high-risk	NN	O	O
patients	NN	O	O
(	NN	O	O
RAEB	NN	O	O
,	NN	O	O
RAEB-t	NN	O	O
)	NN	O	O
,	NN	O	O
clonality	NN	O	O
analysis	NN	O	O
was	NN	O	O
performed	NN	O	O
before	NN	O	O
and	NN	O	O
after	NN	O	O
intensive	NN	O	O
remission-induction	NN	O	O
treatment	NN	O	O
.	NN	O	O

All	NN	O	O
patients	NN	O	O
,	NN	O	O
except	NN	O	O
one	NN	O	O
with	NN	O	O
RA	NN	O	O
,	NN	O	O
had	NN	O	O
predominance	NN	O	O
of	NN	O	O
a	NN	O	O
single	NN	O	O
clone	NN	O	O
in	NN	O	O
their	NN	O	O
granulocytes	NN	O	B-cell_type
and	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

The	NN	O	O
same	NN	O	O
clonal	NN	O	O
pattern	NN	O	O
was	NN	O	O
found	NN	O	O
in	NN	O	O
CD34+	NN	O	B-cell_type
progenitor	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
CD3+	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
were	NN	O	O
polyclonal	NN	O	O
or	NN	O	O
oligoclonal	NN	O	O
in	NN	O	O
14/18	NN	O	O
patients	NN	O	O
.	NN	O	O

X-chromosome	NN	O	B-DNA
inactivation	NN	O	O
patterns	NN	O	O
of	NN	O	O
CD19+	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
highly	NN	O	O
concordant	NN	O	O
with	NN	O	O
CD3+	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
except	NN	O	O
for	NN	O	O
two	NN	O	O
patients	NN	O	O
(	NN	O	O
one	NN	O	O
RA	NN	O	O
,	NN	O	O
one	NN	O	O
CMML	NN	O	O
)	NN	O	O
with	NN	O	O
monoclonal	NN	O	B-cell_type
B	NN	O	I-cell_type
and	NN	O	I-cell_type
polyclonal	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
therefore	NN	O	O
suggesting	NN	O	O
a	NN	O	O
clonal	NN	O	O
mutation	NN	O	O
in	NN	O	O
a	NN	O	O
progenitor	NN	O	B-cell_type
common	NN	O	O
to	NN	O	O
the	NN	O	O
myeloid	NN	O	B-cell_type
and	NN	O	I-cell_type
B-lymphoid	NN	O	I-cell_type
lineages	NN	O	I-cell_type
or	NN	O	O
the	NN	O	O
coexistence	NN	O	O
of	NN	O	O
MDS	NN	O	O
and	NN	O	O
a	NN	O	O
B-cell	NN	O	O
disorder	NN	O	O
in	NN	O	O
these	NN	O	O
particular	NN	O	O
patients	NN	O	O
.	NN	O	O

After	NN	O	O
high-dose	NN	O	O
non-myeloablative	NN	O	O
chemotherapy	NN	O	O
,	NN	O	O
polyclonal	NN	O	O
haemopoiesis	NN	O	O
was	NN	O	O
reinstalled	NN	O	O
in	NN	O	O
the	NN	O	O
mature	NN	O	B-cell_type
myeloid	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
immature	NN	O	O
and	NN	O	O
committed	NN	O	B-cell_type
marrow	NN	O	I-cell_type
progenitors	NN	O	I-cell_type
in	NN	O	O
three	NN	O	O
of	NN	O	O
four	NN	O	O
patients	NN	O	O
achieving	NN	O	O
complete	NN	O	O
haematological	NN	O	O
remission	NN	O	O
.	NN	O	O

Therefore	NN	O	O
we	NN	O	O
conclude	NN	O	O
that	NN	O	O
most	NN	O	O
haematological	NN	O	O
remissions	NN	O	O
in	NN	O	O
MDS	NN	O	O
are	NN	O	O
associated	NN	O	O
with	NN	O	O
restoration	NN	O	O
of	NN	O	O
polyclonal	NN	O	O
haemopoiesis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Prominent	NN	O	O
sex	NN	O	O
steroid	NN	O	O
metabolism	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Steroid	NN	O	O
metabolism	NN	O	O
was	NN	O	O
investigated	NN	O	O
in	NN	O	O
cultured	NN	O	B-cell_line
human	NN	O	I-cell_line
B-lymphoblastoid	NN	O	I-cell_line
cells	NN	O	I-cell_line
(	NN	O	O
B-LCL	NN	O	B-cell_line
)	NN	O	O
,	NN	O	O
and	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
T	NN	O	I-cell_type
and	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Gene	NN	O	O
expression	NN	O	O
was	NN	O	O
examined	NN	O	O
by	NN	O	O
reverse-transcription	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
amplification	NN	O	O
(	NN	O	O
RT-PCR	NN	O	O
)	NN	O	O
.	NN	O	O

Appropriate	NN	O	O
sized	NN	O	O
transcripts	NN	O	B-RNA
were	NN	O	O
detected	NN	O	O
in	NN	O	O
both	NN	O	O
cultured	NN	O	O
and	NN	O	O
fresh	NN	O	O
peripheral	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
for	NN	O	O
CYP11A	NN	O	B-protein
,	NN	O	O
CYP17	NN	O	B-protein
,	NN	O	O
HSD11L	NN	O	B-protein
(	NN	O	O
11beta-hydroxysteroid	NN	O	B-protein
dehydrogenase	NN	O	I-protein
I	NN	O	I-protein
)	NN	O	O
,	NN	O	O
HSD17B1	NN	O	B-protein
(	NN	O	O
17beta-hydroxysteroid	NN	O	B-protein
dehydrogenase	NN	O	I-protein
type	NN	O	I-protein
I	NN	O	I-protein
)	NN	O	O
and	NN	O	O
SRD5A1	NN	O	B-protein
(	NN	O	O
5alpha-reductase	NN	O	B-protein
I	NN	O	I-protein
)	NN	O	O
.	NN	O	O

B-LCL	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
T	NN	O	B-cell_type
and	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
expressed	NN	O	O
CYP11B	NN	O	B-protein
.	NN	O	O

There	NN	O	O
was	NN	O	O
minimal	NN	O	O
expression	NN	O	O
of	NN	O	O
HSD3B1	NN	O	B-protein
and	NN	O	O
HSD3B2	NN	O	B-protein
(	NN	O	O
3beta-hydroxysteroid	NN	O	B-protein
dehydrogenase	NN	O	I-protein
I	NN	O	I-protein
and	NN	O	I-protein
II	NN	O	I-protein
)	NN	O	O
in	NN	O	O
B-LCL	NN	O	B-cell_line
and	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Transcripts	NN	O	O
for	NN	O	O
CYP19	NN	O	B-protein
and	NN	O	O
HSD11K	NN	O	B-protein
were	NN	O	O
not	NN	O	O
detected	NN	O	O
.	NN	O	O

Corresponding	NN	O	O
enzymatic	NN	O	O
activity	NN	O	O
was	NN	O	O
detectable	NN	O	O
only	NN	O	O
for	NN	O	O
17-hydroxysteroid	NN	O	B-protein
dehydrogenase	NN	O	I-protein
and	NN	O	O
5alpha-reductase	NN	O	B-protein
,	NN	O	O
respectively	NN	O	O
producing	NN	O	O
testosterone	NN	O	O
and	NN	O	O
5alpha-dihydrotestosterone	NN	O	O
.	NN	O	O

Steroid	NN	O	O
identities	NN	O	O
were	NN	O	O
confirmed	NN	O	O
by	NN	O	O
gas	NN	O	O
chromatography/mass	NN	O	O
spectrometry	NN	O	O
(	NN	O	O
GC/MS	NN	O	O
)	NN	O	O
.	NN	O	O

One	NN	O	O
metabolite	NN	O	O
thought	NN	O	O
to	NN	O	O
be	NN	O	O
deoxycorticosterone	NN	O	O
was	NN	O	O
identified	NN	O	O
by	NN	O	O
GC/MS	NN	O	O
as	NN	O	O
6alpha-hydroxypregnanolone	NN	O	O
.	NN	O	O

It	NN	O	O
was	NN	O	O
concluded	NN	O	O
that	NN	O	O
sex	NN	O	O
hormone	NN	O	O
metabolism	NN	O	O
,	NN	O	O
including	NN	O	O
androgen	NN	O	O
synthesis	NN	O	O
,	NN	O	O
occurs	NN	O	O
in	NN	O	O
lymphocytes	NN	O	O
,	NN	O	O
and	NN	O	O
may	NN	O	O
modulate	NN	O	O
immune	NN	O	O
response	NN	O	O
.	NN	O	O

-DOCSTART-	O

Human	NN	O	B-cell_type
white	NN	O	I-cell_type
blood	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
hair	NN	O	O
follicles	NN	O	O
are	NN	O	O
good	NN	O	O
sources	NN	O	O
of	NN	O	O
mRNA	NN	O	B-RNA
for	NN	O	O
the	NN	O	O
pterin	NN	O	B-protein
carbinolamine	NN	O	I-protein
dehydratase/dimerization	NN	O	I-protein
cofactor	NN	O	I-protein
of	NN	O	O
HNF1	NN	O	B-protein
for	NN	O	O
mutation	NN	O	O
detection	NN	O	O
.	NN	O	O

Pterin	NN	O	B-protein
carbinolamine	NN	O	I-protein
dehydratase/dimerization	NN	O	I-protein
cofactor	NN	O	I-protein
of	NN	O	O
HNF1	NN	O	B-protein
(	NN	O	O
PCD/DCoH	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
protein	NN	O	O
that	NN	O	O
has	NN	O	O
a	NN	O	O
dual	NN	O	O
function	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
a	NN	O	O
pterin	NN	O	B-protein
4alpha-carbinolamine	NN	O	I-protein
dehydratase	NN	O	I-protein
that	NN	O	O
is	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
regeneration	NN	O	O
of	NN	O	O
the	NN	O	O
cofactor	NN	O	B-protein
tetrahydrobiopterin	NN	O	I-protein
during	NN	O	O
the	NN	O	O
phenylalanine	NN	O	O
hydroxylase-	NN	O	O
catalyzed	NN	O	O
hydroxylation	NN	O	O
of	NN	O	O
phenylalanine	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
it	NN	O	O
is	NN	O	O
the	NN	O	O
dimerization	NN	O	O
cofactor	NN	O	O
of	NN	O	O
HNF1	NN	O	B-protein
that	NN	O	O
is	NN	O	O
able	NN	O	O
to	NN	O	O
activate	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
HNF1	NN	O	B-protein
.	NN	O	O

Deficiencies	NN	O	O
in	NN	O	O
the	NN	O	O
gene	NN	O	O
for	NN	O	O
this	NN	O	O
dual	NN	O	B-protein
functional	NN	O	I-protein
protein	NN	O	I-protein
result	NN	O	O
in	NN	O	O
hyperphenylalaninemia	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
report	NN	O	O
for	NN	O	O
the	NN	O	O
first	NN	O	O
time	NN	O	O
that	NN	O	O
the	NN	O	O
PCD/DCoH	NN	O	B-RNA
mRNA	NN	O	I-RNA
is	NN	O	O
present	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
white	NN	O	I-cell_type
blood	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
hair	NN	O	O
follicles	NN	O	O
.	NN	O	O

Taking	NN	O	O
advantage	NN	O	O
of	NN	O	O
this	NN	O	O
finding	NN	O	O
,	NN	O	O
a	NN	O	O
sensitive	NN	O	O
,	NN	O	O
rapid	NN	O	O
and	NN	O	O
convenient	NN	O	O
method	NN	O	O
for	NN	O	O
screening	NN	O	O
mutations	NN	O	O
occurring	NN	O	O
in	NN	O	O
the	NN	O	O
coding	NN	O	O
region	NN	O	O
of	NN	O	O
this	NN	O	O
gene	NN	O	O
has	NN	O	O
been	NN	O	O
described	NN	O	O
.	NN	O	O

Copyright	NN	O	O
1998	NN	O	O
Academic	NN	O	O
Press	NN	O	O
.	NN	O	O

-DOCSTART-	O

Biochemical	NN	O	O
characterization	NN	O	O
of	NN	O	O
the	NN	O	O
NF-Y	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
complex	NN	O	I-protein
during	NN	O	O
B	NN	O	O
lymphocyte	NN	O	O
development	NN	O	O
.	NN	O	O

The	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
NF-Y	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
tissue-specific	NN	O	O
major	NN	O	B-DNA
histocompatibility	NN	O	I-DNA
complex	NN	O	I-DNA
class	NN	O	I-DNA
II	NN	O	I-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
the	NN	O	O
biochemical	NN	O	O
properties	NN	O	O
of	NN	O	O
the	NN	O	O
heterotrimeric	NN	O	B-protein
NF-Y	NN	O	I-protein
complex	NN	O	I-protein
have	NN	O	O
been	NN	O	O
characterized	NN	O	O
during	NN	O	O
stage-specific	NN	O	O
B-cell	NN	O	O
development	NN	O	O
,	NN	O	O
and	NN	O	O
in	NN	O	O
several	NN	O	O
class	NN	O	B-cell_line
II-	NN	O	I-cell_line
mutant	NN	O	I-cell_line
B-cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
which	NN	O	O
represent	NN	O	O
distinct	NN	O	O
bare	NN	O	O
lymphocyte	NN	O	O
syndrome	NN	O	O
class	NN	O	O
II	NN	O	O
genetic	NN	O	O
complementation	NN	O	O
groups	NN	O	O
.	NN	O	O

The	NN	O	O
NF-Y	NN	O	B-protein
complex	NN	O	I-protein
derived	NN	O	O
from	NN	O	O
class	NN	O	B-cell_type
II+	NN	O	I-cell_type
mature	NN	O	I-cell_type
B-cells	NN	O	I-cell_type
bound	NN	O	O
with	NN	O	O
high	NN	O	O
affinity	NN	O	O
to	NN	O	O
anion	NN	O	B-protein
exchangers	NN	O	I-protein
,	NN	O	O
and	NN	O	O
eluted	NN	O	O
as	NN	O	O
an	NN	O	O
intact	NN	O	O
trimeric	NN	O	B-protein
complex	NN	O	I-protein
,	NN	O	O
whereas	NN	O	O
,	NN	O	O
NF-Y	NN	O	B-protein
derived	NN	O	O
from	NN	O	O
class	NN	O	B-cell_type
II-	NN	O	I-cell_type
plasma	NN	O	I-cell_type
B-cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
from	NN	O	O
bare	NN	O	B-cell_line
lymphocyte	NN	O	I-cell_line
syndrome	NN	O	I-cell_line
group	NN	O	I-cell_line
II	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
RJ2.2.5	NN	O	B-cell_line
and	NN	O	O
RM3	NN	O	B-cell_line
,	NN	O	O
dissociated	NN	O	O
into	NN	O	O
discrete	NN	O	O
NF-YA	NN	O	B-protein
and	NN	O	O
NF-YB	NN	O	B-protein
:	NN	O	I-protein
C	NN	O	I-protein
subunit	NN	O	B-protein
fractions	NN	O	I-protein
.	NN	O	O

Recombination	NN	O	O
of	NN	O	O
the	NN	O	O
MPC11	NN	O	B-protein
plasma	NN	O	I-protein
B-cell	NN	O	I-protein
derived	NN	O	I-protein
NF-Y	NN	O	I-protein
A	NN	O	I-protein
:	NN	O	I-protein
B	NN	O	I-protein
:	NN	O	I-protein
C	NN	O	I-protein
complex	NN	O	I-protein
with	NN	O	O
the	NN	O	O
low	NN	O	O
molecular	NN	O	O
mass	NN	O	O
protein	NN	O	O
fraction	NN	O	O
,	NN	O	O
NF-Y-associated	NN	O	B-protein
factors	NN	O	I-protein
(	NN	O	O
YAFs	NN	O	B-protein
)	NN	O	O
,	NN	O	O
derived	NN	O	O
from	NN	O	O
mature	NN	O	O
A20	NN	O	O
B-cell	NN	O	O
nuclei	NN	O	O
,	NN	O	O
conferred	NN	O	O
high	NN	O	O
affinity	NN	O	O
anion	NN	O	O
exchange	NN	O	O
binding	NN	O	O
to	NN	O	O
NF-Y	NN	O	B-protein
as	NN	O	O
an	NN	O	O
intact	NN	O	B-protein
trimeric	NN	O	I-protein
complex	NN	O	I-protein
.	NN	O	O

Recombination	NN	O	O
of	NN	O	O
the	NN	O	O
native	NN	O	B-protein
NF-YA	NN	O	I-protein
:	NN	O	I-protein
B	NN	O	I-protein
:	NN	O	I-protein
C	NN	O	I-protein
complex	NN	O	I-protein
with	NN	O	O
the	NN	O	O
transcriptional	NN	O	B-protein
cofactor	NN	O	I-protein
,	NN	O	O
PC4	NN	O	B-protein
,	NN	O	O
likewise	NN	O	O
conferred	NN	O	O
high	NN	O	B-protein
affinity	NN	O	I-protein
NF-Y	NN	O	I-protein
binding	NN	O	O
to	NN	O	O
anion	NN	O	B-protein
exchangers	NN	O	I-protein
,	NN	O	O
and	NN	O	O
stabilized	NN	O	O
NF-Y	NN	O	B-protein
interaction	NN	O	O
with	NN	O	O
CCAAT-box	NN	O	B-DNA
DNA	NN	O	I-DNA
motifs	NN	O	I-DNA
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

Interaction	NN	O	O
between	NN	O	O
PC4	NN	O	B-protein
and	NN	O	O
NF-Y	NN	O	B-protein
was	NN	O	O
mapped	NN	O	O
to	NN	O	O
the	NN	O	O
C-terminal	NN	O	B-protein
region	NN	O	I-protein
of	NN	O	O
PC4	NN	O	B-protein
,	NN	O	O
and	NN	O	O
the	NN	O	O
subunit	NN	O	B-protein
interaction	NN	O	I-protein
subdomain	NN	O	I-protein
of	NN	O	O
the	NN	O	O
highly	NN	O	B-protein
conserved	NN	O	I-protein
DNA	NN	O	I-protein
binding-subunit	NN	O	I-protein
interaction	NN	O	I-protein
domain	NN	O	I-protein
(	NN	O	O
DBD	NN	O	B-protein
)	NN	O	O
of	NN	O	O
NF-YA	NN	O	B-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
in	NN	O	O
class	NN	O	B-cell_type
II+	NN	O	I-cell_type
mature	NN	O	I-cell_type
B-cells	NN	O	I-cell_type
NF-Y	NN	O	B-protein
is	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
protein	NN	O	O
cofactor	NN	O	O
,	NN	O	O
PC4	NN	O	B-protein
,	NN	O	O
which	NN	O	O
may	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
NF-Y-mediated	NN	O	O
transcriptional	NN	O	O
control	NN	O	O
of	NN	O	O
class	NN	O	B-DNA
II	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Arrest	NN	O	O
of	NN	O	O
B	NN	O	O
lymphocyte	NN	O	O
terminal	NN	O	O
differentiation	NN	O	O
by	NN	O	O
CD40	NN	O	O
signaling	NN	O	O
:	NN	O	O
mechanism	NN	O	O
for	NN	O	O
lack	NN	O	O
of	NN	O	O
antibody-secreting	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
germinal	NN	O	O
centers	NN	O	O
.	NN	O	O

Despite	NN	O	O
extensive	NN	O	O
research	NN	O	O
,	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
CD40	NN	O	B-protein
signaling	NN	O	O
in	NN	O	O
B	NN	O	O
cell	NN	O	O
terminal	NN	O	O
differentiation	NN	O	O
remains	NN	O	O
controversial	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
CD40	NN	O	B-protein
engagement	NN	O	O
arrests	NN	O	O
B	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
prior	NN	O	O
to	NN	O	O
plasma	NN	O	O
cell	NN	O	O
formation	NN	O	O
.	NN	O	O

This	NN	O	O
arrest	NN	O	O
is	NN	O	O
manifested	NN	O	O
at	NN	O	O
a	NN	O	O
molecular	NN	O	O
level	NN	O	O
as	NN	O	O
a	NN	O	O
reduction	NN	O	O
in	NN	O	O
mRNA	NN	O	O
levels	NN	O	O
of	NN	O	O
secretory	NN	O	O
immunoglobulin	NN	O	O
gene	NN	O	O
products	NN	O	O
such	NN	O	O
as	NN	O	O
mu	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
and	NN	O	O
J	NN	O	O
chain	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
loss	NN	O	O
of	NN	O	O
the	NN	O	O
transcriptional	NN	O	B-protein
regulator	NN	O	I-protein
BLIMP-1	NN	O	B-protein
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
the	NN	O	O
inhibition	NN	O	O
of	NN	O	O
B	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
by	NN	O	O
CD40	NN	O	B-protein
engagement	NN	O	O
could	NN	O	O
not	NN	O	O
be	NN	O	O
overcome	NN	O	O
by	NN	O	O
either	NN	O	O
mitogens	NN	O	B-protein
or	NN	O	O
cytokines	NN	O	B-protein
,	NN	O	O
but	NN	O	O
could	NN	O	O
be	NN	O	O
reversed	NN	O	O
by	NN	O	O
antibodies	NN	O	O
that	NN	O	O
interfere	NN	O	O
with	NN	O	O
the	NN	O	O
CD40	NN	O	B-protein
/	NN	O	O
gp39	NN	O	B-protein
interaction	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
secretory	NN	O	O
immunoglobulin	NN	O	O
is	NN	O	O
not	NN	O	O
produced	NN	O	O
by	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
that	NN	O	O
are	NN	O	O
actively	NN	O	O
engaged	NN	O	O
by	NN	O	O
gp39-expressing	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
positively	NN	O	O
charged	NN	O	O
alpha-lipoic	NN	O	O
acid	NN	O	O
analogue	NN	O	O
with	NN	O	O
increased	NN	O	O
cellular	NN	O	O
uptake	NN	O	O
and	NN	O	O
more	NN	O	O
potent	NN	O	O
immunomodulatory	NN	O	O
activity	NN	O	O
.	NN	O	O

alpha-Lipoic	NN	O	O
acid	NN	O	O
(	NN	O	O
LA	NN	O	O
)	NN	O	O
is	NN	O	O
taken	NN	O	O
up	NN	O	O
by	NN	O	O
cells	NN	O	O
and	NN	O	O
reduced	NN	O	O
to	NN	O	O
its	NN	O	O
potent	NN	O	O
dithiol	NN	O	O
form	NN	O	O
,	NN	O	O
dihydrolipoate	NN	O	O
(	NN	O	O
DHLA	NN	O	O
)	NN	O	O
,	NN	O	O
much	NN	O	O
of	NN	O	O
which	NN	O	O
is	NN	O	O
rapidly	NN	O	O
effluxed	NN	O	O
out	NN	O	O
from	NN	O	O
cells	NN	O	O
.	NN	O	O

To	NN	O	O
improve	NN	O	O
retention	NN	O	O
in	NN	O	O
cells	NN	O	O
,	NN	O	O
the	NN	O	O
LA	NN	O	O
molecule	NN	O	O
was	NN	O	O
modified	NN	O	O
to	NN	O	O
confer	NN	O	O
a	NN	O	O
positive	NN	O	O
charge	NN	O	O
at	NN	O	O
physiological	NN	O	O
pH	NN	O	O
.	NN	O	O

N	NN	O	O
,	NN	O	O
N-dimethyl	NN	O	O
,	NN	O	O
N'-2-amidoethyl-lipoate	NN	O	O
was	NN	O	O
synthesized	NN	O	O
.	NN	O	O

The	NN	O	O
protonated	NN	O	O
form	NN	O	O
of	NN	O	O
the	NN	O	O
new	NN	O	O
molecule	NN	O	O
is	NN	O	O
referred	NN	O	O
to	NN	O	O
as	NN	O	O
LA-Plus	NN	O	O
.	NN	O	O

The	NN	O	O
uptake	NN	O	O
of	NN	O	O
LA-Plus	NN	O	O
by	NN	O	O
human	NN	O	B-cell_type
Wurzburg	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
higher	NN	O	O
compared	NN	O	O
to	NN	O	O
that	NN	O	O
of	NN	O	O
LA	NN	O	O
.	NN	O	O

Several-fold	NN	O	O
higher	NN	O	O
amounts	NN	O	O
of	NN	O	O
DHLA-Plus	NN	O	O
,	NN	O	O
the	NN	O	O
corresponding	NN	O	O
reduced	NN	O	O
form	NN	O	O
of	NN	O	O
LA-Plus	NN	O	O
,	NN	O	O
were	NN	O	O
detected	NN	O	O
in	NN	O	O
LA-Plus	NN	O	O
treated	NN	O	O
cells	NN	O	O
compared	NN	O	O
to	NN	O	O
the	NN	O	O
amount	NN	O	O
of	NN	O	O
DHLA	NN	O	O
found	NN	O	O
in	NN	O	O
cells	NN	O	O
treated	NN	O	O
with	NN	O	O
LA	NN	O	O
.	NN	O	O

At	NN	O	O
100	NN	O	O
microM	NN	O	O
,	NN	O	O
LA	NN	O	O
did	NN	O	O
not	NN	O	O
but	NN	O	O
LA-Plus	NN	O	O
inhibited	NN	O	O
H2O2	NN	O	O
induced	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
and	NN	O	O
NF-kappaB	NN	O	B-protein
directed	NN	O	O
IL-2	NN	O	B-protein
receptor	NN	O	I-protein
expression	NN	O	O
.	NN	O	O

Both	NN	O	O
LA	NN	O	O
and	NN	O	O
LA-Plus	NN	O	O
synergised	NN	O	O
with	NN	O	O
selenium	NN	O	O
in	NN	O	O
inhibiting	NN	O	O
H2O2	NN	O	O
induced	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
.	NN	O	O

At	NN	O	O
150	NN	O	O
microM	NN	O	O
LA-Plus	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
LA	NN	O	O
,	NN	O	O
inhibited	NN	O	O
TNFalpha	NN	O	B-protein
induced	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
.	NN	O	O

At	NN	O	O
5	NN	O	O
microM	NN	O	O
LA-Plus	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
LA	NN	O	O
,	NN	O	O
protected	NN	O	O
against	NN	O	O
both	NN	O	O
spontaneous	NN	O	O
and	NN	O	O
etoposide	NN	O	O
induced	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
rat	NN	O	B-cell_type
thymocytes	NN	O	I-cell_type
.	NN	O	O

LA-Plus	NN	O	O
is	NN	O	O
thus	NN	O	O
an	NN	O	O
improved	NN	O	O
form	NN	O	O
of	NN	O	O
LA	NN	O	O
with	NN	O	O
increased	NN	O	O
therapeutic	NN	O	O
potential	NN	O	O
.	NN	O	O

Copyright	NN	O	O
1998	NN	O	O
Academic	NN	O	O
Press	NN	O	O
.	NN	O	O

-DOCSTART-	O

Carrier	NN	O	O
identification	NN	O	O
in	NN	O	O
X-linked	NN	O	O
immunodeficiency	NN	O	O
diseases	NN	O	O
.	NN	O	O

OBJECTIVE	NN	O	O
:	NN	O	O
Carrier	NN	O	O
identification	NN	O	O
in	NN	O	O
X-linked	NN	O	O
immunodeficiency	NN	O	O
disorders	NN	O	O
can	NN	O	O
be	NN	O	O
based	NN	O	O
on	NN	O	O
the	NN	O	O
demonstration	NN	O	O
of	NN	O	O
non-random	NN	O	O
X	NN	O	O
inactivation	NN	O	O
(	NN	O	O
NRXI	NN	O	O
)	NN	O	O
in	NN	O	O
affected	NN	O	B-cell_type
blood	NN	O	I-cell_type
cell	NN	O	I-cell_type
lineages	NN	O	I-cell_type
when	NN	O	O
growth	NN	O	O
is	NN	O	O
impaired	NN	O	O
in	NN	O	O
cells	NN	O	O
expressing	NN	O	O
the	NN	O	O
abnormal	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
examined	NN	O	O
the	NN	O	O
utility	NN	O	O
of	NN	O	O
seeking	NN	O	O
evidence	NN	O	O
of	NN	O	O
NRXI	NN	O	O
to	NN	O	O
test	NN	O	O
the	NN	O	O
carrier	NN	O	O
status	NN	O	O
of	NN	O	O
women	NN	O	O
in	NN	O	O
families	NN	O	O
affected	NN	O	O
by	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
and	NN	O	O
X-linked	NN	O	O
hypogammaglobulinaemia	NN	O	O
(	NN	O	O
XLH	NN	O	O
)	NN	O	O
,	NN	O	O
to	NN	O	O
identify	NN	O	O
as	NN	O	O
carriers	NN	O	O
the	NN	O	O
mothers	NN	O	O
of	NN	O	O
boys	NN	O	O
with	NN	O	O
SCID	NN	O	O
or	NN	O	O
hypogammaglobulinaemia	NN	O	O
whose	NN	O	O
phenotype	NN	O	O
suggested	NN	O	O
X-linkage	NN	O	O
and	NN	O	O
to	NN	O	O
infer	NN	O	O
X-linkage	NN	O	O
in	NN	O	O
boys	NN	O	O
with	NN	O	O
SCID	NN	O	O
or	NN	O	O
hypogammaglobulinaemia	NN	O	O
whose	NN	O	O
disease	NN	O	O
was	NN	O	O
not	NN	O	O
clearly	NN	O	O
X-linked	NN	O	O
on	NN	O	O
the	NN	O	O
basis	NN	O	O
either	NN	O	O
of	NN	O	O
family	NN	O	O
history	NN	O	O
or	NN	O	O
clinical	NN	O	O
and	NN	O	O
immunological	NN	O	O
characteristics	NN	O	O
.	NN	O	O

METHODOLOGY	NN	O	O
:	NN	O	O
A	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction-based	NN	O	O
method	NN	O	O
was	NN	O	O
used	NN	O	O
to	NN	O	O
amplify	NN	O	O
a	NN	O	O
polymorphic	NN	O	O
CAG	NN	O	B-DNA
repeat	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
first	NN	O	B-DNA
exon	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
androgen	NN	O	B-DNA
receptor	NN	O	I-DNA
gene	NN	O	I-DNA
after	NN	O	O
selective	NN	O	O
digestion	NN	O	O
of	NN	O	O
the	NN	O	O
active	NN	O	B-DNA
X	NN	O	I-DNA
chromosome	NN	O	I-DNA
with	NN	O	O
a	NN	O	O
methylation-sensitive	NN	O	B-protein
enzyme	NN	O	I-protein
,	NN	O	O
HpaII	NN	O	B-protein
to	NN	O	O
distinguish	NN	O	O
between	NN	O	O
the	NN	O	O
paternal	NN	O	B-DNA
and	NN	O	I-DNA
maternal	NN	O	I-DNA
alleles	NN	O	I-DNA
and	NN	O	O
to	NN	O	O
identify	NN	O	O
their	NN	O	O
methylation	NN	O	O
status	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
Heterozygosity	NN	O	O
was	NN	O	O
found	NN	O	O
in	NN	O	O
24	NN	O	O
of	NN	O	O
31	NN	O	O
female	NN	O	O
subjects	NN	O	O
(	NN	O	O
77	NN	O	O
%	NN	O	O
)	NN	O	O
.	NN	O	O

As	NN	O	O
anticipated	NN	O	O
,	NN	O	O
NRXI	NN	O	O
could	NN	O	O
be	NN	O	O
demonstrated	NN	O	O
in	NN	O	O
all	NN	O	O
lymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
studied	NN	O	O
from	NN	O	O
obligate	NN	O	O
carriers	NN	O	O
of	NN	O	O
XSCID	NN	O	O
and	NN	O	O
an	NN	O	O
obligate	NN	O	O
carrier	NN	O	O
of	NN	O	O
XLH	NN	O	O
but	NN	O	O
not	NN	O	O
on	NN	O	O
a	NN	O	O
carrier	NN	O	O
of	NN	O	O
X-linked	NN	O	O
immunodeficiency	NN	O	O
with	NN	O	O
hyper-IgM	NN	O	B-protein
.	NN	O	O

The	NN	O	O
finding	NN	O	O
of	NN	O	O
NRXI	NN	O	O
in	NN	O	O
the	NN	O	O
mother	NN	O	O
of	NN	O	O
a	NN	O	O
boy	NN	O	O
with	NN	O	O
a	NN	O	O
SCID	NN	O	O
variant	NN	O	O
showed	NN	O	O
her	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
carrier	NN	O	O
of	NN	O	O
XSCID	NN	O	O
and	NN	O	O
establishes	NN	O	O
that	NN	O	O
her	NN	O	O
son	NN	O	O
has	NN	O	O
XSCID	NN	O	O
,	NN	O	O
not	NN	O	O
otherwise	NN	O	O
evident	NN	O	O
from	NN	O	O
available	NN	O	O
data	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
:	NN	O	O
This	NN	O	O
PCR	NN	O	O
assay	NN	O	O
provides	NN	O	O
a	NN	O	O
rapid	NN	O	O
method	NN	O	O
for	NN	O	O
carrier	NN	O	O
detection	NN	O	O
of	NN	O	O
X-linked	NN	O	O
immunodeficiencies	NN	O	O
,	NN	O	O
and	NN	O	O
has	NN	O	O
allowed	NN	O	O
us	NN	O	O
to	NN	O	O
expand	NN	O	O
the	NN	O	O
phenotype	NN	O	O
of	NN	O	O
XSCID	NN	O	O

-DOCSTART-	O

Leukocyte-endothelial	NN	O	O
interaction	NN	O	O
is	NN	O	O
augmented	NN	O	O
by	NN	O	O
high	NN	O	O
glucose	NN	O	O
concentrations	NN	O	O
and	NN	O	O
hyperglycemia	NN	O	O
in	NN	O	O
a	NN	O	O
NF-kB-dependent	NN	O	O
fashion	NN	O	O
.	NN	O	O

We	NN	O	O
addressed	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
hyperglycemia	NN	O	O
in	NN	O	O
leukocyte-endothelium	NN	O	O
interaction	NN	O	O
under	NN	O	O
flow	NN	O	O
conditions	NN	O	O
by	NN	O	O
exposing	NN	O	O
human	NN	O	B-cell_type
umbilical	NN	O	I-cell_type
vein	NN	O	I-cell_type
endothelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
for	NN	O	O
24	NN	O	O
h	NN	O	O
to	NN	O	O
normal	NN	O	O
(	NN	O	O
5	NN	O	O
mM	NN	O	O
)	NN	O	O
,	NN	O	O
high	NN	O	O
concentration	NN	O	O
of	NN	O	O
glucose	NN	O	O
(	NN	O	O
30	NN	O	O
mM	NN	O	O
)	NN	O	O
,	NN	O	O
advanced	NN	O	O
glycosylation	NN	O	O
end	NN	O	O
product-albumin	NN	O	B-protein
(	NN	O	O
100	NN	O	O
microg/ml	NN	O	O
)	NN	O	O
,	NN	O	O
or	NN	O	O
hyperglycemic	NN	O	O
(	NN	O	O
174-316	NN	O	O
mg/dl	NN	O	O
)	NN	O	O
sera	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
diabetes	NN	O	O
and	NN	O	O
abnormal	NN	O	O
hemoglobin	NN	O	O
A1c	NN	O	O
(	NN	O	O
8.1+/-1.4	NN	O	O
%	NN	O	O
)	NN	O	O
.	NN	O	O

At	NN	O	O
the	NN	O	O
end	NN	O	O
of	NN	O	O
incubation	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
perfused	NN	O	O
with	NN	O	O
total	NN	O	O
leukocyte	NN	O	O
suspension	NN	O	O
in	NN	O	O
a	NN	O	O
parallel	NN	O	O
plate	NN	O	O
flow	NN	O	O
chamber	NN	O	O
under	NN	O	O
laminar	NN	O	O
flow	NN	O	O
(	NN	O	O
1.5	NN	O	O
dyn/cm2	NN	O	O
)	NN	O	O
.	NN	O	O

Rolling	NN	O	O
and	NN	O	O
adherent	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
evaluated	NN	O	O
by	NN	O	O
digital	NN	O	O
image	NN	O	O
processing	NN	O	O
.	NN	O	O

Results	NN	O	O
showed	NN	O	O
that	NN	O	O
30	NN	O	O
mM	NN	O	O
glucose	NN	O	O
significantly	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.01	NN	O	O
)	NN	O	O
increased	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
adherent	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
to	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
respect	NN	O	O
to	NN	O	O
control	NN	O	O
(	NN	O	O
5	NN	O	O
mM	NN	O	O
glucose	NN	O	O
;	NN	O	O
151+/-19	NN	O	O
versus	NN	O	O
33+/-8	NN	O	O
cells/mm2	NN	O	O
)	NN	O	O
.	NN	O	O

A	NN	O	O
similar	NN	O	O
response	NN	O	O
was	NN	O	O
induced	NN	O	O
by	NN	O	O
endothelial	NN	O	O
stimulation	NN	O	O
with	NN	O	O
IL-1beta	NN	O	B-protein
,	NN	O	O
here	NN	O	O
used	NN	O	O
as	NN	O	O
positive	NN	O	O
control	NN	O	O
(	NN	O	O
195+/-20	NN	O	O
cells/mm2	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
number	NN	O	O
of	NN	O	O
rolling	NN	O	B-cell_type
cells	NN	O	I-cell_type
on	NN	O	O
endothelial	NN	O	O
surface	NN	O	O
was	NN	O	O
not	NN	O	O
affected	NN	O	O
by	NN	O	O
high	NN	O	O
glucose	NN	O	O
level	NN	O	O
.	NN	O	O

Stable	NN	O	O
adhesion	NN	O	O
of	NN	O	O
leukocytes	NN	O	B-cell_type
to	NN	O	O
glucose-treated	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
to	NN	O	O
IL-1beta-stimulated	NN	O	B-cell_type
endothelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
preceded	NN	O	O
by	NN	O	O
short	NN	O	O
interaction	NN	O	O
of	NN	O	O
leukocytes	NN	O	B-cell_type
with	NN	O	O
the	NN	O	O
endothelial	NN	O	O
surface	NN	O	O
.	NN	O	O

The	NN	O	O
distance	NN	O	O
travelled	NN	O	O
by	NN	O	O
leukocytes	NN	O	B-cell_type
before	NN	O	O
arrest	NN	O	O
on	NN	O	O
30	NN	O	O
mM	NN	O	O
glucose	NN	O	O
,	NN	O	O
or	NN	O	O
on	NN	O	O
IL-1beta-treated	NN	O	B-cell_type
endothelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
was	NN	O	O
significantly	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.01	NN	O	O
)	NN	O	O
higher	NN	O	O
than	NN	O	O
that	NN	O	O
observed	NN	O	O
for	NN	O	O
leukocytes	NN	O	B-cell_type
adhering	NN	O	O
on	NN	O	O
control	NN	O	O
endothelium	NN	O	O
(	NN	O	O
30	NN	O	O
mM	NN	O	O
glucose	NN	O	O
:	NN	O	O
76.7+/-3.5	NN	O	O
;	NN	O	O
IL1beta	NN	O	O
:	NN	O	O
69.7+/-4	NN	O	O
versus	NN	O	O
5	NN	O	O
mM	NN	O	O
glucose	NN	O	O
:	NN	O	O
21.5+/-5	NN	O	O
microm	NN	O	O
)	NN	O	O
.	NN	O	O

Functional	NN	O	O
blocking	NN	O	O
of	NN	O	O
E-selectin	NN	O	B-protein
,	NN	O	O
intercellular	NN	O	B-protein
cell	NN	O	I-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
,	NN	O	O
and	NN	O	O
vascular	NN	O	B-protein
cell	NN	O	I-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
on	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
the	NN	O	O
corresponding	NN	O	O
mouse	NN	O	B-protein
mAb	NN	O	I-protein
significantly	NN	O	O
inhibited	NN	O	O
glucose-induced	NN	O	O
increase	NN	O	O
in	NN	O	O
leukocyte	NN	O	O
adhesion	NN	O	O
(	NN	O	O
67+/-16	NN	O	O
,	NN	O	O
83+/-12	NN	O	O
,	NN	O	O
62+/-8	NN	O	O
versus	NN	O	O
144+/-21	NN	O	O
cells/	NN	O	O
mm2	NN	O	O
)	NN	O	O
.	NN	O	O

Confocal	NN	O	O
fluorescence	NN	O	O
microscopy	NN	O	O
studies	NN	O	O
showed	NN	O	O
that	NN	O	O
30	NN	O	O
mM	NN	O	O
glucose	NN	O	O
induced	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
endothelial	NN	O	O
surface	NN	O	O
expression	NN	O	O
of	NN	O	O
E-selectin	NN	O	B-protein
,	NN	O	O
intercellular	NN	O	B-protein
cell	NN	O	I-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
,	NN	O	O
and	NN	O	O
vascular	NN	O	B-protein
cell	NN	O	I-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
.	NN	O	O

Electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
of	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
umbilical	NN	O	I-cell_type
vein	NN	O	I-cell_type
endothelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
HUVEC	NN	O	O
)	NN	O	O
exposed	NN	O	O
for	NN	O	O
1	NN	O	O
h	NN	O	O
to	NN	O	O
30	NN	O	O
mM	NN	O	O
glucose	NN	O	O
revealed	NN	O	O
an	NN	O	O
intense	NN	O	O
NF-kB	NN	O	B-protein
activation	NN	O	O
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
HUVEC	NN	O	O
exposed	NN	O	O
to	NN	O	O
high	NN	O	O
glucose	NN	O	O
with	NN	O	O
the	NN	O	O
NF-kB	NN	O	B-protein
inhibitors	NN	O	I-protein
pyrrolidinedithiocarbamate	NN	O	O
(	NN	O	O
100	NN	O	O
microM	NN	O	O
)	NN	O	O
and	NN	O	O
tosyl-phe-chloromethylketone	NN	O	O
(	NN	O	O
25	NN	O	O
microM	NN	O	O
)	NN	O	O
significantly	NN	O	O
reduced	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.05	NN	O	O
)	NN	O	O
leukocyte	NN	O	O
adhesion	NN	O	O
in	NN	O	O
respect	NN	O	O
to	NN	O	O
HUVEC	NN	O	O
treated	NN	O	O
with	NN	O	O
glucose	NN	O	O
alone	NN	O	O
.	NN	O	O

A	NN	O	O
significant	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.01	NN	O	O
)	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
on	NN	O	O
glucose-induced	NN	O	O
leukocyte	NN	O	O
adhesion	NN	O	O
was	NN	O	O
observed	NN	O	O
after	NN	O	O
blocking	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
activity	NN	O	O
with	NN	O	O
staurosporine	NN	O	O
(	NN	O	O
5	NN	O	O
nM	NN	O	O
)	NN	O	O
.	NN	O	O

When	NN	O	O
HUVEC	NN	O	O
were	NN	O	O
treated	NN	O	O
with	NN	O	O
specific	NN	O	O
antisense	NN	O	O
oligodesoxynucleotides	NN	O	O
against	NN	O	O
PKCalpha	NN	O	B-protein
and	NN	O	I-protein
PKCepsilon	NN	O	I-protein
isoforms	NN	O	I-protein
before	NN	O	O
the	NN	O	O
addition	NN	O	O
of	NN	O	O
30	NN	O	O
mM	NN	O	O
glucose	NN	O	O
,	NN	O	O
a	NN	O	O
significant	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.05	NN	O	O
)	NN	O	O
reduction	NN	O	O
in	NN	O	O
the	NN	O	O
adhesion	NN	O	O
was	NN	O	O
also	NN	O	O
seen	NN	O	O
.	NN	O	O

Advanced	NN	O	O
glycosylation	NN	O	O
end	NN	O	O
product-albumin	NN	O	B-protein
significantly	NN	O	O
increased	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
adhering	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
in	NN	O	O
respect	NN	O	O
to	NN	O	O
native	NN	O	B-protein
albumin	NN	O	I-protein
used	NN	O	O
as	NN	O	O
control	NN	O	O
(	NN	O	O
110+/-16	NN	O	O
versus	NN	O	O
66+/-7	NN	O	O
,	NN	O	O
P	NN	O	O
<	NN	O	O
0.01	NN	O	O
)	NN	O	O
.	NN	O	O

Sera	NN	O	O
from	NN	O	O
diabetic	NN	O	O
patients	NN	O	O
significantly	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.01	NN	O	O
)	NN	O	O
enhanced	NN	O	O
leukocyte	NN	O	O
adhesion	NN	O	O
as	NN	O	O
compared	NN	O	O
with	NN	O	O
controls	NN	O	O
,	NN	O	O
despite	NN	O	O
normal	NN	O	O
levels	NN	O	O
of	NN	O	O
IL-1beta	NN	O	B-protein
and	NN	O	O
TNFalpha	NN	O	B-protein
in	NN	O	O
these	NN	O	O
sera	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
high	NN	O	O
glucose	NN	O	O
concentration	NN	O	O
and	NN	O	O
hyperglycemia	NN	O	O
promote	NN	O	O
leukocyte	NN	O	O
adhesion	NN	O	O
to	NN	O	O
the	NN	O	O
endothelium	NN	O	O
through	NN	O	O
upregulation	NN	O	O
of	NN	O	O
cell	NN	O	O
surface	NN	O	O
expression	NN	O	O
of	NN	O	O
adhesive	NN	O	O
proteins	NN	O	O
,	NN	O	O
possibly	NN	O	O
depending	NN	O	O
on	NN	O	O
NF-kB	NN	O	B-protein
activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Ikaros	NN	O	B-protein
in	NN	O	O
hemopoietic	NN	O	O
lineage	NN	O	O
determination	NN	O	O
and	NN	O	O
homeostasis	NN	O	O
.	NN	O	O

Studies	NN	O	O
on	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
that	NN	O	O
control	NN	O	O
hemopoietic	NN	O	O
differentiation	NN	O	O
have	NN	O	O
focused	NN	O	O
on	NN	O	O
signaling	NN	O	O
cascades	NN	O	O
and	NN	O	O
nuclear	NN	O	O
effectors	NN	O	O
that	NN	O	O
drive	NN	O	O
this	NN	O	O
complex	NN	O	O
developmental	NN	O	O
system	NN	O	O
in	NN	O	O
a	NN	O	O
regulated	NN	O	O
fashion	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
review	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
Ikaros	NN	O	B-protein
,	NN	O	O
the	NN	O	O
founding	NN	O	O
member	NN	O	O
of	NN	O	O
a	NN	O	O
unique	NN	O	O
family	NN	O	O
of	NN	O	O
zinc	NN	O	B-protein
finger	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
in	NN	O	O
this	NN	O	O
developmental	NN	O	O
process	NN	O	O
.	NN	O	O

Studies	NN	O	O
on	NN	O	O
an	NN	O	O
Ikaros	NN	O	B-protein
null	NN	O	O
mutation	NN	O	O
have	NN	O	O
revealed	NN	O	O
an	NN	O	O
essential	NN	O	O
role	NN	O	O
for	NN	O	O
this	NN	O	O
factor	NN	O	O
in	NN	O	O
lymphoid	NN	O	O
cell	NN	O	O
fate	NN	O	O
determination	NN	O	O
and	NN	O	O
at	NN	O	O
subsequent	NN	O	O
branch	NN	O	O
points	NN	O	O
of	NN	O	O
the	NN	O	O
T	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
pathway	NN	O	O
.	NN	O	O

Differences	NN	O	O
in	NN	O	O
the	NN	O	O
phenotypes	NN	O	O
of	NN	O	O
a	NN	O	O
null	NN	O	O
and	NN	O	O
a	NN	O	O
dominant	NN	O	B-DNA
negative	NN	O	I-DNA
(	NN	O	I-DNA
DN	NN	O	I-DNA
)	NN	O	I-DNA
Ikaros	NN	O	I-DNA
mutation	NN	O	I-DNA
provide	NN	O	O
insight	NN	O	O
into	NN	O	O
a	NN	O	O
regulatory	NN	O	O
network	NN	O	O
through	NN	O	O
which	NN	O	O
Ikaros	NN	O	B-protein
proteins	NN	O	I-protein
exert	NN	O	O
their	NN	O	O
effects	NN	O	O
in	NN	O	O
development	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
a	NN	O	O
comparative	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
hemopoietic	NN	O	B-cell_type
stem	NN	O	I-cell_type
cell	NN	O	I-cell_type
and	NN	O	O
precursor	NN	O	O
compartment	NN	O	O
resulting	NN	O	O
from	NN	O	O
the	NN	O	O
two	NN	O	O
Ikaros	NN	O	B-DNA
mutations	NN	O	I-DNA
reveals	NN	O	O
a	NN	O	O
profound	NN	O	O
yet	NN	O	O
not	NN	O	O
absolute	NN	O	O
requirement	NN	O	O
for	NN	O	O
Ikaros	NN	O	B-protein

-DOCSTART-	O

Induction	NN	O	O
of	NN	O	O
early	NN	O	B-protein
B	NN	O	I-protein
cell	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
EBF	NN	O	B-protein
)	NN	O	O
and	NN	O	O
multiple	NN	O	B-DNA
B	NN	O	I-DNA
lineage	NN	O	I-DNA
genes	NN	O	I-DNA
by	NN	O	O
the	NN	O	O
basic	NN	O	B-protein
helix-loop-helix	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
E12	NN	O	B-protein
.	NN	O	O

The	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
encoded	NN	O	O
by	NN	O	O
the	NN	O	O
E2A	NN	O	B-protein
and	NN	O	O
early	NN	O	B-protein
B	NN	O	I-protein
cell	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
EBF	NN	O	B-protein
)	NN	O	O
genes	NN	O	O
are	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
proper	NN	O	O
development	NN	O	O
of	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
B	NN	O	B-cell_type
lineage	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
E2A-	NN	O	O
and	NN	O	O
EBF-deficient	NN	O	O
mice	NN	O	O
has	NN	O	O
made	NN	O	O
it	NN	O	O
difficult	NN	O	O
to	NN	O	O
determine	NN	O	O
the	NN	O	O
function	NN	O	O
or	NN	O	O
relationship	NN	O	O
between	NN	O	O
these	NN	O	O
proteins	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
the	NN	O	O
identification	NN	O	O
of	NN	O	O
a	NN	O	O
novel	NN	O	O
model	NN	O	O
system	NN	O	O
in	NN	O	O
which	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
E2A	NN	O	B-protein
and	NN	O	O
EBF	NN	O	B-protein
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
multiple	NN	O	O
B	NN	O	O
lineage	NN	O	O
traits	NN	O	O
can	NN	O	O
be	NN	O	O
studied	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
conversion	NN	O	O
of	NN	O	O
70Z/3	NN	O	B-cell_line
pre-B	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
to	NN	O	O
cells	NN	O	O
with	NN	O	O
a	NN	O	O
macrophage-like	NN	O	O
phenotype	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
loss	NN	O	O
of	NN	O	O
E2A	NN	O	B-protein
and	NN	O	O
EBF	NN	O	B-protein
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
ectopic	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
E2A	NN	O	B-protein
protein	NN	O	O
E12	NN	O	B-protein
in	NN	O	O
this	NN	O	O
macrophage	NN	O	B-cell_line
line	NN	O	I-cell_line
results	NN	O	O
in	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
many	NN	O	B-DNA
B	NN	O	I-DNA
lineage	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
including	NN	O	O
EBF	NN	O	B-protein
,	NN	O	O
IL7Ralpha	NN	O	B-protein
,	NN	O	O
lambda5	NN	O	B-protein
,	NN	O	O
and	NN	O	O
Rag-1	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
ability	NN	O	O
to	NN	O	O
induce	NN	O	O
kappa	NN	O	B-protein
light	NN	O	I-protein
chain	NN	O	I-protein
in	NN	O	O
response	NN	O	O
to	NN	O	O
mitogen	NN	O	O
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
EBF	NN	O	B-protein
may	NN	O	O
be	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
critical	NN	O	O
functions	NN	O	O
of	NN	O	O
E12	NN	O	B-protein
in	NN	O	O
regulating	NN	O	O
the	NN	O	O
B	NN	O	O
lineage	NN	O	O
phenotype	NN	O	O
since	NN	O	O
expression	NN	O	O
of	NN	O	O
EBF	NN	O	B-protein
alone	NN	O	O
leads	NN	O	O
to	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
a	NN	O	O
subset	NN	O	O
of	NN	O	O
E12	NN	O	B-protein
-inducible	NN	O	O
traits	NN	O	O
.	NN	O	O

Our	NN	O	O
data	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
,	NN	O	O
in	NN	O	O
the	NN	O	O
context	NN	O	O
of	NN	O	O
this	NN	O	O
macrophage	NN	O	B-cell_line
line	NN	O	I-cell_line
,	NN	O	O
E12	NN	O	B-protein
induces	NN	O	O
expression	NN	O	O
of	NN	O	O
EBF	NN	O	B-protein
and	NN	O	O
together	NN	O	O
these	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
coordinately	NN	O	O
regulate	NN	O	O
numerous	NN	O	O
B	NN	O	B-DNA
lineage-associated	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Activation-induced	NN	O	O
down-regulation	NN	O	O
of	NN	O	O
retinoid	NN	O	B-protein
receptor	NN	O	I-protein
RXRalpha	NN	O	I-protein
expression	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Role	NN	O	O
of	NN	O	O
cell	NN	O	O
cycle	NN	O	O
regulation	NN	O	O
.	NN	O	O

A	NN	O	O
5.4-kilobase	NN	O	B-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
which	NN	O	O
is	NN	O	O
down-regulated	NN	O	O
after	NN	O	O
treatment	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
PBMCs	NN	O	B-cell_type
)	NN	O	O
with	NN	O	O
various	NN	O	O
T	NN	O	O
cell-activating	NN	O	O
agents	NN	O	O
,	NN	O	O
was	NN	O	O
isolated	NN	O	O
using	NN	O	O
an	NN	O	O
mRNA	NN	O	O
differential	NN	O	O
display	NN	O	O
method	NN	O	O
.	NN	O	O

Nucleotide	NN	O	O
sequence	NN	O	O
analysis	NN	O	O
identified	NN	O	O
the	NN	O	O
5	NN	O	B-RNA
'	NN	O	I-RNA
end	NN	O	I-RNA
of	NN	O	O
this	NN	O	O
RNA	NN	O	B-RNA
as	NN	O	O
human	NN	O	B-RNA
retinoid	NN	O	I-RNA
receptor	NN	O	I-RNA
RXRalpha	NN	O	I-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
report	NN	O	O
the	NN	O	O
nucleotide	NN	O	O
sequence	NN	O	O
of	NN	O	O
3.6	NN	O	B-DNA
kilobases	NN	O	I-DNA
of	NN	O	O
this	NN	O	O
RNA	NN	O	O
,	NN	O	O
which	NN	O	O
represents	NN	O	O
the	NN	O	O
3	NN	O	B-RNA
'	NN	O	I-RNA
end	NN	O	I-RNA
of	NN	O	O
RXRalpha	NN	O	B-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
the	NN	O	O
sequence	NN	O	O
of	NN	O	O
which	NN	O	O
has	NN	O	O
not	NN	O	O
been	NN	O	O
previously	NN	O	O
described	NN	O	O
.	NN	O	O

Activated	NN	O	O
PBMCs	NN	O	B-cell_type
also	NN	O	O
expressed	NN	O	O
lower	NN	O	O
levels	NN	O	O
of	NN	O	O
RXRalpha	NN	O	B-protein
protein	NN	O	O
,	NN	O	O
and	NN	O	O
a	NN	O	O
DNA	NN	O	O
binding	NN	O	O
assay	NN	O	O
showed	NN	O	O
that	NN	O	O
the	NN	O	O
activation-induced	NN	O	O
loss	NN	O	O
of	NN	O	O
RXRalpha	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
protein	NN	O	O
expression	NN	O	O
correlated	NN	O	O
with	NN	O	O
the	NN	O	O
loss	NN	O	O
of	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
of	NN	O	O
this	NN	O	O
protein	NN	O	O
.	NN	O	O

We	NN	O	O
present	NN	O	O
evidence	NN	O	O
that	NN	O	O
the	NN	O	O
transition	NN	O	O
from	NN	O	O
G0/G1	NN	O	O
to	NN	O	O
S	NN	O	O
phase	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
results	NN	O	O
in	NN	O	O
the	NN	O	O
down-regulation	NN	O	O
of	NN	O	O
RXRalpha	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
that	NN	O	O
cell	NN	O	O
cycle	NN	O	O
inhibitors	NN	O	O
,	NN	O	O
which	NN	O	O
block	NN	O	O
the	NN	O	O
cells	NN	O	O
in	NN	O	O
G1	NN	O	O
phase	NN	O	O
,	NN	O	O
prevent	NN	O	O
this	NN	O	O
down-regulation	NN	O	O
.	NN	O	O

The	NN	O	O
decrease	NN	O	O
in	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
RXRalpha	NN	O	B-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
regulated	NN	O	O
at	NN	O	O
the	NN	O	O
post-transcriptional	NN	O	O
level	NN	O	O
and	NN	O	O
involved	NN	O	O
new	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
.	NN	O	O

These	NN	O	O
observations	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
RXRalpha	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
T	NN	O	O
lymphocytes	NN	O	O
are	NN	O	O
coupled	NN	O	O
to	NN	O	O
cell	NN	O	O
cycle	NN	O	O
progression	NN	O	O
,	NN	O	O
and	NN	O	O
there	NN	O	O
is	NN	O	O
tight	NN	O	O
regulatory	NN	O	O
control	NN	O	O
of	NN	O	O
RXRalpha	NN	O	B-protein
expression	NN	O	O
during	NN	O	O
the	NN	O	O
transition	NN	O	O
from	NN	O	O
G0/G1	NN	O	O
to	NN	O	O
S	NN	O	O
phase	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
.	NN	O	O

-DOCSTART-	O

Epidemiology	NN	O	O
and	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
AIDS-related	NN	O	O
lymphomas	NN	O	O
.	NN	O	O

Among	NN	O	O
patients	NN	O	O
with	NN	O	O
congenital	NN	O	O
and	NN	O	O
acquired	NN	O	O
immunodeficiencies	NN	O	O
,	NN	O	O
non-Hodgkin	NN	O	O
's	NN	O	O
lymphoma	NN	O	O
(	NN	O	O
NHLs	NN	O	O
)	NN	O	O
are	NN	O	O
the	NN	O	O
most	NN	O	O
common	NN	O	O
tumors	NN	O	O
of	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
setting	NN	O	O
of	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
(	NN	O	O
HIV	NN	O	O
)	NN	O	O
infection	NN	O	O
,	NN	O	O
as	NN	O	O
many	NN	O	O
as	NN	O	O
10	NN	O	O
%	NN	O	O
to	NN	O	O
20	NN	O	O
%	NN	O	O
of	NN	O	O
people	NN	O	O
ultimately	NN	O	O
developed	NN	O	O
NHLs	NN	O	O
.	NN	O	O

These	NN	O	O
tumors	NN	O	O
are	NN	O	O
clinically	NN	O	O
aggressive	NN	O	O
,	NN	O	O
frequently	NN	O	O
involve	NN	O	O
extranodal	NN	O	O
sites	NN	O	O
,	NN	O	O
and	NN	O	O
often	NN	O	O
exhibit	NN	O	O
unique	NN	O	O
features	NN	O	O
that	NN	O	O
distinguish	NN	O	O
them	NN	O	O
from	NN	O	O
NHL	NN	O	O
arising	NN	O	O
in	NN	O	O
individuals	NN	O	O
with	NN	O	O
other	NN	O	O
forms	NN	O	O
of	NN	O	O
immunosuppression	NN	O	O
.	NN	O	O

Important	NN	O	O
in	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
HIV-associated	NN	O	O
NHL	NN	O	O
are	NN	O	O
cytokines	NN	O	B-protein
and	NN	O	O
other	NN	O	O
factors	NN	O	O
that	NN	O	O
induce	NN	O	O
B-cell	NN	O	O
proliferation	NN	O	O
and	NN	O	O
increase	NN	O	O
the	NN	O	O
likelihood	NN	O	O
of	NN	O	O
mutations	NN	O	O
of	NN	O	O
c-myc	NN	O	B-DNA
,	NN	O	O
bcl-6	NN	O	B-DNA
,	NN	O	O
and	NN	O	O
other	NN	O	O
tumor-suppressor	NN	O	B-DNA
genes	NN	O	I-DNA
with	NN	O	O
carcinogenic	NN	O	O
potential	NN	O	O
.	NN	O	O

Specific	NN	O	O
forms	NN	O	O
of	NN	O	O
HIV-associated	NN	O	O
NHL	NN	O	O
are	NN	O	O
linked	NN	O	O
to	NN	O	O
expression	NN	O	O
of	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
-latent	NN	O	I-protein
proteins	NN	O	I-protein
;	NN	O	O
the	NN	O	O
newly	NN	O	O
described	NN	O	O
DNA	NN	O	O
virus	NN	O	O
,	NN	O	O
Karposi	NN	O	O
's	NN	O	O
sarcoma-associated	NN	O	O
herpesvirus/human	NN	O	O
herpesvirus-8	NN	O	O
(	NN	O	O
KSHV/HHV-8	NN	O	O
)	NN	O	O
;	NN	O	O
and	NN	O	O
perhaps	NN	O	O
HIV	NN	O	O
.	NN	O	O

Elucidation	NN	O	O
of	NN	O	O
the	NN	O	O
factors	NN	O	O
that	NN	O	O
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
high	NN	O	O
incidence	NN	O	O
of	NN	O	O
NHL	NN	O	O
among	NN	O	O
patients	NN	O	O
infected	NN	O	O
with	NN	O	O
HIV	NN	O	O
provides	NN	O	O
insights	NN	O	O
into	NN	O	O
important	NN	O	O
elements	NN	O	O
of	NN	O	O
lymphomagenesis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Matrix	NN	O	B-protein
metalloproteinase	NN	O	I-protein
expression	NN	O	O
in	NN	O	O
human	NN	O	O
breast	NN	O	O
cancer	NN	O	O
:	NN	O	O
an	NN	O	O
immunohistochemical	NN	O	O
study	NN	O	O
including	NN	O	O
correlation	NN	O	O
with	NN	O	O
cathepsin	NN	O	B-protein
D	NN	O	I-protein
,	NN	O	O
type	NN	O	B-protein
IV	NN	O	I-protein
collagen	NN	O	I-protein
,	NN	O	O
laminin	NN	O	B-protein
,	NN	O	O
fibronectin	NN	O	B-protein
,	NN	O	O
EGFR	NN	O	B-protein
,	NN	O	O
c-erbB-2	NN	O	B-protein
oncoprotein	NN	O	I-protein
,	NN	O	O
p53	NN	O	B-protein
,	NN	O	O
steroid	NN	O	O
receptors	NN	O	O
status	NN	O	O
and	NN	O	O
proliferative	NN	O	O
indices	NN	O	O
.	NN	O	O

Matrix	NN	O	B-protein
metalloproteinase	NN	O	I-protein
s	NN	O	O
(	NN	O	O
MMPs	NN	O	B-protein
)	NN	O	O
are	NN	O	O
a	NN	O	O
group	NN	O	O
of	NN	O	O
enzymes	NN	O	O
thought	NN	O	O
to	NN	O	O
be	NN	O	O
responsible	NN	O	O
for	NN	O	O
both	NN	O	O
normal	NN	O	O
connective	NN	O	O
tissue	NN	O	O
matrix	NN	O	O
remodelling	NN	O	O
and	NN	O	O
accelerated	NN	O	O
breakdown	NN	O	O
associated	NN	O	O
with	NN	O	O
tumour	NN	O	O
development	NN	O	O
.	NN	O	O

The	NN	O	O
current	NN	O	O
study	NN	O	O
aimed	NN	O	O
to	NN	O	O
investigate	NN	O	O
the	NN	O	O
immunohistochemical	NN	O	O
expression	NN	O	O
of	NN	O	O
matrix	NN	O	B-protein
metalloproteinase	NN	O	I-protein
3	NN	O	I-protein
(	NN	O	O
MMP-3	NN	O	B-protein
,	NN	O	O
stromelysin-1	NN	O	B-protein
)	NN	O	O
in	NN	O	O
correlation	NN	O	O
with	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
Basement	NN	O	B-protein
Membrane	NN	O	I-protein
(	NN	O	I-protein
BM	NN	O	I-protein
)	NN	O	I-protein
antigen	NN	O	I-protein
(	NN	O	O
type	NN	O	B-protein
IV	NN	O	I-protein
collagen	NN	O	I-protein
,	NN	O	O
laminin	NN	O	B-protein
)	NN	O	O
,	NN	O	O
fibronectin	NN	O	B-protein
,	NN	O	O
cathepsin	NN	O	B-protein
D	NN	O	I-protein
,	NN	O	O
p53	NN	O	B-protein
,	NN	O	O
c-erbB-2	NN	O	B-protein
,	NN	O	O
proliferative	NN	O	O
activity	NN	O	O
(	NN	O	O
Ki-67	NN	O	B-protein
,	NN	O	O
PCNA	NN	O	B-protein
)	NN	O	O
,	NN	O	O
steroid	NN	O	O
receptor	NN	O	O
content	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
to	NN	O	O
the	NN	O	O
other	NN	O	O
conventional	NN	O	O
clinicopathological	NN	O	O
parameters	NN	O	O
in	NN	O	O
breast	NN	O	O
cancer	NN	O	O
.	NN	O	O

This	NN	O	O
study	NN	O	O
was	NN	O	O
performed	NN	O	O
on	NN	O	O
a	NN	O	O
series	NN	O	O
of	NN	O	O
frozen	NN	O	O
and	NN	O	O
paraffin	NN	O	O
sections	NN	O	O
from	NN	O	O
84	NN	O	O
breast	NN	O	O
cancer	NN	O	O
specimens	NN	O	O
by	NN	O	O
immunohistochemistry	NN	O	O
using	NN	O	O
the	NN	O	O
monoclonal	NN	O	B-protein
antibody	NN	O	I-protein
MMP-3	NN	O	I-protein
(	NN	O	O
Ab-1	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Stromelysin-1	NN	O	B-protein
(	NN	O	O
ST1	NN	O	B-protein
)	NN	O	O
was	NN	O	O
observed	NN	O	O
in	NN	O	O
about	NN	O	O
10	NN	O	O
%	NN	O	O
of	NN	O	O
epithelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
control	NN	O	O
groups	NN	O	O
(	NN	O	O
cases	NN	O	O
of	NN	O	O
fibrocystic	NN	O	O
and	NN	O	O
benign	NN	O	O
proliferative	NN	O	O
breast	NN	O	O
disease	NN	O	O
)	NN	O	O
,	NN	O	O
while	NN	O	O
expression	NN	O	O
(	NN	O	O
>	NN	O	O
10	NN	O	O
%	NN	O	O
of	NN	O	O
expression	NN	O	O
)	NN	O	O
was	NN	O	O
detected	NN	O	O
in	NN	O	O
89.7	NN	O	O
%	NN	O	O
of	NN	O	O
tumours	NN	O	O
.	NN	O	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
ST1	NN	O	B-protein
in	NN	O	O
carcinoma	NN	O	O
cells	NN	O	O
was	NN	O	O
strongly	NN	O	O
associated	NN	O	O
with	NN	O	O
its	NN	O	O
presence	NN	O	O
in	NN	O	O
the	NN	O	O
stroma	NN	O	O
(	NN	O	O
p	NN	O	O
<	NN	O	O
0.001	NN	O	O
)	NN	O	O
.	NN	O	O

A	NN	O	O
significantly	NN	O	O
positive	NN	O	O
correlation	NN	O	O
was	NN	O	O
found	NN	O	O
between	NN	O	O
ST1	NN	O	B-protein
expression	NN	O	O
,	NN	O	O
and	NN	O	O
p53	NN	O	B-protein
tumour	NN	O	O
suppressor	NN	O	O
gene	NN	O	O
product	NN	O	O
(	NN	O	O
p	NN	O	O
=	NN	O	O
0.004	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
a	NN	O	O
relationship	NN	O	O
with	NN	O	O
c-erbB-2	NN	O	B-protein
protein	NN	O	O
and	NN	O	O
progesterone	NN	O	O
receptor	NN	O	O
status	NN	O	O
was	NN	O	O
also	NN	O	O
indicated	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
ST1	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
breast	NN	O	O
cancer	NN	O	O
tissue	NN	O	O
is	NN	O	O
irrespective	NN	O	O
of	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
extracellular	NN	O	O
matrix	NN	O	O
component	NN	O	O
,	NN	O	O
the	NN	O	O
proteolytic	NN	O	B-protein
enzyme	NN	O	I-protein
cathepsin	NN	O	B-protein
D	NN	O	I-protein
and	NN	O	O
the	NN	O	O
growth	NN	O	O
fraction	NN	O	O
of	NN	O	O
the	NN	O	O
tumour	NN	O	O
,	NN	O	O
and	NN	O	O
that	NN	O	O
it	NN	O	O
could	NN	O	O
be	NN	O	O
a	NN	O	O
potential	NN	O	O
new	NN	O	O
prognostic	NN	O	O
marker	NN	O	O
in	NN	O	O
breast	NN	O	O
cancer	NN	O	O
.	NN	O	O

-DOCSTART-	O

Use	NN	O	O
of	NN	O	O
transfected	NN	O	B-cell_line
liver	NN	O	I-cell_line
cells	NN	O	I-cell_line
to	NN	O	O
evaluate	NN	O	O
potential	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
alcohol-induced	NN	O	O
liver	NN	O	O
injury	NN	O	O
[	NN	O	O
see	NN	O	O
comments	NN	O	O
]	NN	O	O

There	NN	O	O
is	NN	O	O
increased	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
proinflammatory	NN	O	B-protein
cytokine	NN	O	I-protein
,	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
TNF	NN	O	B-protein
)	NN	O	O
in	NN	O	O
alcoholic	NN	O	O
liver	NN	O	O
disease	NN	O	O
(	NN	O	O
ALD	NN	O	O
)	NN	O	O
.	NN	O	O

Hepatic	NN	O	O
neutrophil	NN	O	O
infiltration	NN	O	O
is	NN	O	O
a	NN	O	O
principal	NN	O	O
injurious	NN	O	O
manifestation	NN	O	O
of	NN	O	O
ALD	NN	O	O
.	NN	O	O

TNF	NN	O	B-protein
can	NN	O	O
induce	NN	O	O
cellular	NN	O	O
oxidative	NN	O	O
injury	NN	O	O
directly	NN	O	O
,	NN	O	O
and	NN	O	O
indirectly	NN	O	O
by	NN	O	O
inducing	NN	O	O
neutrophil	NN	O	O
chemotactic	NN	O	O
factor	NN	O	O
(	NN	O	O
IL-8	NN	O	B-protein
)	NN	O	O
production	NN	O	O
by	NN	O	O
hepatocytes	NN	O	O
.	NN	O	O

IL-8	NN	O	B-protein
activates	NN	O	O
and	NN	O	O
chemotactically	NN	O	O
attracts	NN	O	O
neutrophils	NN	O	O
to	NN	O	O
the	NN	O	O
liver	NN	O	O
where	NN	O	O
they	NN	O	O
release	NN	O	O
oxidizing	NN	O	O
substances	NN	O	O
.	NN	O	O

Patients	NN	O	O
with	NN	O	O
ALD	NN	O	O
also	NN	O	O
have	NN	O	O
decreased	NN	O	O
protective	NN	O	B-protein
factors	NN	O	I-protein
for	NN	O	O
cellular	NN	O	O
oxidative	NN	O	O
injury	NN	O	O
.	NN	O	O

Manganous	NN	O	B-protein
superoxide	NN	O	I-protein
dismutase	NN	O	I-protein
(	NN	O	O
MnSOD	NN	O	B-protein
)	NN	O	O
is	NN	O	O
an	NN	O	O
antioxidant	NN	O	B-protein
protective	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

The	NN	O	O
objectives	NN	O	O
of	NN	O	O
these	NN	O	O
studies	NN	O	O
were	NN	O	O
to	NN	O	O
investigate	NN	O	O
mechanisms	NN	O	O
for	NN	O	O
induction	NN	O	O
of	NN	O	O
an	NN	O	O
injurious	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	O
IL-8	NN	O	B-protein
)	NN	O	O
and	NN	O	O
a	NN	O	O
protective	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	O
MnSOD	NN	O	B-protein
)	NN	O	O
in	NN	O	O
the	NN	O	O
HepG2	NN	O	B-cell_line
human	NN	O	I-cell_line
hepatoma	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
the	NN	O	O
first	NN	O	O
set	NN	O	O
of	NN	O	O
experiments	NN	O	O
,	NN	O	O
IL-8	NN	O	B-DNA
gene	NN	O	I-DNA
reporter	NN	O	I-DNA
constructs	NN	O	I-DNA
were	NN	O	O
used	NN	O	O
to	NN	O	O
transiently	NN	O	O
transfect	NN	O	O
a	NN	O	O
derivative	NN	O	O
(	NN	O	O
MVh2E1-9	NN	O	B-cell_line
)	NN	O	O
of	NN	O	O
the	NN	O	O
HepG2	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
which	NN	O	O
expresses	NN	O	O
P-4502E1	NN	O	B-protein
and	NN	O	O
metabolizes	NN	O	O
ethanol	NN	O	O
.	NN	O	O

Inactivation	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-DNA
and	NN	O	I-DNA
3'NF-IL-6	NN	O	I-DNA
DNA	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
decreased	NN	O	O
IL-8	NN	O	B-DNA
gene	NN	O	I-DNA
transcriptional	NN	O	O
activation	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
TNF	NN	O	B-protein
while	NN	O	O
inactivation	NN	O	O
of	NN	O	O
the	NN	O	O
5'NF-IL-6	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
increased	NN	O	O
IL-8	NN	O	B-DNA
gene	NN	O	I-DNA
transcriptional	NN	O	O
activity	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
TNF	NN	O	B-protein
.	NN	O	O

This	NN	O	O
system	NN	O	O
may	NN	O	O
be	NN	O	O
useful	NN	O	O
to	NN	O	O
assess	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
ethanol	NN	O	O
on	NN	O	O
TNF	NN	O	B-protein
-induced	NN	O	O
hepatocyte	NN	O	O
IL-8	NN	O	B-protein
production	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
second	NN	O	O
set	NN	O	O
of	NN	O	O
experiments	NN	O	O
,	NN	O	O
HepG2	NN	O	B-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
cultured	NN	O	O
in	NN	O	O
25	NN	O	O
to	NN	O	O
100	NN	O	O
mmol	NN	O	O
concentrations	NN	O	O
of	NN	O	O
ethanol	NN	O	O
.	NN	O	O

Both	NN	O	O
TNF	NN	O	B-protein
and	NN	O	O
ethanol	NN	O	O
increased	NN	O	O
HepG2	NN	O	B-cell_line
cell	NN	O	I-cell_line
MnSOD	NN	O	B-protein
activity	NN	O	O
in	NN	O	O
short-term	NN	O	O
(	NN	O	O
72	NN	O	O
hr	NN	O	O
)	NN	O	O
cultures	NN	O	O
with	NN	O	O
ethanol	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
after	NN	O	O
long-term	NN	O	O
(	NN	O	O
10	NN	O	O
weeks	NN	O	O
)	NN	O	O
culture	NN	O	O
with	NN	O	O
ethanol	NN	O	O
,	NN	O	O
there	NN	O	O
was	NN	O	O
no	NN	O	O
induction	NN	O	O
of	NN	O	O
MnSOD	NN	O	B-protein
by	NN	O	O
ethanol	NN	O	O
and	NN	O	O
there	NN	O	O
was	NN	O	O
a	NN	O	O
diminished	NN	O	O
induction	NN	O	O
of	NN	O	O
MnSOD	NN	O	B-protein
in	NN	O	O
response	NN	O	O
to	NN	O	O
TNF	NN	O	B-protein
.	NN	O	O

Further	NN	O	O
studies	NN	O	O
are	NN	O	O
needed	NN	O	O
to	NN	O	O
assess	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
this	NN	O	O
diminished	NN	O	O
induction	NN	O	O
of	NN	O	O
MnSOD	NN	O	B-protein
with	NN	O	O
chronic	NN	O	O
ethanol	NN	O	O
culture	NN	O	O
on	NN	O	O
HepG2	NN	O	B-cell_line
cell	NN	O	I-cell_line
susceptibility	NN	O	O
to	NN	O	O
TNF	NN	O	B-cell_line
cytotoxicity	NN	O	I-cell_line
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
transfected	NN	O	O
liver	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
can	NN	O	O
be	NN	O	O
used	NN	O	O
to	NN	O	O
evaluate	NN	O	O
mechanisms	NN	O	O
for	NN	O	O
increased	NN	O	O
injurious	NN	O	B-protein
factors	NN	O	I-protein
and	NN	O	O
decreased	NN	O	O
protective	NN	O	B-protein
factors	NN	O	I-protein
in	NN	O	O
alcoholic	NN	O	O
liver	NN	O	O
injury	NN	O	O
.	NN	O	O

-DOCSTART-	O

Tissue	NN	O	O
factor	NN	O	O
transcription	NN	O	O
driven	NN	O	O
by	NN	O	O
Egr-1	NN	O	B-protein
is	NN	O	O
a	NN	O	O
critical	NN	O	O
mechanism	NN	O	O
of	NN	O	O
murine	NN	O	B-protein
pulmonary	NN	O	I-protein
fibrin	NN	O	I-protein
deposition	NN	O	O
in	NN	O	O
hypoxia	NN	O	O
.	NN	O	O

Local	NN	O	O
hypoxemia	NN	O	O
and	NN	O	O
stasis	NN	O	O
trigger	NN	O	O
thrombosis	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
demonstrated	NN	O	O
previously	NN	O	O
that	NN	O	O
in	NN	O	O
a	NN	O	O
murine	NN	O	O
model	NN	O	O
of	NN	O	O
normobaric	NN	O	O
hypoxia	NN	O	O
pulmonary	NN	O	O
fibrin	NN	O	B-protein
deposition	NN	O	O
is	NN	O	O
a	NN	O	O
result	NN	O	O
of	NN	O	O
expression	NN	O	O
of	NN	O	O
tissue	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
especially	NN	O	O
in	NN	O	O
oxygen-deprived	NN	O	B-protein
mononuclear	NN	O	I-protein
phagocytes	NN	O	I-protein
(	NN	O	O
MPs	NN	O	B-protein
)	NN	O	O
.	NN	O	O

We	NN	O	O
now	NN	O	O
show	NN	O	O
that	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
early-growth-response	NN	O	I-protein
gene	NN	O	I-protein
product	NN	O	I-protein
(	NN	O	O
Egr-1	NN	O	B-protein
)	NN	O	O
is	NN	O	O
rapidly	NN	O	O
activated	NN	O	O
in	NN	O	O
hypoxia	NN	O	O
,	NN	O	O
both	NN	O	O
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
in	NN	O	O
vivo	NN	O	O
,	NN	O	O
and	NN	O	O
is	NN	O	O
responsible	NN	O	O
for	NN	O	O
transcription	NN	O	O
and	NN	O	O
expression	NN	O	O
of	NN	O	O
tissue	NN	O	B-protein
factor	NN	O	I-protein
in	NN	O	O
hypoxic	NN	O	O
lung	NN	O	O
.	NN	O	O

MPs	NN	O	B-protein
and	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
subjected	NN	O	O
to	NN	O	O
hypoxia	NN	O	O
(	NN	O	O
pO2	NN	O	O
approximately	NN	O	O
13	NN	O	O
torr	NN	O	O
)	NN	O	O
had	NN	O	O
increased	NN	O	O
levels	NN	O	O
of	NN	O	O
tissue	NN	O	B-RNA
factor	NN	O	I-RNA
transcripts	NN	O	I-RNA
(	NN	O	O
approximately	NN	O	O
18-fold	NN	O	O
)	NN	O	O
and	NN	O	O
an	NN	O	O
increased	NN	O	O
rate	NN	O	O
of	NN	O	O
transcription	NN	O	O
(	NN	O	O
approximately	NN	O	O
15-fold	NN	O	O
)	NN	O	O
,	NN	O	O
based	NN	O	O
on	NN	O	O
nuclear	NN	O	O
run-on	NN	O	O
analysis	NN	O	O
.	NN	O	O

Gel-shift	NN	O	O
analysis	NN	O	O
of	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
from	NN	O	O
hypoxic	NN	O	B-protein
MPs	NN	O	I-protein
and	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
demonstrated	NN	O	O
increased	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
at	NN	O	O
the	NN	O	O
serum	NN	O	B-DNA
response	NN	O	I-DNA
region	NN	O	I-DNA
(	NN	O	O
SRR	NN	O	O
;	NN	O	O
-111/+14	NN	O	O
bp	NN	O	O
)	NN	O	O
of	NN	O	O
the	NN	O	O
tissue	NN	O	B-DNA
factor	NN	O	I-DNA
promoter	NN	O	I-DNA
at	NN	O	O
Egr-1	NN	O	B-DNA
motifs	NN	O	I-DNA
.	NN	O	O

Using	NN	O	O
32P-labeled	NN	O	O
Egr	NN	O	O
consensus	NN	O	O
oligonucleotide	NN	O	O
,	NN	O	O
we	NN	O	O
observed	NN	O	O
induction	NN	O	O
of	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
in	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
from	NN	O	O
hypoxic	NN	O	O
lung	NN	O	O
and	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
because	NN	O	O
of	NN	O	O
activation	NN	O	O
of	NN	O	O
Egr-1	NN	O	B-protein
,	NN	O	O
by	NN	O	O
means	NN	O	O
of	NN	O	O
supershift	NN	O	O
analysis	NN	O	O
.	NN	O	O

Transient	NN	O	O
transfection	NN	O	O
of	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
chimeric	NN	O	B-DNA
plasmids	NN	O	I-DNA
containing	NN	O	O
wild-type	NN	O	B-protein
or	NN	O	I-protein
mutant	NN	O	I-protein
SRR	NN	O	I-protein
from	NN	O	O
the	NN	O	O
tissue	NN	O	B-DNA
factor	NN	O	I-DNA
promoter	NN	O	I-DNA
showed	NN	O	O
that	NN	O	O
intact	NN	O	B-DNA
Sp1	NN	O	I-DNA
sites	NN	O	I-DNA
are	NN	O	O
necessary	NN	O	O
for	NN	O	O
basal	NN	O	O
promoter	NN	O	O
activity	NN	O	O
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
integrity	NN	O	O
of	NN	O	O
Egr-1	NN	O	B-DNA
sites	NN	O	I-DNA
was	NN	O	O
required	NN	O	O
for	NN	O	O
hypoxia-enhanced	NN	O	O
expression	NN	O	O
.	NN	O	O

A	NN	O	O
central	NN	O	O
role	NN	O	O
for	NN	O	O
Egr-1	NN	O	B-protein
in	NN	O	O
hypoxia-mediated	NN	O	O
tissue	NN	O	O
factor	NN	O	O
expression	NN	O	O
was	NN	O	O
confirmed	NN	O	O
by	NN	O	O
experiments	NN	O	O
with	NN	O	O
homozygous	NN	O	O
Egr-1	NN	O	B-protein
null	NN	O	O
mice	NN	O	O
;	NN	O	O
wild-type	NN	O	O
mice	NN	O	O
subjected	NN	O	O
to	NN	O	O
oxygen	NN	O	B-protein
deprivation	NN	O	I-protein
expressed	NN	O	I-protein
tissue	NN	O	I-protein
factor	NN	O	I-protein
and	NN	O	O
showed	NN	O	O
fibrin	NN	O	B-protein
deposition	NN	O	O
,	NN	O	O
but	NN	O	O
hypoxic	NN	O	O
homozygous	NN	O	O
Egr-1	NN	O	B-protein
null	NN	O	O
mice	NN	O	O
displayed	NN	O	O
neither	NN	O	O
tissue	NN	O	B-protein
factor	NN	O	I-protein
nor	NN	O	O
fibrin	NN	O	B-protein
.	NN	O	O

These	NN	O	O
data	NN	O	O
delineate	NN	O	O
a	NN	O	O
novel	NN	O	O
biology	NN	O	O
for	NN	O	O
hypoxia-induced	NN	O	O
fibrin	NN	O	B-protein
deposition	NN	O	O
,	NN	O	O
in	NN	O	O
which	NN	O	O
oxygen	NN	O	O
deprivation-induced	NN	O	O
activation	NN	O	O
of	NN	O	O
Egr-1	NN	O	B-protein
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
expression	NN	O	O
of	NN	O	O
tissue	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
has	NN	O	O
an	NN	O	O
unexpected	NN	O	O
and	NN	O	O
central	NN	O	O
role	NN	O	O
.	NN	O	O

-DOCSTART-	O

Kinetics	NN	O	O
of	NN	O	O
cytokine	NN	O	O
and	NN	O	O
NFAT	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
human	NN	O	B-cell_line
interleukin-2-dependent	NN	O	I-cell_line
T	NN	O	I-cell_line
lymphoblasts	NN	O	I-cell_line
stimulated	NN	O	O
via	NN	O	O
T-cell	NN	O	B-protein
receptor	NN	O	I-protein
.	NN	O	O

T	NN	O	B-cell_type
cells	NN	O	I-cell_type
respond	NN	O	O
to	NN	O	O
mitogenic	NN	O	O
or	NN	O	O
antigenic	NN	O	O
stimulation	NN	O	O
by	NN	O	O
proliferation	NN	O	O
and	NN	O	O
by	NN	O	O
turning	NN	O	O
on	NN	O	O
cytokine	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
have	NN	O	O
analysed	NN	O	O
the	NN	O	O
kinetics	NN	O	O
and	NN	O	O
nature	NN	O	O
of	NN	O	O
cytokine	NN	O	O
production	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood-derived	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphoblasts	NN	O	I-cell_type
stimulated	NN	O	O
with	NN	O	O
anti-CD3	NN	O	B-protein
antibodies	NN	O	I-protein
or	NN	O	O
Lens	NN	O	B-protein
culinaris	NN	O	I-protein
lectin	NN	O	I-protein
(	NN	O	O
LCL	NN	O	B-protein
)	NN	O	O
.	NN	O	O

T	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
purified	NN	O	O
from	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
PBMC	NN	O	B-cell_type
)	NN	O	O
and	NN	O	O
primarily	NN	O	O
activated	NN	O	O
with	NN	O	O
anti-CD3	NN	O	B-protein
antibodies	NN	O	I-protein
and	NN	O	O
cultured	NN	O	O
in	NN	O	O
the	NN	O	O
presence	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

Anti-CD3-restimulated	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
mainly	NN	O	O
CD8+	NN	O	O
)	NN	O	O
produced	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
interferon-gamma	NN	O	B-protein
(	NN	O	O
IFN-gamma	NN	O	B-protein
)	NN	O	O
and	NN	O	O
tumour	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
and	NN	O	O
low	NN	O	O
levels	NN	O	O
of	NN	O	O
IL-4	NN	O	B-RNA
and	NN	O	I-RNA
IL-10	NN	O	I-RNA
transcripts	NN	O	I-RNA
and	NN	O	O
proteins	NN	O	O
.	NN	O	O

No	NN	O	O
IL-6	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
was	NN	O	O
observed	NN	O	O
.	NN	O	O

In	NN	O	O
LCL-stimulated	NN	O	B-cell_type
cells	NN	O	I-cell_type
the	NN	O	O
cytokine	NN	O	O
production	NN	O	O
pattern	NN	O	O
was	NN	O	O
very	NN	O	O
similar	NN	O	O
.	NN	O	O

Steady-state	NN	O	O
mRNA	NN	O	O
levels	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
IL-10	NN	O	B-protein
and	NN	O	O
IFN-gamma	NN	O	B-protein
peaked	NN	O	O
at	NN	O	O
3	NN	O	O
hr	NN	O	O
after	NN	O	O
anti-CD3	NN	O	O
stimulation	NN	O	O
and	NN	O	O
declined	NN	O	O
rapidly	NN	O	O
thereafter	NN	O	O
.	NN	O	O

The	NN	O	O
kinetics	NN	O	O
of	NN	O	O
TNF-alpha	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
was	NN	O	O
faster	NN	O	O
,	NN	O	O
being	NN	O	O
at	NN	O	O
its	NN	O	O
peak	NN	O	O
level	NN	O	O
1	NN	O	O
hr	NN	O	O
after	NN	O	O
stimulation	NN	O	O
.	NN	O	O

Anti-CD3-stimulated	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
was	NN	O	O
down-regulated	NN	O	O
by	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
inhibitor	NN	O	O
,	NN	O	O
whereas	NN	O	O
IL-10	NN	O	B-DNA
,	NN	O	I-DNA
IFN-gamma	NN	O	I-DNA
and	NN	O	I-DNA
TNF-alpha	NN	O	I-DNA
genes	NN	O	I-DNA
were	NN	O	O
readily	NN	O	O
induced	NN	O	O
independent	NN	O	O
of	NN	O	O
ongoing	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
.	NN	O	O

T-cell	NN	O	B-protein
receptor	NN	O	I-protein
stimulation	NN	O	O
also	NN	O	O
induced	NN	O	O
a	NN	O	O
very	NN	O	O
rapid	NN	O	O
expression	NN	O	O
of	NN	O	O
c-jun	NN	O	B-DNA
,	NN	O	I-DNA
c-fos	NN	O	I-DNA
and	NN	O	I-DNA
NFATc1	NN	O	I-DNA
(	NN	O	I-DNA
NFATc	NN	O	I-DNA
)	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
the	NN	O	O
gene	NN	O	O
products	NN	O	O
of	NN	O	O
which	NN	O	O
are	NN	O	O
involved	NN	O	O
in	NN	O	O
cytokine	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

In	NN	O	O
conclusion	NN	O	O
,	NN	O	O
the	NN	O	O
cytokines	NN	O	B-protein
synthesized	NN	O	O
by	NN	O	O
IL-2-dependent	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
predominantly	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
IFN-gamma	NN	O	B-protein
and	NN	O	O
TNF-alpha	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

An	NN	O	O
animal	NN	O	O
model	NN	O	O
to	NN	O	O
study	NN	O	O
local	NN	O	O
oxidation	NN	O	O
of	NN	O	O
LDL	NN	O	O
and	NN	O	O
its	NN	O	O
biological	NN	O	O
effects	NN	O	O
in	NN	O	O
the	NN	O	O
arterial	NN	O	O
wall	NN	O	O
.	NN	O	O

Oxidized	NN	O	O
LDL	NN	O	O
(	NN	O	O
oxLDL	NN	O	O
)	NN	O	O
is	NN	O	O
present	NN	O	O
in	NN	O	O
atherosclerotic	NN	O	O
lesions	NN	O	O
and	NN	O	O
is	NN	O	O
believed	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
atherogenesis	NN	O	O
.	NN	O	O

Mainly	NN	O	O
on	NN	O	O
the	NN	O	O
basis	NN	O	O
of	NN	O	O
cell	NN	O	O
culture	NN	O	O
studies	NN	O	O
,	NN	O	O
oxLDL	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
produce	NN	O	O
many	NN	O	O
biological	NN	O	O
effects	NN	O	O
that	NN	O	O
influence	NN	O	O
the	NN	O	O
atherosclerotic	NN	O	O
process	NN	O	O
.	NN	O	O

To	NN	O	O
study	NN	O	O
LDL	NN	O	O
oxidation	NN	O	O
in	NN	O	O
vivo	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
established	NN	O	O
a	NN	O	O
model	NN	O	O
in	NN	O	O
which	NN	O	O
Sprague-Dawley	NN	O	O
rats	NN	O	O
are	NN	O	O
given	NN	O	O
a	NN	O	O
single	NN	O	O
injection	NN	O	O
of	NN	O	O
unmodified	NN	O	O
human	NN	O	O
LDL	NN	O	O
(	NN	O	O
>	NN	O	O
or	NN	O	O
=	NN	O	O
4	NN	O	O
mg/kg	NN	O	O
body	NN	O	O
weight	NN	O	O
)	NN	O	O
.	NN	O	O

Within	NN	O	O
6	NN	O	O
hours	NN	O	O
,	NN	O	O
an	NN	O	O
accumulation	NN	O	O
of	NN	O	O
apolipoprotein	NN	O	O
B	NN	O	O
and	NN	O	O
epitopes	NN	O	B-protein
present	NN	O	O
on	NN	O	O
oxLDL	NN	O	O
are	NN	O	O
detected	NN	O	O
in	NN	O	O
the	NN	O	O
arterial	NN	O	O
endothelium	NN	O	O
and	NN	O	O
media	NN	O	O
.	NN	O	O

The	NN	O	O
presence	NN	O	O
of	NN	O	O
oxLDL	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
in	NN	O	O
the	NN	O	O
endothelium	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
endothelial	NN	O	O
expression	NN	O	O
of	NN	O	O
intercellular	NN	O	B-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
.	NN	O	O

Injection	NN	O	O
of	NN	O	O
LDL	NN	O	O
enriched	NN	O	O
with	NN	O	O
the	NN	O	O
antioxidant	NN	O	O
probucol	NN	O	O
resulted	NN	O	O
in	NN	O	O
arterial	NN	O	O
accumulation	NN	O	O
of	NN	O	O
apolipoprotein	NN	O	O
B	NN	O	O
,	NN	O	O
but	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
oxLDL-specific	NN	O	O
epitopes	NN	O	O
was	NN	O	O
reduced	NN	O	O
at	NN	O	O
24	NN	O	O
hours	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
this	NN	O	O
simple	NN	O	O
model	NN	O	O
has	NN	O	O
the	NN	O	O
potential	NN	O	O
to	NN	O	O
analyze	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
behind	NN	O	O
and	NN	O	O
biological	NN	O	O
effects	NN	O	O
of	NN	O	O
LDL	NN	O	O
oxidation	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

-DOCSTART-	O

Molecular	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
promoter	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
gp34	NN	O	B-DNA
gene	NN	O	I-DNA
that	NN	O	O
is	NN	O	O
trans-activated	NN	O	O
by	NN	O	O
an	NN	O	O
oncoprotein	NN	O	B-protein
Tax	NN	O	I-protein
of	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
.	NN	O	O

We	NN	O	O
investigated	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanism	NN	O	O
of	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
gp34	NN	O	B-DNA
gene	NN	O	I-DNA
by	NN	O	O
the	NN	O	O
Tax	NN	O	B-protein
oncoprotein	NN	O	I-protein
of	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
(	NN	O	O
HTLV-I	NN	O	O
)	NN	O	O
.	NN	O	O

gp34	NN	O	B-protein
is	NN	O	O
a	NN	O	O
type	NN	O	B-protein
II	NN	O	I-protein
transmembrane	NN	O	I-protein
molecule	NN	O	I-protein
belonging	NN	O	O
to	NN	O	O
the	NN	O	O
tumor	NN	O	O
necrosis	NN	O	O
factor	NN	O	O
family	NN	O	O
and	NN	O	O
is	NN	O	O
constitutively	NN	O	O
expressed	NN	O	O
on	NN	O	O
HTLV-I-producing	NN	O	B-cell_type
cells	NN	O	I-cell_type
but	NN	O	O
not	NN	O	O
normal	NN	O	B-cell_type
resting	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
transcriptional	NN	O	O
regulatory	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
gp34	NN	O	B-DNA
gene	NN	O	I-DNA
was	NN	O	O
activated	NN	O	O
by	NN	O	O
HTLV-I	NN	O	O
Tax	NN	O	B-protein
in	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
Jurkat	NN	O	I-cell_line
,	NN	O	O
in	NN	O	O
which	NN	O	O
endogenous	NN	O	O
gp34	NN	O	B-protein
is	NN	O	O
induced	NN	O	O
by	NN	O	O
Tax	NN	O	B-protein
.	NN	O	O

Sequence	NN	O	O
analysis	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
two	NN	O	O
NF-kappaB-like	NN	O	B-DNA
elements	NN	O	I-DNA
(	NN	O	O
1	NN	O	O
and	NN	O	O
2	NN	O	O
)	NN	O	O
were	NN	O	O
present	NN	O	O
in	NN	O	O
the	NN	O	O
regulatory	NN	O	B-DNA
region	NN	O	I-DNA
.	NN	O	O

Both	NN	O	O
NF-kappaB-like	NN	O	B-DNA
elements	NN	O	I-DNA
were	NN	O	O
able	NN	O	O
to	NN	O	O
bind	NN	O	O
to	NN	O	O
NF-kappaB	NN	O	B-protein
or	NN	O	O
its	NN	O	O
related	NN	O	O
factor	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
in	NN	O	O
a	NN	O	O
Tax	NN	O	B-protein
-dependent	NN	O	O
manner	NN	O	O
.	NN	O	O

Chloramphenicol	NN	O	B-protein
acetyltransferase	NN	O	I-protein
assays	NN	O	O
indicated	NN	O	O
that	NN	O	O
NF-kappaB-like	NN	O	B-DNA
element	NN	O	I-DNA
1	NN	O	I-DNA
was	NN	O	O
Tax	NN	O	B-protein
-responsive	NN	O	O
,	NN	O	O
although	NN	O	O
the	NN	O	O
activity	NN	O	O
was	NN	O	O
lower	NN	O	O
than	NN	O	O
that	NN	O	O
the	NN	O	O
native	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

NF-kappaB	NN	O	B-protein
-like	NN	O	O
element	NN	O	O
2	NN	O	O
elevated	NN	O	O
promoter	NN	O	O
activity	NN	O	O
when	NN	O	O
combined	NN	O	O
with	NN	O	O
NF-kappaB-like	NN	O	B-DNA
element	NN	O	I-DNA
1	NN	O	I-DNA
,	NN	O	O
indicating	NN	O	O
cooperative	NN	O	O
function	NN	O	O
of	NN	O	O
the	NN	O	O
elements	NN	O	O
for	NN	O	O
maximum	NN	O	O
promoter	NN	O	O
function	NN	O	O
.	NN	O	O

Unlike	NN	O	O
typical	NN	O	O
NF-kappaB	NN	O	B-DNA
elements	NN	O	I-DNA
,	NN	O	O
the	NN	O	O
NF-kappaB-like	NN	O	B-DNA
elements	NN	O	I-DNA
in	NN	O	O
gp34	NN	O	B-protein
were	NN	O	O
not	NN	O	O
activated	NN	O	O
by	NN	O	O
treatment	NN	O	O
of	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
despite	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-protein
-like	NN	O	O
binding	NN	O	O
activity	NN	O	O
.	NN	O	O

Chloramphenicol	NN	O	B-protein
acetyltransferase	NN	O	I-protein
reporter	NN	O	O
assays	NN	O	O
using	NN	O	O
the	NN	O	O
region	NN	O	O
upstream	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappaB-like	NN	O	B-DNA
elements	NN	O	I-DNA
identified	NN	O	O
an	NN	O	O
upstream	NN	O	O
region	NN	O	O
that	NN	O	O
reduced	NN	O	O
transcription	NN	O	O
from	NN	O	O
cognate	NN	O	B-DNA
and	NN	O	I-DNA
noncognate	NN	O	I-DNA
core	NN	O	I-DNA
promoters	NN	O	I-DNA
in	NN	O	O
a	NN	O	O
Tax-independent	NN	O	O
manner	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
imply	NN	O	O
complex	NN	O	O
regulation	NN	O	O
of	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
gp34	NN	O	B-DNA
gene	NN	O	I-DNA
and	NN	O	O
suggest	NN	O	O
implication	NN	O	O
of	NN	O	O
gp34	NN	O	B-protein
in	NN	O	O
proliferation	NN	O	O
of	NN	O	O
HTLV-I	NN	O	O
infected	NN	O	O
T	NN	O	O
cells	NN	O	O
.	NN	O	O

-DOCSTART-	O

beta-Amyloid	NN	O	B-protein
fibrils	NN	O	I-protein
activate	NN	O	O
parallel	NN	O	O
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
pathways	NN	O	O
in	NN	O	O
microglia	NN	O	B-cell_type
and	NN	O	O
THP1	NN	O	B-cell_line
monocytes	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
senile	NN	O	O
plaques	NN	O	O
of	NN	O	O
Alzheimer	NN	O	O
's	NN	O	O
disease	NN	O	O
are	NN	O	O
foci	NN	O	O
of	NN	O	O
local	NN	O	O
inflammatory	NN	O	O
responses	NN	O	O
,	NN	O	O
as	NN	O	O
evidenced	NN	O	O
by	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
acute	NN	O	B-protein
phase	NN	O	I-protein
proteins	NN	O	I-protein
and	NN	O	O
oxidative	NN	O	O
damage	NN	O	O
.	NN	O	O

Fibrillar	NN	O	B-protein
forms	NN	O	I-protein
of	NN	O	O
beta-amyloid	NN	O	B-protein
(	NN	O	O
Abeta	NN	O	B-protein
)	NN	O	O
,	NN	O	O
which	NN	O	O
are	NN	O	O
the	NN	O	O
primary	NN	O	O
constituents	NN	O	O
of	NN	O	O
senile	NN	O	O
plaques	NN	O	O
,	NN	O	O
have	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
activate	NN	O	O
tyrosine	NN	O	O
kinase-dependent	NN	O	O
signal	NN	O	O
transduction	NN	O	O
cascades	NN	O	O
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
inflammatory	NN	O	O
responses	NN	O	O
in	NN	O	O
microglia	NN	O	B-cell_type
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
downstream	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
mediating	NN	O	O
Abeta	NN	O	B-protein
-induced	NN	O	O
inflammatory	NN	O	O
events	NN	O	O
are	NN	O	O
not	NN	O	O
well	NN	O	O
characterized	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
that	NN	O	O
exposure	NN	O	O
of	NN	O	O
primary	NN	O	B-cell_type
rat	NN	O	I-cell_type
microglia	NN	O	I-cell_type
and	NN	O	O
human	NN	O	B-cell_line
THP1	NN	O	I-cell_line
monocytes	NN	O	I-cell_line
to	NN	O	O
fibrillar	NN	O	B-protein
Abeta	NN	O	I-protein
results	NN	O	O
in	NN	O	O
the	NN	O	O
tyrosine	NN	O	B-protein
kinase	NN	O	I-protein
-dependent	NN	O	O
activation	NN	O	O
of	NN	O	O
two	NN	O	O
parallel	NN	O	O
signal	NN	O	O
transduction	NN	O	O
cascades	NN	O	O
involving	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	I-protein
MAPK	NN	O	I-protein
)	NN	O	I-protein
superfamily	NN	O	I-protein
.	NN	O	O

Abeta	NN	O	B-protein
stimulated	NN	O	O
the	NN	O	O
rapid	NN	O	O
,	NN	O	O
transient	NN	O	O
activation	NN	O	O
of	NN	O	O
extracellular	NN	O	B-protein
signal-regulated	NN	O	I-protein
kinase	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	O
ERK1	NN	O	B-protein
)	NN	O	O
and	NN	O	O
ERK2	NN	O	B-protein
in	NN	O	O
microglia	NN	O	B-cell_type
and	NN	O	O
ERK2	NN	O	B-protein
in	NN	O	O
THP1	NN	O	B-cell_line
monocytes	NN	O	I-cell_line
.	NN	O	O

A	NN	O	O
second	NN	O	O
superfamily	NN	O	B-protein
member	NN	O	I-protein
,	NN	O	O
p38	NN	O	B-protein
MAPK	NN	O	I-protein
,	NN	O	O
was	NN	O	O
also	NN	O	O
activated	NN	O	O
with	NN	O	O
similar	NN	O	O
kinetics	NN	O	O
.	NN	O	O

Scavenger	NN	O	B-protein
receptor	NN	O	I-protein
and	NN	O	O
receptor	NN	O	B-protein
for	NN	O	I-protein
advanced	NN	O	I-protein
glycated	NN	O	I-protein
end	NN	O	I-protein
products	NN	O	I-protein
(	NN	O	O
RAGE	NN	O	B-protein
)	NN	O	O
ligands	NN	O	O
failed	NN	O	O
to	NN	O	O
activate	NN	O	O
ERK	NN	O	B-protein
and	NN	O	O
p38	NN	O	B-protein
MAPK	NN	O	I-protein
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
significant	NN	O	O
increases	NN	O	O
in	NN	O	O
protein	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
,	NN	O	O
demonstrating	NN	O	O
that	NN	O	O
scavenger	NN	O	B-protein
receptors	NN	O	I-protein
and	NN	O	O
RAGE	NN	O	B-protein
are	NN	O	O
not	NN	O	O
linked	NN	O	O
to	NN	O	O
these	NN	O	O
pathways	NN	O	O
.	NN	O	O

Importantly	NN	O	O
,	NN	O	O
the	NN	O	O
stress-activated	NN	O	B-protein
protein	NN	O	I-protein
kinases	NN	O	I-protein
(	NN	O	O
SAPKs	NN	O	B-protein
)	NN	O	O
were	NN	O	O
not	NN	O	O
significantly	NN	O	O
activated	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
Abeta	NN	O	B-protein
.	NN	O	O

Downstream	NN	O	O
effectors	NN	O	O
of	NN	O	O
the	NN	O	O
MAPK	NN	O	O
signal	NN	O	O
transduction	NN	O	O
cascades	NN	O	O
include	NN	O	O
MAPKAP	NN	O	B-protein
kinases	NN	O	I-protein
,	NN	O	O
such	NN	O	O
as	NN	O	O
RSK1	NN	O	B-protein
and	NN	O	O
RSK2	NN	O	B-protein
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

Exposure	NN	O	O
of	NN	O	O
microglia	NN	O	B-cell_type
and	NN	O	O
THP1	NN	O	B-cell_line
monocytes	NN	O	I-cell_line
to	NN	O	O
Abeta	NN	O	B-protein
resulted	NN	O	O
in	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
RSK1	NN	O	B-protein
and	NN	O	O
RSK2	NN	O	B-protein
and	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
cAMP	NN	O	B-protein
response	NN	O	I-protein
element-binding	NN	O	I-protein
protein	NN	O	I-protein
at	NN	O	O
Ser133	NN	O	O
,	NN	O	O
providing	NN	O	O
a	NN	O	O
mechanism	NN	O	O
for	NN	O	O
Abeta	NN	O	B-protein
-induced	NN	O	O
changes	NN	O	O
in	NN	O	O
gene	NN	O	O
expression	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
cellular	NN	O	B-protein
retinoic	NN	O	I-protein
acid	NN	O	I-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
CRABP	NN	O	B-protein
II	NN	O	I-protein
)	NN	O	O
during	NN	O	O
human	NN	O	B-cell_type
monocyte	NN	O	I-cell_type
differentiation	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

Cellular	NN	O	B-protein
retinoic	NN	O	I-protein
acid	NN	O	I-protein
binding	NN	O	I-protein
proteins	NN	O	I-protein
(	NN	O	O
CRABP	NN	O	B-protein
)	NN	O	O
are	NN	O	O
low	NN	O	B-protein
molecular	NN	O	I-protein
weight	NN	O	I-protein
proteins	NN	O	I-protein
whose	NN	O	O
precise	NN	O	O
function	NN	O	O
remains	NN	O	O
unknown	NN	O	O
.	NN	O	O

They	NN	O	O
bind	NN	O	O
retinoids	NN	O	O
and	NN	O	O
may	NN	O	O
thereby	NN	O	O
modulate	NN	O	O
the	NN	O	O
intracellular	NN	O	O
steady-state	NN	O	O
concentration	NN	O	O
of	NN	O	O
retinoids	NN	O	O
.	NN	O	O

Whereas	NN	O	O
CRABP	NN	O	B-protein
I	NN	O	I-protein
is	NN	O	O
ubiquitously	NN	O	O
expressed	NN	O	O
,	NN	O	O
CRABP	NN	O	B-protein
II	NN	O	I-protein
is	NN	O	O
mainly	NN	O	O
detected	NN	O	O
in	NN	O	O
various	NN	O	O
cell	NN	O	O
types	NN	O	O
of	NN	O	O
the	NN	O	O
skin	NN	O	O
.	NN	O	O

By	NN	O	O
representative	NN	O	O
difference	NN	O	O
analysis	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
CRABP	NN	O	B-protein
II	NN	O	I-protein
is	NN	O	O
also	NN	O	O
strongly	NN	O	O
expressed	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocyte-derived	NN	O	I-cell_type
macrophages	NN	O	I-cell_type
(	NN	O	O
MAC	NN	O	B-cell_type
)	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
freshly	NN	O	B-cell_type
isolated	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
(	NN	O	O
MO	NN	O	B-cell_type
)	NN	O	O
.	NN	O	O

The	NN	O	O
CRABP	NN	O	B-RNA
II	NN	O	I-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
gradually	NN	O	O
upregulated	NN	O	O
during	NN	O	O
differentiation	NN	O	O
from	NN	O	O
MO	NN	O	B-cell_type
to	NN	O	O
MAC	NN	O	B-cell_type
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
2	NN	O	O
%	NN	O	O
serum	NN	O	O
.	NN	O	O

Adherence	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
important	NN	O	O
for	NN	O	O
MO	NN	O	B-cell_type
differentiation	NN	O	O
,	NN	O	O
induced	NN	O	O
CRABP	NN	O	B-protein
II	NN	O	I-protein
expression	NN	O	O
,	NN	O	O
but	NN	O	O
the	NN	O	O
addition	NN	O	O
of	NN	O	O
10	NN	O	O
(	NN	O	O
-7	NN	O	O
)	NN	O	O
M	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
inhibited	NN	O	O
the	NN	O	O
upregulation	NN	O	O
of	NN	O	O
CRABP	NN	O	B-protein
II	NN	O	I-protein
expression	NN	O	O
during	NN	O	O
MO/MAC	NN	O	O
differentiation	NN	O	O
.	NN	O	O

As	NN	O	O
MO	NN	O	B-cell_type
can	NN	O	O
differentiate	NN	O	O
along	NN	O	O
the	NN	O	O
classical	NN	O	O
pathway	NN	O	O
not	NN	O	O
only	NN	O	O
to	NN	O	O
MAC	NN	O	B-cell_type
but	NN	O	O
also	NN	O	O
to	NN	O	O
dendritic	NN	O	O
cells	NN	O	O
we	NN	O	O
analyzed	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
CRABP	NN	O	B-protein
II	NN	O	I-protein
in	NN	O	O
MO-derived	NN	O	B-cell_type
dendritic	NN	O	I-cell_type
cells	NN	O	I-cell_type
cultured	NN	O	O
with	NN	O	O
10	NN	O	O
%	NN	O	O
FCS	NN	O	B-protein
,	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
and	NN	O	O
GM-CSF	NN	O	B-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
MAC	NN	O	B-cell_type
,	NN	O	O
MO-derived	NN	O	B-cell_type
dendritic	NN	O	I-cell_type
cells	NN	O	I-cell_type
showed	NN	O	O
an	NN	O	O
extremely	NN	O	O
low	NN	O	O
expression	NN	O	O
of	NN	O	O
CRABP	NN	O	B-protein
II	NN	O	I-protein
.	NN	O	O

From	NN	O	O
these	NN	O	O
results	NN	O	O
we	NN	O	O
conclude	NN	O	O
(	NN	O	O
1	NN	O	O
)	NN	O	O
that	NN	O	O
the	NN	O	O
availability	NN	O	O
and	NN	O	O
the	NN	O	O
metabolism	NN	O	O
of	NN	O	O
retinoids	NN	O	O
may	NN	O	O
be	NN	O	O
different	NN	O	O
in	NN	O	O
MAC	NN	O	B-cell_type
compared	NN	O	O
to	NN	O	O
MO	NN	O	B-cell_type
and	NN	O	O
dendritic	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
that	NN	O	O
this	NN	O	O
may	NN	O	O
influence	NN	O	O
differentiation	NN	O	O
and	NN	O	O
activation	NN	O	O
of	NN	O	O
those	NN	O	O
cells	NN	O	O
.	NN	O	O

-DOCSTART-	O

Transcription	NN	O	B-protein
factor	NN	O	I-protein
B-cell-specific	NN	O	I-protein
activator	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
BSAP	NN	O	B-protein
)	NN	O	O
is	NN	O	O
differentially	NN	O	O
expressed	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
in	NN	O	O
subsets	NN	O	O
of	NN	O	O
B-cell	NN	O	O
lymphomas	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
encodes	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
BSAP	NN	O	B-protein
(	NN	O	O
B-cell-specific	NN	O	B-protein
activator	NN	O	I-protein
protein	NN	O	I-protein
)	NN	O	O
,	NN	O	O
which	NN	O	O
plays	NN	O	O
a	NN	O	O
key	NN	O	O
role	NN	O	O
in	NN	O	O
B-lymphocyte	NN	O	O
development	NN	O	O
.	NN	O	O

Despite	NN	O	O
its	NN	O	O
known	NN	O	O
involvement	NN	O	O
in	NN	O	O
a	NN	O	O
rare	NN	O	O
subtype	NN	O	O
of	NN	O	O
non-Hodgkin	NN	O	O
's	NN	O	O
lymphoma	NN	O	O
(	NN	O	O
NHL	NN	O	O
)	NN	O	O
,	NN	O	O
a	NN	O	O
detailed	NN	O	O
examination	NN	O	O
of	NN	O	O
BSAP	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
NHL	NN	O	O
has	NN	O	O
not	NN	O	O
been	NN	O	O
previously	NN	O	O
reported	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
analyzed	NN	O	O
normal	NN	O	O
and	NN	O	O
malignant	NN	O	O
lymphoid	NN	O	O
tissues	NN	O	O
and	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
including	NN	O	O
102	NN	O	O
cases	NN	O	O
of	NN	O	O
B-cell	NN	O	O
NHL	NN	O	O
,	NN	O	O
23	NN	O	O
cases	NN	O	O
of	NN	O	O
T-	NN	O	O
and	NN	O	O
null-cell	NN	O	O
NHL	NN	O	O
,	NN	O	O
and	NN	O	O
18	NN	O	O
cases	NN	O	O
of	NN	O	O
Hodgkin	NN	O	O
's	NN	O	O
disease	NN	O	O
.	NN	O	O

Normal	NN	O	O
lymphoid	NN	O	O
tissues	NN	O	O
showed	NN	O	O
strong	NN	O	O
nuclear	NN	O	O
BSAP	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
mantle	NN	O	B-cell_type
zone	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
less	NN	O	O
intense	NN	O	O
reactivity	NN	O	O
in	NN	O	O
follicular	NN	O	B-cell_type
center	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
no	NN	O	O
expression	NN	O	O
in	NN	O	O
cells	NN	O	O
of	NN	O	O
the	NN	O	O
T-cell-rich	NN	O	O
zones	NN	O	O
.	NN	O	O

Monocytoid	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
showed	NN	O	O
weak	NN	O	O
expression	NN	O	O
,	NN	O	O
whereas	NN	O	O
plasma	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
extrafollicular	NN	O	B-cell_type
large	NN	O	I-cell_type
transformed	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
negative	NN	O	O
.	NN	O	O

Of	NN	O	O
the	NN	O	O
102	NN	O	O
B-cell	NN	O	O
NHLs	NN	O	O
,	NN	O	O
83	NN	O	O
(	NN	O	O
81	NN	O	O
%	NN	O	O
)	NN	O	O
demonstrated	NN	O	O
BSAP	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

All	NN	O	O
of	NN	O	O
the	NN	O	O
13	NN	O	O
(	NN	O	O
100	NN	O	O
%	NN	O	O
)	NN	O	O
B-cell	NN	O	O
chronic	NN	O	O
lymphocytic	NN	O	O
leukemias	NN	O	O
(	NN	O	O
B-CLLs	NN	O	O
)	NN	O	O
,	NN	O	O
21	NN	O	O
of	NN	O	O
(	NN	O	O
100	NN	O	O
%	NN	O	O
)	NN	O	O
mantle	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
MCLs	NN	O	B-cell_type
)	NN	O	O
,	NN	O	O
and	NN	O	O
20	NN	O	O
of	NN	O	O
21	NN	O	O
(	NN	O	O
95	NN	O	O
%	NN	O	O
)	NN	O	O
follicular	NN	O	O
lymphomas	NN	O	O
(	NN	O	O
FLs	NN	O	O
)	NN	O	O
were	NN	O	O
positive	NN	O	O
.	NN	O	O

Moderate	NN	O	O
staining	NN	O	O
intensities	NN	O	O
were	NN	O	O
found	NN	O	O
in	NN	O	O
most	NN	O	O
B-CLL	NN	O	O
and	NN	O	O
FL	NN	O	O
cases	NN	O	O
,	NN	O	O
whereas	NN	O	O
most	NN	O	O
MCLs	NN	O	B-cell_type
showed	NN	O	O
strong	NN	O	O
reactions	NN	O	O
,	NN	O	O
paralleling	NN	O	O
the	NN	O	O
strong	NN	O	O
reactivity	NN	O	O
of	NN	O	O
nonmalignant	NN	O	B-cell_type
mantle	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Eight	NN	O	O
of	NN	O	O
12	NN	O	O
(	NN	O	O
67	NN	O	O
%	NN	O	O
)	NN	O	O
marginal	NN	O	O
zone	NN	O	O
lymphoma	NN	O	O
cases	NN	O	O
showed	NN	O	O
negative	NN	O	O
or	NN	O	O
low	NN	O	O
BSAP	NN	O	B-protein
levels	NN	O	O
,	NN	O	O
and	NN	O	O
17	NN	O	O
of	NN	O	O
24	NN	O	O
(	NN	O	O
71	NN	O	O
%	NN	O	O
)	NN	O	O
large	NN	O	O
B-cell	NN	O	O
lymphomas	NN	O	O
displayed	NN	O	O
moderate	NN	O	O
to	NN	O	O
strong	NN	O	O
expression	NN	O	O
.	NN	O	O

None	NN	O	O
of	NN	O	O
the	NN	O	O
23	NN	O	O
T-	NN	O	O
and	NN	O	O
null-cell	NN	O	O
lymphomas	NN	O	O
reacted	NN	O	O
with	NN	O	O
the	NN	O	O
BSAP	NN	O	B-protein
antisera	NN	O	O
,	NN	O	O
whereas	NN	O	O
in	NN	O	O
Hodgkin	NN	O	O
's	NN	O	O
disease	NN	O	O
,	NN	O	O
2	NN	O	O
of	NN	O	O
4	NN	O	O
(	NN	O	O
50	NN	O	O
%	NN	O	O
)	NN	O	O
nodular	NN	O	O
lymphocytic	NN	O	O
predominance	NN	O	O
and	NN	O	O
5	NN	O	O
of	NN	O	O
14	NN	O	O
(	NN	O	O
36	NN	O	O
%	NN	O	O
)	NN	O	O
classical	NN	O	O
cases	NN	O	O
showed	NN	O	O
weak	NN	O	O
nuclear	NN	O	O
or	NN	O	O
nucleolar	NN	O	O
BSAP	NN	O	B-protein
reactions	NN	O	O
in	NN	O	O
a	NN	O	O
fraction	NN	O	O
of	NN	O	O
the	NN	O	O
tumor	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Western	NN	O	O
blot	NN	O	O
analysis	NN	O	O
showed	NN	O	O
a	NN	O	O
52-kD	NN	O	O
BSAP	NN	O	B-protein
band	NN	O	O
in	NN	O	O
B-cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
non-B-cell	NN	O	B-cell_line
or	NN	O	I-cell_line
plasma	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
BSAP	NN	O	B-protein
expression	NN	O	O
is	NN	O	O
largely	NN	O	O
restricted	NN	O	O
to	NN	O	O
lymphomas	NN	O	O
of	NN	O	O
B-cell	NN	O	B-cell_type
lineage	NN	O	I-cell_type
and	NN	O	O
that	NN	O	O
BSAP	NN	O	B-protein
expression	NN	O	O
varies	NN	O	O
in	NN	O	O
B-cell	NN	O	B-cell_type
subsets	NN	O	I-cell_type
and	NN	O	O
subtypes	NN	O	O
of	NN	O	O
B-cell	NN	O	O
NHL	NN	O	O
.	NN	O	O

The	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
BSAP	NN	O	B-protein
,	NN	O	O
especially	NN	O	O
those	NN	O	O
found	NN	O	O
in	NN	O	O
large-cell	NN	O	B-cell_type
lymphomas	NN	O	I-cell_type
and	NN	O	O
in	NN	O	O
some	NN	O	O
follicular	NN	O	B-cell_type
lymphomas	NN	O	I-cell_type
,	NN	O	O
may	NN	O	O
be	NN	O	O
a	NN	O	O
consequence	NN	O	O
of	NN	O	O
deregulated	NN	O	O
gene	NN	O	O
expression	NN	O	O
and	NN	O	O
suggest	NN	O	O
a	NN	O	O
possible	NN	O	O
involvement	NN	O	O
of	NN	O	O
PAX-5	NN	O	B-DNA
in	NN	O	O
certain	NN	O	O
B-cell	NN	O	O
malignancies	NN	O	O
.	NN	O	O

This	NN	O	O
is	NN	O	O
a	NN	O	O
US	NN	O	O
government	NN	O	O
work	NN	O	O
.	NN	O	O

There	NN	O	O
are	NN	O	O
no	NN	O	O
restrictions	NN	O	O
on	NN	O	O
its	NN	O	O
use	NN	O	O
.	NN	O	O

-DOCSTART-	O

Mitogen	NN	O	O
and	NN	O	O
growth	NN	O	O
factor-induced	NN	O	O
activation	NN	O	O
of	NN	O	O
a	NN	O	O
STAT-like	NN	O	B-protein
molecule	NN	O	I-protein
in	NN	O	O
channel	NN	O	B-cell_type
catfish	NN	O	I-cell_type
lymphoid	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

This	NN	O	O
article	NN	O	O
describes	NN	O	O
the	NN	O	O
identification	NN	O	O
of	NN	O	O
a	NN	O	O
putative	NN	O	B-protein
STAT	NN	O	I-protein
molecule	NN	O	I-protein
in	NN	O	O
the	NN	O	O
channel	NN	O	O
catfish	NN	O	O
(	NN	O	O
Ictalurus	NN	O	O
punctatus	NN	O	O
)	NN	O	O
,	NN	O	O
the	NN	O	O
first	NN	O	O
report	NN	O	O
of	NN	O	O
such	NN	O	O
a	NN	O	O
molecule	NN	O	O
in	NN	O	O
a	NN	O	O
'lower	NN	O	O
'	NN	O	O
vertebrate	NN	O	O
.	NN	O	O

A	NN	O	O
monoclonal	NN	O	B-protein
antibody	NN	O	I-protein
against	NN	O	O
human	NN	O	B-protein
STAT6	NN	O	I-protein
recognizes	NN	O	O
an	NN	O	O
approximately	NN	O	O
100	NN	O	B-protein
kDa	NN	O	I-protein
molecule	NN	O	I-protein
that	NN	O	O
becomes	NN	O	O
activated	NN	O	O
and	NN	O	O
translocates	NN	O	O
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
upon	NN	O	O
both	NN	O	O
growth	NN	O	O
factor	NN	O	O
and	NN	O	O
mitogen	NN	O	O
stimulation	NN	O	O
of	NN	O	O
catfish	NN	O	O
leukocytes	NN	O	O
.	NN	O	O

This	NN	O	O
presumed	NN	O	O
catfish	NN	O	B-protein
STAT	NN	O	I-protein
binds	NN	O	O
the	NN	O	O
mammalian	NN	O	B-DNA
interferon-gamma	NN	O	I-DNA
activation	NN	O	I-DNA
site	NN	O	I-DNA
,	NN	O	O
a	NN	O	O
known	NN	O	O
motif	NN	O	O
of	NN	O	O
mammalian	NN	O	O
STAT	NN	O	O
binding	NN	O	O
,	NN	O	O
as	NN	O	O
shown	NN	O	O
by	NN	O	O
electromobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
.	NN	O	O

Purification	NN	O	O
of	NN	O	O
the	NN	O	O
proteins	NN	O	O
present	NN	O	O
in	NN	O	O
these	NN	O	O
DNA	NN	O	B-protein
complexes	NN	O	I-protein
confirms	NN	O	O
that	NN	O	O
the	NN	O	O
catfish	NN	O	O
reactive	NN	O	O
molecule	NN	O	O
binds	NN	O	O
to	NN	O	O
the	NN	O	O
interferon-gamma	NN	O	B-DNA
activation	NN	O	I-DNA
site	NN	O	I-DNA
sequence	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
STAT	NN	O	B-protein
molecules	NN	O	I-protein
have	NN	O	O
been	NN	O	O
highly	NN	O	O
conserved	NN	O	O
in	NN	O	O
vertebrate	NN	O	O
evolution	NN	O	O
.	NN	O	O

-DOCSTART-	O

Isolation	NN	O	O
and	NN	O	O
analysis	NN	O	O
of	NN	O	O
a	NN	O	O
T	NN	O	B-cell_line
cell	NN	O	I-cell_line
clone	NN	O	I-cell_line
variant	NN	O	I-cell_line
exhibiting	NN	O	O
constitutively	NN	O	O
phosphorylated	NN	O	B-protein
Ser133	NN	O	I-protein
cAMP	NN	O	I-protein
response	NN	O	I-protein
element-binding	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

In	NN	O	O
driving	NN	O	O
T	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
,	NN	O	O
IL-2	NN	O	B-protein
stimulates	NN	O	O
a	NN	O	O
new	NN	O	O
program	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
that	NN	O	O
includes	NN	O	O
proliferating	NN	O	B-protein
cell	NN	O	I-protein
nuclear	NN	O	I-protein
antigen	NN	O	I-protein
(	NN	O	O
PCNA	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
requisite	NN	O	O
processivity	NN	O	O
factor	NN	O	O
for	NN	O	O
DNA	NN	O	B-protein
polymerase	NN	O	I-protein
delta	NN	O	I-protein
.	NN	O	O

PCNA	NN	O	B-protein
transcription	NN	O	O
is	NN	O	O
regulated	NN	O	O
in	NN	O	O
part	NN	O	O
through	NN	O	O
tandem	NN	O	O
CRE	NN	O	B-DNA
sequences	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
promoter	NN	O	B-protein
and	NN	O	I-protein
CRE	NN	O	I-protein
binding	NN	O	I-protein
proteins	NN	O	I-protein
;	NN	O	O
IL-2	NN	O	B-protein
stimulates	NN	O	O
CREB	NN	O	O
phosphorylation	NN	O	O
in	NN	O	O
the	NN	O	O
resting	NN	O	B-cell_line
cloned	NN	O	I-cell_line
T	NN	O	I-cell_line
lymphocyte	NN	O	I-cell_line
,	NN	O	O
L2	NN	O	B-cell_line
.	NN	O	O

After	NN	O	O
culturing	NN	O	O
L2	NN	O	B-cell_line
cells	NN	O	I-cell_line
for	NN	O	O
greater	NN	O	O
than	NN	O	O
91	NN	O	O
days	NN	O	O
,	NN	O	O
we	NN	O	O
consistently	NN	O	O
isolate	NN	O	O
a	NN	O	O
stable	NN	O	O
variant	NN	O	O
that	NN	O	O
exhibits	NN	O	O
constitutive	NN	O	O
CREB	NN	O	O
phosphorylation	NN	O	O
.	NN	O	O

L2	NN	O	B-cell_line
and	NN	O	I-cell_line
L2	NN	O	I-cell_line
variant	NN	O	I-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
tested	NN	O	O
for	NN	O	O
IL-2	NN	O	B-protein
responsiveness	NN	O	O
and	NN	O	O
rapamycin	NN	O	O
sensitivity	NN	O	O
with	NN	O	O
respect	NN	O	O
to	NN	O	O
specific	NN	O	O
kinase	NN	O	O
activity	NN	O	O
,	NN	O	O
PCNA	NN	O	O
expression	NN	O	O
and	NN	O	O
proliferation	NN	O	O
.	NN	O	O

In	NN	O	O
L2	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
IL-2	NN	O	B-protein
stimulated	NN	O	O
and	NN	O	O
rapamycin	NN	O	O
inhibited	NN	O	O
the	NN	O	O
following	NN	O	O
:	NN	O	O
cAMP-independent	NN	O	O
CREB	NN	O	B-protein
kinase	NN	O	I-protein
activity	NN	O	O
,	NN	O	O
PCNA	NN	O	O
expression	NN	O	O
and	NN	O	O
proliferation	NN	O	O
.	NN	O	O

In	NN	O	O
L2	NN	O	B-cell_line
variant	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
CREB	NN	O	B-protein
kinase	NN	O	I-protein
activity	NN	O	O
was	NN	O	O
constitutively	NN	O	O
high	NN	O	O
;	NN	O	O
IL-2	NN	O	B-protein
stimulated	NN	O	O
and	NN	O	O
rapamycin	NN	O	O
blocked	NN	O	O
PCNA	NN	O	O
expression	NN	O	O
and	NN	O	O
proliferation	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
IL-2	NN	O	B-protein
induces	NN	O	O
a	NN	O	O
rapamycin-sensitive	NN	O	O
,	NN	O	O
cAMP-independent	NN	O	O
CREB	NN	O	B-protein
kinase	NN	O	I-protein
activity	NN	O	O
in	NN	O	O
L2	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

However	NN	O	O
,	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
CREB	NN	O	B-protein
alone	NN	O	O
is	NN	O	O
not	NN	O	O
sufficient	NN	O	O
to	NN	O	O
drive	NN	O	O
PCNA	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
L2	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Serotonin	NN	O	O
derivative	NN	O	O
,	NN	O	O
N-	NN	O	O
(	NN	O	O
p-coumaroyl	NN	O	O
)	NN	O	O
serotonin	NN	O	O
,	NN	O	O
inhibits	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
TNF-alpha	NN	O	B-protein
,	NN	O	O
IL-1alpha	NN	O	B-protein
,	NN	O	O
IL-1beta	NN	O	B-protein
,	NN	O	O
and	NN	O	O
IL-6	NN	O	B-protein
by	NN	O	O
endotoxin-stimulated	NN	O	B-cell_type
human	NN	O	I-cell_type
blood	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
have	NN	O	O
reported	NN	O	O
that	NN	O	O
N-	NN	O	O
(	NN	O	O
p-coumaroyl	NN	O	O
)	NN	O	O
serotonin	NN	O	O
(	NN	O	O
CS	NN	O	O
)	NN	O	O
and	NN	O	O
its	NN	O	O
derivatives	NN	O	O
with	NN	O	O
antioxidative	NN	O	O
activity	NN	O	O
are	NN	O	O
present	NN	O	O
in	NN	O	O
safflower	NN	O	O
seeds	NN	O	O
.	NN	O	O

As	NN	O	O
reactive	NN	O	O
oxygen	NN	O	O
species	NN	O	O
(	NN	O	O
ROS	NN	O	O
)	NN	O	O
are	NN	O	O
implicated	NN	O	O
in	NN	O	O
the	NN	O	O
signaling	NN	O	O
of	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
,	NN	O	O
we	NN	O	O
examined	NN	O	O
whether	NN	O	O
CS	NN	O	O
has	NN	O	O
a	NN	O	O
suppressive	NN	O	O
effect	NN	O	O
on	NN	O	O
inflammatory	NN	O	B-protein
cytokine	NN	O	I-protein
generation	NN	O	O
from	NN	O	O
human	NN	O	B-cell_type
monocyte	NN	O	I-cell_type
s	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

CS	NN	O	O
at	NN	O	O
50-200	NN	O	O
microM	NN	O	O
reduced	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
TNF	NN	O	B-protein
)	NN	O	O
,	NN	O	O
interleukin-1	NN	O	B-protein
(	NN	O	O
IL-1	NN	O	B-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
IL-6	NN	O	B-protein
activities	NN	O	O
in	NN	O	O
the	NN	O	O
culture	NN	O	O
supernatants	NN	O	O
from	NN	O	O
LPS-stimulated	NN	O	B-cell_line
human	NN	O	I-cell_line
blood	NN	O	I-cell_line
monocytes	NN	O	I-cell_line
without	NN	O	O
cytotoxicity	NN	O	O
.	NN	O	O

ELISA	NN	O	O
assay	NN	O	O
revealed	NN	O	O
that	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
TNF-alpha	NN	O	B-protein
,	NN	O	O
IL-1alpha	NN	O	B-protein
,	NN	O	O
IL-1beta	NN	O	B-protein
,	NN	O	O
and	NN	O	O
IL-6	NN	O	B-protein
was	NN	O	O
inhibited	NN	O	O
by	NN	O	O
CS	NN	O	O
.	NN	O	O

Northern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
showed	NN	O	O
that	NN	O	O
LPS-induced	NN	O	O
expression	NN	O	O
of	NN	O	O
these	NN	O	O
cytokine	NN	O	B-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
monocytes	NN	O	B-cell_type
was	NN	O	O
suppressed	NN	O	O
by	NN	O	O
CS	NN	O	O
.	NN	O	O

NF-kappaB	NN	O	O
activation	NN	O	O
was	NN	O	O
also	NN	O	O
inhibited	NN	O	O
by	NN	O	O
CS	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
indicate	NN	O	O
that	NN	O	O
CS	NN	O	O
has	NN	O	O
a	NN	O	O
suppressive	NN	O	O
effect	NN	O	O
on	NN	O	O
proinflammatory	NN	O	O
cytokine	NN	O	O
production	NN	O	O
from	NN	O	O
monocytes	NN	O	B-cell_type
,	NN	O	O
and	NN	O	O
this	NN	O	O
effect	NN	O	O
is	NN	O	O
based	NN	O	O
in	NN	O	O
part	NN	O	O
on	NN	O	O
the	NN	O	O
suppression	NN	O	O
of	NN	O	O
cytokine	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
through	NN	O	O
inhibition	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Elevated	NN	O	O
expression	NN	O	O
of	NN	O	O
differentiation	NN	O	B-RNA
inhibitory	NN	O	I-RNA
factor	NN	O	I-RNA
nm23	NN	O	I-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
monoblastic	NN	O	O
crisis	NN	O	O
of	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
chronic	NN	O	O
myelogenous	NN	O	O
leukemia	NN	O	O
.	NN	O	O

Differentiation	NN	O	B-protein
inhibitory	NN	O	I-protein
factor	NN	O	I-protein
nm23	NN	O	B-DNA
gene	NN	O	I-DNA
has	NN	O	O
been	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
expressed	NN	O	O
in	NN	O	O
high	NN	O	O
quantities	NN	O	O
in	NN	O	O
acute	NN	O	O
myelogenous	NN	O	O
leukemia	NN	O	O
(	NN	O	O
AML	NN	O	O
)	NN	O	O
,	NN	O	O
especially	NN	O	O
in	NN	O	O
acute	NN	O	O
monocytic	NN	O	O
leukemia	NN	O	O
(	NN	O	O
AML-M5	NN	O	O
)	NN	O	O
and	NN	O	O
is	NN	O	O
suggested	NN	O	O
as	NN	O	O
a	NN	O	O
new	NN	O	O
prognostic	NN	O	O
factor	NN	O	O
in	NN	O	O
AML-M5	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
an	NN	O	O
example	NN	O	O
of	NN	O	O
elevated	NN	O	O
expression	NN	O	O
of	NN	O	O
nm23	NN	O	B-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
chronic	NN	O	O
myelogenous	NN	O	O
leukemia	NN	O	O
(	NN	O	O
CML	NN	O	O
)	NN	O	O
who	NN	O	O
developed	NN	O	O
monoblastic	NN	O	O
crisis	NN	O	O
.	NN	O	O

Relative	NN	O	O
levels	NN	O	O
of	NN	O	O
nm23-H1	NN	O	B-RNA
and	NN	O	I-RNA
-H2	NN	O	I-RNA
mRNA	NN	O	I-RNA
extracted	NN	O	O
from	NN	O	O
the	NN	O	O
patient	NN	O	O
's	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
bone	NN	O	B-cell_type
marrow	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
measured	NN	O	O
by	NN	O	O
quantitative	NN	O	O
reverse	NN	O	O
transcriptase	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
.	NN	O	O

The	NN	O	O
level	NN	O	O
of	NN	O	O
nm23-H1	NN	O	B-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
CML	NN	O	B-cell_line
cells	NN	O	I-cell_line
at	NN	O	O
the	NN	O	O
chronic	NN	O	O
phase	NN	O	O
was	NN	O	O
as	NN	O	O
high	NN	O	O
as	NN	O	O
that	NN	O	O
in	NN	O	O
bone	NN	O	B-cell_type
marrow	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
healthy	NN	O	O
volunteers	NN	O	O
.	NN	O	O

The	NN	O	O
mRNA	NN	O	O
level	NN	O	O
of	NN	O	O
nm23-H2	NN	O	B-protein
was	NN	O	O
slightly	NN	O	O
below	NN	O	O
the	NN	O	O
normal	NN	O	O
level	NN	O	O
.	NN	O	O

At	NN	O	O
blastic	NN	O	O
crisis	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
expression	NN	O	O
of	NN	O	O
both	NN	O	O
nm23-H1	NN	O	B-RNA
and	NN	O	I-RNA
-H2	NN	O	I-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
elevated	NN	O	O
to	NN	O	O
about	NN	O	O
three	NN	O	O
to	NN	O	O
nine	NN	O	O
times	NN	O	O
of	NN	O	O
that	NN	O	O
at	NN	O	O
the	NN	O	O
chronic	NN	O	O
phase	NN	O	O
.	NN	O	O

Proliferated	NN	O	B-cell_type
blastic	NN	O	I-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
positive	NN	O	O
for	NN	O	O
non-specific	NN	O	B-protein
esterase	NN	O	I-protein
,	NN	O	O
and	NN	O	O
the	NN	O	O
serum	NN	O	O
lysozyme	NN	O	O
level	NN	O	O
was	NN	O	O
elevated	NN	O	O
and	NN	O	O
diagnosed	NN	O	O
as	NN	O	O
monoblastic	NN	O	O
crisis	NN	O	O
.	NN	O	O

The	NN	O	O
patient	NN	O	O
received	NN	O	O
combined	NN	O	O
chemotherapy	NN	O	O
but	NN	O	O
response	NN	O	O
was	NN	O	O
partial	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
are	NN	O	O
compatible	NN	O	O
with	NN	O	O
our	NN	O	O
previous	NN	O	O
report	NN	O	O
that	NN	O	O
nm23	NN	O	B-DNA
gene	NN	O	I-DNA
is	NN	O	O
overexpressed	NN	O	O
in	NN	O	O
monocytic	NN	O	O
leukemia	NN	O	O
.	NN	O	O

-DOCSTART-	O

Increased	NN	O	O
transcription	NN	O	O
decreases	NN	O	O
the	NN	O	O
spontaneous	NN	O	O
mutation	NN	O	O
rate	NN	O	O
at	NN	O	O
the	NN	O	O
thymidine	NN	O	B-DNA
kinase	NN	O	I-DNA
locus	NN	O	I-DNA
in	NN	O	O
human	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Transcription	NN	O	O
increases	NN	O	O
DNA	NN	O	O
repair	NN	O	O
efficiency	NN	O	O
and	NN	O	O
modulates	NN	O	O
the	NN	O	O
distribution	NN	O	O
of	NN	O	O
certain	NN	O	O
types	NN	O	O
of	NN	O	O
DNA	NN	O	O
damage	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
increased	NN	O	O
transcription	NN	O	O
level	NN	O	O
stimulates	NN	O	O
spontaneous	NN	O	O
mutation	NN	O	O
rate	NN	O	O
in	NN	O	O
yeast	NN	O	O
.	NN	O	O

We	NN	O	O
explored	NN	O	O
whether	NN	O	O
transcription	NN	O	O
level	NN	O	O
affects	NN	O	O
spontaneous	NN	O	O
mutation	NN	O	O
rate	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
first	NN	O	O
developed	NN	O	O
two	NN	O	O
thymidine	NN	O	B-cell_line
kinase	NN	O	I-cell_line
(	NN	O	I-cell_line
tk	NN	O	I-cell_line
)	NN	O	I-cell_line
inducible	NN	O	I-cell_line
human	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
using	NN	O	O
the	NN	O	O
Gal4-Estrogen	NN	O	O
receptor	NN	O	O
system	NN	O	O
.	NN	O	O

In	NN	O	O
our	NN	O	O
TK6i-G3	NN	O	B-cell_line
and	NN	O	I-cell_line
G9	NN	O	I-cell_line
tk	NN	O	I-cell_line
heterozygous	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
the	NN	O	O
active	NN	O	O
tk	NN	O	B-DNA
allele	NN	O	I-DNA
is	NN	O	O
linked	NN	O	O
to	NN	O	O
an	NN	O	O
inducible	NN	O	O
promoter	NN	O	B-DNA
element	NN	O	I-DNA
.	NN	O	O

Tk	NN	O	B-RNA
mRNA	NN	O	I-RNA
is	NN	O	O
induced	NN	O	O
following	NN	O	O
treatment	NN	O	O
with	NN	O	O
estrogen	NN	O	O
.	NN	O	O

Spontaneous	NN	O	O
mutation	NN	O	O
rate	NN	O	O
was	NN	O	O
significantly	NN	O	O
decreased	NN	O	O
in	NN	O	O
human	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
after	NN	O	O
induction	NN	O	O
in	NN	O	O
contrast	NN	O	O
to	NN	O	O
the	NN	O	O
report	NN	O	O
in	NN	O	O
yeast	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
humans	NN	O	O
may	NN	O	O
have	NN	O	O
evolved	NN	O	O
different	NN	O	O
or	NN	O	O
additional	NN	O	O
mechanisms	NN	O	O
to	NN	O	O
deal	NN	O	O
with	NN	O	O
transcription	NN	O	O
related	NN	O	O
spontaneous	NN	O	O
mutagenesis	NN	O	O
.	NN	O	O

Copyright	NN	O	O
1998	NN	O	O
Elsevier	NN	O	O
Science	NN	O	O
B.V	NN	O	O
.	NN	O	O

All	NN	O	O
rights	NN	O	O
reserved	NN	O	O
.	NN	O	O

-DOCSTART-	O

Mutation	NN	O	O
of	NN	O	O
BCL-6	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
normal	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
the	NN	O	O
process	NN	O	O
of	NN	O	O
somatic	NN	O	O
hypermutation	NN	O	O
of	NN	O	O
Ig	NN	O	B-DNA
genes	NN	O	I-DNA
.	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
are	NN	O	O
hypermutated	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
that	NN	O	O
are	NN	O	O
the	NN	O	O
precursors	NN	O	O
to	NN	O	O
memory	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
mutations	NN	O	O
are	NN	O	O
linked	NN	O	O
to	NN	O	O
transcription	NN	O	O
initiation	NN	O	O
,	NN	O	O
but	NN	O	O
non-Ig	NN	O	B-DNA
promoters	NN	O	I-DNA
are	NN	O	O
permissible	NN	O	O
for	NN	O	O
the	NN	O	O
mutation	NN	O	O
process	NN	O	O
;	NN	O	O
thus	NN	O	O
,	NN	O	O
other	NN	O	O
genes	NN	O	O
expressed	NN	O	O
in	NN	O	O
mutating	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
may	NN	O	O
also	NN	O	O
be	NN	O	O
subject	NN	O	O
to	NN	O	O
somatic	NN	O	O
hypermutation	NN	O	O
.	NN	O	O

Significant	NN	O	O
mutations	NN	O	O
were	NN	O	O
not	NN	O	O
observed	NN	O	O
in	NN	O	O
c-MYC	NN	O	B-DNA
,	NN	O	I-DNA
S14	NN	O	I-DNA
,	NN	O	I-DNA
or	NN	O	I-DNA
alpha-fetoprotein	NN	O	I-DNA
(	NN	O	I-DNA
AFP	NN	O	I-DNA
)	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
but	NN	O	O
BCL-6	NN	O	B-protein
was	NN	O	O
highly	NN	O	O
mutated	NN	O	O
in	NN	O	O
a	NN	O	O
large	NN	O	O
proportion	NN	O	O
of	NN	O	O
memory	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
normal	NN	O	O
individuals	NN	O	O
.	NN	O	O

The	NN	O	O
mutation	NN	O	O
pattern	NN	O	O
was	NN	O	O
similar	NN	O	O
to	NN	O	O
that	NN	O	O
of	NN	O	O
Ig	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

CD28-mediated	NN	O	O
activation	NN	O	O
in	NN	O	O
CD45RA+	NN	O	B-cell_type
and	NN	O	I-cell_type
CD45RO+	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
:	NN	O	O
enhanced	NN	O	O
levels	NN	O	O
of	NN	O	O
reactive	NN	O	O
oxygen	NN	O	O
intermediates	NN	O	O
and	NN	O	O
c-Rel	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
in	NN	O	O
CD45RA+	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
have	NN	O	O
analyzed	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
complete	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
(	NN	O	O
anti-CD3	NN	O	O
plus	NN	O	O
anti-CD28	NN	O	O
)	NN	O	O
on	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
in	NN	O	O
CD45RA+	NN	O	B-cell_type
(	NN	O	I-cell_type
naive	NN	O	I-cell_type
)	NN	O	I-cell_type
and	NN	O	I-cell_type
CD45RO+	NN	O	I-cell_type
(	NN	O	I-cell_type
memory/effector	NN	O	I-cell_type
)	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Long	NN	O	O
exposure	NN	O	O
(	NN	O	O
24	NN	O	O
h	NN	O	O
)	NN	O	O
induced	NN	O	O
stronger	NN	O	O
NF-kappaB	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
in	NN	O	O
CD45RA+	NN	O	B-cell_type
cells	NN	O	I-cell_type
than	NN	O	O
in	NN	O	O
CD45RO+	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
c-Rel	NN	O	I-protein
protein	NN	O	I-protein
indicated	NN	O	O
that	NN	O	O
after	NN	O	O
anti-CD3+anti-CD28	NN	O	O
stimulation	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
c-Rel	NN	O	B-protein
was	NN	O	O
higher	NN	O	O
in	NN	O	O
CD45RA+	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
the	NN	O	O
cytoplasmic	NN	O	B-protein
inhibitor	NN	O	I-protein
IkappaBalpha	NN	O	B-protein
indicated	NN	O	O
that	NN	O	O
anti-CD3+anti-CD28	NN	O	O
stimulation	NN	O	O
induced	NN	O	O
a	NN	O	O
long-lasting	NN	O	O
degradation	NN	O	O
in	NN	O	O
CD45RA+	NN	O	B-cell_type
cells	NN	O	I-cell_type
but	NN	O	O
in	NN	O	O
CD45RO+	NN	O	B-cell_type
cells	NN	O	I-cell_type
the	NN	O	O
degradation	NN	O	O
process	NN	O	O
was	NN	O	O
more	NN	O	O
rapid	NN	O	O
.	NN	O	O

Because	NN	O	O
the	NN	O	O
CD28	NN	O	B-protein
costimulus	NN	O	O
is	NN	O	O
known	NN	O	O
to	NN	O	O
induce	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
reactive	NN	O	O
oxygen	NN	O	O
intermediates	NN	O	O
(	NN	O	O
ROIs	NN	O	O
)	NN	O	O
,	NN	O	O
the	NN	O	O
intracellular	NN	O	O
ROI	NN	O	O
levels	NN	O	O
in	NN	O	O
CD45RA+	NN	O	B-cell_type
and	NN	O	I-cell_type
CD45RO+	NN	O	I-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
compared	NN	O	O
by	NN	O	O
flow	NN	O	O
cytometry	NN	O	O
.	NN	O	O

ROIs	NN	O	O
were	NN	O	O
produced	NN	O	O
in	NN	O	O
both	NN	O	O
cell	NN	O	O
types	NN	O	O
,	NN	O	O
but	NN	O	O
more	NN	O	O
strongly	NN	O	O
in	NN	O	O
CD45RA+	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
data	NN	O	O
presented	NN	O	O
in	NN	O	O
this	NN	O	O
study	NN	O	O
further	NN	O	O
emphasize	NN	O	O
the	NN	O	O
differences	NN	O	O
between	NN	O	O
CD45RA+	NN	O	B-cell_type
and	NN	O	I-cell_type
CD45RO+	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
in	NN	O	O
ROI-dependent	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
.	NN	O	O

-DOCSTART-	O

Non-Hodgkin	NN	O	O
's	NN	O	O
lymphoma	NN	O	O
involving	NN	O	O
bilateral	NN	O	O
breasts	NN	O	O
[	NN	O	O
see	NN	O	O
comments	NN	O	O
]	NN	O	O

We	NN	O	O
describe	NN	O	O
here	NN	O	O
two	NN	O	O
cases	NN	O	O
of	NN	O	O
diffuse	NN	O	O
large	NN	O	O
cell	NN	O	O
type	NN	O	O
non-Hodgkin	NN	O	O
's	NN	O	O
lymphoma	NN	O	O
affecting	NN	O	O
the	NN	O	O
bilateral	NN	O	O
breasts	NN	O	O
.	NN	O	O

The	NN	O	O
contralateral	NN	O	O
tumor	NN	O	O
in	NN	O	O
one	NN	O	O
case	NN	O	O
appeared	NN	O	O
17	NN	O	O
months	NN	O	O
after	NN	O	O
the	NN	O	O
first	NN	O	O
mastectomy	NN	O	O
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
bilateral	NN	O	O
tumors	NN	O	O
occurred	NN	O	O
concurrently	NN	O	O
in	NN	O	O
the	NN	O	O
other	NN	O	O
patient	NN	O	O
who	NN	O	O
was	NN	O	O
pregnant	NN	O	O
and	NN	O	O
showed	NN	O	O
widespread	NN	O	O
dissemination	NN	O	O
at	NN	O	O
initial	NN	O	O
presentation	NN	O	O
.	NN	O	O

Lymphoma	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
both	NN	O	O
cases	NN	O	O
showed	NN	O	O
the	NN	O	O
mature	NN	O	O
B-cell	NN	O	O
immunophenotype	NN	O	O
and	NN	O	O
had	NN	O	O
rearrangements	NN	O	O
of	NN	O	O
the	NN	O	O
BCL6	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

Both	NN	O	O
patients	NN	O	O
developed	NN	O	O
progressive	NN	O	O
disease	NN	O	O
despite	NN	O	O
chemo-radiotherapy	NN	O	O
and	NN	O	O
died	NN	O	O
of	NN	O	O
leukemic	NN	O	O
manifestations	NN	O	O
.	NN	O	O

There	NN	O	O
were	NN	O	O
no	NN	O	O
apparent	NN	O	O
pathological	NN	O	O
features	NN	O	O
of	NN	O	O
lymphomas	NN	O	O
of	NN	O	O
mucosa-associated	NN	O	O
lymphoid	NN	O	O
tissue	NN	O	O
origin	NN	O	O

-DOCSTART-	O

Biochemical	NN	O	O
characterization	NN	O	O
of	NN	O	O
MIP-1	NN	O	B-protein
alpha	NN	O	I-protein
nuclear	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

A	NN	O	O
family	NN	O	O
of	NN	O	O
hematopoietic	NN	O	B-protein
specific	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
,	NN	O	O
MIP-1	NN	O	B-protein
alpha	NN	O	I-protein
nuclear	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	I-protein
MNP	NN	O	I-protein
)	NN	O	I-protein
family	NN	O	I-protein
,	NN	O	O
has	NN	O	O
recently	NN	O	O
been	NN	O	O
identified	NN	O	O
.	NN	O	O

They	NN	O	O
are	NN	O	O
intimately	NN	O	O
involved	NN	O	O
in	NN	O	O
regulating	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
huMIP-1	NN	O	B-DNA
alpha	NN	O	I-DNA
gene	NN	O	I-DNA
in	NN	O	O
monocytes	NN	O	B-cell_type
,	NN	O	O
T-cells	NN	O	B-cell_type
,	NN	O	O
and	NN	O	O
transformed	NN	O	B-cell_type
B-cells	NN	O	I-cell_type
.	NN	O	O

One	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
family	NN	O	O
(	NN	O	O
MNP-1	NN	O	B-protein
)	NN	O	O
is	NN	O	O
essential	NN	O	O
for	NN	O	O
promoter	NN	O	O
activity	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
B-cells	NN	O	B-cell_type
,	NN	O	O
while	NN	O	O
another	NN	O	O
(	NN	O	O
MNP-2	NN	O	B-protein
)	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
full	NN	O	O
promotor	NN	O	O
activity	NN	O	O
in	NN	O	O
T-cells	NN	O	B-cell_type
.	NN	O	O

A	NN	O	O
third	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
family	NN	O	O
(	NN	O	O
MNP-3	NN	O	B-protein
)	NN	O	O
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
PMA	NN	O	B-cell_line
induced	NN	O	I-cell_line
HL60	NN	O	I-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
probably	NN	O	O
has	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
monocyte	NN	O	O
differentiation	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
communication	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
by	NN	O	O
two	NN	O	O
techniques	NN	O	O
that	NN	O	O
MNP-1	NN	O	B-protein
and	NN	O	O
MNP-2	NN	O	B-protein
are	NN	O	O
distinct	NN	O	O
but	NN	O	O
related	NN	O	O
factors	NN	O	O
,	NN	O	O
and	NN	O	O
we	NN	O	O
present	NN	O	O
further	NN	O	O
evidence	NN	O	O
to	NN	O	O
show	NN	O	O
that	NN	O	O
MNP-1	NN	O	B-protein
acts	NN	O	O
as	NN	O	O
a	NN	O	O
heterodimer	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Bcl-3	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
are	NN	O	O
induced	NN	O	O
by	NN	O	O
granulocyte-macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
and	NN	O	O
erythropoietin	NN	O	B-protein
in	NN	O	O
proliferating	NN	O	B-cell_type
human	NN	O	I-cell_type
erythroid	NN	O	I-cell_type
precursors	NN	O	I-cell_type
.	NN	O	O

Bcl-3	NN	O	B-protein
is	NN	O	O
a	NN	O	O
proto-oncogene	NN	O	B-DNA
involved	NN	O	O
in	NN	O	O
the	NN	O	O
chromosomal	NN	O	O
translocation	NN	O	O
t	NN	O	B-DNA
(	NN	O	I-DNA
14	NN	O	I-DNA
;	NN	O	I-DNA
19	NN	O	I-DNA
)	NN	O	I-DNA
found	NN	O	O
in	NN	O	O
some	NN	O	O
patients	NN	O	O
with	NN	O	O
chronic	NN	O	O
lymphocytic	NN	O	O
leukemia	NN	O	O
.	NN	O	O

It	NN	O	O
shares	NN	O	O
structural	NN	O	O
similarities	NN	O	O
with	NN	O	O
and	NN	O	O
is	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
IkappaB	NN	O	B-protein
family	NN	O	I-protein
of	NN	O	O
proteins	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
,	NN	O	O
involvement	NN	O	O
of	NN	O	O
Bcl-3	NN	O	B-protein
in	NN	O	O
hematopoietic	NN	O	O
growth	NN	O	O
factor-stimulated	NN	O	O
erythroid	NN	O	O
proliferation	NN	O	O
and	NN	O	O
differentiation	NN	O	O
was	NN	O	O
examined	NN	O	O
.	NN	O	O

In	NN	O	O
TF-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
an	NN	O	O
erythroleukemia	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
granulocyte-macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
GM-CSF	NN	O	B-protein
)	NN	O	O
and	NN	O	O
erythropoietin	NN	O	B-protein
(	NN	O	O
Epo	NN	O	B-protein
)	NN	O	O
greatly	NN	O	O
enhanced	NN	O	O
Bcl-3	NN	O	B-protein
expression	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
in	NN	O	O
association	NN	O	O
with	NN	O	O
stimulation	NN	O	O
of	NN	O	O
proliferation	NN	O	O
.	NN	O	O

Bcl-3	NN	O	B-protein
protein	NN	O	O
was	NN	O	O
also	NN	O	O
highly	NN	O	O
expressed	NN	O	O
in	NN	O	O
early	NN	O	B-cell_type
burst-forming	NN	O	I-cell_type
unit-erythroid	NN	O	I-cell_type
(	NN	O	I-cell_type
BFU-E	NN	O	I-cell_type
)	NN	O	I-cell_type
-derived	NN	O	I-cell_type
erythroid	NN	O	I-cell_type
precursors	NN	O	I-cell_type
(	NN	O	O
day	NN	O	O
7	NN	O	O
)	NN	O	O
and	NN	O	O
decreased	NN	O	O
during	NN	O	O
maturation	NN	O	O
(	NN	O	O
days	NN	O	O
10	NN	O	O
and	NN	O	O
14	NN	O	O
)	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
Bcl-3	NN	O	B-protein
is	NN	O	O
involved	NN	O	O
in	NN	O	O
normal	NN	O	O
erythroid	NN	O	O
proliferation	NN	O	O
.	NN	O	O

In	NN	O	O
these	NN	O	O
hematopoietic	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
Bcl-3	NN	O	B-protein
was	NN	O	O
hyperphosphorylated	NN	O	O
.	NN	O	O

GM-CSF	NN	O	B-protein
and	NN	O	O
Epo	NN	O	B-protein
modulated	NN	O	O
the	NN	O	O
subcellular	NN	O	O
localization	NN	O	O
of	NN	O	O
Bcl-3	NN	O	B-protein
.	NN	O	O

Upon	NN	O	O
stimulation	NN	O	O
of	NN	O	O
TF-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
GM-CSF	NN	O	B-protein
or	NN	O	O
Epo	NN	O	B-protein
,	NN	O	O
the	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
Bcl-3	NN	O	B-protein
was	NN	O	O
dramatically	NN	O	O
enhanced	NN	O	O
.	NN	O	O

Overexpression	NN	O	O
of	NN	O	O
Bcl-3	NN	O	B-protein
in	NN	O	O
TF-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
by	NN	O	O
transient	NN	O	O
transfection	NN	O	O
along	NN	O	O
with	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-protein
factors	NN	O	I-protein
p50	NN	O	I-protein
or	NN	O	I-protein
p52	NN	O	I-protein
resulted	NN	O	O
in	NN	O	O
significant	NN	O	O
induction	NN	O	O
of	NN	O	O
an	NN	O	O
human	NN	O	B-DNA
immunodeficiency	NN	O	I-DNA
virus-type	NN	O	I-DNA
1	NN	O	I-DNA
(	NN	O	I-DNA
HIV-1	NN	O	I-DNA
)	NN	O	I-DNA
kappaB-TATA-luceriferase	NN	O	I-DNA
reporter	NN	O	I-DNA
plasmid	NN	O	I-DNA
,	NN	O	O
demonstrating	NN	O	O
that	NN	O	O
Bcl-3	NN	O	B-protein
has	NN	O	O
a	NN	O	O
positive	NN	O	O
role	NN	O	O
in	NN	O	O
transactivation	NN	O	O
of	NN	O	O
kappaB-containing	NN	O	B-DNA
genes	NN	O	I-DNA
in	NN	O	O
erythroid	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Stimulation	NN	O	O
with	NN	O	O
GM-CSF	NN	O	B-protein
enhanced	NN	O	O
c-myb	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

Bcl-3	NN	O	B-protein
in	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
of	NN	O	O
TF-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
bound	NN	O	O
to	NN	O	O
a	NN	O	O
kappaB	NN	O	B-DNA
enhancer	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
c-myb	NN	O	B-DNA
promoter	NN	O	I-DNA
together	NN	O	O
with	NN	O	O
NF-kappaB2/p52	NN	O	B-protein
and	NN	O	O
this	NN	O	O
binding	NN	O	O
activity	NN	O	O
was	NN	O	O
enhanced	NN	O	O
by	NN	O	O
GM-CSF	NN	O	B-protein
stimulation	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
cotransfection	NN	O	O
of	NN	O	O
Bcl-3	NN	O	B-protein
with	NN	O	O
p52	NN	O	B-protein
or	NN	O	O
p50	NN	O	B-protein
in	NN	O	O
TF-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
resulted	NN	O	O
in	NN	O	O
significant	NN	O	O
activation	NN	O	O
of	NN	O	O
a	NN	O	O
c-myb	NN	O	B-DNA
kappaB-TATA-luceriferase	NN	O	I-DNA
reporter	NN	O	I-DNA
plasmid	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
Bcl-3	NN	O	B-protein
may	NN	O	O
participate	NN	O	O
in	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
certain	NN	O	O
kappaB-containing	NN	O	B-DNA
genes	NN	O	I-DNA
involved	NN	O	O
in	NN	O	O
hematopoiesis	NN	O	O
,	NN	O	O
including	NN	O	O
c-myb	NN	O	B-DNA
.	NN	O	O

Copyright	NN	O	O
1998	NN	O	O
by	NN	O	O
The	NN	O	O
American	NN	O	O
Society	NN	O	O
of	NN	O	O
Hematology	NN	O	O
.	NN	O	O

-DOCSTART-	O

5-Lipoxygenase	NN	O	B-protein
compartmentalization	NN	O	O
in	NN	O	O
granulocytic	NN	O	O
cells	NN	O	O
is	NN	O	O
modulated	NN	O	O
by	NN	O	O
an	NN	O	O
internal	NN	O	B-protein
bipartite	NN	O	I-protein
nuclear	NN	O	I-protein
localizing	NN	O	I-protein
sequence	NN	O	I-protein
and	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
complex	NN	O	I-protein
formation	NN	O	O
.	NN	O	O

A	NN	O	O
region	NN	O	O
of	NN	O	O
basic	NN	O	O
amino	NN	O	O
acids	NN	O	O
spanning	NN	O	O
residues	NN	O	B-protein
639-656	NN	O	I-protein
in	NN	O	O
the	NN	O	O
human	NN	O	B-protein
5-lipoxygenase	NN	O	I-protein
sequence	NN	O	I-protein
resembles	NN	O	O
a	NN	O	O
consensus	NN	O	B-protein
bipartite	NN	O	I-protein
nuclear	NN	O	I-protein
localizing	NN	O	I-protein
sequence	NN	O	I-protein
.	NN	O	O

A	NN	O	O
synthetic	NN	O	O
peptide	NN	O	O
consisting	NN	O	O
of	NN	O	O
the	NN	O	O
Kaposi	NN	O	B-protein
fibroblast	NN	O	I-protein
growth	NN	O	I-protein
factor	NN	O	I-protein
signal	NN	O	I-protein
sequence	NN	O	I-protein
fused	NN	O	O
to	NN	O	O
the	NN	O	O
5-lipoxygenase639-656	NN	O	B-protein
bipartite	NN	O	I-protein
nuclear	NN	O	I-protein
localizing	NN	O	I-protein
sequence	NN	O	I-protein
has	NN	O	O
a	NN	O	O
prominent	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
on	NN	O	O
5-lipoxygenase	NN	O	B-protein
catalysis	NN	O	O
in	NN	O	O
granulocytic	NN	O	O
HL-60	NN	O	B-cell_line
cells	NN	O	I-cell_line
activated	NN	O	O
by	NN	O	O
calcium	NN	O	O
ionophor	NN	O	O
A23187	NN	O	O
.	NN	O	O

Recombinant	NN	O	O
5-lipoxygenase	NN	O	B-protein
was	NN	O	O
not	NN	O	O
affected	NN	O	O
by	NN	O	O
the	NN	O	O
peptide	NN	O	O
.	NN	O	O

The	NN	O	O
peptide	NN	O	O
also	NN	O	O
inhibited	NN	O	O
redistribution	NN	O	O
of	NN	O	O
5-lipoxygenase	NN	O	B-protein
from	NN	O	O
the	NN	O	O
cytosol	NN	O	O
to	NN	O	O
the	NN	O	O
nuclear	NN	O	O
membrane	NN	O	O
of	NN	O	O
HL-60	NN	O	B-cell_line
cells	NN	O	I-cell_line
stimulated	NN	O	O
by	NN	O	O
A23187	NN	O	O
.	NN	O	O

5-Lipoxygenase	NN	O	B-protein
protein	NN	O	I-protein
was	NN	O	O
detected	NN	O	O
in	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappaB	NN	O	I-protein
(	NN	O	I-protein
NF-kappaB	NN	O	I-protein
)	NN	O	I-protein
p65	NN	O	I-protein
subunit	NN	O	I-protein
immunoprecipitate	NN	O	O
fractions	NN	O	O
prepared	NN	O	O
from	NN	O	O
HL-60	NN	O	B-cell_line
cell	NN	O	I-cell_line
lysates	NN	O	O
.	NN	O	O

The	NN	O	O
amount	NN	O	O
of	NN	O	O
5-lipoxygenase	NN	O	B-protein
protein	NN	O	I-protein
coimmunoprecipitated	NN	O	O
by	NN	O	O
NF-kappaB	NN	O	O
antiserum	NN	O	O
was	NN	O	O
increased	NN	O	O
following	NN	O	O
A23187	NN	O	O
stimulation	NN	O	O
.	NN	O	O

In	NN	O	O
cells	NN	O	O
treated	NN	O	O
with	NN	O	O
agents	NN	O	O
that	NN	O	O
block	NN	O	O
5-lipoxygenase	NN	O	B-protein
translocation	NN	O	O
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
,	NN	O	O
5-lipoxygenase	NN	O	B-protein
protein	NN	O	O
appearing	NN	O	O
in	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-protein
immunoprecipitate	NN	O	O
was	NN	O	O
diminished	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
implicate	NN	O	O
an	NN	O	O
internal	NN	O	B-protein
bipartite	NN	O	I-protein
nuclear	NN	O	I-protein
localizing	NN	O	I-protein
sequence	NN	O	I-protein
as	NN	O	O
a	NN	O	O
regulatory	NN	O	O
domain	NN	O	O
that	NN	O	O
modulates	NN	O	O
5-lipoxygenase	NN	O	B-protein
redistribution	NN	O	O
and	NN	O	O
catalysis	NN	O	O
in	NN	O	O
granulocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Additionally	NN	O	O
,	NN	O	O
our	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
molecular	NN	O	O
determinants	NN	O	O
which	NN	O	O
govern	NN	O	O
5-lipoxygenase	NN	O	O
and	NN	O	O
NF-kappaB	NN	O	O
redistribution	NN	O	O
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
may	NN	O	O
be	NN	O	O
coordinately	NN	O	O
controlled	NN	O	O
in	NN	O	O
granulocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Copyright	NN	O	O
1998	NN	O	O
Academic	NN	O	O
Press	NN	O	O
.	NN	O	O

-DOCSTART-	O

Inhibition	NN	O	O
of	NN	O	O
CD28/CD3-mediated	NN	O	O
costimulation	NN	O	O
of	NN	O	O
naive	NN	O	B-cell_type
and	NN	O	I-cell_type
memory	NN	O	I-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
by	NN	O	O
intracellular	NN	O	O
incorporation	NN	O	O
of	NN	O	O
polyclonal	NN	O	B-protein
antibodies	NN	O	I-protein
specific	NN	O	O
for	NN	O	O
the	NN	O	O
activator	NN	O	B-protein
protein-1	NN	O	I-protein
transcriptional	NN	O	I-protein
complex	NN	O	I-protein
.	NN	O	O

A	NN	O	O
number	NN	O	O
of	NN	O	O
indirect	NN	O	O
methods	NN	O	O
have	NN	O	O
been	NN	O	O
utilized	NN	O	O
in	NN	O	O
demonstrating	NN	O	O
activator	NN	O	B-protein
protein-1	NN	O	I-protein
transcription	NN	O	O
factor	NN	O	O
function	NN	O	O
in	NN	O	O
IL-2	NN	O	O
promoter	NN	O	O
activity	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
there	NN	O	O
has	NN	O	O
been	NN	O	O
no	NN	O	O
direct	NN	O	O
demonstration	NN	O	O
that	NN	O	O
activator	NN	O	B-protein
protein-1	NN	O	I-protein
is	NN	O	O
involved	NN	O	O
in	NN	O	O
CD28-dependent	NN	O	O
costimulation	NN	O	O
of	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
in	NN	O	O
freshly	NN	O	O
isolated	NN	O	O
naive	NN	O	B-cell_type
and	NN	O	I-cell_type
memory	NN	O	I-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

To	NN	O	O
address	NN	O	O
this	NN	O	O
issue	NN	O	O
,	NN	O	O
the	NN	O	O
method	NN	O	O
of	NN	O	O
scrape	NN	O	O
loading	NN	O	O
was	NN	O	O
applied	NN	O	O
to	NN	O	O
purified	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Since	NN	O	O
scrape	NN	O	O
loading	NN	O	O
relies	NN	O	O
on	NN	O	O
adherent	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
(	NN	O	I-cell_type
PB-T	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
immobilized	NN	O	O
on	NN	O	O
the	NN	O	O
nonspecific	NN	O	O
cell	NN	O	O
attachment	NN	O	O
factor	NN	O	O
poly-L-lysine	NN	O	O
.	NN	O	O

Cells	NN	O	O
scraped	NN	O	O
off	NN	O	O
poly-L-lysine	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
Ig	NN	O	B-protein
FITC	NN	O	I-protein
efficiently	NN	O	O
incorporated	NN	O	O
Ig	NN	O	O
,	NN	O	O
with	NN	O	O
relatively	NN	O	O
uniform	NN	O	O
fluorescence	NN	O	O
.	NN	O	O

T	NN	O	O
cells	NN	O	O
retained	NN	O	O
their	NN	O	O
physical	NN	O	O
parameters	NN	O	O
as	NN	O	O
measured	NN	O	O
by	NN	O	O
forward	NN	O	O
and	NN	O	O
side	NN	O	O
light	NN	O	O
scatter	NN	O	O
,	NN	O	O
and	NN	O	O
functional	NN	O	O
activity	NN	O	O
as	NN	O	O
measured	NN	O	O
by	NN	O	O
costimulation	NN	O	O
of	NN	O	O
proliferation	NN	O	O
and	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
after	NN	O	O
being	NN	O	O
scraped	NN	O	O
off	NN	O	O
this	NN	O	O
substrate	NN	O	O
.	NN	O	O

CD28/CD3-costimulated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
produced	NN	O	O
intracellular	NN	O	O
IL-2	NN	O	B-protein
from	NN	O	O
all	NN	O	O
subsets	NN	O	O
measured	NN	O	O
(	NN	O	O
CD4+	NN	O	B-cell_type
,	NN	O	O
CD4-	NN	O	B-cell_type
,	NN	O	O
CD45RO+	NN	O	B-cell_type
,	NN	O	O
and	NN	O	O
CD45RO-	NN	O	B-cell_type
)	NN	O	O
.	NN	O	O

IL-2	NN	O	B-protein
production	NN	O	O
and	NN	O	O
intracellular	NN	O	O
accumulation	NN	O	O
in	NN	O	O
nonscraped	NN	O	O
PB-T	NN	O	B-cell_type
cells	NN	O	I-cell_type
activated	NN	O	O
with	NN	O	O
CD28/CD3	NN	O	O
coligation	NN	O	O
were	NN	O	O
skewed	NN	O	O
favoring	NN	O	O
CD45RO+	NN	O	B-cell_type
and	NN	O	I-cell_type
CD4+	NN	O	I-cell_type
subsets	NN	O	I-cell_type
,	NN	O	O
as	NN	O	O
was	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
in	NN	O	O
scraped	NN	O	O
PB-T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
intracellular	NN	O	O
incorporation	NN	O	O
of	NN	O	O
Abs	NN	O	O
specific	NN	O	O
for	NN	O	O
c-Fos	NN	O	B-protein
and	NN	O	I-protein
c-Jun	NN	O	I-protein
family	NN	O	I-protein
members	NN	O	I-protein
by	NN	O	O
scrape	NN	O	O
loading	NN	O	O
inhibited	NN	O	O
the	NN	O	O
production	NN	O	O
and	NN	O	O
intracellular	NN	O	O
accumulation	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
within	NN	O	O
6	NN	O	O
h	NN	O	O
of	NN	O	O
costimulation	NN	O	O
with	NN	O	O
PMA/ionomycin	NN	O	O
,	NN	O	O
or	NN	O	O
costimulation	NN	O	O
by	NN	O	O
CD28	NN	O	O
and	NN	O	O
CD3	NN	O	O
ligation	NN	O	O
.	NN	O	O

Scrape	NN	O	O
loading	NN	O	O
thus	NN	O	O
provides	NN	O	O
an	NN	O	O
efficient	NN	O	O
mechanism	NN	O	O
for	NN	O	O
intracellular	NN	O	O
incorporation	NN	O	O
of	NN	O	O
macromolecules	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
first	NN	O	O
direct	NN	O	O
evidence	NN	O	O
that	NN	O	O
c-Fos	NN	O	B-protein
and	NN	O	O
c-Jun	NN	O	B-protein
are	NN	O	O
involved	NN	O	O
in	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
within	NN	O	O
its	NN	O	O
correct	NN	O	O
chromosomal	NN	O	O
context	NN	O	O
,	NN	O	O
in	NN	O	O
resting	NN	O	O
human	NN	O	O
T	NN	O	O
lymphocyte	NN	O	O
subpopulations	NN	O	O
.	NN	O	O

-DOCSTART-	O

Differential	NN	O	O
expression	NN	O	O
of	NN	O	O
Nur77	NN	O	B-protein
family	NN	O	I-protein
members	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
T-lymphotropic	NN	O	I-cell_type
virus	NN	O	I-cell_type
type	NN	O	I-cell_type
1-infected	NN	O	I-cell_type
cells	NN	O	I-cell_type
:	NN	O	O
transactivation	NN	O	O
of	NN	O	O
the	NN	O	O
TR3/nur77	NN	O	B-DNA
gene	NN	O	I-DNA
by	NN	O	O
Tax	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

We	NN	O	O
analyzed	NN	O	O
the	NN	O	O
differential	NN	O	O
expression	NN	O	O
and	NN	O	O
regulation	NN	O	O
of	NN	O	O
three	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
Nur77	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
family	NN	O	I-protein
by	NN	O	O
the	NN	O	O
human	NN	O	O
T-lymphotropic	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
(	NN	O	O
HTLV-1	NN	O	O
)	NN	O	O
Tax	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

We	NN	O	O
have	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
in	NN	O	O
both	NN	O	O
HTLV-1-infected	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
Tax-expressing	NN	O	B-cell_line
JPX-9	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
TR3/nur77	NN	O	B-protein
is	NN	O	O
highly	NN	O	O
expressed	NN	O	O
,	NN	O	O
whereas	NN	O	O
neither	NN	O	O
NOR-1	NN	O	O
nor	NN	O	O
NOT	NN	O	O
expression	NN	O	O
is	NN	O	O
detectable	NN	O	O
.	NN	O	O

Transient	NN	O	O
transfection	NN	O	O
analysis	NN	O	O
further	NN	O	O
confirmed	NN	O	O
the	NN	O	O
Tax	NN	O	B-protein
transactivation	NN	O	O
of	NN	O	O
the	NN	O	O
TR3/nur77	NN	O	B-protein
promoter	NN	O	O
but	NN	O	O
not	NN	O	O
the	NN	O	O
NOR-1	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
different	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
luciferase	NN	O	B-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
driven	NN	O	O
by	NN	O	O
the	NN	O	O
NGFI-B	NN	O	B-DNA
(	NN	O	I-DNA
rat	NN	O	I-DNA
homolog	NN	O	I-DNA
of	NN	O	I-DNA
TR3/Nur77	NN	O	I-DNA
)	NN	O	I-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
NBRE	NN	O	B-DNA
)	NN	O	O
provided	NN	O	O
evidence	NN	O	O
that	NN	O	O
Tax	NN	O	B-protein
-mediated	NN	O	O
transactivation	NN	O	O
resulted	NN	O	O
in	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
a	NN	O	O
functional	NN	O	O
protein	NN	O	O
.	NN	O	O

Cotransfection	NN	O	O
assays	NN	O	O
with	NN	O	O
the	NN	O	O
TR3/nur77	NN	O	B-DNA
promoter	NN	O	I-DNA
sequence	NN	O	I-DNA
or	NN	O	O
the	NN	O	O
NBRE	NN	O	B-DNA
binding	NN	O	O
motif	NN	O	O
together	NN	O	O
with	NN	O	O
a	NN	O	O
series	NN	O	O
of	NN	O	O
Tax	NN	O	B-protein
mutants	NN	O	O
have	NN	O	O
shown	NN	O	O
that	NN	O	O
Tax	NN	O	B-protein
-induced	NN	O	O
TR3/nur77	NN	O	B-protein
expression	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
CREB/ATF-related	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Negative	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
heat	NN	O	O
shock	NN	O	O
transcriptional	NN	O	O
response	NN	O	O
by	NN	O	O
HSBP1	NN	O	B-protein
.	NN	O	O

In	NN	O	O
response	NN	O	O
to	NN	O	O
stress	NN	O	O
,	NN	O	O
heat	NN	O	B-protein
shock	NN	O	I-protein
factor	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	O
HSF1	NN	O	B-protein
)	NN	O	O
acquires	NN	O	O
rapid	NN	O	O
DNA	NN	O	O
binding	NN	O	O
and	NN	O	O
transient	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
while	NN	O	O
undergoing	NN	O	O
conformational	NN	O	O
transition	NN	O	O
from	NN	O	O
an	NN	O	O
inert	NN	O	O
non-DNA-binding	NN	O	B-protein
monomer	NN	O	I-protein
to	NN	O	O
active	NN	O	O
functional	NN	O	O
trimers	NN	O	O
.	NN	O	O

Attenuation	NN	O	O
of	NN	O	O
the	NN	O	O
inducible	NN	O	O
transcriptional	NN	O	O
response	NN	O	O
occurs	NN	O	O
during	NN	O	O
heat	NN	O	O
shock	NN	O	O
or	NN	O	O
upon	NN	O	O
recovery	NN	O	O
at	NN	O	O
non-stress	NN	O	O
conditions	NN	O	O
and	NN	O	O
involves	NN	O	O
dissociation	NN	O	O
of	NN	O	O
the	NN	O	O
HSF1	NN	O	B-protein
trimer	NN	O	I-protein
and	NN	O	O
loss	NN	O	O
of	NN	O	O
activity	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
used	NN	O	O
the	NN	O	O
hydrophobic	NN	O	B-protein
repeats	NN	O	I-protein
of	NN	O	O
the	NN	O	O
HSF1	NN	O	B-protein
trimerization	NN	O	O
domain	NN	O	O
in	NN	O	O
the	NN	O	O
yeast	NN	O	O
two-hybrid	NN	O	O
protein	NN	O	O
interaction	NN	O	O
assay	NN	O	O
to	NN	O	O
identify	NN	O	O
heat	NN	O	B-protein
shock	NN	O	I-protein
factor	NN	O	I-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	O
HSBP1	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
novel	NN	O	O
,	NN	O	O
conserved	NN	O	O
,	NN	O	O
76-amino-acid	NN	O	B-protein
protein	NN	O	I-protein
that	NN	O	O
contains	NN	O	O
two	NN	O	O
extended	NN	O	O
arrays	NN	O	O
of	NN	O	O
hydrophobic	NN	O	B-protein
repeats	NN	O	I-protein
that	NN	O	O
interact	NN	O	O
with	NN	O	O
the	NN	O	O
HSF1	NN	O	B-protein
heptad	NN	O	I-protein
repeats	NN	O	I-protein
.	NN	O	O

HSBP1	NN	O	B-protein
is	NN	O	O
nuclear-localized	NN	O	O
and	NN	O	O
interacts	NN	O	O
in	NN	O	O
vivo	NN	O	O
with	NN	O	O
the	NN	O	O
active	NN	O	O
trimeric	NN	O	O
state	NN	O	O
of	NN	O	O
HSF1	NN	O	B-protein
that	NN	O	O
appears	NN	O	O
during	NN	O	O
heat	NN	O	O
shock	NN	O	O
.	NN	O	O

During	NN	O	O
attenuation	NN	O	O
of	NN	O	O
HSF1	NN	O	B-protein
to	NN	O	O
the	NN	O	O
inert	NN	O	O
monomer	NN	O	O
,	NN	O	O
HSBP1	NN	O	B-protein
associates	NN	O	O
with	NN	O	O
Hsp70	NN	O	O
.	NN	O	O

HSBP1	NN	O	B-protein
negatively	NN	O	O
affects	NN	O	O
HSF1	NN	O	B-protein
DNA-binding	NN	O	O
activity	NN	O	O
,	NN	O	O
and	NN	O	O
overexpression	NN	O	O
of	NN	O	O
HSBP1	NN	O	B-protein
in	NN	O	O
mammalian	NN	O	B-cell_type
cells	NN	O	I-cell_type
represses	NN	O	O
the	NN	O	O
transactivation	NN	O	O
activity	NN	O	O
of	NN	O	O
HSF1	NN	O	B-protein
.	NN	O	O

To	NN	O	O
establish	NN	O	O
a	NN	O	O
biological	NN	O	O
role	NN	O	O
for	NN	O	O
HSBP1	NN	O	B-protein
,	NN	O	O
the	NN	O	O
homologous	NN	O	O
Caenorhabditis	NN	O	B-protein
elegans	NN	O	I-protein
protein	NN	O	I-protein
was	NN	O	O
overexpressed	NN	O	O
in	NN	O	O
body	NN	O	B-cell_type
wall	NN	O	I-cell_type
muscle	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
was	NN	O	O
shown	NN	O	O
to	NN	O	O
block	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
heat	NN	O	O
shock	NN	O	O
response	NN	O	O
from	NN	O	O
a	NN	O	O
heat	NN	O	B-DNA
shock	NN	O	I-DNA
promoter-reporter	NN	O	I-DNA
construct	NN	O	I-DNA
.	NN	O	O

Alteration	NN	O	O
in	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
HSBP1	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
C.	NN	O	O
elegans	NN	O	O
has	NN	O	O
severe	NN	O	O
effects	NN	O	O
on	NN	O	O
survival	NN	O	O
of	NN	O	O
the	NN	O	O
animals	NN	O	O
after	NN	O	O
thermal	NN	O	O
and	NN	O	O
chemical	NN	O	O
stress	NN	O	O
,	NN	O	O
consistent	NN	O	O
with	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
HSBP1	NN	O	B-protein
as	NN	O	O
a	NN	O	O
negative	NN	O	O
regulator	NN	O	O
of	NN	O	O
the	NN	O	O
heat	NN	O	O
shock	NN	O	O
response	NN	O	O
.	NN	O	O

-DOCSTART-	O

Long-range	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
cytokine	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

Most	NN	O	O
studies	NN	O	O
on	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
cytokine	NN	O	O
gene	NN	O	O
expression	NN	O	O
have	NN	O	O
involved	NN	O	O
the	NN	O	O
functional	NN	O	O
analysis	NN	O	O
of	NN	O	O
proximal	NN	O	B-DNA
promoters	NN	O	I-DNA
.	NN	O	O

Recent	NN	O	O
work	NN	O	O
has	NN	O	O
identified	NN	O	O
distal	NN	O	B-DNA
elements	NN	O	I-DNA
that	NN	O	O
mediate	NN	O	O
long-range	NN	O	O
cytokine	NN	O	O
gene	NN	O	O
regulation	NN	O	O
and	NN	O	O
has	NN	O	O
implicated	NN	O	O
chromatin	NN	O	O
reorganization	NN	O	O
in	NN	O	O
regulation	NN	O	O
of	NN	O	O
cytokine	NN	O	B-DNA
gene	NN	O	I-DNA
loci	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
studies	NN	O	O
have	NN	O	O
begun	NN	O	O
to	NN	O	O
elucidate	NN	O	O
the	NN	O	O
basis	NN	O	O
for	NN	O	O
cell-specificity	NN	O	O
and	NN	O	O
high-level	NN	O	O
expression	NN	O	O
of	NN	O	O
cytokine	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
signaling	NN	O	O
complex	NN	O	O
of	NN	O	O
Ca2+-calmodulin-dependent	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
IV	NN	O	I-protein
and	NN	O	O
protein	NN	O	B-protein
phosphatase	NN	O	I-protein
2A	NN	O	I-protein
[	NN	O	O
see	NN	O	O
comments	NN	O	O
]	NN	O	O

Stimulation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
results	NN	O	O
in	NN	O	O
a	NN	O	O
rapid	NN	O	O
increase	NN	O	O
in	NN	O	O
intracellular	NN	O	O
calcium	NN	O	O
concentration	NN	O	O
(	NN	O	O
[	NN	O	O
Ca2+	NN	O	O
]	NN	O	O
i	NN	O	O
)	NN	O	O
that	NN	O	O
parallels	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
Ca2+-calmodulin-dependent	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
IV	NN	O	I-protein
(	NN	O	O
CaMKIV	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
nuclear	NN	O	B-protein
enzyme	NN	O	I-protein
that	NN	O	O
can	NN	O	O
phosphorylate	NN	O	O
and	NN	O	O
activate	NN	O	O
the	NN	O	O
cyclic	NN	O	B-protein
adenosine	NN	O	I-protein
monophosphate	NN	O	I-protein
(	NN	O	I-protein
cAMP	NN	O	I-protein
)	NN	O	I-protein
response	NN	O	I-protein
element-binding	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
CREB	NN	O	B-protein
)	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
inactivation	NN	O	O
of	NN	O	O
CaMKIV	NN	O	B-protein
occurs	NN	O	O
despite	NN	O	O
the	NN	O	O
sustained	NN	O	O
increase	NN	O	O
in	NN	O	O
[	NN	O	O
Ca2+	NN	O	O
]	NN	O	O
i	NN	O	O
that	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
.	NN	O	O

A	NN	O	O
stable	NN	O	O
and	NN	O	O
stoichiometric	NN	O	O
complex	NN	O	O
of	NN	O	O
CaMKIV	NN	O	B-protein
with	NN	O	O
protein	NN	O	B-protein
serine-threonine	NN	O	I-protein
phosphatase	NN	O	I-protein
2A	NN	O	I-protein
(	NN	O	O
PP2A	NN	O	B-protein
)	NN	O	O
was	NN	O	O
identified	NN	O	O
in	NN	O	O
which	NN	O	O
PP2A	NN	O	B-protein
dephosphorylates	NN	O	O
CaMKIV	NN	O	B-protein
and	NN	O	O
functions	NN	O	O
as	NN	O	O
a	NN	O	O
negative	NN	O	O
regulator	NN	O	O
of	NN	O	O
CaMKIV	NN	O	B-protein
signaling	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
inhibition	NN	O	O
of	NN	O	O
PP2A	NN	O	B-protein
activity	NN	O	O
by	NN	O	O
small	NN	O	O
t	NN	O	O
antigen	NN	O	O
enhanced	NN	O	O
activation	NN	O	O
of	NN	O	O
CREB	NN	O	B-protein
-mediated	NN	O	O
transcription	NN	O	O
by	NN	O	O
CaMKIV	NN	O	B-protein
.	NN	O	O

These	NN	O	O
findings	NN	O	O
reveal	NN	O	O
an	NN	O	O
intracellular	NN	O	O
signaling	NN	O	O
mechanism	NN	O	O
whereby	NN	O	O
a	NN	O	O
protein	NN	O	B-protein
serine-threonine	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	O
CaMKIV	NN	O	B-protein
)	NN	O	O
is	NN	O	O
regulated	NN	O	O
by	NN	O	O
a	NN	O	O
tightly	NN	O	O
associated	NN	O	O
protein	NN	O	B-protein
serine-threonine	NN	O	I-protein
phosphatase	NN	O	I-protein
(	NN	O	O
PP2A	NN	O	B-protein
)	NN	O	O
.	NN	O	O

-DOCSTART-	O

Hypoxia	NN	O	O
down-regulates	NN	O	O
MCP-1	NN	O	B-protein
expression	NN	O	O
:	NN	O	O
implications	NN	O	O
for	NN	O	O
macrophage	NN	O	O
distribution	NN	O	O
in	NN	O	O
tumors	NN	O	O
.	NN	O	O

Monocyte	NN	O	B-protein
chemoattractant	NN	O	I-protein
protein	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	O
MCP-1	NN	O	B-protein
)	NN	O	O
is	NN	O	O
likely	NN	O	O
to	NN	O	O
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
macrophage	NN	O	B-cell_type
infiltrate	NN	O	I-cell_type
in	NN	O	O
human	NN	O	O
ovarian	NN	O	O
carcinomas	NN	O	O
.	NN	O	O

Although	NN	O	O
MCP-1	NN	O	B-protein
is	NN	O	O
predominantly	NN	O	O
expressed	NN	O	O
by	NN	O	O
the	NN	O	O
tumor	NN	O	O
parenchyma	NN	O	O
,	NN	O	O
macrophages	NN	O	O
accumulate	NN	O	O
at	NN	O	O
highest	NN	O	O
density	NN	O	O
in	NN	O	O
necrotic	NN	O	O
regions	NN	O	O
,	NN	O	O
which	NN	O	O
are	NN	O	O
associated	NN	O	O
with	NN	O	O
low	NN	O	O
oxygen	NN	O	O
tensions	NN	O	O
.	NN	O	O

Tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
can	NN	O	O
stimulate	NN	O	O
MCP-1	NN	O	B-protein
production	NN	O	O
and	NN	O	O
is	NN	O	O
also	NN	O	O
present	NN	O	O
within	NN	O	O
ovarian	NN	O	O
carcinomas	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
investigated	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
hypoxia	NN	O	O
both	NN	O	O
on	NN	O	O
MCP-1	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
ovarian	NN	O	B-cell_line
cancer	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
and	NN	O	O
monocyte	NN	O	O
migration	NN	O	O
.	NN	O	O

Hypoxia	NN	O	O
down-regulated	NN	O	O
TNF-alpha-induced	NN	O	B-RNA
MCP-1	NN	O	I-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
protein	NN	O	O
production	NN	O	O
by	NN	O	O
ovarian	NN	O	B-cell_type
cancer	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
effect	NN	O	O
was	NN	O	O
mimicked	NN	O	O
by	NN	O	O
cobalt	NN	O	O
chloride	NN	O	O
and	NN	O	O
desferrioxamine	NN	O	O
,	NN	O	O
consistent	NN	O	O
with	NN	O	O
a	NN	O	O
specific	NN	O	O
oxygen-sensing	NN	O	O
mechanism	NN	O	O
.	NN	O	O

Unlike	NN	O	O
antioxidants	NN	O	O
,	NN	O	O
hypoxia	NN	O	O
did	NN	O	O
not	NN	O	O
inhibit	NN	O	O
nuclear	NN	O	O
factor	NN	O	O
KB	NN	O	O
mobilization	NN	O	O
.	NN	O	O

Monocyte	NN	O	O
migration	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
MCP-1	NN	O	B-protein
was	NN	O	O
also	NN	O	O
diminished	NN	O	O
under	NN	O	O
hypoxic	NN	O	O
conditions	NN	O	O
.	NN	O	O

Down-regulation	NN	O	O
of	NN	O	O
MCP-1	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
the	NN	O	O
inhibition	NN	O	O
of	NN	O	O
monocyte	NN	O	O
migration	NN	O	O
are	NN	O	O
independent	NN	O	O
effects	NN	O	O
of	NN	O	O
hypoxia	NN	O	O
that	NN	O	O
may	NN	O	O
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
distribution	NN	O	O
of	NN	O	O
macrophages	NN	O	B-cell_type
within	NN	O	O
ovarian	NN	O	O
tumors	NN	O	O
.	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
inflammatory	NN	O	O
bowel	NN	O	O
disease	NN	O	O
[	NN	O	O
see	NN	O	O
comments	NN	O	O
]	NN	O	O

BACKGROUND	NN	O	O
:	NN	O	O
Expression	NN	O	O
of	NN	O	O
pro-inflammatory	NN	O	B-protein
cytokines	NN	O	I-protein
is	NN	O	O
increased	NN	O	O
in	NN	O	O
the	NN	O	O
intestinal	NN	O	O
lamina	NN	O	O
propria	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
inflammatory	NN	O	O
bowel	NN	O	O
disease	NN	O	O
(	NN	O	O
IBD	NN	O	O
)	NN	O	O
.	NN	O	O

Nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
)	NN	O	O
controls	NN	O	O
transcription	NN	O	O
of	NN	O	O
inflammation	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

On	NN	O	O
activation	NN	O	O
,	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
is	NN	O	O
rapidly	NN	O	O
released	NN	O	O
from	NN	O	O
its	NN	O	O
cytoplasmic	NN	O	B-protein
inhibitor	NN	O	I-protein
(	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
)	NN	O	O
,	NN	O	O
transmigrates	NN	O	O
into	NN	O	O
the	NN	O	O
nucleus	NN	O	O
,	NN	O	O
and	NN	O	O
binds	NN	O	O
to	NN	O	O
DNA	NN	O	B-DNA
response	NN	O	I-DNA
elements	NN	O	I-DNA
in	NN	O	O
gene	NN	O	O
promoter	NN	O	O
regions	NN	O	O
.	NN	O	O

AIMS	NN	O	O
:	NN	O	O
To	NN	O	O
investigate	NN	O	O
whether	NN	O	O
increased	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
is	NN	O	O
important	NN	O	O
in	NN	O	O
IBD	NN	O	O
and	NN	O	O
may	NN	O	O
be	NN	O	O
down-regulated	NN	O	O
by	NN	O	O
anti-inflammatory	NN	O	O
treatment	NN	O	O
.	NN	O	O

METHODS	NN	O	O
:	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
was	NN	O	O
determined	NN	O	O
by	NN	O	O
western	NN	O	O
blot	NN	O	O
assessment	NN	O	O
and	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
in	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
of	NN	O	O
colonic	NN	O	O
biopsy	NN	O	O
samples	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
lamina	NN	O	B-cell_type
propria	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
Nuclear	NN	O	O
levels	NN	O	O
of	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
p65	NN	O	B-protein
are	NN	O	O
increased	NN	O	O
in	NN	O	O
lamina	NN	O	O
propria	NN	O	O
biopsy	NN	O	O
specimens	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
Crohn	NN	O	O
's	NN	O	O
disease	NN	O	O
in	NN	O	O
comparison	NN	O	O
with	NN	O	O
patients	NN	O	O
with	NN	O	O
ulcerative	NN	O	O
colitis	NN	O	O
and	NN	O	O
controls	NN	O	O
.	NN	O	O

Increased	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
was	NN	O	O
detected	NN	O	O
in	NN	O	O
lamina	NN	O	B-cell_type
propria	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
patients	NN	O	O
with	NN	O	O
active	NN	O	O
IBD	NN	O	O
.	NN	O	O

Corticosteroids	NN	O	O
strongly	NN	O	O
inhibit	NN	O	O
intestinal	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
activation	NN	O	O
in	NN	O	O
IBD	NN	O	O
in	NN	O	O
vivo	NN	O	O
and	NN	O	O
in	NN	O	O
vitro	NN	O	O
by	NN	O	O
stabilising	NN	O	O
the	NN	O	O
cytosolic	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
against	NN	O	O
activation	NN	O	O
induced	NN	O	O
degradation	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
:	NN	O	O
In	NN	O	O
both	NN	O	O
IBDs	NN	O	O
,	NN	O	O
but	NN	O	O
particularly	NN	O	O
Crohn	NN	O	O
's	NN	O	O
disease	NN	O	O
,	NN	O	O
increased	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
may	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
inflammatory	NN	O	O
response	NN	O	O
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
activation	NN	O	O
may	NN	O	O
represent	NN	O	O
a	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
steroids	NN	O	O
exert	NN	O	O
an	NN	O	O
anti-inflammatory	NN	O	O
effect	NN	O	O
in	NN	O	O
IBD	NN	O	O

-DOCSTART-	O

Insufficient	NN	O	O
glycemic	NN	O	O
control	NN	O	O
increases	NN	O	O
nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
binding	NN	O	O
activity	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
isolated	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
type	NN	O	O
1	NN	O	O
diabetes	NN	O	O
.	NN	O	O

OBJECTIVE	NN	O	O
:	NN	O	O
The	NN	O	O
redox-sensitive	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
nuclear	NN	O	I-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
is	NN	O	O
believed	NN	O	O
to	NN	O	O
contribute	NN	O	O
to	NN	O	O
late	NN	O	O
diabetic	NN	O	O
complications	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
unknown	NN	O	O
whether	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
is	NN	O	O
influenced	NN	O	O
by	NN	O	O
glycemic	NN	O	O
control	NN	O	O
.	NN	O	O

RESEARCH	NN	O	O
DESIGN	NN	O	O
AND	NN	O	O
METHODS	NN	O	O
:	NN	O	O
To	NN	O	O
determine	NN	O	O
whether	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
is	NN	O	O
activated	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
insufficient	NN	O	O
glycemic	NN	O	O
control	NN	O	O
(	NN	O	O
HbA1c	NN	O	O
>	NN	O	O
10	NN	O	O
%	NN	O	O
)	NN	O	O
,	NN	O	O
we	NN	O	O
developed	NN	O	O
a	NN	O	O
tissue	NN	O	O
culture-independent	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
(	NN	O	O
EMSA	NN	O	O
)	NN	O	O
-based	NN	O	O
semiquantitative	NN	O	O
detection	NN	O	O
system	NN	O	O
that	NN	O	O
allowed	NN	O	O
us	NN	O	O
to	NN	O	O
determine	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
in	NN	O	O
ex	NN	O	O
vivo-isolated	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
PBMCs	NN	O	B-cell_type
)	NN	O	O
.	NN	O	O

We	NN	O	O
included	NN	O	O
43	NN	O	O
patients	NN	O	O
with	NN	O	O
type	NN	O	O
1	NN	O	O
diabetes	NN	O	O
in	NN	O	O
this	NN	O	O
cross-sectional	NN	O	O
study	NN	O	O
.	NN	O	O

10	NN	O	O
of	NN	O	O
those	NN	O	O
received	NN	O	O
the	NN	O	O
antioxidant	NN	O	O
thioctic	NN	O	O
acid	NN	O	O
(	NN	O	O
600	NN	O	O
mg/day	NN	O	O
p.o.	NN	O	O
)	NN	O	O
for	NN	O	O
2	NN	O	O
weeks	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
Monocytes	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
HbA1c	NN	O	O
levels	NN	O	O
>	NN	O	O
10	NN	O	O
%	NN	O	O
demonstrated	NN	O	O
significantly	NN	O	O
higher	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
an	NN	O	O
EMSA	NN	O	O
and	NN	O	O
a	NN	O	O
stronger	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
staining	NN	O	O
in	NN	O	O
immunohistochemistry	NN	O	O
than	NN	O	O
monocytes	NN	O	B-cell_type
of	NN	O	O
patients	NN	O	O
with	NN	O	O
HbA1c	NN	O	O
levels	NN	O	O
of	NN	O	O
6-8	NN	O	O
%	NN	O	O
.	NN	O	O

The	NN	O	O
increase	NN	O	O
in	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
an	NN	O	O
increase	NN	O	O
in	NN	O	O
plasmatic	NN	O	O
markers	NN	O	O
of	NN	O	O
lipid	NN	O	O
peroxidation	NN	O	O
.	NN	O	O

Treatment	NN	O	O
with	NN	O	O
the	NN	O	O
antioxidant	NN	O	O
thioctic	NN	O	O
acid	NN	O	O
decreased	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
binding	NN	O	O
activity	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
:	NN	O	O
Hyperglycemia	NN	O	O
induces	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	O
factor	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
ex	NN	O	O
vivo-isolated	NN	O	O
PBMCs	NN	O	B-cell_type
of	NN	O	O
patients	NN	O	O
with	NN	O	O
type	NN	O	O
1	NN	O	O
diabetes	NN	O	O
.	NN	O	O

NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
is	NN	O	O
at	NN	O	O
least	NN	O	O
partially	NN	O	O
dependent	NN	O	O
on	NN	O	O
oxidative	NN	O	O
stress	NN	O	O
,	NN	O	O
since	NN	O	O
the	NN	O	O
antioxidant	NN	O	O
thioctic	NN	O	O
acid	NN	O	O
significantly	NN	O	O
lowered	NN	O	O
the	NN	O	O
extent	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
binding	NN	O	O
activity	NN	O	O
.	NN	O	O

-DOCSTART-	O

Synergistic	NN	O	O
activation	NN	O	O
of	NN	O	O
MAP	NN	O	B-protein
kinase	NN	O	I-protein
(	NN	O	O
ERK1/2	NN	O	B-protein
)	NN	O	O
by	NN	O	O
erythropoietin	NN	O	B-protein
and	NN	O	O
stem	NN	O	B-protein
cell	NN	O	I-protein
factor	NN	O	I-protein
is	NN	O	O
essential	NN	O	O
for	NN	O	O
expanded	NN	O	O
erythropoiesis	NN	O	O
.	NN	O	O

Stem	NN	O	B-protein
cell	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
SCF	NN	O	B-protein
)	NN	O	O
and	NN	O	O
erythropoietin	NN	O	B-protein
(	NN	O	O
EPO	NN	O	B-protein
)	NN	O	O
work	NN	O	O
synergistically	NN	O	O
to	NN	O	O
support	NN	O	O
erythropoiesis	NN	O	O
,	NN	O	O
but	NN	O	O
the	NN	O	O
mechanism	NN	O	O
for	NN	O	O
this	NN	O	O
synergism	NN	O	O
is	NN	O	O
unknown	NN	O	O
.	NN	O	O

By	NN	O	O
using	NN	O	O
purified	NN	O	B-cell_line
human	NN	O	I-cell_line
erythroid	NN	O	I-cell_line
colony-forming	NN	O	I-cell_line
cells	NN	O	I-cell_line
(	NN	O	O
ECFC	NN	O	B-cell_line
)	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
found	NN	O	O
that	NN	O	O
SCF	NN	O	B-protein
and	NN	O	O
EPO	NN	O	B-protein
synergistically	NN	O	O
activate	NN	O	O
MAP	NN	O	B-protein
kinase	NN	O	I-protein
(	NN	O	O
MAPK	NN	O	B-protein
,	NN	O	O
ERK1/2	NN	O	B-protein
)	NN	O	O
,	NN	O	O
which	NN	O	O
correlates	NN	O	O
with	NN	O	O
the	NN	O	O
cell	NN	O	O
growth	NN	O	O
and	NN	O	O
thus	NN	O	O
may	NN	O	O
be	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
synergistic	NN	O	O
effects	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
PD98059	NN	O	O
and	NN	O	O
wortmannin	NN	O	O
,	NN	O	O
inhibitors	NN	O	O
of	NN	O	O
MEK	NN	O	B-protein
and	NN	O	O
PI-3	NN	O	B-protein
kinase	NN	O	I-protein
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
inhibited	NN	O	O
the	NN	O	O
synergistic	NN	O	O
activation	NN	O	O
of	NN	O	O
MAPK	NN	O	B-protein
and	NN	O	O
also	NN	O	O
the	NN	O	O
cell	NN	O	O
growth	NN	O	O
,	NN	O	O
further	NN	O	O
supporting	NN	O	O
this	NN	O	O
conclusion	NN	O	O
.	NN	O	O

Wortmannin	NN	O	O
only	NN	O	O
inhibits	NN	O	O
MAPK	NN	O	B-protein
activation	NN	O	O
induced	NN	O	O
by	NN	O	O
EPO	NN	O	B-protein
but	NN	O	O
not	NN	O	O
that	NN	O	O
by	NN	O	O
SCF	NN	O	B-protein
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
SCF	NN	O	B-protein
and	NN	O	O
EPO	NN	O	B-protein
may	NN	O	O
activate	NN	O	O
MAPK	NN	O	B-protein
through	NN	O	O
different	NN	O	O
pathways	NN	O	O
,	NN	O	O
which	NN	O	O
would	NN	O	O
facilitate	NN	O	O
synergy	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
EPO	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
SCF	NN	O	B-protein
,	NN	O	O
led	NN	O	O
to	NN	O	O
activation	NN	O	O
of	NN	O	O
STAT5	NN	O	B-protein
,	NN	O	O
whereas	NN	O	O
SCF	NN	O	B-protein
and	NN	O	O
wortmannin	NN	O	O
had	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
EPO	NN	O	B-protein
-induced	NN	O	O
STAT5	NN	O	B-protein
activation	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
STAT5	NN	O	B-protein
is	NN	O	O
not	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
synergistic	NN	O	O
action	NN	O	O
of	NN	O	O
SCF	NN	O	B-protein
and	NN	O	O
EPO	NN	O	B-protein
.	NN	O	O

Together	NN	O	O
,	NN	O	O
the	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
synergistic	NN	O	O
activation	NN	O	O
of	NN	O	O
MAPK	NN	O	B-protein
by	NN	O	O
SCF	NN	O	B-protein
and	NN	O	O
EPO	NN	O	B-protein
is	NN	O	O
essential	NN	O	O
for	NN	O	O
expanded	NN	O	O
erythropoiesis	NN	O	O
.	NN	O	O

Copyright	NN	O	O
1998	NN	O	O
by	NN	O	O
The	NN	O	O
American	NN	O	O
Society	NN	O	O
of	NN	O	O
Hematology	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
molecular	NN	O	O
and	NN	O	O
phenotypic	NN	O	O
profile	NN	O	O
of	NN	O	O
primary	NN	O	O
central	NN	O	O
nervous	NN	O	O
system	NN	O	O
lymphoma	NN	O	O
identifies	NN	O	O
distinct	NN	O	O
categories	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
and	NN	O	O
is	NN	O	O
consistent	NN	O	O
with	NN	O	O
histogenetic	NN	O	O
derivation	NN	O	O
from	NN	O	O
germinal	NN	O	B-cell_type
center-related	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Primary	NN	O	O
central	NN	O	O
nervous	NN	O	O
system	NN	O	O
lymphoma	NN	O	O
(	NN	O	O
PCNSL	NN	O	O
)	NN	O	O
is	NN	O	O
a	NN	O	O
major	NN	O	O
cause	NN	O	O
of	NN	O	O
morbidity	NN	O	O
and	NN	O	O
mortality	NN	O	O
among	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
(	NN	O	O
HIV	NN	O	O
)	NN	O	O
-infected	NN	O	O
individuals	NN	O	O
.	NN	O	O

The	NN	O	O
precise	NN	O	O
histogenetic	NN	O	O
derivation	NN	O	O
and	NN	O	O
the	NN	O	O
molecular	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
PCNSL	NN	O	O
is	NN	O	O
poorly	NN	O	O
understood	NN	O	O
.	NN	O	O

In	NN	O	O
an	NN	O	O
attempt	NN	O	O
to	NN	O	O
clarify	NN	O	O
the	NN	O	O
histogenesis	NN	O	O
and	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
these	NN	O	O
lymphomas	NN	O	O
,	NN	O	O
49	NN	O	O
PCNSL	NN	O	O
(	NN	O	O
26	NN	O	O
acquired	NN	O	O
immunodeficiency	NN	O	O
syndrome	NN	O	O
[	NN	O	O
AIDS	NN	O	O
]	NN	O	O
-related	NN	O	O
and	NN	O	O
23	NN	O	O
AIDS-unrelated	NN	O	O
)	NN	O	O
were	NN	O	O
analyzed	NN	O	O
for	NN	O	O
multiple	NN	O	O
biologic	NN	O	O
markers	NN	O	O
,	NN	O	O
which	NN	O	O
are	NN	O	O
known	NN	O	O
to	NN	O	O
bear	NN	O	O
histogenetic	NN	O	O
and	NN	O	O
pathogenetic	NN	O	O
significance	NN	O	O
for	NN	O	O
mature	NN	O	O
B-cell	NN	O	O
neoplasms	NN	O	O
.	NN	O	O

PCNSL	NN	O	O
associated	NN	O	O
frequently	NN	O	O
(	NN	O	O
50.0	NN	O	O
%	NN	O	O
)	NN	O	O
with	NN	O	O
mutations	NN	O	O
of	NN	O	O
BCL-6	NN	O	B-DNA
5	NN	O	I-DNA
'	NN	O	I-DNA
noncoding	NN	O	I-DNA
regions	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
are	NN	O	O
regarded	NN	O	O
as	NN	O	O
a	NN	O	O
marker	NN	O	O
of	NN	O	O
B-cell	NN	O	O
transition	NN	O	O
through	NN	O	O
the	NN	O	O
germinal	NN	O	O
center	NN	O	O
(	NN	O	O
GC	NN	O	O
)	NN	O	O
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
BCL-6	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
restricted	NN	O	O
to	NN	O	O
GC	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
throughout	NN	O	O
physiologic	NN	O	O
B-cell	NN	O	O
maturation	NN	O	O
,	NN	O	O
was	NN	O	O
detected	NN	O	O
in	NN	O	O
100	NN	O	O
%	NN	O	O
AIDS-unrelated	NN	O	O
PCNSL	NN	O	O
and	NN	O	O
in	NN	O	O
56.2	NN	O	O
%	NN	O	O
AIDS-related	NN	O	O
cases	NN	O	O
.	NN	O	O

Notably	NN	O	O
,	NN	O	O
among	NN	O	O
AIDS-related	NN	O	O
PCNSL	NN	O	O
,	NN	O	O
expression	NN	O	O
of	NN	O	O
BCL-6	NN	O	B-protein
was	NN	O	O
mutually	NN	O	O
exclusive	NN	O	O
with	NN	O	O
expression	NN	O	O
of	NN	O	O
Epstein-Barr	NN	O	B-protein
virus	NN	O	I-protein
(	NN	O	I-protein
EBV	NN	O	I-protein
)	NN	O	I-protein
-encoded	NN	O	I-protein
latent	NN	O	I-protein
membrane	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	I-protein
LMP	NN	O	I-protein
)	NN	O	I-protein
-1	NN	O	I-protein
and	NN	O	O
,	NN	O	O
with	NN	O	O
few	NN	O	O
exceptions	NN	O	O
,	NN	O	O
also	NN	O	O
of	NN	O	O
BCL-2	NN	O	B-protein
.	NN	O	O

All	NN	O	O
but	NN	O	O
one	NN	O	O
PCNSL	NN	O	O
expressed	NN	O	O
hMSH2	NN	O	O
,	NN	O	O
which	NN	O	O
among	NN	O	O
mature	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
selectively	NN	O	O
stains	NN	O	O
GC	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
PCNSL	NN	O	O
may	NN	O	O
be	NN	O	O
frequently	NN	O	O
related	NN	O	O
to	NN	O	O
GC	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
may	NN	O	O
be	NN	O	O
segregated	NN	O	O
into	NN	O	O
two	NN	O	O
major	NN	O	O
biologic	NN	O	O
categories	NN	O	O
based	NN	O	O
on	NN	O	O
the	NN	O	O
expression	NN	O	O
pattern	NN	O	O
of	NN	O	O
BCL-6	NN	O	B-protein
,	NN	O	O
LMP-1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
BCL-2	NN	O	B-protein
.	NN	O	O

BCL-6	NN	O	B-protein
(	NN	O	O
+	NN	O	O
)	NN	O	O
/	NN	O	O
LMP-1	NN	O	B-protein
(	NN	O	O
-	NN	O	O
)	NN	O	O
/	NN	O	O
BCL-2	NN	O	B-protein
(	NN	O	O
-	NN	O	O
)	NN	O	O
PCNSL	NN	O	O
occur	NN	O	O
both	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
and	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
HIV	NN	O	O
infection	NN	O	O
and	NN	O	O
consistently	NN	O	O
display	NN	O	O
a	NN	O	O
large	NN	O	O
noncleaved	NN	O	O
cell	NN	O	O
morphology	NN	O	O
.	NN	O	O

Conversely	NN	O	O
,	NN	O	O
BCL-6	NN	O	B-protein
(	NN	O	O
-	NN	O	O
)	NN	O	O
/	NN	O	O
LMP-1	NN	O	B-protein
(	NN	O	O
+	NN	O	O
)	NN	O	O
/	NN	O	O
BCL-2	NN	O	B-protein
(	NN	O	O
+	NN	O	O
)	NN	O	O
PCNSL	NN	O	O
are	NN	O	O
restricted	NN	O	O
to	NN	O	O
HIV-infected	NN	O	O
hosts	NN	O	O
and	NN	O	O
are	NN	O	O
represented	NN	O	O
by	NN	O	O
lymphomas	NN	O	O
with	NN	O	O
immunoblastic	NN	O	O
features	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
are	NN	O	O
relevant	NN	O	O
for	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
and	NN	O	O
histogenesis	NN	O	O
of	NN	O	O
PCNSL	NN	O	O
and	NN	O	O
may	NN	O	O
be	NN	O	O
helpful	NN	O	O
to	NN	O	O
segregate	NN	O	O
distinct	NN	O	O
biologic	NN	O	O
and	NN	O	O
prognostic	NN	O	O
categories	NN	O	O
of	NN	O	O
these	NN	O	O
lymphomas	NN	O	O
.	NN	O	O

Copyright	NN	O	O
1998	NN	O	O
by	NN	O	O
The	NN	O	O
American	NN	O	O
Society	NN	O	O
of	NN	O	O
Hematology	NN	O	O
.	NN	O	O

-DOCSTART-	O

Antioxidant	NN	O	O
regulation	NN	O	O
of	NN	O	O
phorbol	NN	O	O
ester-induced	NN	O	O
adhesion	NN	O	O
of	NN	O	O
human	NN	O	B-cell_line
Jurkat	NN	O	I-cell_line
T-cells	NN	O	I-cell_line
to	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Regulation	NN	O	O
of	NN	O	O
adhesion	NN	O	O
molecule	NN	O	O
expression	NN	O	O
and	NN	O	O
function	NN	O	O
by	NN	O	O
reactive	NN	O	O
oxygen	NN	O	O
species	NN	O	O
via	NN	O	O
specific	NN	O	O
redox	NN	O	O
sensitive	NN	O	O
mechanisms	NN	O	O
have	NN	O	O
been	NN	O	O
reported	NN	O	O
.	NN	O	O

The	NN	O	O
effects	NN	O	O
of	NN	O	O
clinically	NN	O	O
safe	NN	O	O
antioxidants	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
adhesion	NN	O	O
molecule	NN	O	O
expression	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
endothelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
ECV	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
adherence	NN	O	O
of	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
to	NN	O	O
ECV	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
investigated	NN	O	O
.	NN	O	O

The	NN	O	O
thiol	NN	O	O
antioxidant	NN	O	O
,	NN	O	O
alpha-lipoate	NN	O	O
,	NN	O	O
at	NN	O	O
clinically	NN	O	O
relevant	NN	O	O
doses	NN	O	O
down-regulated	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
-induced	NN	O	O
adhesion	NN	O	O
molecule	NN	O	O
expression	NN	O	O
and	NN	O	O
cell-cell	NN	O	O
adhesion	NN	O	O
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
PMA-induced	NN	O	O
ICAM-1	NN	O	O
and	NN	O	O
VCAM-1	NN	O	O
expression	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
PMA-induced	NN	O	O
adhesion	NN	O	O
of	NN	O	O
Jurkat	NN	O	B-cell_line
T-cells	NN	O	I-cell_line
to	NN	O	O
ECV	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
alpha-lipoate	NN	O	O
was	NN	O	O
dose	NN	O	O
dependent	NN	O	O
(	NN	O	O
50-250	NN	O	O
microM	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
effect	NN	O	O
was	NN	O	O
significant	NN	O	O
for	NN	O	O
ICAM-1	NN	O	B-protein
(	NN	O	O
p	NN	O	O
<	NN	O	O
.01	NN	O	O
)	NN	O	O
and	NN	O	O
VCAM-1	NN	O	B-protein
(	NN	O	O
p	NN	O	O
<	NN	O	O
.01	NN	O	O
)	NN	O	O
expression	NN	O	O
in	NN	O	O
cells	NN	O	O
pretreated	NN	O	O
with	NN	O	O
100	NN	O	O
microM	NN	O	O
alpha-lipoate	NN	O	O
compared	NN	O	O
to	NN	O	O
PMA-activated	NN	O	B-cell_type
untreated	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
PMA-induced	NN	O	O
adhesion	NN	O	O
molecule	NN	O	O
expression	NN	O	O
and	NN	O	O
cell-cell	NN	O	O
adhesion	NN	O	O
was	NN	O	O
more	NN	O	O
pronounced	NN	O	O
when	NN	O	O
a	NN	O	O
combination	NN	O	O
of	NN	O	O
antioxidants	NN	O	O
,	NN	O	O
alpha-lipoate	NN	O	O
and	NN	O	O
alpha-tocopherol	NN	O	O
,	NN	O	O
were	NN	O	O
used	NN	O	O
compared	NN	O	O
to	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
either	NN	O	O
of	NN	O	O
these	NN	O	O
antioxidant	NN	O	O
alone	NN	O	O
.	NN	O	O

The	NN	O	O
regulation	NN	O	O
of	NN	O	O
adhesion	NN	O	O
molecule	NN	O	O
expression	NN	O	O
and	NN	O	O
function	NN	O	O
by	NN	O	O
low	NN	O	O
concentration	NN	O	O
of	NN	O	O
antioxidants	NN	O	O
investigated	NN	O	O
does	NN	O	O
not	NN	O	O
appear	NN	O	O
to	NN	O	O
be	NN	O	O
NF-kappaB	NN	O	B-protein
regulated	NN	O	O
or	NN	O	O
transcription	NN	O	O
dependent	NN	O	O
because	NN	O	O
no	NN	O	O
change	NN	O	O
in	NN	O	O
the	NN	O	O
mRNA	NN	O	O
response	NN	O	O
was	NN	O	O
observed	NN	O	O
.	NN	O	O

Protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
(	NN	O	O
PKC	NN	O	B-protein
)	NN	O	O
has	NN	O	O
been	NN	O	O
suggested	NN	O	O
to	NN	O	O
regulate	NN	O	O
PMA-induced	NN	O	O
adhesion	NN	O	O
molecule	NN	O	O
expression	NN	O	O
by	NN	O	O
post-transcriptional	NN	O	O
stabilization	NN	O	O
of	NN	O	O
adhesion	NN	O	B-RNA
molecule	NN	O	I-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

Alpha-lipoate	NN	O	O
pretreatment	NN	O	O
did	NN	O	O
not	NN	O	O
influence	NN	O	O
the	NN	O	O
response	NN	O	O
of	NN	O	O
PKC	NN	O	B-protein
activity	NN	O	O
to	NN	O	O
PMA	NN	O	O
.	NN	O	O

Oxidants	NN	O	O
are	NN	O	O
known	NN	O	O
to	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
cell	NN	O	O
adhesion	NN	O	O
processes	NN	O	O
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
ECV	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
PMA	NN	O	O
induced	NN	O	O
generation	NN	O	O
of	NN	O	O
intracellular	NN	O	O
oxidants	NN	O	O
.	NN	O	O

Alpha-lipoate	NN	O	O
(	NN	O	O
100	NN	O	O
or	NN	O	O
250	NN	O	O
microM	NN	O	O
)	NN	O	O
treatment	NN	O	O
decreased	NN	O	O
PMA-induced	NN	O	O
generation	NN	O	O
of	NN	O	O
intracellular	NN	O	O
oxidants	NN	O	O
.	NN	O	O

The	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
of	NN	O	O
low	NN	O	O
concentration	NN	O	O
of	NN	O	O
alpha-lipaote	NN	O	O
alone	NN	O	O
or	NN	O	O
in	NN	O	O
combination	NN	O	O
with	NN	O	O
alpha-tocopherol	NN	O	O
on	NN	O	O
agonist-induced	NN	O	O
adhesion	NN	O	O
processes	NN	O	O
observed	NN	O	O
in	NN	O	O
this	NN	O	O
study	NN	O	O
may	NN	O	O
be	NN	O	O
of	NN	O	O
potential	NN	O	O
therapeutic	NN	O	O
value	NN	O	O
.	NN	O	O

-DOCSTART-	O

Binding	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
to	NN	O	O
CD4	NN	O	B-protein
and	NN	O	I-protein
CXCR4	NN	O	I-protein
receptors	NN	O	I-protein
differentially	NN	O	O
regulates	NN	O	O
expression	NN	O	O
of	NN	O	O
inflammatory	NN	O	B-DNA
genes	NN	O	I-DNA
and	NN	O	O
activates	NN	O	O
the	NN	O	O
MEK	NN	O	B-protein
/ERK	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
shown	NN	O	O
that	NN	O	O
binding	NN	O	O
of	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
(	NN	O	O
HIV-1	NN	O	O
)	NN	O	O
virions	NN	O	O
to	NN	O	O
CD4	NN	O	B-protein
receptors	NN	O	I-protein
stimulates	NN	O	O
association	NN	O	O
of	NN	O	O
Lck	NN	O	B-protein
with	NN	O	O
Raf-1	NN	O	B-protein
and	NN	O	O
results	NN	O	O
in	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
Raf-1	NN	O	B-protein
kinase	NN	O	O
in	NN	O	O
a	NN	O	O
Ras-independent	NN	O	O
manner	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
HIV-1	NN	O	B-protein
envelope	NN	O	I-protein
glycoproteins	NN	O	I-protein
of	NN	O	O
both	NN	O	O
T-cell-tropic	NN	O	O
and	NN	O	O
macrophagetropic	NN	O	O
strains	NN	O	O
rapidly	NN	O	O
activate	NN	O	O
the	NN	O	O
ERK/mitogen-activated	NN	O	O
protein	NN	O	O
(	NN	O	O
MAP	NN	O	O
)	NN	O	O
kinase	NN	O	O
pathway	NN	O	O
and	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
(	NN	O	O
AP-1	NN	O	B-protein
,	NN	O	O
NF-kappaB	NN	O	B-protein
,	NN	O	O
and	NN	O	O
C/EBP	NN	O	B-protein
)	NN	O	O
and	NN	O	O
stimulate	NN	O	O
expression	NN	O	O
of	NN	O	O
cytokine	NN	O	B-DNA
and	NN	O	I-DNA
chemokine	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
activation	NN	O	O
of	NN	O	O
this	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
requires	NN	O	O
functional	NN	O	O
CD4	NN	O	B-protein
receptors	NN	O	I-protein
and	NN	O	O
is	NN	O	O
independent	NN	O	O
of	NN	O	O
binding	NN	O	O
to	NN	O	O
CXCR4	NN	O	B-protein
.	NN	O	O

Binding	NN	O	O
of	NN	O	O
the	NN	O	O
natural	NN	O	O
ligand	NN	O	O
stromal	NN	O	B-protein
cell-derived	NN	O	I-protein
factor	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	O
SDF-1	NN	O	B-protein
)	NN	O	O
to	NN	O	O
CXCR4	NN	O	B-protein
,	NN	O	O
which	NN	O	O
inhibits	NN	O	O
entry	NN	O	O
of	NN	O	O
T-cell-tropic	NN	O	O
HIV-1	NN	O	O
,	NN	O	O
activates	NN	O	O
also	NN	O	O
the	NN	O	O
ERK/	NN	O	O
MAP	NN	O	B-protein
kinase	NN	O	I-protein
pathway	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
SDF-1	NN	O	B-protein
did	NN	O	O
not	NN	O	O
affect	NN	O	O
the	NN	O	O
CD4-mediated	NN	O	O
expression	NN	O	O
of	NN	O	O
cytokine	NN	O	B-DNA
and	NN	O	I-DNA
chemokine	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
results	NN	O	O
provide	NN	O	O
firm	NN	O	O
molecular	NN	O	O
evidence	NN	O	O
that	NN	O	O
binding	NN	O	O
of	NN	O	O
HIV-1	NN	O	B-protein
envelope	NN	O	I-protein
glycoproteins	NN	O	I-protein
to	NN	O	O
CD4	NN	O	B-protein
receptor	NN	O	I-protein
initiates	NN	O	O
a	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
independent	NN	O	O
of	NN	O	O
the	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
chemokine	NN	O	B-protein
receptor	NN	O	I-protein
that	NN	O	O
leads	NN	O	O
to	NN	O	O
the	NN	O	O
aberrant	NN	O	O
expression	NN	O	O
of	NN	O	O
inflammatory	NN	O	B-DNA
genes	NN	O	I-DNA
and	NN	O	O
may	NN	O	O
contribute	NN	O	O
significantly	NN	O	O
to	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
to	NN	O	O
deregulation	NN	O	O
of	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
.	NN	O	O

-DOCSTART-	O

Antigen	NN	O	O
receptor	NN	O	O
signaling	NN	O	O
induces	NN	O	O
MAP	NN	O	O
kinase-mediated	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
degradation	NN	O	O
of	NN	O	O
the	NN	O	O
BCL-6	NN	O	B-protein
transcription	NN	O	B-protein
factor	NN	O	I-protein
.	NN	O	O

The	NN	O	O
bcl-6	NN	O	B-DNA
proto-oncogene	NN	O	I-DNA
encodes	NN	O	O
a	NN	O	O
POZ/zinc	NN	O	B-protein
finger	NN	O	I-protein
transcriptional	NN	O	I-protein
repressor	NN	O	I-protein
expressed	NN	O	O
in	NN	O	O
germinal	NN	O	B-cell_type
center	NN	O	I-cell_type
(	NN	O	I-cell_type
GC	NN	O	I-cell_type
)	NN	O	I-cell_type
B	NN	O	I-cell_type
and	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
required	NN	O	O
for	NN	O	O
GC	NN	O	O
formation	NN	O	O
and	NN	O	O
antibody	NN	O	O
affinity	NN	O	O
maturation	NN	O	O
.	NN	O	O

Deregulation	NN	O	O
of	NN	O	O
bcl-6	NN	O	B-DNA
expression	NN	O	O
by	NN	O	O
chromosomal	NN	O	O
rearrangements	NN	O	O
and	NN	O	O
point	NN	O	O
mutations	NN	O	O
of	NN	O	O
the	NN	O	O
bcl-6	NN	O	B-DNA
promoter	NN	O	I-DNA
region	NN	O	I-DNA
are	NN	O	O
implicated	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
B-cell	NN	O	O
lymphoma	NN	O	O
.	NN	O	O

The	NN	O	O
signals	NN	O	O
regulating	NN	O	O
bcl-6	NN	O	B-DNA
expression	NN	O	O
are	NN	O	O
not	NN	O	O
known	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
antigen	NN	O	O
receptor	NN	O	O
activation	NN	O	O
leads	NN	O	O
to	NN	O	O
BCL-6	NN	O	B-protein
phosphorylation	NN	O	O
by	NN	O	O
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	O
MAPK	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Phosphorylation	NN	O	O
,	NN	O	O
in	NN	O	O
turn	NN	O	O
,	NN	O	O
targets	NN	O	O
BCL-6	NN	O	B-protein
for	NN	O	O
rapid	NN	O	O
degradation	NN	O	O
by	NN	O	O
the	NN	O	O
ubiquitin	NN	O	B-protein
/	NN	O	O
proteasome	NN	O	B-protein
pathway	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
indicate	NN	O	O
that	NN	O	O
BCL-6	NN	O	B-protein
expression	NN	O	O
is	NN	O	O
directly	NN	O	O
controlled	NN	O	O
by	NN	O	O
the	NN	O	O
antigen	NN	O	O
receptor	NN	O	O
via	NN	O	O
MAPK	NN	O	B-protein
activation	NN	O	O
.	NN	O	O

This	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
may	NN	O	O
be	NN	O	O
crucial	NN	O	O
for	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
B-cell	NN	O	O
differentiation	NN	O	O
and	NN	O	O
antibody	NN	O	O
response	NN	O	O
and	NN	O	O
has	NN	O	O
implications	NN	O	O
for	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
other	NN	O	O
POZ/zinc	NN	O	B-protein
finger	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
in	NN	O	O
other	NN	O	O
tissues	NN	O	O
.	NN	O	O

-DOCSTART-	O

Glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
are	NN	O	O
differentially	NN	O	O
expressed	NN	O	O
in	NN	O	O
the	NN	O	O
cells	NN	O	O
and	NN	O	O
tissues	NN	O	O
of	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
.	NN	O	O

Cytosolic	NN	O	B-protein
glucocorticoid	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
GR	NN	O	B-protein
)	NN	O	O
binding	NN	O	O
studies	NN	O	O
on	NN	O	O
immune	NN	O	O
tissues	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
thymus	NN	O	O
exhibits	NN	O	O
three	NN	O	O
to	NN	O	O
four	NN	O	O
times	NN	O	O
higher	NN	O	O
levels	NN	O	O
of	NN	O	O
GR	NN	O	B-protein
protein	NN	O	I-protein
than	NN	O	O
the	NN	O	O
spleen	NN	O	O
.	NN	O	O

High	NN	O	O
levels	NN	O	O
of	NN	O	O
GR	NN	O	B-protein
are	NN	O	O
consistent	NN	O	O
with	NN	O	O
the	NN	O	O
exquisite	NN	O	O
sensitivity	NN	O	O
of	NN	O	O
the	NN	O	O
thymus	NN	O	O
to	NN	O	O
glucocorticoid	NN	O	O
exposure	NN	O	O
.	NN	O	O

Nevertheless	NN	O	O
,	NN	O	O
whole	NN	O	O
cell	NN	O	O
binding	NN	O	O
studies	NN	O	O
reveal	NN	O	O
similar	NN	O	O
levels	NN	O	O
of	NN	O	O
GR	NN	O	B-protein
in	NN	O	O
immature	NN	O	B-cell_type
thymic	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
more	NN	O	O
mature	NN	O	O
,	NN	O	O
splenic	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
whole	NN	O	O
cell	NN	O	O
binding	NN	O	O
techniques	NN	O	O
indicate	NN	O	O
that	NN	O	O
neutrophils	NN	O	B-cell_type
(	NN	O	O
which	NN	O	O
represent	NN	O	O
roughly	NN	O	O
30	NN	O	O
%	NN	O	O
of	NN	O	O
splenic	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
)	NN	O	O
exhibit	NN	O	O
higher	NN	O	O
GR	NN	O	B-protein
than	NN	O	O
both	NN	O	O
T	NN	O	B-cell_type
and	NN	O	I-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
further	NN	O	O
contradicting	NN	O	O
results	NN	O	O
from	NN	O	O
cytosolic	NN	O	O
binding	NN	O	O
studies	NN	O	O
.	NN	O	O

To	NN	O	O
address	NN	O	O
these	NN	O	O
inconsistencies	NN	O	O
,	NN	O	O
GR	NN	O	B-protein
protein	NN	O	I-protein
was	NN	O	O
assessed	NN	O	O
in	NN	O	O
immune	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	I-cell_type
tissues	NN	O	I-cell_type
using	NN	O	O
cytosolic	NN	O	O
radioligand	NN	O	O
binding	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
immunocytochemistry	NN	O	O
.	NN	O	O

Consistent	NN	O	O
with	NN	O	O
previous	NN	O	O
cytosolic	NN	O	O
receptor	NN	O	O
binding	NN	O	O
studies	NN	O	O
on	NN	O	O
immune	NN	O	O
tissue	NN	O	O
homogenates	NN	O	O
,	NN	O	O
thymic	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
found	NN	O	O
to	NN	O	O
have	NN	O	O
higher	NN	O	O
levels	NN	O	O
of	NN	O	O
GR	NN	O	B-protein
than	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
isolated	NN	O	O
from	NN	O	O
the	NN	O	O
spleen	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
neutrophils	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
have	NN	O	O
fewer	NN	O	O
GR	NN	O	B-protein
than	NN	O	O
lymphocytes	NN	O	B-cell_type
and	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
a	NN	O	O
meaningful	NN	O	O
relationship	NN	O	O
between	NN	O	O
receptor	NN	O	O
expression	NN	O	O
and	NN	O	O
known	NN	O	O
sensitivity	NN	O	O
to	NN	O	O
glucocorticoids	NN	O	O
.	NN	O	O

-DOCSTART-	O

Functional	NN	O	O
replacement	NN	O	O
of	NN	O	O
the	NN	O	O
mouse	NN	O	B-DNA
E2A	NN	O	I-DNA
gene	NN	O	I-DNA
with	NN	O	O
a	NN	O	O
human	NN	O	B-DNA
HEB	NN	O	I-DNA
cDNA	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
mammalian	NN	O	B-DNA
E2A	NN	O	I-DNA
,	NN	O	I-DNA
HEB	NN	O	I-DNA
,	NN	O	I-DNA
and	NN	O	I-DNA
E2-2	NN	O	I-DNA
genes	NN	O	I-DNA
encode	NN	O	O
a	NN	O	O
unique	NN	O	O
class	NN	O	O
of	NN	O	O
basic	NN	O	B-protein
helix-loop-helix	NN	O	I-protein
(	NN	O	I-protein
bHLH	NN	O	I-protein
)	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
that	NN	O	O
are	NN	O	O
evolutionarily	NN	O	O
conserved	NN	O	O
and	NN	O	O
essential	NN	O	O
for	NN	O	O
embryonic	NN	O	O
and	NN	O	O
postnatal	NN	O	O
development	NN	O	O
.	NN	O	O

While	NN	O	O
the	NN	O	O
structural	NN	O	O
and	NN	O	O
functional	NN	O	O
similarities	NN	O	O
among	NN	O	O
the	NN	O	O
gene	NN	O	B-protein
products	NN	O	I-protein
are	NN	O	O
well	NN	O	O
demonstrated	NN	O	O
,	NN	O	O
it	NN	O	O
is	NN	O	O
not	NN	O	O
clear	NN	O	O
why	NN	O	O
deletion	NN	O	O
of	NN	O	O
E2A	NN	O	B-DNA
,	NN	O	O
but	NN	O	O
not	NN	O	O
HEB	NN	O	B-DNA
or	NN	O	O
E2-2	NN	O	B-DNA
,	NN	O	O
leads	NN	O	O
to	NN	O	O
a	NN	O	O
complete	NN	O	O
arrest	NN	O	O
in	NN	O	O
B-lymphocyte	NN	O	O
development	NN	O	O
.	NN	O	O

To	NN	O	O
understand	NN	O	O
the	NN	O	O
molecular	NN	O	O
basis	NN	O	O
of	NN	O	O
the	NN	O	O
functional	NN	O	O
specificity	NN	O	O
between	NN	O	O
E2A	NN	O	B-DNA
and	NN	O	O
HEB	NN	O	B-DNA
/	NN	O	O
E2-2	NN	O	B-DNA
in	NN	O	O
mammalian	NN	O	O
development	NN	O	O
,	NN	O	O
we	NN	O	O
generated	NN	O	O
and	NN	O	O
tested	NN	O	O
a	NN	O	O
panel	NN	O	O
of	NN	O	O
E2A	NN	O	B-DNA
knockin	NN	O	O
mutations	NN	O	O
including	NN	O	O
subtle	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
E12	NN	O	B-DNA
and	NN	O	I-DNA
E47	NN	O	I-DNA
exons	NN	O	I-DNA
and	NN	O	O
substitution	NN	O	O
of	NN	O	O
both	NN	O	O
E12	NN	O	B-DNA
and	NN	O	I-DNA
E47	NN	O	I-DNA
exons	NN	O	I-DNA
with	NN	O	O
a	NN	O	O
human	NN	O	B-DNA
HEB	NN	O	I-DNA
cDNA	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
find	NN	O	O
that	NN	O	O
the	NN	O	O
alternatively	NN	O	O
spliced	NN	O	O
E12	NN	O	B-protein
and	NN	O	I-protein
E47	NN	O	I-protein
bHLH	NN	O	I-protein
proteins	NN	O	I-protein
of	NN	O	O
the	NN	O	O
E2A	NN	O	B-DNA
gene	NN	O	I-DNA
play	NN	O	O
similar	NN	O	O
and	NN	O	O
additive	NN	O	O
roles	NN	O	O
in	NN	O	O
supporting	NN	O	O
B	NN	O	B-cell_type
lymphopoiesis	NN	O	I-cell_type
.	NN	O	O

Further	NN	O	O
,	NN	O	O
we	NN	O	O
find	NN	O	O
that	NN	O	O
HEB	NN	O	B-DNA
driven	NN	O	O
by	NN	O	O
the	NN	O	O
endogenous	NN	O	B-DNA
E2A	NN	O	I-DNA
promoter	NN	O	I-DNA
can	NN	O	O
functionally	NN	O	O
replace	NN	O	O
E2A	NN	O	B-DNA
in	NN	O	O
supporting	NN	O	O
B-cell	NN	O	O
commitment	NN	O	O
and	NN	O	O
differentiation	NN	O	O
toward	NN	O	O
completion	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
the	NN	O	O
postnatal	NN	O	O
lethality	NN	O	O
associated	NN	O	O
with	NN	O	O
E2A	NN	O	B-DNA
disruption	NN	O	O
is	NN	O	O
fully	NN	O	O
rescued	NN	O	O
by	NN	O	O
the	NN	O	O
addition	NN	O	O
of	NN	O	O
HEB	NN	O	B-DNA
.	NN	O	O

This	NN	O	O
study	NN	O	O
suggests	NN	O	O
that	NN	O	O
the	NN	O	O
functional	NN	O	O
divergence	NN	O	O
among	NN	O	O
E12	NN	O	B-DNA
,	NN	O	O
E47	NN	O	B-DNA
,	NN	O	O
and	NN	O	O
HEB	NN	O	B-DNA
in	NN	O	O
different	NN	O	O
cell	NN	O	O
types	NN	O	O
is	NN	O	O
partially	NN	O	O
defined	NN	O	O
by	NN	O	O
the	NN	O	O
context	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

Biased	NN	O	O
dependency	NN	O	O
of	NN	O	O
CD80	NN	O	B-protein
versus	NN	O	O
CD86	NN	O	B-protein
in	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
regulating	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
IL-2	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
addition	NN	O	O
to	NN	O	O
the	NN	O	O
signals	NN	O	O
obtained	NN	O	O
by	NN	O	O
ligation	NN	O	O
of	NN	O	O
the	NN	O	O
TCR	NN	O	O
,	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
need	NN	O	O
additional	NN	O	O
,	NN	O	O
co-stimulatory	NN	O	O
signals	NN	O	O
to	NN	O	O
be	NN	O	O
activated	NN	O	O
.	NN	O	O

One	NN	O	O
such	NN	O	O
co-stimulatory	NN	O	O
signal	NN	O	O
is	NN	O	O
delivered	NN	O	O
when	NN	O	O
CD28	NN	O	B-protein
on	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
binds	NN	O	O
to	NN	O	O
CD80	NN	O	B-protein
or	NN	O	O
CD86	NN	O	B-protein
on	NN	O	O
antigen-presenting	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
APC	NN	O	B-cell_type
)	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
analyzed	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
CD80	NN	O	B-protein
and	NN	O	O
CD86	NN	O	B-protein
to	NN	O	O
co-stimulate	NN	O	B-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
activated	NN	O	O
by	NN	O	O
superantigen	NN	O	O
.	NN	O	O

Using	NN	O	O
the	NN	O	O
Raji	NN	O	B-cell_line
B	NN	O	I-cell_line
cell	NN	O	I-cell_line
lymphoma	NN	O	I-cell_line
,	NN	O	O
which	NN	O	O
express	NN	O	O
similar	NN	O	O
levels	NN	O	O
of	NN	O	O
CD80	NN	O	B-protein
and	NN	O	O
CD86	NN	O	B-protein
,	NN	O	O
it	NN	O	O
was	NN	O	O
found	NN	O	O
that	NN	O	O
T	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
was	NN	O	O
mainly	NN	O	O
co-stimulated	NN	O	O
by	NN	O	O
CD80	NN	O	B-protein
.	NN	O	O

To	NN	O	O
further	NN	O	O
characterize	NN	O	O
the	NN	O	O
consequences	NN	O	O
of	NN	O	O
this	NN	O	O
biased	NN	O	O
co-stimulatory	NN	O	O
dependency	NN	O	O
,	NN	O	O
we	NN	O	O
employed	NN	O	O
a	NN	O	O
well-defined	NN	O	O
system	NN	O	O
of	NN	O	O
transfected	NN	O	O
CHO	NN	O	B-cell_line
cells	NN	O	I-cell_line
expressing	NN	O	O
human	NN	O	B-protein
MHC	NN	O	I-protein
class	NN	O	I-protein
II	NN	O	I-protein
together	NN	O	O
with	NN	O	O
CD80	NN	O	B-protein
,	NN	O	O
CD86	NN	O	B-protein
or	NN	O	O
CD80	NN	O	B-protein
and	NN	O	O
CD86	NN	O	B-protein
.	NN	O	O

Proliferation	NN	O	O
of	NN	O	O
freshly	NN	O	O
prepared	NN	O	O
CD4+	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
required	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
either	NN	O	O
CD80	NN	O	B-protein
or	NN	O	O
CD86	NN	O	B-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
reached	NN	O	O
only	NN	O	O
suboptimal	NN	O	O
levels	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
CD86	NN	O	B-protein
but	NN	O	O
optimal	NN	O	O
levels	NN	O	O
with	NN	O	O
CD80	NN	O	B-protein
.	NN	O	O

To	NN	O	O
analyze	NN	O	O
IL-2	NN	O	B-protein
transcriptional	NN	O	O
activity	NN	O	O
in	NN	O	O
CD80	NN	O	B-protein
and	NN	O	O
CD86	NN	O	B-protein
co-stimulated	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
we	NN	O	O
used	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
transfected	NN	O	O
with	NN	O	O
luciferase	NN	O	B-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
constructs	NN	O	I-DNA
.	NN	O	O

CD80	NN	O	B-protein
induced	NN	O	O
higher	NN	O	O
levels	NN	O	O
of	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
-enhancer	NN	O	O
activity	NN	O	O
compared	NN	O	O
to	NN	O	O
CD86	NN	O	B-protein
.	NN	O	O

Furthermore	NN	O	O
,	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
regulating	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter-enhancer	NN	O	I-DNA
region	NN	O	I-DNA
including	NN	O	O
activation	NN	O	B-protein
protein-1	NN	O	I-protein
,	NN	O	O
CD28	NN	O	B-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
and	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappaB	NN	O	I-protein
were	NN	O	O
4-8	NN	O	O
times	NN	O	O
higher	NN	O	O
after	NN	O	O
CD80	NN	O	B-protein
compared	NN	O	O
to	NN	O	O
CD86	NN	O	B-protein
ligation	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
eventual	NN	O	O
appearance	NN	O	O
of	NN	O	O
CD80	NN	O	B-protein
on	NN	O	O
recently	NN	O	O
activated	NN	O	O
CD86+	NN	O	B-cell_type
APC	NN	O	I-cell_type
is	NN	O	O
important	NN	O	O
for	NN	O	O
the	NN	O	O
superinduction	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
and	NN	O	O
to	NN	O	O
support	NN	O	O
vigorous	NN	O	O
T	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Duplication	NN	O	O
of	NN	O	O
the	NN	O	O
DR3	NN	O	B-DNA
gene	NN	O	I-DNA
on	NN	O	O
human	NN	O	B-DNA
chromosome	NN	O	I-DNA
1p36	NN	O	I-DNA
and	NN	O	O
its	NN	O	O
deletion	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
neuroblastoma	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
human	NN	O	B-DNA
DR3	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
whose	NN	O	O
product	NN	O	O
is	NN	O	O
also	NN	O	O
known	NN	O	O
as	NN	O	O
Wsl-1/APO-3/TRAMP/LARD	NN	O	B-protein
,	NN	O	O
encodes	NN	O	O
a	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-related	NN	O	I-protein
receptor	NN	O	I-protein
that	NN	O	O
is	NN	O	O
expressed	NN	O	O
primarily	NN	O	O
on	NN	O	O
the	NN	O	O
surface	NN	O	O
of	NN	O	O
thymocytes	NN	O	B-cell_type
and	NN	O	O
lymphocytes	NN	O	B-cell_type
.	NN	O	O

DR3	NN	O	B-protein
is	NN	O	O
capable	NN	O	O
of	NN	O	O
inducing	NN	O	O
both	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
and	NN	O	O
apoptosis	NN	O	O
when	NN	O	O
overexpressed	NN	O	O
in	NN	O	O
mammalian	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
although	NN	O	O
its	NN	O	O
ligand	NN	O	O
has	NN	O	O
not	NN	O	O
yet	NN	O	O
been	NN	O	O
identified	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
DR3	NN	O	B-DNA
gene	NN	O	I-DNA
locus	NN	O	I-DNA
is	NN	O	O
tandemly	NN	O	O
duplicated	NN	O	O
on	NN	O	O
human	NN	O	B-DNA
chromosome	NN	O	I-DNA
band	NN	O	I-DNA
1p36.2-p36.3	NN	O	I-DNA
and	NN	O	O
that	NN	O	O
these	NN	O	O
genes	NN	O	O
are	NN	O	O
hemizygously	NN	O	O
deleted	NN	O	O
and/or	NN	O	O
translocated	NN	O	O
to	NN	O	O
another	NN	O	O
chromosome	NN	O	O
in	NN	O	O
neuroblastoma	NN	O	B-cell_line
(	NN	O	I-cell_line
NB	NN	O	I-cell_line
)	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
with	NN	O	O
amplified	NN	O	O
MYCN	NN	O	O
.	NN	O	O

Duplication	NN	O	O
of	NN	O	O
at	NN	O	O
least	NN	O	O
a	NN	O	O
portion	NN	O	O
of	NN	O	O
the	NN	O	O
DR3	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	O
including	NN	O	O
the	NN	O	O
extracellular	NN	O	B-protein
and	NN	O	I-protein
transmembrane	NN	O	I-protein
regions	NN	O	I-protein
but	NN	O	O
not	NN	O	O
the	NN	O	O
cytoplasmic	NN	O	B-protein
domain	NN	O	I-protein
,	NN	O	O
was	NN	O	O
demonstrated	NN	O	O
by	NN	O	O
both	NN	O	O
fluorescence	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
and	NN	O	O
genomic	NN	O	O
Southern	NN	O	O
blotting	NN	O	O
.	NN	O	O

In	NN	O	O
most	NN	O	O
NB	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
both	NN	O	O
the	NN	O	O
DR3	NN	O	B-DNA
and	NN	O	I-DNA
the	NN	O	I-DNA
DR3L	NN	O	I-DNA
sequences	NN	O	I-DNA
are	NN	O	O
simultaneously	NN	O	O
deleted	NN	O	O
and/or	NN	O	O
translocated	NN	O	O
to	NN	O	O
another	NN	O	O
chromosome	NN	O	B-DNA
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
DR3/	NN	O	B-protein
Wsl-1	NN	O	I-protein
protein	NN	O	I-protein
expression	NN	O	O
is	NN	O	O
quite	NN	O	O
variable	NN	O	O
among	NN	O	O
these	NN	O	O
NB	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
with	NN	O	O
very	NN	O	O
low	NN	O	O
or	NN	O	O
undetectable	NN	O	O
levels	NN	O	O
in	NN	O	O
7	NN	O	O
of	NN	O	O
17	NN	O	O
NB	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line

-DOCSTART-	O

Differential	NN	O	O
protection	NN	O	O
of	NN	O	O
normal	NN	O	B-cell_type
and	NN	O	I-cell_type
malignant	NN	O	I-cell_type
human	NN	O	I-cell_type
myeloid	NN	O	I-cell_type
progenitors	NN	O	I-cell_type
(	NN	O	O
CFU-GM	NN	O	B-cell_type
)	NN	O	O
from	NN	O	O
Ara-C	NN	O	O
toxicity	NN	O	O
using	NN	O	O
cycloheximide	NN	O	O
.	NN	O	O

Cycloheximide	NN	O	O
,	NN	O	O
a	NN	O	O
reversible	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
inhibitor	NN	O	O
,	NN	O	O
is	NN	O	O
thought	NN	O	O
to	NN	O	O
block	NN	O	O
DNA	NN	O	O
replication	NN	O	O
in	NN	O	O
normal	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
preventing	NN	O	O
synthesis	NN	O	O
of	NN	O	O
a	NN	O	O
labile	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

In	NN	O	O
animal	NN	O	O
systems	NN	O	O
,	NN	O	O
cycloheximide	NN	O	O
protects	NN	O	O
normal	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
cytotoxic	NN	O	O
S-phase	NN	O	O
specific	NN	O	O
agents	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
cytosine	NN	O	O
arabinoside	NN	O	O
(	NN	O	O
Ara-C	NN	O	O
)	NN	O	O
.	NN	O	O

Malignant	NN	O	B-cell_type
cells	NN	O	I-cell_type
appear	NN	O	O
not	NN	O	O
to	NN	O	O
be	NN	O	O
susceptible	NN	O	O
to	NN	O	O
cycloheximide-induced	NN	O	O
cycle	NN	O	O
arrest	NN	O	O
and	NN	O	O
,	NN	O	O
subsequently	NN	O	O
,	NN	O	O
may	NN	O	O
not	NN	O	O
be	NN	O	O
protected	NN	O	O
from	NN	O	O
Ara-C	NN	O	O
cytotoxicity	NN	O	O
.	NN	O	O

The	NN	O	O
effect	NN	O	O
of	NN	O	O
cycloheximide	NN	O	O
on	NN	O	O
granulocyte/macrophage	NN	O	B-cell_type
progenitors	NN	O	I-cell_type
(	NN	O	O
CFU-GM	NN	O	B-cell_type
)	NN	O	O
after	NN	O	O
in	NN	O	O
vitro	NN	O	O
Ara-C	NN	O	O
exposure	NN	O	O
was	NN	O	O
examined	NN	O	O
using	NN	O	O
normal	NN	O	O
human	NN	O	O
bone	NN	O	O
marrow	NN	O	O
,	NN	O	O
malignant	NN	O	B-cell_type
progenitors	NN	O	I-cell_type
from	NN	O	O
patients	NN	O	O
with	NN	O	O
chronic	NN	O	O
myelogenous	NN	O	O
leukemia	NN	O	O
(	NN	O	O
CML	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
clonogenic	NN	O	B-cell_line
cells	NN	O	I-cell_line
from	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
acute	NN	O	I-cell_line
nonlymphocytic	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
HL-60	NN	O	I-cell_line
and	NN	O	I-cell_line
KG-1	NN	O	I-cell_line
.	NN	O	O

Mononuclear	NN	O	B-cell_line
or	NN	O	I-cell_line
clonogenic	NN	O	I-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
incubated	NN	O	O
for	NN	O	O
one	NN	O	O
hour	NN	O	O
with	NN	O	O
cycloheximide	NN	O	O
,	NN	O	O
followed	NN	O	O
by	NN	O	O
the	NN	O	O
addition	NN	O	O
,	NN	O	O
for	NN	O	O
three	NN	O	O
or	NN	O	O
17	NN	O	O
hours	NN	O	O
,	NN	O	O
of	NN	O	O
Ara-C	NN	O	O
before	NN	O	O
being	NN	O	O
plated	NN	O	O
in	NN	O	O
a	NN	O	O
methylcellulose	NN	O	O
culture	NN	O	O
system	NN	O	O
.	NN	O	O

CFU-GM	NN	O	O
survival	NN	O	O
was	NN	O	O
significantly	NN	O	O
increase	NN	O	O
if	NN	O	O
normal	NN	O	O
cells	NN	O	O
were	NN	O	O
treated	NN	O	O
with	NN	O	O
cycloheximide	NN	O	O
before	NN	O	O
Ara-C	NN	O	O
exposure	NN	O	O
.	NN	O	O

Similar	NN	O	O
cycloheximide	NN	O	O
pretreatment	NN	O	O
of	NN	O	O
CML	NN	O	B-cell_type
progenitors	NN	O	I-cell_type
and	NN	O	O
clonogenic	NN	O	B-cell_line
HL-60	NN	O	I-cell_line
and	NN	O	I-cell_line
KG-1	NN	O	I-cell_line
cells	NN	O	I-cell_line
failed	NN	O	O
to	NN	O	O
protect	NN	O	O
CFU-GM	NN	O	B-cell_type
from	NN	O	O
Ara-C-induced	NN	O	O
cytotoxicity	NN	O	O
.	NN	O	O

-DOCSTART-	O

Primary	NN	O	O
cortisol	NN	O	O
resistance	NN	O	O
accompanied	NN	O	O
by	NN	O	O
a	NN	O	O
reduction	NN	O	O
in	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
two	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
same	NN	O	O
family	NN	O	O
.	NN	O	O

This	NN	O	O
report	NN	O	O
describes	NN	O	O
studies	NN	O	O
of	NN	O	O
a	NN	O	O
man	NN	O	O
suspected	NN	O	O
of	NN	O	O
having	NN	O	O
primary	NN	O	O
cortisol	NN	O	O
resistance	NN	O	O
.	NN	O	O

This	NN	O	O
conclusion	NN	O	O
is	NN	O	O
based	NN	O	O
on	NN	O	O
his	NN	O	O
high	NN	O	O
plasma	NN	O	O
cortisol	NN	O	O
levels	NN	O	O
and	NN	O	O
high	NN	O	O
24-h	NN	O	O
urinary	NN	O	O
17-hydroxycorticosteroid	NN	O	O
and	NN	O	O
cortisol	NN	O	O
excretion	NN	O	O
,	NN	O	O
plus	NN	O	O
the	NN	O	O
fact	NN	O	O
that	NN	O	O
he	NN	O	O
had	NN	O	O
no	NN	O	O
manifestations	NN	O	O
of	NN	O	O
Cushing	NN	O	O
's	NN	O	O
syndrome	NN	O	O
.	NN	O	O

Among	NN	O	O
family	NN	O	O
members	NN	O	O
tested	NN	O	O
,	NN	O	O
his	NN	O	O
mother	NN	O	O
also	NN	O	O
had	NN	O	O
hypercortisolemia	NN	O	O
.	NN	O	O

Both	NN	O	O
mother	NN	O	O
and	NN	O	O
son	NN	O	O
had	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
unbound	NN	O	O
plasma	NN	O	O
cortisol	NN	O	O
,	NN	O	O
but	NN	O	O
their	NN	O	O
plasma	NN	O	O
ACTH	NN	O	O
concentrations	NN	O	O
were	NN	O	O
within	NN	O	O
the	NN	O	O
normal	NN	O	O
range	NN	O	O
.	NN	O	O

Both	NN	O	O
were	NN	O	O
partially	NN	O	O
resistant	NN	O	O
to	NN	O	O
dexamethasone	NN	O	O
adrenal	NN	O	O
suppression	NN	O	O
,	NN	O	O
and	NN	O	O
both	NN	O	O
had	NN	O	O
mild	NN	O	O
hypertension	NN	O	O
without	NN	O	O
hypokalemia	NN	O	O
.	NN	O	O

To	NN	O	O
study	NN	O	O
this	NN	O	O
apparent	NN	O	O
end-organ	NN	O	O
resistance	NN	O	O
to	NN	O	O
cortisol	NN	O	O
,	NN	O	O
we	NN	O	O
examined	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
peripheral	NN	O	B-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Using	NN	O	O
whole	NN	O	O
cell	NN	O	O
assays	NN	O	O
,	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
both	NN	O	O
patients	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
have	NN	O	O
reduced	NN	O	O
total	NN	O	O
binding	NN	O	O
capacity	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
these	NN	O	O
two	NN	O	O
patients	NN	O	O
,	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
same	NN	O	O
family	NN	O	O
,	NN	O	O
have	NN	O	O
primary	NN	O	O
cortisol	NN	O	O
resistance	NN	O	O
accompanied	NN	O	O
by	NN	O	O
a	NN	O	O
reduced	NN	O	O
number	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

[	NN	O	O
Glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
and	NN	O	O
response	NN	O	O
to	NN	O	O
polychemotherapy	NN	O	O
in	NN	O	O
acute	NN	O	O
lymphatic	NN	O	O
leukemia	NN	O	O
]	NN	O	O

Glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
GR	NN	O	B-protein
)	NN	O	O
levels	NN	O	O
were	NN	O	O
quantified	NN	O	O
in	NN	O	O
leukemic	NN	O	B-cell_type
blasts	NN	O	I-cell_type
from	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
of	NN	O	O
86	NN	O	O
patients	NN	O	O
with	NN	O	O
acute	NN	O	O
lymphoblastic	NN	O	O
leukemia	NN	O	O
.	NN	O	O

The	NN	O	O
subsequent	NN	O	O
achievement	NN	O	O
of	NN	O	O
complete	NN	O	O
remission	NN	O	O
after	NN	O	O
combination	NN	O	O
chemotherapy	NN	O	O
was	NN	O	O
correlated	NN	O	O
with	NN	O	O
high	NN	O	O
receptor	NN	O	O
levels	NN	O	O
.	NN	O	O

Forty-seven	NN	O	O
of	NN	O	O
50	NN	O	O
patients	NN	O	O
with	NN	O	O
leukemic	NN	O	B-cell_type
cells	NN	O	I-cell_type
containing	NN	O	O
more	NN	O	O
than	NN	O	O
6	NN	O	O
,	NN	O	O
000	NN	O	O
receptor	NN	O	O
sites	NN	O	O
and	NN	O	O
22	NN	O	O
of	NN	O	O
36	NN	O	O
patients	NN	O	O
with	NN	O	O
cells	NN	O	O
containing	NN	O	O
less	NN	O	O
than	NN	O	O
6	NN	O	O
,	NN	O	O
000	NN	O	O
receptor	NN	O	O
sites	NN	O	O
achieved	NN	O	O
remission	NN	O	O
.	NN	O	O

The	NN	O	O
study	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
leukemic	NN	O	B-cell_type
cells	NN	O	I-cell_type
may	NN	O	O
predict	NN	O	O
response	NN	O	O
to	NN	O	O
combination	NN	O	O
chemotherapy	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
acute	NN	O	O
lymphoblastic	NN	O	O
leukemia	NN	O	O
.	NN	O	O

-DOCSTART-	O

Specific	NN	O	O
uptake	NN	O	O
of	NN	O	O
1	NN	O	O
,	NN	O	O
25-dihydroxycholecalciferol	NN	O	O
by	NN	O	O
human	NN	O	B-cell_type
chronic	NN	O	I-cell_type
myeloid	NN	O	I-cell_type
leukemia	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
have	NN	O	O
examined	NN	O	O
mononuclear	NN	O	B-cell_line
cell	NN	O	I-cell_line
preparations	NN	O	I-cell_line
from	NN	O	O
patients	NN	O	O
with	NN	O	O
chronic	NN	O	O
myeloid	NN	O	O
leukemia	NN	O	O
[	NN	O	O
CML	NN	O	O
]	NN	O	O
for	NN	O	O
binding	NN	O	O
of	NN	O	O
and	NN	O	O
response	NN	O	O
to	NN	O	O
1	NN	O	O
,	NN	O	O
25-dihydroxycholecalciferol	NN	O	O
[	NN	O	O
1	NN	O	O
,	NN	O	O
25-	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
]	NN	O	O
.	NN	O	O

Whole	NN	O	O
cells	NN	O	O
specifically	NN	O	O
took	NN	O	O
up	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
-1	NN	O	O
,	NN	O	O
25-	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
with	NN	O	O
high	NN	O	O
affinity	NN	O	O
(	NN	O	O
Kd	NN	O	O
3.6	NN	O	O
X	NN	O	O
10	NN	O	O
(	NN	O	O
-11	NN	O	O
)	NN	O	O
M	NN	O	O
)	NN	O	O
and	NN	O	O
low	NN	O	O
capacity	NN	O	O
.	NN	O	O

Subcellular	NN	O	O
fractionation	NN	O	O
of	NN	O	O
labeled	NN	O	B-cell_line
cells	NN	O	I-cell_line
showed	NN	O	O
that	NN	O	O
binding	NN	O	O
was	NN	O	O
restricted	NN	O	O
to	NN	O	O
cytosols	NN	O	O
and	NN	O	O
nuclei	NN	O	O
.	NN	O	O

Sucrose	NN	O	O
gradient	NN	O	O
centrifugation	NN	O	O
of	NN	O	O
cells	NN	O	O
preincubated	NN	O	O
with	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
-1	NN	O	O
,	NN	O	O
25-	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
revealed	NN	O	O
a	NN	O	O
single	NN	O	O
3.6S	NN	O	O
peak	NN	O	O
which	NN	O	O
was	NN	O	O
totally	NN	O	O
displaced	NN	O	O
with	NN	O	O
100-fold	NN	O	O
excess	NN	O	O
nonradioactive	NN	O	O
hormone	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
we	NN	O	O
were	NN	O	O
unable	NN	O	O
to	NN	O	O
demonstrate	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
by	NN	O	O
postlabeling	NN	O	O
standard	NN	O	O
cytosol	NN	O	O
preparations	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
cytosols	NN	O	O
prepared	NN	O	O
from	NN	O	O
a	NN	O	O
mixture	NN	O	O
of	NN	O	O
CML	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
1	NN	O	B-cell_line
,	NN	O	I-cell_line
25-	NN	O	I-cell_line
(	NN	O	I-cell_line
OH	NN	O	I-cell_line
)	NN	O	I-cell_line
2D3	NN	O	I-cell_line
receptor-positive	NN	O	I-cell_line
T47D	NN	O	I-cell_line
(	NN	O	O
human	NN	O	O
breast	NN	O	O
cancer	NN	O	O
)	NN	O	O
cells	NN	O	O
had	NN	O	O
less	NN	O	O
than	NN	O	O
10	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
binding	NN	O	O
measured	NN	O	O
in	NN	O	O
T47D	NN	O	B-cell_line
cytosol	NN	O	O
alone	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
binding	NN	O	O
in	NN	O	O
T47D	NN	O	B-cell_line
cytosols	NN	O	O
were	NN	O	O
not	NN	O	O
reduced	NN	O	O
if	NN	O	O
the	NN	O	O
receptors	NN	O	O
were	NN	O	O
occupied	NN	O	O
with	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
-1	NN	O	O
,	NN	O	O
25-	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
prior	NN	O	O
to	NN	O	O
the	NN	O	O
addition	NN	O	O
of	NN	O	O
the	NN	O	O
CML	NN	O	B-cell_line
cytosols	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
CML	NN	O	B-cell_line
cells	NN	O	I-cell_line
appear	NN	O	O
to	NN	O	O
contain	NN	O	O
both	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
and	NN	O	O
an	NN	O	O
unknown	NN	O	O
substance	NN	O	O
which	NN	O	O
prevents	NN	O	O
its	NN	O	O
detection	NN	O	O
following	NN	O	O
the	NN	O	O
preparation	NN	O	O
of	NN	O	O
cytosol	NN	O	O
.	NN	O	O

Cells	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
CML	NN	O	O
in	NN	O	O
the	NN	O	O
chronic	NN	O	O
phase	NN	O	O
specifically	NN	O	O
bound	NN	O	O
more	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
18.0	NN	O	O
+/-	NN	O	O
3.2	NN	O	O
(	NN	O	O
S.E.	NN	O	O
)	NN	O	O
fmol/10	NN	O	O
(	NN	O	O
7	NN	O	O
)	NN	O	O
cells	NN	O	O
]	NN	O	O
than	NN	O	O
did	NN	O	O
those	NN	O	O
in	NN	O	O
acute	NN	O	O
myeloid	NN	O	O
transformation	NN	O	O
[	NN	O	O
7.2	NN	O	O
+/-	NN	O	O
1.5	NN	O	O
]	NN	O	O
or	NN	O	O
than	NN	O	O
did	NN	O	O
cells	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
acute	NN	O	O
myeloid	NN	O	O
leukemia	NN	O	O
[	NN	O	O
2.6	NN	O	O
+/-	NN	O	O
0.8	NN	O	O
]	NN	O	O
.	NN	O	O

Only	NN	O	O
cells	NN	O	O
from	NN	O	O
the	NN	O	O
first	NN	O	O
group	NN	O	O
of	NN	O	O
patients	NN	O	O
responded	NN	O	O
to	NN	O	O
the	NN	O	O
addition	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
by	NN	O	O
differentiating	NN	O	O
along	NN	O	O
the	NN	O	O
monocyte-macrophage	NN	O	O
pathway	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
the	NN	O	O
differentiation-induction	NN	O	O
effect	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
is	NN	O	O
likely	NN	O	O
to	NN	O	O
depend	NN	O	O
on	NN	O	O
adequate	NN	O	O
levels	NN	O	O
of	NN	O	O
receptor	NN	O	O
and	NN	O	O
that	NN	O	O
intact	NN	O	B-cell_type
cells	NN	O	I-cell_type
rather	NN	O	O
than	NN	O	O
cytosol	NN	O	O
preparations	NN	O	O
should	NN	O	O
be	NN	O	O
studied	NN	O	O
before	NN	O	O
cells	NN	O	O
of	NN	O	O
a	NN	O	O
particular	NN	O	O
tissue	NN	O	O
are	NN	O	O
designated	NN	O	O
as	NN	O	O
receptor	NN	O	O
negative	NN	O	O
.	NN	O	O

-DOCSTART-	O

Effect	NN	O	O
of	NN	O	O
cell	NN	O	O
cycle	NN	O	O
position	NN	O	O
on	NN	O	O
dexamethasone	NN	O	O
binding	NN	O	O
by	NN	O	O
mouse	NN	O	B-cell_line
and	NN	O	I-cell_line
human	NN	O	I-cell_line
lymphoid	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
:	NN	O	O
correlation	NN	O	O
between	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
dexamethasone	NN	O	O
binding	NN	O	O
during	NN	O	O
S	NN	O	O
phase	NN	O	O
and	NN	O	O
dexamethasone	NN	O	O
sensitivity	NN	O	O
.	NN	O	O

We	NN	O	O
determined	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
cell	NN	O	O
cycle	NN	O	O
position	NN	O	O
on	NN	O	O
the	NN	O	O
amount	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
that	NN	O	O
was	NN	O	O
specifically	NN	O	O
bound	NN	O	O
by	NN	O	O
mouse	NN	O	B-cell_line
and	NN	O	I-cell_line
human	NN	O	I-cell_line
lymphoid	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
that	NN	O	O
were	NN	O	O
either	NN	O	O
sensitive	NN	O	O
or	NN	O	O
resistant	NN	O	O
to	NN	O	O
growth	NN	O	O
inhibition	NN	O	O
by	NN	O	O
dexamethasone	NN	O	O
were	NN	O	O
compared	NN	O	O
.	NN	O	O

Exponentially	NN	O	B-cell_type
growing	NN	O	I-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
separated	NN	O	O
by	NN	O	O
centrifugal	NN	O	O
elutriation	NN	O	O
into	NN	O	O
fractions	NN	O	O
that	NN	O	O
corresponded	NN	O	O
to	NN	O	O
different	NN	O	O
positions	NN	O	O
in	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
.	NN	O	O

The	NN	O	O
cell	NN	O	O
cycle	NN	O	O
phase	NN	O	O
distribution	NN	O	O
of	NN	O	O
each	NN	O	O
fraction	NN	O	O
was	NN	O	O
estimated	NN	O	O
by	NN	O	O
flow	NN	O	O
cytometry	NN	O	O
and	NN	O	O
autoradiography	NN	O	O
.	NN	O	O

The	NN	O	O
amount	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
bound	NN	O	O
per	NN	O	O
cell	NN	O	O
in	NN	O	O
each	NN	O	O
fraction	NN	O	O
was	NN	O	O
measured	NN	O	O
by	NN	O	O
a	NN	O	O
whole	NN	O	O
cell	NN	O	O
binding	NN	O	O
assay	NN	O	O
.	NN	O	O

In	NN	O	O
three	NN	O	O
dexamethasone-sensitive	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
(	NN	O	O
two	NN	O	O
mouse	NN	O	O
and	NN	O	O
one	NN	O	O
human	NN	O	O
)	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
amount	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
bound	NN	O	O
per	NN	O	O
cell	NN	O	O
increased	NN	O	O
2-4-fold	NN	O	O
between	NN	O	O
G1	NN	O	O
phase	NN	O	O
and	NN	O	O
S	NN	O	O
phase	NN	O	O
,	NN	O	O
and	NN	O	O
then	NN	O	O
decreased	NN	O	O
during	NN	O	O
G2/M	NN	O	O
phase	NN	O	O
.	NN	O	O

Results	NN	O	O
were	NN	O	O
the	NN	O	O
same	NN	O	O
when	NN	O	O
the	NN	O	O
amount	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
bound	NN	O	O
per	NN	O	O
milligram	NN	O	O
of	NN	O	O
cell	NN	O	B-protein
protein	NN	O	I-protein
was	NN	O	O
measured	NN	O	O
.	NN	O	O

Binding	NN	O	O
affinity	NN	O	O
was	NN	O	O
the	NN	O	O
same	NN	O	O
during	NN	O	O
G1	NN	O	O
phase	NN	O	O
and	NN	O	O
S	NN	O	O
phase	NN	O	O
,	NN	O	O
but	NN	O	O
the	NN	O	O
proportion	NN	O	O
of	NN	O	O
bound	NN	O	O
dexamethasone	NN	O	O
that	NN	O	O
translocated	NN	O	O
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
was	NN	O	O
greater	NN	O	O
during	NN	O	O
S	NN	O	O
phase	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
amount	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
bound	NN	O	O
per	NN	O	O
cell	NN	O	O
by	NN	O	O
three	NN	O	O
dexamethasone-resistant	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
(	NN	O	O
two	NN	O	O
mouse	NN	O	O
and	NN	O	O
one	NN	O	O
human	NN	O	O
)	NN	O	O
did	NN	O	O
not	NN	O	O
increase	NN	O	O
during	NN	O	O
S	NN	O	O
phase	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
cell	NN	O	O
cycle	NN	O	O
changes	NN	O	O
in	NN	O	O
dexamethasone	NN	O	O
binding	NN	O	O
are	NN	O	O
not	NN	O	O
simply	NN	O	O
related	NN	O	O
to	NN	O	O
changes	NN	O	O
in	NN	O	O
cell	NN	O	B-protein
protein	NN	O	I-protein
or	NN	O	O
cell	NN	O	O
volume	NN	O	O
during	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
.	NN	O	O

An	NN	O	O
increase	NN	O	O
in	NN	O	O
dexamethasone	NN	O	O
binding	NN	O	O
during	NN	O	O
S	NN	O	O
phase	NN	O	O
may	NN	O	O
be	NN	O	O
required	NN	O	O
for	NN	O	O
dexamethasone	NN	O	O
to	NN	O	O
inhibit	NN	O	O
cell	NN	O	O
growth	NN	O	O
,	NN	O	O
and	NN	O	O
a	NN	O	O
failure	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
binding	NN	O	O
to	NN	O	O
increase	NN	O	O
during	NN	O	O
S	NN	O	O
phase	NN	O	O
might	NN	O	O
represent	NN	O	O
a	NN	O	O
new	NN	O	O
mechanism	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
resistance	NN	O	O
in	NN	O	O
lymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Presence	NN	O	O
and	NN	O	O
steroid	NN	O	O
inducibility	NN	O	O
of	NN	O	O
glutamine	NN	O	B-protein
synthetase	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
leukemic	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Glutamine	NN	O	B-protein
synthetase	NN	O	I-protein
(	NN	O	O
EC	NN	O	O
6.3.1.2	NN	O	O
;	NN	O	O
GS	NN	O	O
)	NN	O	O
is	NN	O	O
present	NN	O	O
in	NN	O	O
lymphoblasts	NN	O	B-cell_type
from	NN	O	O
patients	NN	O	O
with	NN	O	O
acute	NN	O	O
lymphoblastic	NN	O	O
leukemia	NN	O	O
(	NN	O	O
ALL	NN	O	O
)	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
in	NN	O	O
normal	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
16	NN	O	O
out	NN	O	O
of	NN	O	O
20	NN	O	O
ALL	NN	O	O
patients	NN	O	O
studied	NN	O	O
exposure	NN	O	O
of	NN	O	O
the	NN	O	O
cells	NN	O	O
to	NN	O	O
physiological	NN	O	O
concentrations	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
in	NN	O	O
vitro	NN	O	O
increased	NN	O	O
enzyme	NN	O	B-protein
activity	NN	O	O
above	NN	O	O
the	NN	O	O
control	NN	O	O
levels	NN	O	O
.	NN	O	O

The	NN	O	O
increase	NN	O	O
was	NN	O	O
specific	NN	O	O
for	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
ligands	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
magnitude	NN	O	O
of	NN	O	O
glucocorticoid-mediated	NN	O	O
increase	NN	O	O
of	NN	O	O
GS	NN	O	O
activity	NN	O	O
and	NN	O	O
the	NN	O	O
cellular	NN	O	O
levels	NN	O	O
of	NN	O	O
specific	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
assayed	NN	O	O
in	NN	O	O
the	NN	O	O
same	NN	O	O
cell	NN	O	B-cell_type
specimen	NN	O	I-cell_type
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
the	NN	O	O
basal	NN	O	O
levels	NN	O	O
of	NN	O	O
the	NN	O	O
enzyme	NN	O	B-protein
measured	NN	O	O
in	NN	O	O
cells	NN	O	O
prior	NN	O	O
to	NN	O	O
exposure	NN	O	O
to	NN	O	O
dexamethasone	NN	O	O
correlated	NN	O	O
negatively	NN	O	O
with	NN	O	O
receptor	NN	O	O
density	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
suggested	NN	O	O
that	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
steroid-inducible	NN	O	B-protein
GS	NN	O	I-protein
in	NN	O	O
ALL	NN	O	B-cell_type
cells	NN	O	I-cell_type
may	NN	O	O
prove	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
marker	NN	O	O
for	NN	O	O
functional	NN	O	B-protein
receptor	NN	O	I-protein
sites	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Binding	NN	O	O
of	NN	O	O
progestins	NN	O	O
to	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
.	NN	O	O

Correlation	NN	O	O
to	NN	O	O
their	NN	O	O
glucocorticoid-like	NN	O	O
effects	NN	O	O
on	NN	O	O
in	NN	O	O
vitro	NN	O	O
functions	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
mononuclear	NN	O	I-cell_type
leukocytes	NN	O	I-cell_type
.	NN	O	O

A	NN	O	O
number	NN	O	O
of	NN	O	O
physiological	NN	O	O
and	NN	O	O
synthetic	NN	O	O
progestins	NN	O	O
were	NN	O	O
tested	NN	O	O
for	NN	O	O
their	NN	O	O
ability	NN	O	O
to	NN	O	O
compete	NN	O	O
with	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
dexamethasone	NN	O	O
for	NN	O	O
the	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
of	NN	O	O
human	NN	O	B-cell_type
mononuclear	NN	O	I-cell_type
leukocytes	NN	O	I-cell_type
and	NN	O	O
their	NN	O	O
ability	NN	O	O
to	NN	O	O
elicit	NN	O	O
glucocorticoid-like	NN	O	O
effects	NN	O	O
on	NN	O	O
the	NN	O	O
same	NN	O	O
cells	NN	O	O
.	NN	O	O

As	NN	O	O
compared	NN	O	O
to	NN	O	O
the	NN	O	O
reference	NN	O	O
compound	NN	O	O
dexamethasone	NN	O	O
(	NN	O	O
relative	NN	O	O
receptor	NN	O	O
binding	NN	O	O
affinity	NN	O	O
defined	NN	O	O
as	NN	O	O
100	NN	O	O
%	NN	O	O
)	NN	O	O
,	NN	O	O
two	NN	O	O
potent	NN	O	O
synthetic	NN	O	O
progestins	NN	O	O
with	NN	O	O
a	NN	O	O
pregnane-type	NN	O	O
structure	NN	O	O
,	NN	O	O
megestrol	NN	O	O
acetate	NN	O	O
and	NN	O	O
medroxyprogesterone	NN	O	O
acetate	NN	O	O
,	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
display	NN	O	O
a	NN	O	O
considerable	NN	O	O
binding	NN	O	O
affinity	NN	O	O
towards	NN	O	O
the	NN	O	O
receptor	NN	O	O
(	NN	O	O
46	NN	O	O
and	NN	O	O
42	NN	O	O
%	NN	O	O
,	NN	O	O
respectively	NN	O	O
)	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
the	NN	O	O
naturally	NN	O	O
occurring	NN	O	O
ligand	NN	O	O
,	NN	O	O
cortisol	NN	O	O
,	NN	O	O
to	NN	O	O
the	NN	O	O
receptor	NN	O	O
was	NN	O	O
clearly	NN	O	O
lower	NN	O	O
(	NN	O	O
25	NN	O	O
%	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
effective	NN	O	O
binding	NN	O	O
of	NN	O	O
medroxyprogesterone	NN	O	O
acetate	NN	O	O
to	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
was	NN	O	O
confirmed	NN	O	O
by	NN	O	O
direct	NN	O	O
binding	NN	O	O
studies	NN	O	O
utilizing	NN	O	O
a	NN	O	O
tritiated	NN	O	O
derivative	NN	O	O
of	NN	O	O
this	NN	O	O
steroid	NN	O	O
.	NN	O	O

No	NN	O	O
evidence	NN	O	O
for	NN	O	O
the	NN	O	O
existence	NN	O	O
of	NN	O	O
a	NN	O	O
specific	NN	O	O
progesterone	NN	O	B-protein
receptor	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
was	NN	O	O
obtained	NN	O	O
as	NN	O	O
judged	NN	O	O
by	NN	O	O
the	NN	O	O
results	NN	O	O
of	NN	O	O
competition	NN	O	O
experiments	NN	O	O
where	NN	O	O
a	NN	O	O
progesterone	NN	O	O
receptor-specific	NN	O	O
ligand	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
Org	NN	O	O
2058	NN	O	O
was	NN	O	O
used	NN	O	O
.	NN	O	O

Medroxyprogesterone	NN	O	O
acetate	NN	O	O
and	NN	O	O
megestrol	NN	O	O
acetate	NN	O	O
also	NN	O	O
induced	NN	O	O
glucocorticoid-like	NN	O	O
effects	NN	O	O
on	NN	O	O
the	NN	O	O
lymphocyte	NN	O	B-cell_type
functions	NN	O	O
.	NN	O	O

These	NN	O	O
included	NN	O	O
inhibition	NN	O	O
of	NN	O	O
the	NN	O	O
proliferative	NN	O	O
responses	NN	O	O
to	NN	O	O
the	NN	O	O
T-cell	NN	O	B-protein
mitogens	NN	O	I-protein
concanavalin	NN	O	B-protein
A	NN	O	I-protein
and	NN	O	O
phytohaemagglutinin	NN	O	B-protein
and	NN	O	O
an	NN	O	O
enhanced	NN	O	O
accumulation	NN	O	O
of	NN	O	O
immunoglobulin	NN	O	B-cell_type
secreting	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
pokeweed	NN	O	B-cell_line
mitogen-stimulated	NN	O	I-cell_line
cultures	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
progestin	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
a	NN	O	O
radiosensitive	NN	O	O
(	NN	O	O
suppressor	NN	O	O
)	NN	O	O
subpopulation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
the	NN	O	O
synthetic	NN	O	O
progestins	NN	O	O
related	NN	O	O
structurally	NN	O	O
to	NN	O	O
19-nortestosterone	NN	O	O
,	NN	O	O
norethisterone	NN	O	O
and	NN	O	O
d-norgestrel	NN	O	O
,	NN	O	O
were	NN	O	O
virtually	NN	O	O
devoid	NN	O	O
of	NN	O	O
binding	NN	O	O
affinity	NN	O	O
towards	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
nor	NN	O	O
did	NN	O	O
they	NN	O	O
measurably	NN	O	O
influence	NN	O	O
the	NN	O	O
in	NN	O	O
vitro	NN	O	O
lymphocyte	NN	O	B-cell_type
functions	NN	O	O
.	NN	O	O

These	NN	O	O
studies	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
certain	NN	O	O
progestins	NN	O	O
in	NN	O	O
common	NN	O	O
clinical	NN	O	O
use	NN	O	O
probably	NN	O	O
possess	NN	O	O
inherent	NN	O	O
glucocorticoid	NN	O	O
activity	NN	O	O
and	NN	O	O
suggest	NN	O	O
that	NN	O	O
side	NN	O	O
effects	NN	O	O
attributable	NN	O	O
to	NN	O	O
this	NN	O	O
character	NN	O	O
(	NN	O	O
e.g.	NN	O	O
suppression	NN	O	O
of	NN	O	O
the	NN	O	O
pituitary-adrenal	NN	O	O
axis	NN	O	O
)	NN	O	O
might	NN	O	O
be	NN	O	O
expected	NN	O	O
when	NN	O	O
these	NN	O	O
compounds	NN	O	O
are	NN	O	O
used	NN	O	O
in	NN	O	O
pharmacological	NN	O	O
doses	NN	O	O
.	NN	O	O

-DOCSTART-	O

Metabolic	NN	O	O
and	NN	O	O
ultrastructural	NN	O	O
aspects	NN	O	O
of	NN	O	O
the	NN	O	O
in	NN	O	O
vitro	NN	O	O
lysis	NN	O	O
of	NN	O	O
chronic	NN	O	B-cell_type
lymphocytic	NN	O	I-cell_type
leukemia	NN	O	I-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
glucocorticoids	NN	O	O
.	NN	O	O

Human	NN	O	B-cell_type
chronic	NN	O	I-cell_type
lymphocytic	NN	O	I-cell_type
leukemia	NN	O	I-cell_type
(	NN	O	I-cell_type
CLL	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
like	NN	O	O
prothymocytes	NN	O	B-cell_type
and	NN	O	O
immunoactivated	NN	O	B-cell_type
T-lymphocytes	NN	O	I-cell_type
are	NN	O	O
readily	NN	O	O
lysed	NN	O	O
in	NN	O	O
vitro	NN	O	O
by	NN	O	O
pharmacological	NN	O	O
concentrations	NN	O	O
of	NN	O	O
glucocorticoids	NN	O	O
such	NN	O	O
as	NN	O	O
cortisol	NN	O	O
,	NN	O	O
whereas	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
thymocytes	NN	O	B-cell_type
are	NN	O	O
unaffected	NN	O	O
by	NN	O	O
the	NN	O	O
hormone	NN	O	B-protein
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
metabolic	NN	O	O
and	NN	O	O
ultrastructural	NN	O	O
aspects	NN	O	O
of	NN	O	O
the	NN	O	O
cortisol-induced	NN	O	O
killing	NN	O	O
process	NN	O	O
of	NN	O	O
CLL	NN	O	B-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
recorded	NN	O	O
.	NN	O	O

In	NN	O	O
vitro	NN	O	O
lysis	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
temperature	NN	O	O
dependent	NN	O	O
and	NN	O	O
was	NN	O	O
detected	NN	O	O
only	NN	O	O
after	NN	O	O
6	NN	O	O
to	NN	O	O
8	NN	O	O
hr	NN	O	O
incubation	NN	O	O
with	NN	O	O
cortisol	NN	O	O
by	NN	O	O
means	NN	O	O
of	NN	O	O
the	NN	O	O
trypan	NN	O	O
blue	NN	O	O
exclusion	NN	O	O
test	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
30	NN	O	O
min	NN	O	O
of	NN	O	O
incubation	NN	O	O
with	NN	O	O
cortisol	NN	O	O
at	NN	O	O
either	NN	O	O
37	NN	O	O
degrees	NN	O	O
or	NN	O	O
4	NN	O	O
degrees	NN	O	O
followed	NN	O	O
by	NN	O	O
the	NN	O	O
removal	NN	O	O
of	NN	O	O
the	NN	O	O
hormone	NN	O	B-protein
was	NN	O	O
still	NN	O	O
sufficient	NN	O	O
to	NN	O	O
induce	NN	O	O
the	NN	O	O
lytic	NN	O	O
process	NN	O	O
.	NN	O	O

Ultrastructural	NN	O	O
studies	NN	O	O
demonstrated	NN	O	O
sequential	NN	O	O
changes	NN	O	O
in	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
,	NN	O	O
including	NN	O	O
swelling	NN	O	O
of	NN	O	O
mitochondria	NN	O	O
and	NN	O	O
cytoplasmic	NN	O	O
decompartmentalization	NN	O	O
,	NN	O	O
followed	NN	O	O
by	NN	O	O
loss	NN	O	O
of	NN	O	O
surface	NN	O	O
microvilli	NN	O	O
with	NN	O	O
the	NN	O	O
appearance	NN	O	O
of	NN	O	O
``	NN	O	O
holes	NN	O	O
''	NN	O	O
in	NN	O	O
the	NN	O	O
cell	NN	O	O
membrane	NN	O	O
,	NN	O	O
and	NN	O	O
subsequent	NN	O	O
condensation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-DNA
chromatin	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
large	NN	O	O
holes	NN	O	O
in	NN	O	O
the	NN	O	O
membrane	NN	O	O
appearing	NN	O	O
after	NN	O	O
6	NN	O	O
hr	NN	O	O
of	NN	O	O
incubation	NN	O	O
with	NN	O	O
the	NN	O	O
hormone	NN	O	B-protein
may	NN	O	O
be	NN	O	O
the	NN	O	O
cause	NN	O	O
for	NN	O	O
the	NN	O	O
penetration	NN	O	O
of	NN	O	O
the	NN	O	O
viable	NN	O	O
stain	NN	O	O
into	NN	O	O
the	NN	O	O
dead	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
as	NN	O	O
seen	NN	O	O
by	NN	O	O
light	NN	O	O
microscopy	NN	O	O
.	NN	O	O

Addition	NN	O	O
of	NN	O	O
metabolic	NN	O	O
inhibitors	NN	O	O
including	NN	O	O
actinomycin	NN	O	O
D	NN	O	O
,	NN	O	O
puromycin	NN	O	O
,	NN	O	O
and	NN	O	O
cycloheximide	NN	O	O
following	NN	O	O
administration	NN	O	O
of	NN	O	O
cortisol	NN	O	O
resulted	NN	O	O
in	NN	O	O
inhibition	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	O
lysis	NN	O	O
.	NN	O	O

An	NN	O	O
excess	NN	O	O
of	NN	O	O
an	NN	O	O
antagonist	NN	O	O
such	NN	O	O
as	NN	O	O
cortexolone	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
inhibit	NN	O	O
the	NN	O	O
cortisol-induced	NN	O	O
cytolysis	NN	O	O
of	NN	O	O
the	NN	O	O
CLL	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

It	NN	O	O
is	NN	O	O
suggested	NN	O	O
that	NN	O	O
the	NN	O	O
glucocorticoid-induced	NN	O	O
lysis	NN	O	O
of	NN	O	O
human	NN	O	B-cell_line
CLL	NN	O	I-cell_line
cells	NN	O	I-cell_line
is	NN	O	O
similar	NN	O	O
to	NN	O	O
the	NN	O	O
phenomenon	NN	O	O
observed	NN	O	O
in	NN	O	O
rat	NN	O	B-cell_type
or	NN	O	I-cell_type
murine	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
interaction	NN	O	O
of	NN	O	O
the	NN	O	O
steroid	NN	O	O
molecule	NN	O	O
with	NN	O	O
the	NN	O	O
cytoplasmic	NN	O	B-protein
receptor	NN	O	I-protein
.	NN	O	O

The	NN	O	O
resulting	NN	O	O
complex	NN	O	O
appears	NN	O	O
to	NN	O	O
activate	NN	O	O
specific	NN	O	O
gene	NN	O	B-DNA
(	NN	O	O
s	NN	O	O
)	NN	O	O
the	NN	O	O
products	NN	O	O
of	NN	O	O
which	NN	O	O
eventually	NN	O	O
cause	NN	O	O
cytolysis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
lymphoid	NN	O	O
tumors	NN	O	O
.	NN	O	O

There	NN	O	O
is	NN	O	O
a	NN	O	O
range	NN	O	O
of	NN	O	O
levels	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
numbers	NN	O	O
seen	NN	O	O
in	NN	O	O
the	NN	O	O
various	NN	O	O
subclasses	NN	O	O
of	NN	O	O
acute	NN	O	O
lymphatic	NN	O	O
leukemia	NN	O	O
(	NN	O	O
ALL	NN	O	O
)	NN	O	O
.	NN	O	O

This	NN	O	O
variability	NN	O	O
can	NN	O	O
not	NN	O	O
be	NN	O	O
explained	NN	O	O
by	NN	O	O
the	NN	O	O
known	NN	O	O
correlation	NN	O	O
between	NN	O	O
active	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
and	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
receptors	NN	O	O
,	NN	O	O
since	NN	O	O
the	NN	O	O
tumors	NN	O	O
with	NN	O	O
the	NN	O	O
highest	NN	O	O
growth	NN	O	O
fraction	NN	O	O
(	NN	O	O
i.e.	NN	O	O
,	NN	O	O
Burkitt	NN	O	O
's	NN	O	O
lymphoma	NN	O	O
and	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
)	NN	O	O
tend	NN	O	O
to	NN	O	O
have	NN	O	O
lower	NN	O	O
average	NN	O	O
receptor	NN	O	O
numbers	NN	O	O
than	NN	O	O
do	NN	O	O
tumors	NN	O	O
with	NN	O	O
lower	NN	O	O
growth	NN	O	O
fractions	NN	O	O
such	NN	O	O
as	NN	O	O
common	NN	O	O
ALL	NN	O	O
.	NN	O	O

All	NN	O	O
clinical	NN	O	O
specimens	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
lymphatic	NN	O	O
leukemia	NN	O	O
have	NN	O	O
some	NN	O	O
measurable	NN	O	O
level	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
;	NN	O	O
therefore	NN	O	O
,	NN	O	O
the	NN	O	O
resistance	NN	O	O
seen	NN	O	O
in	NN	O	O
vivo	NN	O	O
can	NN	O	O
not	NN	O	O
be	NN	O	O
explained	NN	O	O
by	NN	O	O
the	NN	O	O
lack	NN	O	O
of	NN	O	O
receptors	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
there	NN	O	O
has	NN	O	O
been	NN	O	O
a	NN	O	O
positive	NN	O	O
correlation	NN	O	O
,	NN	O	O
in	NN	O	O
our	NN	O	O
hands	NN	O	O
,	NN	O	O
with	NN	O	O
receptor	NN	O	O
level	NN	O	O
and	NN	O	O
prognosis	NN	O	O
.	NN	O	O

On	NN	O	O
the	NN	O	O
basis	NN	O	O
of	NN	O	O
in	NN	O	O
vitro	NN	O	O
models	NN	O	O
,	NN	O	O
it	NN	O	O
is	NN	O	O
proposed	NN	O	O
that	NN	O	O
perhaps	NN	O	O
the	NN	O	O
high	NN	O	B-cell_line
receptor	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
(	NN	O	O
i.e.	NN	O	O
,	NN	O	O
common	NN	O	O
ALL	NN	O	O
of	NN	O	O
childhood	NN	O	O
)	NN	O	O
have	NN	O	O
relative	NN	O	O
stability	NN	O	O
of	NN	O	O
their	NN	O	O
genetic	NN	O	O
material	NN	O	O
making	NN	O	O
glucocorticoid-resistant	NN	O	O
mutations	NN	O	O
less	NN	O	O
likely	NN	O	O
to	NN	O	O
occur	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
these	NN	O	O
cells	NN	O	O
than	NN	O	O
in	NN	O	O
low-receptor	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
(	NN	O	O
i.e.	NN	O	O
,	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
)	NN	O	O
.	NN	O	O

This	NN	O	O
greater	NN	O	O
genetic	NN	O	O
variability	NN	O	O
in	NN	O	O
the	NN	O	O
low-receptor	NN	O	B-cell_line
lines	NN	O	I-cell_line
could	NN	O	O
account	NN	O	O
for	NN	O	O
the	NN	O	O
earlier	NN	O	O
emergence	NN	O	O
of	NN	O	O
clinical	NN	O	O
glucocorticoid	NN	O	O
resistance	NN	O	O
in	NN	O	O
these	NN	O	O
patients	NN	O	O
.	NN	O	O

-DOCSTART-	O

[	NN	O	O
Tumor	NN	O	O
histology	NN	O	O
and	NN	O	O
steroid	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
breast	NN	O	O
carcinoma	NN	O	O
]	NN	O	O

In	NN	O	O
Specimens	NN	O	O
of	NN	O	O
115	NN	O	O
patients	NN	O	O
with	NN	O	O
breast	NN	O	O
cancer	NN	O	O
4	NN	O	O
tumorparameters	NN	O	O
(	NN	O	O
tumorsize	NN	O	O
,	NN	O	O
tumorboder	NN	O	O
,	NN	O	O
nucleargrade	NN	O	O
,	NN	O	O
lymphocytic	NN	O	O
stromal	NN	O	O
reaction	NN	O	O
)	NN	O	O
3	NN	O	O
features	NN	O	O
of	NN	O	O
regional	NN	O	O
lymphnodes	NN	O	O
(	NN	O	O
sinushistiocytosis	NN	O	O
,	NN	O	O
T-cellreaction	NN	O	O
,	NN	O	O
lymphnode	NN	O	O
metastases	NN	O	O
)	NN	O	O
and	NN	O	O
estrogen	NN	O	B-protein
and	NN	O	I-protein
progesteron	NN	O	I-protein
receptors	NN	O	I-protein
were	NN	O	O
determined	NN	O	O
.	NN	O	O

A	NN	O	O
strong	NN	O	O
sinushistiocytosis	NN	O	O
and	NN	O	O
T-cellreaction	NN	O	O
could	NN	O	O
be	NN	O	O
verified	NN	O	O
mainly	NN	O	O
in	NN	O	O
metastases	NN	O	O
in	NN	O	O
free	NN	O	O
lymphnodes	NN	O	O
.	NN	O	O

The	NN	O	O
steroid	NN	O	B-protein
receptor	NN	O	I-protein
content	NN	O	O
does	NN	O	O
not	NN	O	O
correlate	NN	O	O
with	NN	O	O
histological	NN	O	O
parameters	NN	O	O

-DOCSTART-	O

Aldosterone-receptor	NN	O	B-protein
deficiency	NN	O	O
in	NN	O	O
pseudohypoaldosteronism	NN	O	O
.	NN	O	O

Pseudohypoaldosteronism	NN	O	O
,	NN	O	O
a	NN	O	O
syndrome	NN	O	O
characterized	NN	O	O
by	NN	O	O
salt	NN	O	O
wasting	NN	O	O
and	NN	O	O
failure	NN	O	O
to	NN	O	O
thrive	NN	O	O
,	NN	O	O
usually	NN	O	O
presents	NN	O	O
in	NN	O	O
infancy	NN	O	O
as	NN	O	O
high	NN	O	O
urinary	NN	O	O
levels	NN	O	O
of	NN	O	O
sodium	NN	O	O
despite	NN	O	O
hyponatremia	NN	O	O
,	NN	O	O
hyperkalemia	NN	O	O
,	NN	O	O
hyperreninemia	NN	O	O
,	NN	O	O
and	NN	O	O
elevated	NN	O	O
aldosterone	NN	O	O
levels	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
investigated	NN	O	O
this	NN	O	O
syndrome	NN	O	O
for	NN	O	O
the	NN	O	O
possibility	NN	O	O
of	NN	O	O
abnormal	NN	O	B-protein
Type	NN	O	I-protein
I	NN	O	I-protein
or	NN	O	O
``	NN	O	B-protein
mineralocorticoid-like	NN	O	I-protein
''	NN	O	I-protein
receptors	NN	O	I-protein
,	NN	O	O
which	NN	O	O
have	NN	O	O
intrinsic	NN	O	O
steroid	NN	O	O
specificity	NN	O	O
indistinguishable	NN	O	O
from	NN	O	O
that	NN	O	O
of	NN	O	O
renal	NN	O	B-protein
mineralocorticoid	NN	O	I-protein
receptors	NN	O	I-protein
and	NN	O	O
are	NN	O	O
found	NN	O	O
in	NN	O	O
many	NN	O	O
tissues	NN	O	O
and	NN	O	O
cells	NN	O	B-cell_type
,	NN	O	O
including	NN	O	O
mononuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
have	NN	O	O
studied	NN	O	O
three	NN	O	O
patients	NN	O	O
with	NN	O	O
pseudohypoaldosteronism	NN	O	O
:	NN	O	O
the	NN	O	O
28-year-old	NN	O	O
index	NN	O	O
case	NN	O	O
in	NN	O	O
Melbourne	NN	O	O
(	NN	O	O
Patient	NN	O	O
1	NN	O	O
)	NN	O	O
and	NN	O	O
two	NN	O	O
siblings	NN	O	O
in	NN	O	O
Munich	NN	O	O
,	NN	O	O
eight	NN	O	O
and	NN	O	O
two	NN	O	O
years	NN	O	O
of	NN	O	O
age	NN	O	O
(	NN	O	O
Patients	NN	O	O
2	NN	O	O
and	NN	O	O
3	NN	O	O
)	NN	O	O
;	NN	O	O
clinically	NN	O	O
,	NN	O	O
Patient	NN	O	O
3	NN	O	O
had	NN	O	O
a	NN	O	O
less	NN	O	O
severe	NN	O	O
case	NN	O	O
than	NN	O	O
his	NN	O	O
sister	NN	O	O
.	NN	O	O

Percoll-separated	NN	O	B-cell_type
control	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
bound	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
aldosterone	NN	O	O
with	NN	O	O
high	NN	O	O
affinity	NN	O	O
(	NN	O	O
Kd	NN	O	O
approximately	NN	O	O
3	NN	O	O
nM	NN	O	O
)	NN	O	O
and	NN	O	O
limited	NN	O	O
capacity	NN	O	O
(	NN	O	O
150	NN	O	O
to	NN	O	O
600	NN	O	O
sites	NN	O	O
per	NN	O	O
cell	NN	O	O
)	NN	O	O
.	NN	O	O

On	NN	O	O
repeated	NN	O	O
examination	NN	O	O
,	NN	O	O
no	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
aldosterone	NN	O	O
binding	NN	O	O
was	NN	O	O
found	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
from	NN	O	O
Patients	NN	O	O
1	NN	O	O
and	NN	O	O
2	NN	O	O
;	NN	O	O
in	NN	O	O
Patient	NN	O	O
3	NN	O	O
,	NN	O	O
the	NN	O	O
levels	NN	O	O
were	NN	O	O
62	NN	O	O
sites	NN	O	O
per	NN	O	O
cell	NN	O	O
,	NN	O	O
more	NN	O	O
than	NN	O	O
2	NN	O	O
S.D.	NN	O	O
below	NN	O	O
those	NN	O	O
of	NN	O	O
the	NN	O	O
control	NN	O	O
.	NN	O	O

Levels	NN	O	O
in	NN	O	O
the	NN	O	O
parents	NN	O	O
of	NN	O	O
the	NN	O	O
Munich	NN	O	O
patients	NN	O	O
(	NN	O	O
first	NN	O	O
cousins	NN	O	O
)	NN	O	O
were	NN	O	O
normal	NN	O	O
.	NN	O	O

It	NN	O	O
appears	NN	O	O
that	NN	O	O
pseudohypoaldosteronism	NN	O	O
is	NN	O	O
caused	NN	O	O
by	NN	O	O
a	NN	O	O
Type	NN	O	O
I	NN	O	O
receptor	NN	O	O
defect	NN	O	O
,	NN	O	O
that	NN	O	O
the	NN	O	O
defect	NN	O	O
may	NN	O	O
be	NN	O	O
complete	NN	O	O
or	NN	O	O
partial	NN	O	O
,	NN	O	O
that	NN	O	O
transmission	NN	O	O
may	NN	O	O
be	NN	O	O
autosomal	NN	O	O
recessive	NN	O	O
,	NN	O	O
and	NN	O	O
that	NN	O	O
the	NN	O	O
study	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
pseudohypoaldosteronism	NN	O	O
may	NN	O	O
indicate	NN	O	O
physiologic	NN	O	O
roles	NN	O	O
for	NN	O	O
Type	NN	O	B-protein
I	NN	O	I-protein
receptors	NN	O	I-protein
in	NN	O	O
nonepithelial	NN	O	O
tissues	NN	O	O
.	NN	O	O

-DOCSTART-	O

Glucocorticoid	NN	O	O
inhibition	NN	O	O
of	NN	O	O
urokinase-like	NN	O	B-protein
plasminogen	NN	O	I-protein
activators	NN	O	I-protein
in	NN	O	O
cultured	NN	O	B-cell_line
human	NN	O	I-cell_line
lymphoblasts	NN	O	I-cell_line
.	NN	O	O

Two	NN	O	O
human	NN	O	B-cell_line
lymphoblast	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
LICR-LON-HMy2	NN	O	B-cell_line
(	NN	O	O
HMy2	NN	O	B-cell_line
cells	NN	O	I-cell_line
)	NN	O	O
and	NN	O	O
GM4672A	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
are	NN	O	O
moderately	NN	O	O
growth	NN	O	O
inhibited	NN	O	O
by	NN	O	O
dexamethasone	NN	O	O
(	NN	O	O
1	NN	O	O
,	NN	O	O
4-pregnadien-9-fluoro-16	NN	O	O
alpha-methyl-11	NN	O	O
beta	NN	O	O
,	NN	O	O
17	NN	O	O
alpha	NN	O	O
,	NN	O	O
21-triol-3	NN	O	O
,	NN	O	O
20-dione	NN	O	O
)	NN	O	O
(	NN	O	O
Dex	NN	O	O
)	NN	O	O
.	NN	O	O

Both	NN	O	O
cell	NN	O	O
types	NN	O	O
secrete	NN	O	O
a	NN	O	O
urokinase	NN	O	B-protein
(	NN	O	I-protein
UK	NN	O	I-protein
)	NN	O	I-protein
-like	NN	O	I-protein
plasminogen	NN	O	I-protein
activator	NN	O	I-protein
(	NN	O	O
PA	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
both	NN	O	O
HMy2	NN	O	B-cell_line
and	NN	O	I-cell_line
GM4672A	NN	O	I-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
Dex	NN	O	O
for	NN	O	O
1-4	NN	O	O
days	NN	O	O
inhibits	NN	O	O
extracellular	NN	O	O
PA	NN	O	B-protein
activity	NN	O	O
in	NN	O	O
a	NN	O	O
concentration-dependent	NN	O	O
manner	NN	O	O
,	NN	O	O
being	NN	O	O
half-maximal	NN	O	O
at	NN	O	O
approximately	NN	O	O
1	NN	O	O
X	NN	O	O
10	NN	O	O
(	NN	O	O
-9	NN	O	O
)	NN	O	O
M	NN	O	O
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
PA	NN	O	B-protein
in	NN	O	O
both	NN	O	O
cell	NN	O	O
types	NN	O	O
is	NN	O	O
specific	NN	O	O
for	NN	O	O
active	NN	O	O
glucocorticoids	NN	O	O
,	NN	O	O
and	NN	O	O
this	NN	O	O
specificity	NN	O	O
parallels	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
various	NN	O	O
steroids	NN	O	O
to	NN	O	O
bind	NN	O	O
to	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
.	NN	O	O

HMy2	NN	O	B-cell_line
cell	NN	O	I-cell_line
PA	NN	O	B-protein
is	NN	O	O
fully	NN	O	O
suppressible	NN	O	O
by	NN	O	O
Dex	NN	O	O
,	NN	O	O
whereas	NN	O	O
up	NN	O	O
to	NN	O	O
one	NN	O	O
third	NN	O	O
of	NN	O	O
the	NN	O	O
activator	NN	O	O
expressed	NN	O	O
by	NN	O	O
GM4672A	NN	O	B-cell_line
cells	NN	O	I-cell_line
is	NN	O	O
resistant	NN	O	O
to	NN	O	O
glucocorticoid	NN	O	O
inhibition	NN	O	O
.	NN	O	O

Mixing	NN	O	O
experiments	NN	O	O
using	NN	O	O
a	NN	O	O
UK	NN	O	O
standard	NN	O	O
and	NN	O	O
conditioned	NN	O	O
media	NN	O	O
from	NN	O	O
Dex-treated	NN	O	B-cell_line
cells	NN	O	I-cell_line
suggest	NN	O	O
an	NN	O	O
absence	NN	O	O
of	NN	O	O
glucocorticoid-inducible	NN	O	B-protein
inhibitors	NN	O	I-protein
to	NN	O	O
UK	NN	O	B-protein
or	NN	O	O
plasmin	NN	O	B-protein
in	NN	O	O
both	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
conditioned	NN	O	O
media	NN	O	O
from	NN	O	O
Dex-treated	NN	O	B-cell_line
GM4672A	NN	O	I-cell_line
cells	NN	O	I-cell_line
inhibits	NN	O	O
a	NN	O	O
portion	NN	O	O
of	NN	O	O
the	NN	O	O
homologous	NN	O	O
cellular	NN	O	O
activator	NN	O	O
in	NN	O	O
conditioned	NN	O	O
media	NN	O	O
from	NN	O	O
control	NN	O	O
GM4672A	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
low	NN	O	O
levels	NN	O	O
of	NN	O	O
glucocorticoid-inducible	NN	O	B-protein
inhibitors	NN	O	I-protein
may	NN	O	O
contribute	NN	O	O
to	NN	O	O
,	NN	O	O
but	NN	O	O
can	NN	O	O
not	NN	O	O
fully	NN	O	O
account	NN	O	O
for	NN	O	O
,	NN	O	O
Dex	NN	O	O
inhibition	NN	O	O
of	NN	O	O
GM4672A	NN	O	B-protein
PA	NN	O	I-protein
activity	NN	O	O
.	NN	O	O

Glucocorticoid-inducible	NN	O	B-protein
inhibitors	NN	O	I-protein
in	NN	O	O
HMy2	NN	O	B-cell_line
cells	NN	O	I-cell_line
are	NN	O	O
either	NN	O	O
totally	NN	O	O
absent	NN	O	O
or	NN	O	O
are	NN	O	O
present	NN	O	O
at	NN	O	O
undetectable	NN	O	O
levels	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
regulation	NN	O	O
of	NN	O	O
UK-like	NN	O	B-protein
PAs	NN	O	I-protein
in	NN	O	O
HMy2	NN	O	B-cell_line
and	NN	O	I-cell_line
GM4672A	NN	O	I-cell_line
cells	NN	O	I-cell_line
differs	NN	O	O
with	NN	O	O
respect	NN	O	O
to	NN	O	O
the	NN	O	O
extent	NN	O	O
to	NN	O	O
which	NN	O	O
glucocorticoids	NN	O	O
inhibit	NN	O	O
constitutively	NN	O	B-protein
expressed	NN	O	I-protein
activator	NN	O	I-protein
levels	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
possible	NN	O	O
contribution	NN	O	O
of	NN	O	O
glucocorticoid-inducible	NN	O	B-protein
inhibitors	NN	O	I-protein
to	NN	O	O
the	NN	O	O
regulatory	NN	O	O
process	NN	O	O
in	NN	O	O
GM4672A	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

Characterization	NN	O	O
of	NN	O	O
aldosterone	NN	O	O
binding	NN	O	O
sites	NN	O	O
in	NN	O	O
circulating	NN	O	B-cell_type
human	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
leukocytes	NN	O	I-cell_type
.	NN	O	O

Aldosterone	NN	O	O
binding	NN	O	O
sites	NN	O	O
in	NN	O	O
human	NN	O	B-cell_line
mononuclear	NN	O	I-cell_line
leukocytes	NN	O	I-cell_line
were	NN	O	O
characterized	NN	O	O
after	NN	O	O
separation	NN	O	O
of	NN	O	O
cells	NN	O	O
from	NN	O	O
blood	NN	O	O
by	NN	O	O
a	NN	O	O
Percoll	NN	O	O
gradient	NN	O	O
.	NN	O	O

After	NN	O	O
washing	NN	O	O
and	NN	O	O
resuspension	NN	O	O
in	NN	O	O
RPMI-1640	NN	O	O
medium	NN	O	O
,	NN	O	O
cells	NN	O	O
were	NN	O	O
incubated	NN	O	O
at	NN	O	O
37	NN	O	O
degrees	NN	O	O
C	NN	O	O
for	NN	O	O
1	NN	O	O
h	NN	O	O
with	NN	O	O
different	NN	O	O
concentrations	NN	O	O
of	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
aldosterone	NN	O	O
plus	NN	O	O
a	NN	O	O
100-fold	NN	O	O
concentration	NN	O	O
of	NN	O	O
RU-26988	NN	O	O
(	NN	O	O
11	NN	O	O
alpha	NN	O	O
,	NN	O	O
17	NN	O	O
alpha-dihydroxy-17	NN	O	O
beta-propynylandrost-1	NN	O	O
,	NN	O	O
4	NN	O	O
,	NN	O	O
6-trien-3-one	NN	O	O
)	NN	O	O
,	NN	O	O
with	NN	O	O
or	NN	O	O
without	NN	O	O
an	NN	O	O
excess	NN	O	O
of	NN	O	O
unlabeled	NN	O	O
aldosterone	NN	O	O
.	NN	O	O

Aldosterone	NN	O	O
binds	NN	O	O
to	NN	O	O
a	NN	O	O
single	NN	O	O
class	NN	O	O
of	NN	O	O
receptors	NN	O	O
with	NN	O	O
an	NN	O	O
affinity	NN	O	O
of	NN	O	O
2.7	NN	O	O
+/-	NN	O	O
0.5	NN	O	O
nM	NN	O	O
(	NN	O	O
means	NN	O	O
+/-	NN	O	O
SD	NN	O	O
,	NN	O	O
n	NN	O	O
=	NN	O	O
14	NN	O	O
)	NN	O	O
and	NN	O	O
a	NN	O	O
capacity	NN	O	O
of	NN	O	O
290	NN	O	O
+/-	NN	O	O
108	NN	O	O
sites/cell	NN	O	O
(	NN	O	O
n	NN	O	O
=	NN	O	O
14	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
specificity	NN	O	O
data	NN	O	O
show	NN	O	O
a	NN	O	O
hierarchy	NN	O	O
of	NN	O	O
affinity	NN	O	O
of	NN	O	O
desoxycorticosterone	NN	O	O
=	NN	O	O
corticosterone	NN	O	O
=	NN	O	O
aldosterone	NN	O	O
greater	NN	O	O
than	NN	O	O
hydrocortisone	NN	O	O
greater	NN	O	O
than	NN	O	O
dexamethasone	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
mononuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
could	NN	O	O
be	NN	O	O
useful	NN	O	O
for	NN	O	O
studying	NN	O	O
the	NN	O	O
physiological	NN	O	O
significance	NN	O	O
of	NN	O	O
these	NN	O	O
mineralocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
and	NN	O	O
their	NN	O	O
regulation	NN	O	O
in	NN	O	O
humans	NN	O	O
.	NN	O	O

-DOCSTART-	O

Mineralocorticoid	NN	O	B-protein
and	NN	O	I-protein
glucocorticoid	NN	O	I-protein
receptors	NN	O	I-protein
in	NN	O	O
circulating	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
primary	NN	O	O
hyperaldosteronism	NN	O	O
.	NN	O	O

Mineralocorticoid	NN	O	B-protein
and	NN	O	I-protein
glucocorticoid	NN	O	I-protein
receptors	NN	O	I-protein
were	NN	O	O
measured	NN	O	O
in	NN	O	O
circulating	NN	O	B-cell_type
mononuclear	NN	O	I-cell_type
leukocytes	NN	O	I-cell_type
in	NN	O	O
5	NN	O	O
patients	NN	O	O
affected	NN	O	O
by	NN	O	O
Conn	NN	O	O
's	NN	O	O
syndrome	NN	O	O
(	NN	O	O
3	NN	O	O
cases	NN	O	O
of	NN	O	O
bilateral	NN	O	O
adrenal	NN	O	O
hyperplasia	NN	O	O
and	NN	O	O
2	NN	O	O
cases	NN	O	O
of	NN	O	O
adenoma	NN	O	O
plus	NN	O	O
unilateral	NN	O	O
hyperplasia	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
number	NN	O	O
of	NN	O	O
the	NN	O	O
binding	NN	O	O
sites	NN	O	O
per	NN	O	O
cell	NN	O	O
resulted	NN	O	O
significantly	NN	O	O
lower	NN	O	O
(	NN	O	O
189	NN	O	O
+/-	NN	O	O
114	NN	O	O
,	NN	O	O
mean	NN	O	O
+/-	NN	O	O
SD	NN	O	O
)	NN	O	O
,	NN	O	O
as	NN	O	O
compared	NN	O	O
with	NN	O	O
the	NN	O	O
normal	NN	O	O
controls	NN	O	O
(	NN	O	O
298	NN	O	O
+/-	NN	O	O
105	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
affinity	NN	O	O
of	NN	O	O
aldosterone	NN	O	O
for	NN	O	O
the	NN	O	O
receptor	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
not	NN	O	O
different	NN	O	O
than	NN	O	O
that	NN	O	O
of	NN	O	O
healthy	NN	O	O
control	NN	O	O
subjects	NN	O	O
.	NN	O	O

The	NN	O	O
capacity	NN	O	O
and	NN	O	O
the	NN	O	O
affinity	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
for	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
ranged	NN	O	O
in	NN	O	O
the	NN	O	O
normal	NN	O	O
values	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
a	NN	O	O
possible	NN	O	O
down-regulation	NN	O	O
of	NN	O	O
mineralocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
humans	NN	O	O
.	NN	O	O

-DOCSTART-	O

Short-term	NN	O	O
and	NN	O	O
long-term	NN	O	O
effects	NN	O	O
of	NN	O	O
estrogen	NN	O	O
on	NN	O	O
lymphoid	NN	O	O
tissues	NN	O	O
and	NN	O	O
lymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
some	NN	O	O
remarks	NN	O	O
on	NN	O	O
the	NN	O	O
significance	NN	O	O
for	NN	O	O
carcinogenesis	NN	O	O
.	NN	O	O

Estrogens	NN	O	O
have	NN	O	O
long	NN	O	O
been	NN	O	O
thought	NN	O	O
to	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
regulating	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
.	NN	O	O

The	NN	O	O
difference	NN	O	O
in	NN	O	O
some	NN	O	O
types	NN	O	O
of	NN	O	O
immune	NN	O	O
responses	NN	O	O
between	NN	O	O
males	NN	O	O
and	NN	O	O
females	NN	O	O
is	NN	O	O
well-known	NN	O	O
,	NN	O	O
as	NN	O	O
is	NN	O	O
the	NN	O	O
pronounced	NN	O	O
thymic	NN	O	O
involution	NN	O	O
induced	NN	O	O
by	NN	O	O
exogenous	NN	O	O
estrogens	NN	O	O
.	NN	O	O

Estrogens	NN	O	O
stimulate	NN	O	O
some	NN	O	O
aspects	NN	O	O
of	NN	O	O
macrophage	NN	O	B-cell_type
activity	NN	O	O
and	NN	O	O
,	NN	O	O
depending	NN	O	O
on	NN	O	O
dose	NN	O	O
and	NN	O	O
mitogen	NN	O	B-protein
,	NN	O	O
inhibit	NN	O	O
or	NN	O	O
stimulate	NN	O	O
lymphocyte	NN	O	O
proliferative	NN	O	O
response	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

Another	NN	O	O
example	NN	O	O
is	NN	O	O
the	NN	O	O
estrogen	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
delayed	NN	O	O
type	NN	O	O
hypersensitivity	NN	O	O
response	NN	O	O
.	NN	O	O

A	NN	O	O
broad	NN	O	O
review	NN	O	O
is	NN	O	O
given	NN	O	O
of	NN	O	O
such	NN	O	O
estrogen	NN	O	O
effects	NN	O	O
on	NN	O	O
lymphoid	NN	O	B-cell_type
tissue	NN	O	I-cell_type
and	NN	O	O
immune	NN	O	O
response	NN	O	O
.	NN	O	O

Most	NN	O	O
of	NN	O	O
the	NN	O	O
studies	NN	O	O
published	NN	O	O
so	NN	O	O
far	NN	O	O
are	NN	O	O
phenomenological	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
recent	NN	O	O
description	NN	O	O
of	NN	O	O
estrogen	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
the	NN	O	O
thymus	NN	O	O
and	NN	O	O
in	NN	O	O
some	NN	O	O
lymphocyte	NN	O	B-cell_line
subpopulations	NN	O	I-cell_line
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
a	NN	O	O
deeper	NN	O	O
understanding	NN	O	O
of	NN	O	O
regulating	NN	O	O
factors	NN	O	O
in	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
,	NN	O	O
open	NN	O	O
the	NN	O	O
possibility	NN	O	O
of	NN	O	O
a	NN	O	O
more	NN	O	O
detailed	NN	O	O
understanding	NN	O	O
of	NN	O	O
the	NN	O	O
estrogen	NN	O	O
mechanism	NN	O	O
of	NN	O	O
interference	NN	O	O
.	NN	O	O

Estrogen	NN	O	O
effects	NN	O	O
in	NN	O	O
adults	NN	O	O
are	NN	O	O
reversible	NN	O	O
.	NN	O	O

After	NN	O	O
treating	NN	O	O
neonatal	NN	O	O
mice	NN	O	O
with	NN	O	O
the	NN	O	O
synthetic	NN	O	O
estrogen	NN	O	O
diethylstilbestrol	NN	O	O
(	NN	O	O
DES	NN	O	O
)	NN	O	O
,	NN	O	O
disturbances	NN	O	O
are	NN	O	O
induced	NN	O	O
in	NN	O	O
lymphocyte	NN	O	B-cell_line
populations	NN	O	I-cell_line
and	NN	O	O
lymphocyte	NN	O	B-cell_type
functions	NN	O	O
which	NN	O	O
are	NN	O	O
permanent	NN	O	O
and	NN	O	O
irreversible	NN	O	O
.	NN	O	O

Lymphocytes	NN	O	B-cell_type
from	NN	O	O
adult	NN	O	O
,	NN	O	O
neonatally	NN	O	O
DES-treated	NN	O	O
female	NN	O	O
mice	NN	O	O
have	NN	O	O
a	NN	O	O
reduced	NN	O	O
mitogen	NN	O	O
response	NN	O	O
to	NN	O	O
ConA	NN	O	B-protein
and	NN	O	O
LPS	NN	O	B-protein
(	NN	O	O
T	NN	O	B-protein
and	NN	O	I-protein
B	NN	O	I-protein
cell	NN	O	I-protein
mitogen	NN	O	I-protein
)	NN	O	O
and	NN	O	O
the	NN	O	O
delayed	NN	O	O
type	NN	O	O
hypersensitivity	NN	O	O
response	NN	O	O
is	NN	O	O
depressed	NN	O	O
.	NN	O	O

A	NN	O	O
detailed	NN	O	O
analysis	NN	O	O
demonstrated	NN	O	O
a	NN	O	O
decreased	NN	O	O
T	NN	O	B-cell_type
helper	NN	O	I-cell_type
cell	NN	O	I-cell_type
population	NN	O	O
.	NN	O	O

The	NN	O	O
activity	NN	O	O
of	NN	O	O
Natural	NN	O	B-cell_type
Killer	NN	O	I-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
permanently	NN	O	O
reduced	NN	O	O
and	NN	O	O
this	NN	O	O
functional	NN	O	O
impairment	NN	O	O
is	NN	O	O
related	NN	O	O
to	NN	O	O
a	NN	O	O
decreased	NN	O	O
number	NN	O	O
of	NN	O	O
these	NN	O	O
cells	NN	O	O
,	NN	O	O
in	NN	O	O
turn	NN	O	O
determined	NN	O	O
at	NN	O	O
the	NN	O	O
bone	NN	O	O
marrow	NN	O	O
level	NN	O	O
.	NN	O	O

The	NN	O	O
same	NN	O	O
animals	NN	O	O
have	NN	O	O
an	NN	O	O
increased	NN	O	O
sensitivity	NN	O	O
to	NN	O	O
chemical	NN	O	O
carcinogens	NN	O	O
(	NN	O	O
methylcholanthrene	NN	O	O
)	NN	O	O
and	NN	O	O
they	NN	O	O
spontaneously	NN	O	O
develop	NN	O	O
epithelial	NN	O	O
changes	NN	O	O
in	NN	O	O
the	NN	O	O
uterine	NN	O	O
cervix	NN	O	O
which	NN	O	O
morphologically	NN	O	O
are	NN	O	O
similar	NN	O	O
to	NN	O	O
adenocarcinoma	NN	O	O
.	NN	O	O

The	NN	O	O
association	NN	O	O
between	NN	O	O
estrogen-associated	NN	O	O
malignancy	NN	O	O
and	NN	O	O
estrogen	NN	O	O
effects	NN	O	O
in	NN	O	O
lymphocyte	NN	O	B-cell_type
functions	NN	O	O
deserves	NN	O	O
further	NN	O	O
study	NN	O	O
.	NN	O	O

-DOCSTART-	O

Drugs	NN	O	O
affecting	NN	O	O
the	NN	O	O
hormonal	NN	O	O
receptors	NN	O	O
of	NN	O	O
normal	NN	O	B-cell_type
and	NN	O	I-cell_type
leukaemic	NN	O	I-cell_type
peripheral	NN	O	I-cell_type
leucocytes	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
authors	NN	O	O
investigated	NN	O	O
the	NN	O	O
behaviour	NN	O	O
of	NN	O	O
steroid	NN	O	O
hormone	NN	O	O
uptake	NN	O	O
in	NN	O	O
leukaemic	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
CML	NN	O	B-cell_line
,	NN	O	O
CLL	NN	O	B-cell_line
,	NN	O	O
AML	NN	O	B-cell_line
,	NN	O	O
ALL	NN	O	B-cell_line
)	NN	O	O
,	NN	O	O
in	NN	O	O
basal	NN	O	O
conditions	NN	O	O
and	NN	O	O
after	NN	O	O
incubation	NN	O	O
with	NN	O	O
drugs	NN	O	O
which	NN	O	O
modify	NN	O	O
the	NN	O	O
cellular	NN	O	O
concentration	NN	O	O
of	NN	O	O
cAMP	NN	O	O
,	NN	O	O
PGE	NN	O	O
and	NN	O	O
PGF	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
demonstrated	NN	O	O
the	NN	O	O
presence	NN	O	O
in	NN	O	O
leukaemic	NN	O	B-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
an	NN	O	O
alteration	NN	O	O
in	NN	O	O
the	NN	O	O
incorporation	NN	O	O
of	NN	O	O
steroid	NN	O	O
hormones	NN	O	O
.	NN	O	O

This	NN	O	O
alteration	NN	O	O
was	NN	O	O
scarcely	NN	O	O
modified	NN	O	O
by	NN	O	O
incubation	NN	O	O
with	NN	O	O
theophylline	NN	O	B-protein
,	NN	O	O
which	NN	O	O
increases	NN	O	O
cellular	NN	O	O
concentration	NN	O	O
of	NN	O	O
cAMP	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
it	NN	O	O
was	NN	O	O
moderately	NN	O	O
counteracted	NN	O	O
by	NN	O	O
thioproline	NN	O	O
and	NN	O	O
was	NN	O	O
evidently	NN	O	O
inhibited	NN	O	O
by	NN	O	O
flurbiprofen	NN	O	O
,	NN	O	O
which	NN	O	O
also	NN	O	O
reduced	NN	O	O
cellular	NN	O	O
concentrations	NN	O	O
of	NN	O	O
prostaglandins	NN	O	O
,	NN	O	O
particularly	NN	O	O
PGE2	NN	O	O
,	NN	O	O
with	NN	O	O
the	NN	O	O
exception	NN	O	O
of	NN	O	O
PGF2	NN	O	O
which	NN	O	O
showed	NN	O	O
a	NN	O	O
poor	NN	O	O
response	NN	O	O
.	NN	O	O

Differences	NN	O	O
were	NN	O	O
observed	NN	O	O
in	NN	O	O
the	NN	O	O
behavior	NN	O	O
of	NN	O	O
hormonal	NN	O	O
uptake	NN	O	O
of	NN	O	O
CML	NN	O	B-cell_line
,	NN	O	O
in	NN	O	O
contrast	NN	O	O
to	NN	O	O
that	NN	O	O
of	NN	O	O
AML	NN	O	B-cell_line
,	NN	O	I-cell_line
CLL	NN	O	I-cell_line
and	NN	O	I-cell_line
ALL	NN	O	I-cell_line
peripheral	NN	O	I-cell_line
leucocytes	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

Human	NN	O	O
breast	NN	O	O
cancer	NN	O	O
and	NN	O	O
impaired	NN	O	O
NK	NN	O	B-cell_type
cell	NN	O	I-cell_type
function	NN	O	O
.	NN	O	O

Recent	NN	O	O
advances	NN	O	O
in	NN	O	O
tumor	NN	O	O
immunology	NN	O	O
have	NN	O	O
led	NN	O	O
to	NN	O	O
the	NN	O	O
discovery	NN	O	O
of	NN	O	O
a	NN	O	O
new	NN	O	O
lymphoid	NN	O	B-cell_type
cell	NN	O	I-cell_type
with	NN	O	O
unique	NN	O	O
antitumor	NN	O	O
activity	NN	O	O
.	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
form	NN	O	O
an	NN	O	O
antitumor	NN	O	O
surveillance	NN	O	O
system	NN	O	O
and	NN	O	O
appear	NN	O	O
to	NN	O	O
be	NN	O	O
vital	NN	O	O
in	NN	O	O
preventing	NN	O	O
tumor	NN	O	O
growth	NN	O	O
and	NN	O	O
metastasis	NN	O	O
in	NN	O	O
animal	NN	O	O
models	NN	O	O
.	NN	O	O

We	NN	O	O
studied	NN	O	O
NK	NN	O	B-cell_type
activity	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
benign	NN	O	O
and	NN	O	O
malignant	NN	O	O
breast	NN	O	O
disease	NN	O	O
,	NN	O	O
using	NN	O	O
a	NN	O	O
chromium-51	NN	O	O
release	NN	O	O
microtiter	NN	O	O
cytotoxicity	NN	O	O
assay	NN	O	O
with	NN	O	O
K562	NN	O	B-cell_line
cells	NN	O	I-cell_line
as	NN	O	O
targets	NN	O	O
.	NN	O	O

Compared	NN	O	O
with	NN	O	O
benign	NN	O	O
controls	NN	O	O
,	NN	O	O
patients	NN	O	O
with	NN	O	O
malignancies	NN	O	O
had	NN	O	O
significantly	NN	O	O
depressed	NN	O	O
NK	NN	O	B-cell_type
-mediated	NN	O	O
lysis	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

Furthermore	NN	O	O
,	NN	O	O
lysis	NN	O	O
in	NN	O	O
those	NN	O	O
with	NN	O	O
advanced	NN	O	O
disease	NN	O	O
(	NN	O	O
stages	NN	O	O
II	NN	O	O
,	NN	O	O
III	NN	O	O
,	NN	O	O
and	NN	O	O
IV	NN	O	O
)	NN	O	O
was	NN	O	O
significantly	NN	O	O
less	NN	O	O
than	NN	O	O
in	NN	O	O
those	NN	O	O
with	NN	O	O
limited	NN	O	O
disease	NN	O	O
(	NN	O	O
stage	NN	O	O
I	NN	O	O
)	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

NK	NN	O	B-cell_type
activity	NN	O	O
was	NN	O	O
not	NN	O	O
correlated	NN	O	O
to	NN	O	O
estrogen	NN	O	O
or	NN	O	O
progesterone	NN	O	O
receptor	NN	O	O
states	NN	O	O
.	NN	O	O

Positive	NN	O	O
correlation	NN	O	O
of	NN	O	O
a	NN	O	O
depressed	NN	O	O
natural	NN	O	O
killer	NN	O	O
activity	NN	O	O
with	NN	O	O
the	NN	O	O
extent	NN	O	O
of	NN	O	O
tumor	NN	O	O
spread	NN	O	O
supports	NN	O	O
the	NN	O	O
concept	NN	O	O
of	NN	O	O
an	NN	O	O
NK	NN	O	B-cell_type
cell	NN	O	I-cell_type
immune	NN	O	O
surveillance	NN	O	O
system	NN	O	O
in	NN	O	O
breast	NN	O	O
cancer	NN	O	O
and	NN	O	O
emphasizes	NN	O	O
its	NN	O	O
importance	NN	O	O
in	NN	O	O
this	NN	O	O
malignancy	NN	O	O
.	NN	O	O

-DOCSTART-	O

Immunosuppressive	NN	O	O
effect	NN	O	O
of	NN	O	O
serum	NN	O	O
progesterone	NN	O	O
during	NN	O	O
pregnancy	NN	O	O
depends	NN	O	O
on	NN	O	O
the	NN	O	O
progesterone	NN	O	O
binding	NN	O	O
capacity	NN	O	O
of	NN	O	O
the	NN	O	O
lymphocytes	NN	O	B-cell_type
.	NN	O	O

Cytotoxic	NN	O	O
activity	NN	O	O
and	NN	O	O
progesterone	NN	O	O
binding	NN	O	O
capacity	NN	O	O
of	NN	O	O
the	NN	O	O
lymphocytes	NN	O	B-cell_type
,	NN	O	O
together	NN	O	O
with	NN	O	O
serum	NN	O	O
progesterone	NN	O	O
concentrations	NN	O	O
,	NN	O	O
were	NN	O	O
determined	NN	O	O
in	NN	O	O
women	NN	O	O
with	NN	O	O
normal	NN	O	O
pregnancy	NN	O	O
or	NN	O	O
with	NN	O	O
a	NN	O	O
clinical	NN	O	O
diagnosis	NN	O	O
of	NN	O	O
threatened	NN	O	O
abortion	NN	O	O
or	NN	O	O
threatened	NN	O	O
premature	NN	O	O
labour	NN	O	O
.	NN	O	O

The	NN	O	O
lymphocytes	NN	O	O
of	NN	O	O
women	NN	O	O
with	NN	O	O
threatened	NN	O	O
abortion	NN	O	O
or	NN	O	O
threatened	NN	O	O
premature	NN	O	O
labour	NN	O	O
showed	NN	O	O
significantly	NN	O	O
higher	NN	O	O
cytotoxic	NN	O	O
activity	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
significantly	NN	O	O
lower	NN	O	O
progesterone	NN	O	O
binding	NN	O	O
capacity	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
than	NN	O	O
did	NN	O	O
lymphocytes	NN	O	B-cell_type
obtained	NN	O	O
from	NN	O	O
the	NN	O	O
healthy	NN	O	O
pregnant	NN	O	O
women	NN	O	O
.	NN	O	O

Significant	NN	O	O
inverse	NN	O	O
correlation	NN	O	O
was	NN	O	O
found	NN	O	O
between	NN	O	O
progesterone	NN	O	O
binding	NN	O	O
capacity	NN	O	O
and	NN	O	O
cytotoxic	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
lymphocytes	NN	O	B-cell_type
(	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
but	NN	O	O
the	NN	O	O
progesterone	NN	O	O
concentration	NN	O	O
of	NN	O	O
the	NN	O	O
pregnancy	NN	O	O
serum	NN	O	O
appeared	NN	O	O
to	NN	O	O
have	NN	O	O
no	NN	O	O
influence	NN	O	O
on	NN	O	O
the	NN	O	O
other	NN	O	O
two	NN	O	O
parameters	NN	O	O
.	NN	O	O

The	NN	O	O
findings	NN	O	O
indicate	NN	O	O
that	NN	O	O
intact	NN	O	O
progesterone	NN	O	O
binding	NN	O	O
capacity	NN	O	O
of	NN	O	O
the	NN	O	O
lymphocytes	NN	O	B-cell_type
is	NN	O	O
an	NN	O	O
essential	NN	O	O
factor	NN	O	O
for	NN	O	O
the	NN	O	O
manifestation	NN	O	O
of	NN	O	O
the	NN	O	O
blocking	NN	O	O
effect	NN	O	O
exerted	NN	O	O
by	NN	O	O
pregnancy	NN	O	O
serum	NN	O	O
on	NN	O	O
lymphocyte	NN	O	B-cell_type
cytotoxicity	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

-DOCSTART-	O

Serum	NN	O	O
sex	NN	O	O
steroid	NN	O	O
and	NN	O	O
peptide	NN	O	O
hormone	NN	O	O
concentrations	NN	O	O
,	NN	O	O
and	NN	O	O
endometrial	NN	O	O
estrogen	NN	O	O
and	NN	O	O
progestin	NN	O	O
receptor	NN	O	O
levels	NN	O	O
during	NN	O	O
administration	NN	O	O
of	NN	O	O
human	NN	O	B-protein
leukocyte	NN	O	I-protein
interferon	NN	O	I-protein
.	NN	O	O

Five	NN	O	O
normally	NN	O	O
cycling	NN	O	O
healthy	NN	O	O
women	NN	O	O
were	NN	O	O
given	NN	O	O
daily	NN	O	O
subcutaneous	NN	O	O
injections	NN	O	O
of	NN	O	O
human	NN	O	B-protein
leukocyte	NN	O	I-protein
interferon	NN	O	I-protein
(	NN	O	O
3	NN	O	O
X	NN	O	O
10	NN	O	O
(	NN	O	O
6	NN	O	O
)	NN	O	O
units/day	NN	O	O
)	NN	O	O
from	NN	O	O
the	NN	O	O
3rd	NN	O	O
through	NN	O	O
23rd	NN	O	O
day	NN	O	O
of	NN	O	O
the	NN	O	O
menstrual	NN	O	O
cycle	NN	O	O
,	NN	O	O
and	NN	O	O
serum	NN	O	O
steroid	NN	O	O
and	NN	O	O
peptide	NN	O	O
hormone	NN	O	O
concentrations	NN	O	O
monitored	NN	O	O
at	NN	O	O
3-day	NN	O	O
intervals	NN	O	O
during	NN	O	O
the	NN	O	O
treatment	NN	O	O
and	NN	O	O
the	NN	O	O
preceding	NN	O	O
control	NN	O	O
cycle	NN	O	O
.	NN	O	O

Concentrations	NN	O	O
of	NN	O	O
cytosol	NN	O	B-protein
and	NN	O	I-protein
nuclear	NN	O	I-protein
estrogen	NN	O	I-protein
receptors	NN	O	I-protein
(	NN	O	O
ERC	NN	O	B-protein
and	NN	O	O
ERN	NN	O	B-protein
,	NN	O	O
respectively	NN	O	O
)	NN	O	O
and	NN	O	O
progestin	NN	O	B-protein
receptors	NN	O	I-protein
(	NN	O	O
PRC	NN	O	B-protein
and	NN	O	O
PRN	NN	O	B-protein
)	NN	O	O
were	NN	O	O
also	NN	O	O
measured	NN	O	O
from	NN	O	O
endometrial	NN	O	O
biopsies	NN	O	O
taken	NN	O	O
on	NN	O	O
the	NN	O	O
24th	NN	O	O
day	NN	O	O
of	NN	O	O
the	NN	O	O
control	NN	O	O
and	NN	O	O
treatment	NN	O	O
cycle	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
an	NN	O	O
extensive	NN	O	O
monitoring	NN	O	O
of	NN	O	O
clinical	NN	O	O
chemical	NN	O	O
and	NN	O	O
hematological	NN	O	O
tests	NN	O	O
from	NN	O	O
the	NN	O	O
blood	NN	O	O
samples	NN	O	O
were	NN	O	O
performed	NN	O	O
.	NN	O	O

Serum	NN	O	O
estradiol	NN	O	O
and	NN	O	O
progesterone	NN	O	O
concentrations	NN	O	O
were	NN	O	O
significantly	NN	O	O
decreased	NN	O	O
during	NN	O	O
the	NN	O	O
treatment	NN	O	O
cycle	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
interferon	NN	O	B-protein
interacts	NN	O	O
in	NN	O	O
vivo	NN	O	O
with	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
both	NN	O	O
FSH	NN	O	O
and	NN	O	O
LH	NN	O	O
.	NN	O	O

No	NN	O	O
significant	NN	O	O
changes	NN	O	O
were	NN	O	O
observed	NN	O	O
in	NN	O	O
the	NN	O	O
serum	NN	O	O
peptide	NN	O	O
hormone	NN	O	O
concentrations	NN	O	O
measured	NN	O	O
(	NN	O	O
FSH	NN	O	O
,	NN	O	O
LH	NN	O	O
,	NN	O	O
prolactin	NN	O	O
,	NN	O	O
insulin	NN	O	O
,	NN	O	O
growth	NN	O	O
hormone	NN	O	O
and	NN	O	O
TSH	NN	O	O
)	NN	O	O
;	NN	O	O
neither	NN	O	O
were	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
endometrial	NN	O	B-protein
ERC	NN	O	I-protein
,	NN	O	I-protein
ERN	NN	O	I-protein
,	NN	O	I-protein
PRC	NN	O	I-protein
and	NN	O	I-protein
PRN	NN	O	I-protein
affected	NN	O	O
by	NN	O	O
interferon	NN	O	B-protein
administration	NN	O	O
.	NN	O	O

As	NN	O	O
expected	NN	O	O
,	NN	O	O
interferon	NN	O	B-protein
administration	NN	O	O
resulted	NN	O	O
in	NN	O	O
decreased	NN	O	O
leukocyte	NN	O	B-cell_type
counts	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
an	NN	O	O
increasing	NN	O	O
tendency	NN	O	O
in	NN	O	O
the	NN	O	O
activities	NN	O	O
of	NN	O	O
serum	NN	O	B-protein
alkaline	NN	O	I-protein
phosphatase	NN	O	I-protein
and	NN	O	O
gamma-glutamyltransferase	NN	O	B-protein
during	NN	O	O
the	NN	O	O
interferon	NN	O	B-protein
therapy	NN	O	O
shows	NN	O	O
that	NN	O	O
interferon	NN	O	B-protein
may	NN	O	O
slightly	NN	O	O
interfere	NN	O	O
with	NN	O	O
the	NN	O	O
liver	NN	O	O
function	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
by	NN	O	O
which	NN	O	O
interferon	NN	O	B-protein
treatment	NN	O	O
may	NN	O	O
affect	NN	O	O
the	NN	O	O
growth	NN	O	O
of	NN	O	O
hormone-dependent	NN	O	O
neoplasms	NN	O	O
could	NN	O	O
be	NN	O	O
the	NN	O	O
interaction	NN	O	O
with	NN	O	O
production	NN	O	O
and/or	NN	O	O
function	NN	O	O
of	NN	O	O
circulating	NN	O	O
hormonal	NN	O	O
compounds	NN	O	O
.	NN	O	O

-DOCSTART-	O

[	NN	O	O
Glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
level	NN	O	O
in	NN	O	O
the	NN	O	O
blood	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
in	NN	O	O
different	NN	O	O
acute	NN	O	O
diseases	NN	O	O
]	NN	O	O

Content	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
cytosol	NN	O	O
of	NN	O	O
blood	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
,	NN	O	O
concentration	NN	O	O
of	NN	O	O
cortisol	NN	O	O
and	NN	O	O
amount	NN	O	O
of	NN	O	O
leukocytes	NN	O	B-cell_type
in	NN	O	O
blood	NN	O	O
were	NN	O	O
studied	NN	O	O
in	NN	O	O
20	NN	O	O
patients	NN	O	O
with	NN	O	O
acute	NN	O	O
impairments	NN	O	O
within	NN	O	O
the	NN	O	O
second	NN	O	O
day	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
.	NN	O	O

Content	NN	O	O
of	NN	O	O
receptors	NN	O	O
in	NN	O	O
cytosol	NN	O	O
of	NN	O	O
blood	NN	O	O
leukocytes	NN	O	O
was	NN	O	O
studied	NN	O	O
using	NN	O	O
3H-triamcinolone	NN	O	O
acetonide	NN	O	O
.	NN	O	O

Distinct	NN	O	O
increase	NN	O	O
in	NN	O	O
amount	NN	O	O
of	NN	O	O
the	NN	O	O
leukocyte	NN	O	B-protein
glucocorticoid	NN	O	I-protein
receptors	NN	O	I-protein
was	NN	O	O
found	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
poisoning	NN	O	O
by	NN	O	O
dichlorethane	NN	O	O
and	NN	O	O
hypnotic	NN	O	O
drugs	NN	O	O
under	NN	O	O
conditions	NN	O	O
of	NN	O	O
acute	NN	O	O
myocardial	NN	O	O
infarction	NN	O	O
.	NN	O	O

In	NN	O	O
acute	NN	O	O
pancreatitis	NN	O	O
content	NN	O	O
of	NN	O	O
the	NN	O	O
leukocyte	NN	O	B-protein
receptors	NN	O	I-protein
was	NN	O	O
not	NN	O	O
altered	NN	O	O
as	NN	O	O
compared	NN	O	O
with	NN	O	O
controls	NN	O	O
.	NN	O	O

Concentration	NN	O	O
of	NN	O	O
endogenous	NN	O	O
cortisol	NN	O	O
was	NN	O	O
increased	NN	O	O
in	NN	O	O
blood	NN	O	O
of	NN	O	O
all	NN	O	O
the	NN	O	O
patients	NN	O	O
,	NN	O	O
except	NN	O	O
of	NN	O	O
the	NN	O	O
cases	NN	O	O
of	NN	O	O
acetate	NN	O	O
intoxication	NN	O	O
.	NN	O	O

Reverse	NN	O	O
correlation	NN	O	O
was	NN	O	O
observed	NN	O	O
between	NN	O	O
concentration	NN	O	O
of	NN	O	O
cortisol	NN	O	O
in	NN	O	O
blood	NN	O	O
and	NN	O	O
content	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	O
receptors	NN	O	O
in	NN	O	O
leukocytes	NN	O	B-cell_type
.	NN	O	O

But	NN	O	O
in	NN	O	O
the	NN	O	O
patients	NN	O	O
with	NN	O	O
acute	NN	O	O
pancreatitis	NN	O	O
the	NN	O	O
decrease	NN	O	O
in	NN	O	O
content	NN	O	O
of	NN	O	O
leukocyte	NN	O	B-protein
glucocorticoid	NN	O	I-protein
receptors	NN	O	I-protein
was	NN	O	O
not	NN	O	O
observed	NN	O	O
although	NN	O	O
there	NN	O	O
was	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
cortisol	NN	O	O
concentration	NN	O	O
in	NN	O	O
blood	NN	O	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
in	NN	O	O
immunological	NN	O	O
processes	NN	O	O
under	NN	O	O
conditions	NN	O	O
of	NN	O	O
purulent	NN	O	O
complications	NN	O	O
and	NN	O	O
possibility	NN	O	O
to	NN	O	O
regulate	NN	O	O
the	NN	O	O
metabolism	NN	O	O
in	NN	O	O
leukocytes	NN	O	B-cell_type

-DOCSTART-	O

Therapeutic	NN	O	O
concentrations	NN	O	O
of	NN	O	O
glucocorticoids	NN	O	O
suppress	NN	O	O
the	NN	O	O
antimicrobial	NN	O	O
activity	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
without	NN	O	O
impairing	NN	O	O
their	NN	O	O
responsiveness	NN	O	O
to	NN	O	O
gamma	NN	O	B-protein
interferon	NN	O	I-protein
.	NN	O	O

By	NN	O	O
exposing	NN	O	O
human	NN	O	B-cell_type
blood-derived	NN	O	I-cell_type
macrophages	NN	O	I-cell_type
and	NN	O	I-cell_type
alveolar	NN	O	I-cell_type
macrophages	NN	O	I-cell_type
in	NN	O	O
vitro	NN	O	O
to	NN	O	O
dexamethasone	NN	O	O
,	NN	O	O
we	NN	O	O
showed	NN	O	O
in	NN	O	O
these	NN	O	O
studies	NN	O	O
that	NN	O	O
glucocorticoids	NN	O	O
markedly	NN	O	O
suppress	NN	O	O
the	NN	O	O
antimicrobial	NN	O	O
activity	NN	O	O
of	NN	O	O
macrophages	NN	O	B-cell_type
but	NN	O	O
not	NN	O	O
macrophage	NN	O	B-cell_type
activation	NN	O	O
by	NN	O	O
lymphokines	NN	O	B-cell_type
.	NN	O	O

As	NN	O	O
little	NN	O	O
as	NN	O	O
2.5	NN	O	O
X	NN	O	O
10	NN	O	O
(	NN	O	O
-8	NN	O	O
)	NN	O	O
mol/liter	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
prevented	NN	O	O
macrophages	NN	O	B-cell_type
from	NN	O	O
inhibiting	NN	O	O
germination	NN	O	O
of	NN	O	O
Aspergillus	NN	O	O
spores	NN	O	O
or	NN	O	O
from	NN	O	O
eliminating	NN	O	O
ingested	NN	O	O
bacteria	NN	O	O
such	NN	O	O
as	NN	O	O
Listeria	NN	O	O
,	NN	O	O
Nocardia	NN	O	O
,	NN	O	O
or	NN	O	O
Salmonella	NN	O	O
.	NN	O	O

Damage	NN	O	O
to	NN	O	O
macrophage	NN	O	B-cell_type
function	NN	O	O
was	NN	O	O
inhibited	NN	O	O
by	NN	O	O
progesterone	NN	O	O
and	NN	O	O
appeared	NN	O	O
to	NN	O	O
be	NN	O	O
receptor-mediated	NN	O	O
.	NN	O	O

In	NN	O	O
accordance	NN	O	O
with	NN	O	O
in	NN	O	O
vivo	NN	O	O
observations	NN	O	O
,	NN	O	O
dexamethasone	NN	O	O
required	NN	O	O
24-36	NN	O	O
h	NN	O	O
to	NN	O	O
suppress	NN	O	O
antimicrobial	NN	O	O
activity	NN	O	O
.	NN	O	O

While	NN	O	O
glucocorticoids	NN	O	O
interfered	NN	O	O
with	NN	O	O
base-line	NN	O	O
activity	NN	O	O
of	NN	O	O
macrophages	NN	O	B-cell_type
,	NN	O	O
dexamethasone	NN	O	O
concentrations	NN	O	O
comparable	NN	O	O
to	NN	O	O
drug	NN	O	O
levels	NN	O	O
in	NN	O	O
patients	NN	O	O
had	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
macrophage	NN	O	B-cell_type
activation	NN	O	O
.	NN	O	O

Proliferating	NN	O	O
lymphocytes	NN	O	B-cell_type
and	NN	O	O
gamma-interferon	NN	O	B-protein
thus	NN	O	O
increased	NN	O	O
the	NN	O	O
antimicrobial	NN	O	O
activity	NN	O	O
of	NN	O	O
phagocytes	NN	O	B-cell_type
exposed	NN	O	O
to	NN	O	O
glucocorticoids	NN	O	O
over	NN	O	O
that	NN	O	O
of	NN	O	O
control	NN	O	O
cells	NN	O	O
.	NN	O	O

Macrophage	NN	O	O
activation	NN	O	O
and	NN	O	O
correction	NN	O	O
of	NN	O	O
the	NN	O	O
dexamethasone	NN	O	O
effect	NN	O	O
by	NN	O	O
gamma-interferon	NN	O	B-protein
,	NN	O	O
however	NN	O	O
,	NN	O	O
was	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
pathogen	NN	O	O
.	NN	O	O

The	NN	O	O
lymphokine	NN	O	B-protein
enhanced	NN	O	O
the	NN	O	O
antimicrobial	NN	O	O
activity	NN	O	O
of	NN	O	O
dexamethasone-treated	NN	O	B-cell_line
macrophages	NN	O	I-cell_line
against	NN	O	O
Listeria	NN	O	O
and	NN	O	O
Salmonella	NN	O	O
but	NN	O	O
not	NN	O	O
against	NN	O	O
Aspergillus	NN	O	O
or	NN	O	O
Nocardia	NN	O	O
.	NN	O	O

Dexamethasone-induced	NN	O	O
damage	NN	O	O
to	NN	O	O
the	NN	O	O
antimicrobial	NN	O	O
activity	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
in	NN	O	O
vitro	NN	O	O
parallels	NN	O	O
observations	NN	O	O
that	NN	O	O
glucocorticoids	NN	O	O
render	NN	O	O
laboratory	NN	O	O
animals	NN	O	O
susceptible	NN	O	O
to	NN	O	O
listeriosis	NN	O	O
and	NN	O	O
aspergillosis	NN	O	O
by	NN	O	O
damaging	NN	O	O
resident	NN	O	O
macrophages	NN	O	B-cell_line
.	NN	O	O

Suppression	NN	O	O
of	NN	O	O
macrophage	NN	O	B-cell_type
antimicrobial	NN	O	O
activity	NN	O	O
should	NN	O	O
thus	NN	O	O
be	NN	O	O
considered	NN	O	O
when	NN	O	O
treating	NN	O	O
patients	NN	O	O
with	NN	O	O
glucocorticoids	NN	O	O
;	NN	O	O
its	NN	O	O
prevention	NN	O	O
by	NN	O	O
gamma-interferon	NN	O	B-protein
might	NN	O	O
be	NN	O	O
beneficial	NN	O	O
for	NN	O	O
some	NN	O	O
but	NN	O	O
not	NN	O	O
all	NN	O	O
pathogens	NN	O	O
.	NN	O	O

-DOCSTART-	O

Interleukin	NN	O	B-protein
2	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
Tac	NN	O	B-protein
antigen	NN	O	I-protein
)	NN	O	O
expression	NN	O	O
in	NN	O	O
HTLV-I-associated	NN	O	O
adult	NN	O	O
T-cell	NN	O	B-cell_type
leukemia	NN	O	O
.	NN	O	O

Interleukin-2	NN	O	B-protein
(	NN	O	O
IL-2	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
lymphokine	NN	O	B-protein
synthesized	NN	O	O
by	NN	O	O
some	NN	O	O
T-cells	NN	O	B-cell_type
following	NN	O	O
activation	NN	O	O
.	NN	O	O

Resting	NN	O	O
T-cells	NN	O	B-cell_type
do	NN	O	O
not	NN	O	O
express	NN	O	O
IL-2	NN	O	B-protein
receptors	NN	O	I-protein
,	NN	O	O
but	NN	O	O
receptors	NN	O	O
are	NN	O	O
rapidly	NN	O	O
expressed	NN	O	O
on	NN	O	O
T-cells	NN	O	B-cell_type
following	NN	O	O
interaction	NN	O	O
of	NN	O	O
antigens	NN	O	O
,	NN	O	O
mitogens	NN	O	O
,	NN	O	O
or	NN	O	O
monoclonal	NN	O	B-protein
antibodies	NN	O	I-protein
with	NN	O	O
the	NN	O	O
antigen-specific	NN	O	B-protein
T-cell	NN	O	I-protein
receptor	NN	O	I-protein
complex	NN	O	I-protein
.	NN	O	O

Using	NN	O	O
anti-Tac	NN	O	B-protein
,	NN	O	O
a	NN	O	O
monoclonal	NN	O	B-protein
antibody	NN	O	I-protein
that	NN	O	O
recognizes	NN	O	O
the	NN	O	O
IL-2	NN	O	B-protein
receptor	NN	O	I-protein
,	NN	O	O
the	NN	O	O
receptor	NN	O	O
has	NN	O	O
been	NN	O	O
purified	NN	O	O
and	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
Mr	NN	O	O
33	NN	O	O
,	NN	O	O
000	NN	O	O
peptide	NN	O	O
that	NN	O	O
is	NN	O	O
posttranslationally	NN	O	O
glycosylated	NN	O	O
to	NN	O	O
a	NN	O	O
Mr	NN	O	O
55	NN	O	O
,	NN	O	O
000	NN	O	O
mature	NN	O	O
form	NN	O	O
.	NN	O	O

Normal	NN	O	O
resting	NN	O	O
T-cells	NN	O	B-cell_type
and	NN	O	O
most	NN	O	O
leukemic	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
populations	NN	O	I-cell_line
do	NN	O	O
not	NN	O	O
express	NN	O	O
IL-2	NN	O	B-protein
receptors	NN	O	I-protein
;	NN	O	O
however	NN	O	O
,	NN	O	O
the	NN	O	O
leukemic	NN	O	B-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
the	NN	O	O
11	NN	O	O
patients	NN	O	O
examined	NN	O	O
who	NN	O	O
had	NN	O	O
human	NN	O	O
T-cell	NN	O	O
lymphotropic	NN	O	O
virus-associated	NN	O	O
adult	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
expressed	NN	O	O
the	NN	O	O
Tac	NN	O	B-protein
antigen	NN	O	I-protein
.	NN	O	O

In	NN	O	O
human	NN	O	B-cell_type
T-cell	NN	O	I-cell_type
lymphotropic	NN	O	I-cell_type
virus-I	NN	O	I-cell_type
infected	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
the	NN	O	O
Mr	NN	O	O
42	NN	O	O
,	NN	O	O
000	NN	O	O
long	NN	O	O
open	NN	O	B-protein
reading	NN	O	I-protein
frame	NN	O	I-protein
protein	NN	O	I-protein
encoded	NN	O	O
in	NN	O	O
part	NN	O	O
by	NN	O	O
the	NN	O	O
pX	NN	O	B-DNA
region	NN	O	I-DNA
of	NN	O	O
this	NN	O	O
virus	NN	O	O
may	NN	O	O
act	NN	O	O
as	NN	O	O
a	NN	O	O
transacting	NN	O	B-protein
transcriptional	NN	O	I-protein
activator	NN	O	I-protein
that	NN	O	O
induces	NN	O	O
IL-2	NN	O	B-DNA
receptor	NN	O	I-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
,	NN	O	O
thus	NN	O	O
providing	NN	O	O
an	NN	O	O
explanation	NN	O	O
for	NN	O	O
the	NN	O	O
constant	NN	O	O
association	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
receptor	NN	O	I-protein
expression	NN	O	O
with	NN	O	O
adult	NN	O	O
T-cell	NN	O	O
lymphotropic	NN	O	O
virus-I	NN	O	O
infection	NN	O	O
of	NN	O	O
lymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
constant	NN	O	O
expression	NN	O	O
of	NN	O	O
large	NN	O	O
numbers	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
receptors	NN	O	I-protein
which	NN	O	O
may	NN	O	O
be	NN	O	O
aberrant	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
uncontrolled	NN	O	O
growth	NN	O	O
of	NN	O	O
adult	NN	O	B-cell_type
T-cell	NN	O	I-cell_type
leukemia	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Two	NN	O	O
patients	NN	O	O
with	NN	O	O
Tac-positive	NN	O	O
adult	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
have	NN	O	O
been	NN	O	O
treated	NN	O	O
with	NN	O	O
the	NN	O	O
anti-Tac	NN	O	B-protein
.	NN	O	O

One	NN	O	O
of	NN	O	O
the	NN	O	O
patients	NN	O	O
had	NN	O	O
6-	NN	O	O
and	NN	O	O
3-mo	NN	O	O
remissions	NN	O	O
of	NN	O	O
his	NN	O	O
leukemia	NN	O	O
following	NN	O	O
two	NN	O	O
courses	NN	O	O
of	NN	O	O
therapy	NN	O	O
with	NN	O	O
this	NN	O	O
monoclonal	NN	O	B-protein
antibody	NN	O	I-protein
directed	NN	O	O
toward	NN	O	O
this	NN	O	O
growth	NN	O	B-protein
factor	NN	O	I-protein
receptor	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Lymphocyte	NN	O	B-cell_type
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
binding	NN	O	O
in	NN	O	O
depressed	NN	O	O
patients	NN	O	O
with	NN	O	O
hypercortisolemia	NN	O	O
.	NN	O	O

Despite	NN	O	O
elevated	NN	O	O
levels	NN	O	O
of	NN	O	O
serum	NN	O	O
and	NN	O	O
urinary	NN	O	O
cortisol	NN	O	O
,	NN	O	O
patients	NN	O	O
with	NN	O	O
depressive	NN	O	O
illness	NN	O	O
manifest	NN	O	O
none	NN	O	O
of	NN	O	O
the	NN	O	O
clinical	NN	O	O
stigmata	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	O
excess	NN	O	O
.	NN	O	O

This	NN	O	O
hypercortisolemia	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
clinical	NN	O	O
effects	NN	O	O
suggests	NN	O	O
a	NN	O	O
state	NN	O	O
of	NN	O	O
hormone	NN	O	O
resistance	NN	O	O
and	NN	O	O
could	NN	O	O
be	NN	O	O
mediated	NN	O	O
by	NN	O	O
alterations	NN	O	O
in	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
.	NN	O	O

Earlier	NN	O	O
studies	NN	O	O
have	NN	O	O
shown	NN	O	O
that	NN	O	O
small	NN	O	O
doses	NN	O	O
of	NN	O	O
glucocorticoids	NN	O	O
cause	NN	O	O
a	NN	O	O
decrease	NN	O	O
in	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
binding	NN	O	O
in	NN	O	O
normal	NN	O	O
human	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

White	NN	O	O
cells	NN	O	O
from	NN	O	O
depressed	NN	O	O
patients	NN	O	O
with	NN	O	O
significant	NN	O	O
hypercortisolemia	NN	O	O
would	NN	O	O
be	NN	O	O
expected	NN	O	O
to	NN	O	O
show	NN	O	O
a	NN	O	O
similar	NN	O	O
change	NN	O	O
in	NN	O	O
receptor	NN	O	O
concentration	NN	O	O
if	NN	O	O
peripheral	NN	O	O
tissues	NN	O	O
are	NN	O	O
adequately	NN	O	O
exposed	NN	O	O
to	NN	O	O
and	NN	O	O
sensitive	NN	O	O
to	NN	O	O
the	NN	O	O
hormone	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
we	NN	O	O
compared	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
dexamethasone	NN	O	O
to	NN	O	O
lymphocytes	NN	O	B-cell_type
from	NN	O	O
normal	NN	O	O
subjects	NN	O	O
and	NN	O	O
depressed	NN	O	O
patients	NN	O	O
with	NN	O	O
hypercortisolemia	NN	O	O
.	NN	O	O

Lymphocytes	NN	O	B-cell_type
from	NN	O	O
normal	NN	O	O
subjects	NN	O	O
had	NN	O	O
a	NN	O	O
mean	NN	O	O
receptor	NN	O	O
concentration	NN	O	O
of	NN	O	O
10.2	NN	O	O
+/-	NN	O	O
0.66	NN	O	O
fm/10	NN	O	O
(	NN	O	O
6	NN	O	O
)	NN	O	O
cells	NN	O	O
(	NN	O	O
S.E.M.	NN	O	O
)	NN	O	O
and	NN	O	O
a	NN	O	O
dissociation	NN	O	O
constant	NN	O	O
of	NN	O	O
4.8	NN	O	O
+/-	NN	O	O
0.47	NN	O	O
nM	NN	O	O
.	NN	O	O

Lymphocytes	NN	O	B-cell_type
from	NN	O	O
depressed	NN	O	O
patients	NN	O	O
with	NN	O	O
abnormal	NN	O	O
0800	NN	O	O
h	NN	O	O
serum	NN	O	O
cortisol	NN	O	O
after	NN	O	O
dexamethasone	NN	O	O
had	NN	O	O
a	NN	O	O
mean	NN	O	O
receptor	NN	O	O
concentration	NN	O	O
of	NN	O	O
8.8	NN	O	O
+/-	NN	O	O
0.75	NN	O	O
fm/10	NN	O	O
(	NN	O	O
6	NN	O	O
)	NN	O	O
cells	NN	O	O
,	NN	O	O
which	NN	O	O
was	NN	O	O
not	NN	O	O
significantly	NN	O	O
different	NN	O	O
from	NN	O	O
that	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
from	NN	O	O
normal	NN	O	O
subjects	NN	O	O
or	NN	O	O
from	NN	O	O
depressed	NN	O	O
subjects	NN	O	O
with	NN	O	O
normal	NN	O	O
post-dexamethasone	NN	O	O
cortisol	NN	O	O
levels	NN	O	O
(	NN	O	O
9.4	NN	O	O
+/-	NN	O	O
0.95	NN	O	O
fm/10	NN	O	O
(	NN	O	O
6	NN	O	O
)	NN	O	O
cells	NN	O	O
)	NN	O	O
.	NN	O	O

Lymphocytes	NN	O	B-cell_type
from	NN	O	O
depressed	NN	O	O
patients	NN	O	O
with	NN	O	O
elevated	NN	O	O
urinary	NN	O	O
free	NN	O	O
cortisol	NN	O	O
excretion	NN	O	O
(	NN	O	O
UFC	NN	O	O
)	NN	O	O
also	NN	O	O
had	NN	O	O
normal	NN	O	O
receptor	NN	O	O
concentration	NN	O	O
and	NN	O	O
binding	NN	O	O
affinity	NN	O	O
for	NN	O	O
dexamethasone	NN	O	O
.	NN	O	O

The	NN	O	O
lack	NN	O	O
of	NN	O	O
a	NN	O	O
change	NN	O	O
in	NN	O	O
lymphocyte	NN	O	B-cell_type
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
concentration	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
cortisol	NN	O	O
excess	NN	O	O
suggests	NN	O	O
the	NN	O	O
possibility	NN	O	O
that	NN	O	O
hypercortisolemia	NN	O	O
in	NN	O	O
depressive	NN	O	O
illness	NN	O	O
represents	NN	O	O
a	NN	O	O
state	NN	O	O
of	NN	O	O
peripheral	NN	O	O
glucocorticoid	NN	O	O
resistance	NN	O	O
.	NN	O	O

-DOCSTART-	O

Structure	NN	O	O
and	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
hormone	NN	O	I-protein
receptor	NN	O	I-protein
.	NN	O	O

The	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
is	NN	O	O
an	NN	O	O
intracellular	NN	O	B-protein
protein	NN	O	I-protein
which	NN	O	O
possesses	NN	O	O
three	NN	O	O
distinct	NN	O	O
domains	NN	O	O
,	NN	O	O
one	NN	O	O
that	NN	O	O
binds	NN	O	O
agonist	NN	O	O
and	NN	O	O
antagonist	NN	O	O
steroids	NN	O	O
,	NN	O	O
one	NN	O	O
that	NN	O	O
binds	NN	O	O
DNA	NN	O	O
,	NN	O	O
and	NN	O	O
one	NN	O	O
that	NN	O	O
binds	NN	O	O
anti-receptor	NN	O	B-protein
antibodies	NN	O	I-protein
and	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
glucocorticoid	NN	O	O
modulation	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

In	NN	O	O
intact	NN	O	O
cells	NN	O	O
,	NN	O	O
receptor	NN	O	O
number	NN	O	O
,	NN	O	O
affinity	NN	O	O
and	NN	O	O
activity	NN	O	O
can	NN	O	O
change	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
factors	NN	O	O
that	NN	O	O
bind	NN	O	O
to	NN	O	O
the	NN	O	O
receptor	NN	O	O
,	NN	O	O
or	NN	O	O
that	NN	O	O
act	NN	O	O
indirectly	NN	O	O
through	NN	O	O
ill-defined	NN	O	O
mechanisms	NN	O	O
which	NN	O	O
may	NN	O	O
include	NN	O	O
resumption	NN	O	O
or	NN	O	O
arrest	NN	O	O
of	NN	O	O
cell	NN	O	O
cycling	NN	O	O
and	NN	O	O
variations	NN	O	O
in	NN	O	O
intracellular	NN	O	O
calcium	NN	O	O
ion	NN	O	O
concentrations	NN	O	O
.	NN	O	O

Some	NN	O	O
of	NN	O	O
these	NN	O	O
factors	NN	O	O
appear	NN	O	O
to	NN	O	O
exert	NN	O	O
their	NN	O	O
effect	NN	O	O
by	NN	O	O
controlling	NN	O	O
critical	NN	O	O
receptor	NN	O	O
properties	NN	O	O
such	NN	O	O
as	NN	O	O
ATP-dependent	NN	O	O
phosphorylation	NN	O	O
,	NN	O	O
integrity	NN	O	O
of	NN	O	O
thiol	NN	O	B-protein
groups	NN	O	I-protein
,	NN	O	O
and	NN	O	O
exposure	NN	O	O
of	NN	O	O
key	NN	O	B-protein
amino	NN	O	I-protein
acid	NN	O	I-protein
residues	NN	O	I-protein
.	NN	O	O

Glucocorticoid	NN	O	O
agonists	NN	O	O
promote	NN	O	O
the	NN	O	O
'transformation	NN	O	O
'	NN	O	O
of	NN	O	O
the	NN	O	O
receptor	NN	O	O
into	NN	O	O
the	NN	O	O
DNA-binding	NN	O	O
state	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
competent	NN	O	O
for	NN	O	O
modulating	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

Glucocorticoid	NN	O	O
antagonists	NN	O	O
are	NN	O	O
steroids	NN	O	O
that	NN	O	O
interact	NN	O	O
with	NN	O	O
the	NN	O	O
receptor	NN	O	O
but	NN	O	O
either	NN	O	O
fail	NN	O	O
to	NN	O	O
produce	NN	O	O
a	NN	O	O
stable	NN	O	B-protein
complex	NN	O	I-protein
or	NN	O	O
produce	NN	O	O
a	NN	O	O
stable	NN	O	B-protein
but	NN	O	I-protein
inefficient	NN	O	I-protein
complex	NN	O	I-protein
.	NN	O	O

Although	NN	O	O
substituent	NN	O	O
groups	NN	O	O
that	NN	O	O
confer	NN	O	O
agonist	NN	O	O
or	NN	O	O
antagonist	NN	O	O
activity	NN	O	O
to	NN	O	O
the	NN	O	O
steroid	NN	O	O
have	NN	O	O
been	NN	O	O
identified	NN	O	O
,	NN	O	O
the	NN	O	O
molecular	NN	O	O
determinants	NN	O	O
of	NN	O	O
this	NN	O	O
difference	NN	O	O
at	NN	O	O
the	NN	O	O
receptor	NN	O	O
level	NN	O	O
remain	NN	O	O
unknown	NN	O	O
.	NN	O	O

Most	NN	O	O
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
in	NN	O	O
vivo	NN	O	O
data	NN	O	O
on	NN	O	O
receptor	NN	O	O
regulation	NN	O	O
can	NN	O	O
be	NN	O	O
accommodated	NN	O	O
by	NN	O	O
postulating	NN	O	O
the	NN	O	O
existence	NN	O	O
of	NN	O	O
an	NN	O	O
intracellular	NN	O	O
cycle	NN	O	O
that	NN	O	O
involves	NN	O	O
five	NN	O	O
states	NN	O	O
of	NN	O	O
the	NN	O	O
receptor	NN	O	O
.	NN	O	O

The	NN	O	O
active	NN	O	O
free	NN	O	O
receptor	NN	O	O
is	NN	O	O
phosphorylated	NN	O	O
,	NN	O	O
reduced	NN	O	O
,	NN	O	O
and	NN	O	O
presumably	NN	O	O
oligomeric	NN	O	O
(	NN	O	O
state	NN	O	O
A	NN	O	O
)	NN	O	O
.	NN	O	O

Following	NN	O	O
binding	NN	O	O
of	NN	O	O
an	NN	O	O
agonist	NN	O	O
(	NN	O	O
state	NN	O	O
B	NN	O	O
)	NN	O	O
,	NN	O	O
it	NN	O	O
can	NN	O	O
become	NN	O	O
transformed	NN	O	O
by	NN	O	O
dissociation	NN	O	O
into	NN	O	O
its	NN	O	O
subunits	NN	O	O
and	NN	O	O
dephosphorylation	NN	O	O
(	NN	O	O
state	NN	O	O
C	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
transformed	NN	O	B-protein
receptor	NN	O	I-protein
then	NN	O	O
interacts	NN	O	O
with	NN	O	O
chromatin	NN	O	B-DNA
(	NN	O	O
state	NN	O	O
D	NN	O	O
)	NN	O	O
.	NN	O	O

Dissociation	NN	O	O
of	NN	O	O
the	NN	O	O
steroid	NN	O	O
and	NN	O	O
oxidation	NN	O	O
of	NN	O	O
receptor	NN	O	B-protein
thiol	NN	O	I-protein
group	NN	O	I-protein
(	NN	O	O
s	NN	O	O
)	NN	O	O
lead	NN	O	O
to	NN	O	O
the	NN	O	O
inactive	NN	O	O
receptor	NN	O	O
form	NN	O	O
(	NN	O	O
state	NN	O	O
E	NN	O	O
)	NN	O	O
.	NN	O	O

Reduction	NN	O	O
and	NN	O	O
rephosphorylation	NN	O	O
of	NN	O	O
the	NN	O	O
receptor	NN	O	O
enable	NN	O	O
it	NN	O	O
to	NN	O	O
bind	NN	O	O
steroids	NN	O	O
again	NN	O	O
so	NN	O	O
that	NN	O	O
the	NN	O	O
cycle	NN	O	O
is	NN	O	O
closed	NN	O	O
.	NN	O	O

-DOCSTART-	O

Thermodynamics	NN	O	O
of	NN	O	O
steroid	NN	O	O
binding	NN	O	O
to	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

The	NN	O	O
thermodynamics	NN	O	O
of	NN	O	O
the	NN	O	O
interaction	NN	O	O
of	NN	O	O
glucocorticoids	NN	O	O
with	NN	O	O
their	NN	O	O
receptor	NN	O	O
were	NN	O	O
studied	NN	O	O
in	NN	O	O
cytosol	NN	O	O
from	NN	O	O
human	NN	O	B-cell_type
lymphoblastoid	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
rate	NN	O	O
and	NN	O	O
affinity	NN	O	O
constants	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
and	NN	O	O
cortisol	NN	O	O
between	NN	O	O
0	NN	O	O
degree	NN	O	O
and	NN	O	O
25	NN	O	O
degrees	NN	O	O
C	NN	O	O
were	NN	O	O
calculated	NN	O	O
by	NN	O	O
curve-fitting	NN	O	O
from	NN	O	O
time-course	NN	O	O
and	NN	O	O
equilibrium	NN	O	O
kinetics	NN	O	O
.	NN	O	O

The	NN	O	O
data	NN	O	O
were	NN	O	O
consistent	NN	O	O
with	NN	O	O
a	NN	O	O
simple	NN	O	O
reversible	NN	O	O
bimolecular	NN	O	O
interaction	NN	O	O
.	NN	O	O

Arrhenius	NN	O	O
and	NN	O	O
Va	NN	O	O
n't	NN	O	O
Hoff	NN	O	O
plots	NN	O	O
were	NN	O	O
curvilinear	NN	O	O
for	NN	O	O
both	NN	O	O
steroids	NN	O	O
.	NN	O	O

At	NN	O	O
equilibrium	NN	O	O
,	NN	O	O
the	NN	O	O
solution	NN	O	O
for	NN	O	O
the	NN	O	O
equation	NN	O	O
delta	NN	O	O
G	NN	O	O
=	NN	O	O
delta	NN	O	O
H	NN	O	O
-	NN	O	O
T	NN	O	O
X	NN	O	O
delta	NN	O	O
S	NN	O	O
(	NN	O	O
eqn.	NN	O	O
1	NN	O	O
)	NN	O	O
was	NN	O	O
(	NN	O	O
in	NN	O	O
kJ	NN	O	O
X	NN	O	O
mol-1	NN	O	O
)	NN	O	O
-47	NN	O	O
=	NN	O	O
36	NN	O	O
-	NN	O	O
83	NN	O	O
(	NN	O	O
dexamethasone	NN	O	O
)	NN	O	O
and	NN	O	O
-42	NN	O	O
=	NN	O	O
-9	NN	O	O
-	NN	O	O
33	NN	O	O
(	NN	O	O
cortisol	NN	O	O
)	NN	O	O
at	NN	O	O
0	NN	O	O
degree	NN	O	O
C	NN	O	O
.	NN	O	O

Enthalpy	NN	O	O
and	NN	O	O
entropy	NN	O	O
changes	NN	O	O
decreased	NN	O	O
quasi-linearly	NN	O	O
with	NN	O	O
temperature	NN	O	O
such	NN	O	O
that	NN	O	O
,	NN	O	O
at	NN	O	O
25	NN	O	O
degrees	NN	O	O
C	NN	O	O
,	NN	O	O
the	NN	O	O
respective	NN	O	O
values	NN	O	O
were	NN	O	O
-50	NN	O	O
=	NN	O	O
-75	NN	O	O
+	NN	O	O
25	NN	O	O
and	NN	O	O
-43	NN	O	O
=	NN	O	O
-48	NN	O	O
+	NN	O	O
5	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
for	NN	O	O
both	NN	O	O
steroids	NN	O	O
,	NN	O	O
the	NN	O	O
interaction	NN	O	O
was	NN	O	O
entropy-driven	NN	O	O
at	NN	O	O
low	NN	O	O
temperature	NN	O	O
and	NN	O	O
became	NN	O	O
entirely	NN	O	O
enthalpy-driven	NN	O	O
at	NN	O	O
20	NN	O	O
degrees	NN	O	O
C	NN	O	O
.	NN	O	O

Thermodynamic	NN	O	O
values	NN	O	O
for	NN	O	O
the	NN	O	O
transition	NN	O	O
state	NN	O	O
were	NN	O	O
calculated	NN	O	O
from	NN	O	O
the	NN	O	O
rate	NN	O	O
constants	NN	O	O
.	NN	O	O

For	NN	O	O
the	NN	O	O
forward	NN	O	O
reaction	NN	O	O
,	NN	O	O
eqn.	NN	O	O
(	NN	O	O
1	NN	O	O
)	NN	O	O
gave	NN	O	O
45	NN	O	O
=	NN	O	O
84	NN	O	O
-	NN	O	O
39	NN	O	O
(	NN	O	O
dexamethasone	NN	O	O
)	NN	O	O
and	NN	O	O
46	NN	O	O
=	NN	O	O
60	NN	O	O
-	NN	O	O
14	NN	O	O
(	NN	O	O
cortisol	NN	O	O
)	NN	O	O
at	NN	O	O
0	NN	O	O
degree	NN	O	O
C	NN	O	O
,	NN	O	O
and	NN	O	O
44	NN	O	O
=	NN	O	O
24	NN	O	O
+	NN	O	O
20	NN	O	O
(	NN	O	O
dexamethasone	NN	O	O
)	NN	O	O
and	NN	O	O
46	NN	O	O
=	NN	O	O
28	NN	O	O
+	NN	O	O
18	NN	O	O
(	NN	O	O
cortisol	NN	O	O
)	NN	O	O
at	NN	O	O
25	NN	O	O
degrees	NN	O	O
C	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
fit	NN	O	O
quite	NN	O	O
well	NN	O	O
with	NN	O	O
a	NN	O	O
two-step	NN	O	O
model	NN	O	O
[	NN	O	O
Ross	NN	O	O
&	NN	O	O
Subramanian	NN	O	O
(	NN	O	O
1981	NN	O	O
)	NN	O	O
Biochemistry	NN	O	O
20	NN	O	O
,	NN	O	O
3096-3102	NN	O	O
]	NN	O	O
proposed	NN	O	O
for	NN	O	O
ligand-protein	NN	O	O
interactions	NN	O	O
,	NN	O	O
which	NN	O	O
involves	NN	O	O
a	NN	O	O
partial	NN	O	O
immobilization	NN	O	O
of	NN	O	O
the	NN	O	O
reacting	NN	O	O
species	NN	O	O
governed	NN	O	O
by	NN	O	O
hydrophobic	NN	O	O
forces	NN	O	O
,	NN	O	O
followed	NN	O	O
by	NN	O	O
stabilization	NN	O	O
of	NN	O	O
the	NN	O	O
complex	NN	O	O
by	NN	O	O
short-range	NN	O	O
interactions	NN	O	O
.	NN	O	O

On	NN	O	O
the	NN	O	O
basis	NN	O	O
of	NN	O	O
this	NN	O	O
model	NN	O	O
,	NN	O	O
an	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
transition-state	NN	O	O
thermodynamics	NN	O	O
led	NN	O	O
to	NN	O	O
the	NN	O	O
conclusion	NN	O	O
that	NN	O	O
no	NN	O	O
more	NN	O	O
than	NN	O	O
half	NN	O	O
of	NN	O	O
the	NN	O	O
steroid	NN	O	O
molecular	NN	O	O
area	NN	O	O
is	NN	O	O
engaged	NN	O	O
in	NN	O	O
the	NN	O	O
binding	NN	O	O
process	NN	O	O
.	NN	O	O

-DOCSTART-	O

Cell	NN	O	O
cycle-related	NN	O	O
changes	NN	O	O
in	NN	O	O
number	NN	O	O
of	NN	O	O
T-lymphocyte	NN	O	B-protein
receptors	NN	O	I-protein
for	NN	O	O
glucocorticoids	NN	O	O
and	NN	O	O
insulin	NN	O	O
.	NN	O	O

Enriched	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
T-lymphocytes	NN	O	I-cell_type
were	NN	O	O
stimulated	NN	O	O
with	NN	O	O
PHA	NN	O	O
and	NN	O	O
examined	NN	O	O
for	NN	O	O
variations	NN	O	O
in	NN	O	O
insulin	NN	O	O
and	NN	O	O
glucocorticoid	NN	O	O
(	NN	O	O
dexamethasone	NN	O	O
)	NN	O	O
receptor	NN	O	O
numbers	NN	O	O
during	NN	O	O
the	NN	O	O
early	NN	O	O
phases	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
.	NN	O	O

Cells	NN	O	O
in	NN	O	O
G0	NN	O	O
,	NN	O	O
G1a	NN	O	O
and	NN	O	O
G1b	NN	O	O
phases	NN	O	O
,	NN	O	O
where	NN	O	O
the	NN	O	O
G1a	NN	O	O
-	NN	O	O
G1b	NN	O	O
transition	NN	O	O
is	NN	O	O
an	NN	O	O
Interleukin	NN	O	B-protein
2	NN	O	I-protein
dependent	NN	O	O
event	NN	O	O
,	NN	O	O
were	NN	O	O
quantitated	NN	O	O
by	NN	O	O
flow	NN	O	O
cytometry	NN	O	O
.	NN	O	O

Few	NN	O	O
but	NN	O	O
significant	NN	O	O
numbers	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
(	NN	O	O
2700/cell	NN	O	O
)	NN	O	O
and	NN	O	O
no	NN	O	O
insulin	NN	O	B-protein
receptors	NN	O	I-protein
(	NN	O	O
-1/cell	NN	O	O
)	NN	O	O
were	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
resting	NN	O	O
(	NN	O	O
G0	NN	O	O
)	NN	O	O
phase	NN	O	O
.	NN	O	O

As	NN	O	O
cells	NN	O	O
entered	NN	O	O
the	NN	O	O
G1a	NN	O	O
phase	NN	O	O
the	NN	O	O
specific	NN	O	O
binding	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
increased	NN	O	O
and	NN	O	O
of	NN	O	O
insulin	NN	O	O
took	NN	O	O
place	NN	O	O
.	NN	O	O

Although	NN	O	O
the	NN	O	O
specific	NN	O	O
binding	NN	O	O
further	NN	O	O
increased	NN	O	O
as	NN	O	O
T-cells	NN	O	B-cell_type
entered	NN	O	O
the	NN	O	O
G1b	NN	O	O
phase	NN	O	O
(	NN	O	O
as	NN	O	O
measured	NN	O	O
at	NN	O	O
44	NN	O	O
h	NN	O	O
of	NN	O	O
incubation	NN	O	O
and	NN	O	O
using	NN	O	O
hydroxyurea-treated	NN	O	B-cell_line
cells	NN	O	I-cell_line
)	NN	O	O
,	NN	O	O
the	NN	O	O
major	NN	O	O
changes	NN	O	O
in	NN	O	O
the	NN	O	O
specific	NN	O	O
binding	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
took	NN	O	O
place	NN	O	O
during	NN	O	O
the	NN	O	O
period	NN	O	O
16	NN	O	O
-	NN	O	O
20	NN	O	O
h	NN	O	O
after	NN	O	O
stimulation	NN	O	O
.	NN	O	O

Based	NN	O	O
on	NN	O	O
these	NN	O	O
findings	NN	O	O
,	NN	O	O
it	NN	O	O
is	NN	O	O
concluded	NN	O	O
that	NN	O	O
both	NN	O	O
receptor	NN	O	O
types	NN	O	O
(	NN	O	O
cell	NN	O	B-protein
membrane	NN	O	I-protein
and	NN	O	I-protein
cytoplasmic	NN	O	I-protein
receptors	NN	O	I-protein
)	NN	O	O
are	NN	O	O
being	NN	O	O
formed	NN	O	O
and	NN	O	O
increased	NN	O	O
at	NN	O	O
G1	NN	O	O
phase	NN	O	O
prior	NN	O	O
to	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
,	NN	O	O
indicating	NN	O	O
the	NN	O	O
importance	NN	O	O
of	NN	O	O
G1	NN	O	O
phase	NN	O	O
in	NN	O	O
immunoregulation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
and	NN	O	O
cortico-sensitivity	NN	O	O
in	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
clonal	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
CM-SM	NN	O	B-cell_line
.	NN	O	O

CM-SM	NN	O	B-cell_line
is	NN	O	O
a	NN	O	O
clonal	NN	O	B-cell_line
line	NN	O	I-cell_line
of	NN	O	O
human	NN	O	B-cell_type
precursor	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
phagocytes	NN	O	I-cell_type
inducible	NN	O	O
to	NN	O	O
macrophage	NN	O	B-cell_type
differentiation	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
the	NN	O	O
tumor	NN	O	O
promoter	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
12-O-tetradecanoyl-phorbol-13-acetate	NN	O	O
(	NN	O	O
TPA	NN	O	O
)	NN	O	O
.	NN	O	O

Untreated	NN	O	O
CM-SM	NN	O	O
cells	NN	O	O
contain	NN	O	O
single	NN	O	O
class	NN	O	O
,	NN	O	O
high-affinity	NN	O	O
(	NN	O	O
KD	NN	O	O
=	NN	O	O
4.0	NN	O	O
X	NN	O	O
10	NN	O	O
(	NN	O	O
-9	NN	O	O
)	NN	O	O
M	NN	O	O
)	NN	O	O
glucocorticoid-specific	NN	O	O
receptor	NN	O	O
sites	NN	O	O
(	NN	O	O
approximately	NN	O	O
60	NN	O	O
,	NN	O	O
000	NN	O	O
per	NN	O	O
cell	NN	O	O
)	NN	O	O
,	NN	O	O
as	NN	O	O
measured	NN	O	O
by	NN	O	O
a	NN	O	O
whole	NN	O	O
cell	NN	O	O
assay	NN	O	O
,	NN	O	O
at	NN	O	O
37	NN	O	O
degrees	NN	O	O
C	NN	O	O
,	NN	O	O
using	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
triamcinolone	NN	O	O
acetonide	NN	O	O
(	NN	O	O
TA	NN	O	O
)	NN	O	O
.	NN	O	O

Exposure	NN	O	O
of	NN	O	O
CM-SM	NN	O	B-cell_line
to	NN	O	O
dexamethasone	NN	O	O
(	NN	O	O
DEX	NN	O	O
)	NN	O	O
produced	NN	O	O
a	NN	O	O
progressive	NN	O	O
,	NN	O	O
dose-	NN	O	O
and	NN	O	O
time-related	NN	O	O
series	NN	O	O
of	NN	O	O
changes	NN	O	O
in	NN	O	O
CM-SM	NN	O	B-cell_line
cell	NN	O	O
growth	NN	O	O
,	NN	O	O
saturation	NN	O	O
density	NN	O	O
,	NN	O	O
morphology	NN	O	O
,	NN	O	O
and	NN	O	O
functional	NN	O	O
properties	NN	O	O
,	NN	O	O
with	NN	O	O
half-maximal	NN	O	O
effects	NN	O	O
at	NN	O	O
about	NN	O	O
10	NN	O	O
(	NN	O	O
-9	NN	O	O
)	NN	O	O
M	NN	O	O
for	NN	O	O
DEX	NN	O	O
.	NN	O	O

TA-receptor	NN	O	O
sites	NN	O	O
rapidly	NN	O	O
decreased	NN	O	O
(	NN	O	O
about	NN	O	O
70	NN	O	O
%	NN	O	O
)	NN	O	O
after	NN	O	O
DEX	NN	O	O
treatment	NN	O	O
,	NN	O	O
without	NN	O	O
any	NN	O	O
apparent	NN	O	O
change	NN	O	O
in	NN	O	O
steroid	NN	O	O
specificity	NN	O	O
and	NN	O	O
affinity	NN	O	O
.	NN	O	O

After	NN	O	O
5	NN	O	O
days	NN	O	O
in	NN	O	O
culture	NN	O	O
with	NN	O	O
a	NN	O	O
saturating	NN	O	O
concentration	NN	O	O
(	NN	O	O
3.6	NN	O	O
X	NN	O	O
10	NN	O	O
(	NN	O	O
-8	NN	O	O
)	NN	O	O
M	NN	O	O
)	NN	O	O
of	NN	O	O
hormone	NN	O	O
,	NN	O	O
the	NN	O	O
cells	NN	O	O
reached	NN	O	O
a	NN	O	O
saturation	NN	O	O
density	NN	O	O
of	NN	O	O
about	NN	O	O
9.0	NN	O	O
X	NN	O	O
10	NN	O	O
(	NN	O	O
6	NN	O	O
)	NN	O	O
viable	NN	O	O
cells/ml	NN	O	O
(	NN	O	O
about	NN	O	O
4.0	NN	O	O
X	NN	O	O
10	NN	O	O
(	NN	O	O
6	NN	O	O
)	NN	O	O
viable	NN	O	O
cells/ml	NN	O	O
in	NN	O	O
the	NN	O	O
controls	NN	O	O
)	NN	O	O
,	NN	O	O
while	NN	O	O
the	NN	O	O
modal	NN	O	O
volume	NN	O	O
of	NN	O	O
the	NN	O	O
resulting	NN	O	O
cell	NN	O	B-cell_line
population	NN	O	I-cell_line
was	NN	O	O
approximately	NN	O	O
60	NN	O	O
%	NN	O	O
,	NN	O	O
as	NN	O	O
compared	NN	O	O
to	NN	O	O
the	NN	O	O
volume	NN	O	O
of	NN	O	O
untreated	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

DEX-treated	NN	O	B-cell_line
cells	NN	O	I-cell_line
appeared	NN	O	O
less	NN	O	O
differentiated	NN	O	O
than	NN	O	O
controls	NN	O	O
,	NN	O	O
as	NN	O	O
assessed	NN	O	O
by	NN	O	O
combined	NN	O	O
morphologic	NN	O	O
,	NN	O	O
antigenic	NN	O	O
,	NN	O	O
and	NN	O	O
cytoenzymatic	NN	O	O
analyses	NN	O	O
.	NN	O	O

DEX	NN	O	O
almost	NN	O	O
completely	NN	O	O
inhibited	NN	O	O
TPA	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
following	NN	O	O
macrophage	NN	O	B-cell_type
functions	NN	O	O
:	NN	O	O
adherency	NN	O	O
to	NN	O	O
the	NN	O	O
culture	NN	O	O
plate	NN	O	O
,	NN	O	O
expression	NN	O	O
of	NN	O	O
lysosomal	NN	O	B-protein
enzymes	NN	O	I-protein
,	NN	O	O
Fc	NN	O	B-protein
and	NN	O	I-protein
C3	NN	O	I-protein
receptors	NN	O	I-protein
,	NN	O	O
and	NN	O	O
stimulation	NN	O	O
of	NN	O	O
phagocytosis	NN	O	O
.	NN	O	O

After	NN	O	O
removal	NN	O	O
of	NN	O	O
DEX	NN	O	O
,	NN	O	O
the	NN	O	O
cells	NN	O	O
,	NN	O	O
within	NN	O	O
a	NN	O	O
few	NN	O	O
passages	NN	O	O
,	NN	O	O
returned	NN	O	O
to	NN	O	O
a	NN	O	O
state	NN	O	O
apparently	NN	O	O
identical	NN	O	O
to	NN	O	O
the	NN	O	O
untreated	NN	O	O
controls	NN	O	O
and	NN	O	O
could	NN	O	O
be	NN	O	O
induced	NN	O	O
to	NN	O	O
macrophage	NN	O	B-cell_type
differentiation	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
TPA	NN	O	O
.	NN	O	O

-DOCSTART-	O

Acute	NN	O	O
lymphoblastic	NN	O	O
leukemia	NN	O	O
in	NN	O	O
children	NN	O	O
:	NN	O	O
current	NN	O	O
status	NN	O	O
,	NN	O	O
controversies	NN	O	O
,	NN	O	O
and	NN	O	O
future	NN	O	O
perspective	NN	O	O
.	NN	O	O

Disease-free	NN	O	O
survival	NN	O	O
(	NN	O	O
DFS	NN	O	O
)	NN	O	O
in	NN	O	O
childhood	NN	O	O
ALL	NN	O	O
is	NN	O	O
60	NN	O	O
%	NN	O	O
,	NN	O	O
and	NN	O	O
survival	NN	O	O
in	NN	O	O
good	NN	O	O
,	NN	O	O
average	NN	O	O
,	NN	O	O
and	NN	O	O
poor	NN	O	O
prognostic	NN	O	O
groups	NN	O	O
defined	NN	O	O
by	NN	O	O
initial	NN	O	O
WBC	NN	O	O
and	NN	O	O
age	NN	O	O
is	NN	O	O
90	NN	O	O
,	NN	O	O
60	NN	O	O
,	NN	O	O
and	NN	O	O
45	NN	O	O
%	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

Additional	NN	O	O
immunological	NN	O	O
,	NN	O	O
morphological	NN	O	O
,	NN	O	O
biochemical	NN	O	O
,	NN	O	O
cytokinetic	NN	O	O
,	NN	O	O
and	NN	O	O
cytogenetic	NN	O	O
factors	NN	O	O
have	NN	O	O
been	NN	O	O
identified	NN	O	O
,	NN	O	O
illustrating	NN	O	O
the	NN	O	O
heterogeneity	NN	O	O
of	NN	O	O
ALL	NN	O	O
and	NN	O	O
its	NN	O	O
derivation	NN	O	O
from	NN	O	O
malignant	NN	O	B-cell_line
clones	NN	O	I-cell_line
at	NN	O	O
various	NN	O	O
stages	NN	O	O
of	NN	O	O
differentiation	NN	O	O
and	NN	O	O
with	NN	O	O
varying	NN	O	O
rates	NN	O	O
of	NN	O	O
proliferation	NN	O	O
.	NN	O	O

Of	NN	O	O
biologic	NN	O	O
importance	NN	O	O
,	NN	O	O
these	NN	O	O
factors	NN	O	O
may	NN	O	O
refine	NN	O	O
further	NN	O	O
the	NN	O	O
characteristic	NN	O	O
features	NN	O	O
of	NN	O	O
clinically-determined	NN	O	O
prognostic	NN	O	O
groups	NN	O	O
.	NN	O	O

Multivariate	NN	O	O
analysis	NN	O	O
of	NN	O	O
large	NN	O	O
prospective	NN	O	O
trials	NN	O	O
with	NN	O	O
homogeneous	NN	O	O
therapy	NN	O	O
will	NN	O	O
be	NN	O	O
required	NN	O	O
to	NN	O	O
determine	NN	O	O
the	NN	O	O
independent	NN	O	O
prognostic	NN	O	O
importance	NN	O	O
of	NN	O	O
these	NN	O	O
factors	NN	O	O
.	NN	O	O

Current	NN	O	O
treatment	NN	O	O
strategies	NN	O	O
in	NN	O	O
ALL	NN	O	O
include	NN	O	O
(	NN	O	O
1	NN	O	O
)	NN	O	O
tailoring	NN	O	O
therapy	NN	O	O
and	NN	O	O
its	NN	O	O
intensity	NN	O	O
to	NN	O	O
prognostic	NN	O	O
groups	NN	O	O
;	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
multiple-drug	NN	O	O
combinations	NN	O	O
in	NN	O	O
induction	NN	O	O
;	NN	O	O
(	NN	O	O
3	NN	O	O
)	NN	O	O
early	NN	O	O
use	NN	O	O
of	NN	O	O
intrathecal	NN	O	O
(	NN	O	O
IT	NN	O	O
)	NN	O	O
methotrexate	NN	O	O
(	NN	O	O
MTX	NN	O	O
)	NN	O	O
;	NN	O	O
(	NN	O	O
4	NN	O	O
)	NN	O	O
CNS	NN	O	O
prophylaxis	NN	O	O
with	NN	O	O
IT	NN	O	O
MTX	NN	O	O
alone	NN	O	O
in	NN	O	O
good	NN	O	O
prognosis	NN	O	O
patients	NN	O	O
and	NN	O	O
combined	NN	O	O
cranial	NN	O	O
radiation	NN	O	O
(	NN	O	O
CXRT	NN	O	O
)	NN	O	O
,	NN	O	O
1800	NN	O	O
rads	NN	O	O
plus	NN	O	O
IT	NN	O	O
MTX	NN	O	O
,	NN	O	O
in	NN	O	O
average	NN	O	O
and	NN	O	O
poor	NN	O	O
prognosis	NN	O	O
patients	NN	O	O
.	NN	O	O

Current	NN	O	O
studies	NN	O	O
show	NN	O	O
a	NN	O	O
CNS	NN	O	O
relapse	NN	O	O
rate	NN	O	O
of	NN	O	O
5	NN	O	O
%	NN	O	O
in	NN	O	O
all	NN	O	O
prognostic	NN	O	O
groups	NN	O	O
.	NN	O	O

Late	NN	O	O
neuropsychological	NN	O	O
defects	NN	O	O
caused	NN	O	O
by	NN	O	O
cranial	NN	O	O
XRT	NN	O	O
and	NN	O	O
IT	NN	O	O
MTX	NN	O	O
have	NN	O	O
prompted	NN	O	O
programs	NN	O	O
designed	NN	O	O
to	NN	O	O
reduce	NN	O	O
the	NN	O	O
potential	NN	O	O
late	NN	O	O
toxicity	NN	O	O
of	NN	O	O
CNS	NN	O	O
prophylaxis	NN	O	O
.	NN	O	O

More	NN	O	O
pronounced	NN	O	O
in	NN	O	O
younger	NN	O	O
children	NN	O	O
,	NN	O	O
these	NN	O	O
abnormalities	NN	O	O
include	NN	O	O
decreased	NN	O	O
IQ	NN	O	O
,	NN	O	O
visual-motor	NN	O	O
incoordination	NN	O	O
,	NN	O	O
poor	NN	O	O
performance	NN	O	O
in	NN	O	O
mathematics	NN	O	O
,	NN	O	O
and	NN	O	O
memory	NN	O	O
dysfunction	NN	O	O
.	NN	O	O

Until	NN	O	O
1980	NN	O	O
,	NN	O	O
more	NN	O	O
intensive	NN	O	O
induction	NN	O	O
,	NN	O	O
consolidation	NN	O	O
,	NN	O	O
and	NN	O	O
maintenance	NN	O	O
therapy	NN	O	O
had	NN	O	O
failed	NN	O	O
to	NN	O	O
prolong	NN	O	O
DFS	NN	O	O
in	NN	O	O
children	NN	O	O
with	NN	O	O
a	NN	O	O
poor	NN	O	O
prognosis	NN	O	O
.	NN	O	O

In	NN	O	O
West	NN	O	O
Germany	NN	O	O
(	NN	O	O
Berlin-Frankfurt-Muenster	NN	O	O
protocol	NN	O	O
)	NN	O	O
a	NN	O	O
70	NN	O	O
to	NN	O	O
75	NN	O	O
%	NN	O	O
DFS	NN	O	O
is	NN	O	O
seen	NN	O	O
in	NN	O	O
all	NN	O	O
patients	NN	O	O
regardless	NN	O	O
of	NN	O	O
initial	NN	O	O
WBC	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
effective	NN	O	O
therapy	NN	O	O
will	NN	O	O
override	NN	O	O
prognostic	NN	O	O
factors	NN	O	O
.	NN	O	O

Ultra-high-dose	NN	O	O
MTX	NN	O	O
,	NN	O	O
without	NN	O	O
cranial	NN	O	O
radiation	NN	O	O
,	NN	O	O
is	NN	O	O
also	NN	O	O
showing	NN	O	O
promise	NN	O	O
in	NN	O	O
poor	NN	O	O
prognosis	NN	O	O
patients	NN	O	O
.	NN	O	O

Other	NN	O	O
issues	NN	O	O
include	NN	O	O
the	NN	O	O
optimal	NN	O	O
duration	NN	O	O
of	NN	O	O
therapy	NN	O	O
,	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
testicular	NN	O	O
biopsies	NN	O	O
,	NN	O	O
and	NN	O	O
prophylactic	NN	O	O
testicular	NN	O	O
radiation	NN	O	O
.	NN	O	O

Recent	NN	O	O
studies	NN	O	O
suggest	NN	O	O
that	NN	O	O
prognostic	NN	O	O
factors	NN	O	O
lose	NN	O	O
their	NN	O	O
significance	NN	O	O
after	NN	O	O
2	NN	O	O
years	NN	O	O
of	NN	O	O
continuous	NN	O	O
complete	NN	O	O
remission	NN	O	O
and	NN	O	O
that	NN	O	O
2	NN	O	O
years	NN	O	O
of	NN	O	O
maintenance	NN	O	O
therapy	NN	O	O
is	NN	O	O
adequate	NN	O	O
.	NN	O	O

Bilateral	NN	O	O
open-wedge	NN	O	O
testicular	NN	O	O
biopsies	NN	O	O
have	NN	O	O
identified	NN	O	O
occult	NN	O	O
testicular	NN	O	O
disease	NN	O	O
in	NN	O	O
8	NN	O	O
to	NN	O	O
10	NN	O	O
%	NN	O	O
of	NN	O	O
males	NN	O	O
.	NN	O	O

A	NN	O	O
unified	NN	O	O
approach	NN	O	O
to	NN	O	O
children	NN	O	O
with	NN	O	O
leukemia/lymphoma	NN	O	O
,	NN	O	O
a	NN	O	O
group	NN	O	O
with	NN	O	O
a	NN	O	O
particularly	NN	O	O
poor	NN	O	O
prognosis	NN	O	O
,	NN	O	O
utilizing	NN	O	O
NHL-type	NN	O	O
therapy	NN	O	O
may	NN	O	O
be	NN	O	O
more	NN	O	O
effective	NN	O	O
than	NN	O	O
conventional	NN	O	O
ALL	NN	O	O
therapy	NN	O	O
.	NN	O	O

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

-DOCSTART-	O

Glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
and	NN	O	O
in	NN	O	O
vitro	NN	O	O
sensitivity	NN	O	O
to	NN	O	O
steroid	NN	O	O
hormones	NN	O	O
in	NN	O	O
human	NN	O	O
lymphoproliferative	NN	O	O
diseases	NN	O	O
and	NN	O	O
myeloid	NN	O	O
leukemia	NN	O	O
.	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
quantitation	NN	O	O
by	NN	O	O
a	NN	O	O
whole-cell	NN	O	O
assay	NN	O	O
and/or	NN	O	O
cytosol	NN	O	O
technique	NN	O	O
and	NN	O	O
the	NN	O	O
in	NN	O	O
vitro	NN	O	O
sensitivity	NN	O	O
to	NN	O	O
steroids	NN	O	O
have	NN	O	O
been	NN	O	O
assessed	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
normal	NN	O	O
donors	NN	O	O
and	NN	O	O
patients	NN	O	O
with	NN	O	O
chronic	NN	O	O
lymphatic	NN	O	O
leukemia	NN	O	O
(	NN	O	O
CLL	NN	O	O
)	NN	O	O
,	NN	O	O
acute	NN	O	O
lymphoblastic	NN	O	O
leukemia	NN	O	O
(	NN	O	O
ALL	NN	O	O
)	NN	O	O
,	NN	O	O
lymphosarcoma	NN	O	O
cell	NN	O	O
leukemia	NN	O	O
(	NN	O	O
LSCL	NN	O	O
)	NN	O	O
,	NN	O	O
acute	NN	O	O
nonlymphatic	NN	O	O
leukemia	NN	O	O
(	NN	O	O
ANLL	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
chronic	NN	O	O
myeloid	NN	O	O
leukemia	NN	O	O
(	NN	O	O
CML	NN	O	O
)	NN	O	O
.	NN	O	O

Within	NN	O	O
the	NN	O	O
lymphoproliferative	NN	O	O
diseases	NN	O	O
,	NN	O	O
ALL	NN	O	B-cell_line
cells	NN	O	I-cell_line
exhibited	NN	O	O
the	NN	O	O
highest	NN	O	O
GR	NN	O	B-protein
concentration	NN	O	O
(	NN	O	O
regardless	NN	O	O
of	NN	O	O
the	NN	O	O
method	NN	O	O
used	NN	O	O
)	NN	O	O
and	NN	O	O
the	NN	O	O
highest	NN	O	O
in	NN	O	O
vitro	NN	O	O
inhibition	NN	O	O
of	NN	O	O
spontaneous	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
thymidine	NN	O	O
(	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
TdR	NN	O	O
)	NN	O	O
uptake	NN	O	O
by	NN	O	O
glucocorticoids	NN	O	O
.	NN	O	O

A	NN	O	O
significant	NN	O	O
relationship	NN	O	O
between	NN	O	O
GR	NN	O	B-protein
concentration	NN	O	O
(	NN	O	O
whole-cell	NN	O	O
assay	NN	O	O
)	NN	O	O
and	NN	O	O
in	NN	O	O
vitro	NN	O	O
sensitivity	NN	O	O
to	NN	O	O
dexamethasone	NN	O	O
was	NN	O	O
also	NN	O	O
found	NN	O	O
.	NN	O	O

On	NN	O	O
the	NN	O	O
contrary	NN	O	O
,	NN	O	O
CLL	NN	O	B-cell_line
cells	NN	O	I-cell_line
presented	NN	O	O
the	NN	O	O
highest	NN	O	O
sensitivity	NN	O	O
to	NN	O	O
glucocorticoids	NN	O	O
in	NN	O	O
PHA-stimulated	NN	O	B-cell_line
cell	NN	O	I-cell_line
cultures	NN	O	I-cell_line
.	NN	O	O

Cells	NN	O	O
from	NN	O	O
the	NN	O	O
only	NN	O	O
two	NN	O	O
ALL	NN	O	O
patients	NN	O	O
who	NN	O	O
did	NN	O	O
not	NN	O	O
undergo	NN	O	O
a	NN	O	O
remission	NN	O	O
after	NN	O	O
glucocorticoid-inclusive	NN	O	O
chemotherapy	NN	O	O
had	NN	O	O
both	NN	O	O
the	NN	O	O
lowest	NN	O	O
in	NN	O	O
vitro	NN	O	O
sensitivity	NN	O	O
to	NN	O	O
dexamethasone	NN	O	O
and	NN	O	O
the	NN	O	O
lowest	NN	O	O
GR	NN	O	O
concentration	NN	O	O
with	NN	O	O
whole-cell	NN	O	O
assay	NN	O	O
.	NN	O	O

Concerning	NN	O	O
myeloid	NN	O	O
leukemia	NN	O	O
,	NN	O	O
ANLL	NN	O	O
patients	NN	O	O
had	NN	O	O
GR	NN	O	B-protein
concentrations	NN	O	O
slightly	NN	O	O
higher	NN	O	O
than	NN	O	O
those	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
ALL	NN	O	O
group	NN	O	O
but	NN	O	O
exhibited	NN	O	O
the	NN	O	O
lowest	NN	O	O
degree	NN	O	O
of	NN	O	O
inhibition	NN	O	O
of	NN	O	O
spontaneous	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
TdR	NN	O	O
uptake	NN	O	O
by	NN	O	O
dexamethasone	NN	O	O
(	NN	O	O
stimulatory	NN	O	O
effects	NN	O	O
occurred	NN	O	O
in	NN	O	O
some	NN	O	O
cases	NN	O	O
)	NN	O	O
.	NN	O	O

CML	NN	O	B-cell_line
cells	NN	O	I-cell_line
exhibited	NN	O	O
an	NN	O	O
inhibition	NN	O	O
degree	NN	O	O
by	NN	O	O
in	NN	O	O
vitro	NN	O	O
glucocorticoids	NN	O	O
significantly	NN	O	O
higher	NN	O	O
than	NN	O	O
that	NN	O	O
of	NN	O	O
ANLL	NN	O	B-cell_line
cells	NN	O	I-cell_line
but	NN	O	O
not	NN	O	O
different	NN	O	O
from	NN	O	O
that	NN	O	O
of	NN	O	O
lymphoproliferative	NN	O	O
diseases	NN	O	O
.	NN	O	O

No	NN	O	O
clear	NN	O	O
relationship	NN	O	O
among	NN	O	O
GR	NN	O	O
pattern	NN	O	O
,	NN	O	O
in	NN	O	O
vitro	NN	O	O
cell	NN	O	O
sensitivity	NN	O	O
to	NN	O	O
glucocorticoids	NN	O	O
,	NN	O	O
and	NN	O	O
clinicohematologic	NN	O	O
parameters	NN	O	O
was	NN	O	O
observed	NN	O	O
in	NN	O	O
myeloid	NN	O	O
leukemia-bearing	NN	O	O
patients	NN	O	O
.	NN	O	O

-DOCSTART-	O

Glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
and	NN	O	O
in	NN	O	O
vitro	NN	O	O
corticosensitivity	NN	O	O
of	NN	O	O
peanut-positive	NN	O	B-cell_line
and	NN	O	I-cell_line
peanut-negative	NN	O	I-cell_line
human	NN	O	I-cell_line
thymocyte	NN	O	I-cell_line
subpopulations	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
6	NN	O	O
human	NN	O	O
thymus	NN	O	O
glands	NN	O	O
,	NN	O	O
the	NN	O	O
immature	NN	O	O
subset	NN	O	O
of	NN	O	O
thymocytes	NN	O	B-cell_type
was	NN	O	O
separated	NN	O	O
from	NN	O	O
the	NN	O	O
more	NN	O	O
mature	NN	O	O
one	NN	O	O
,	NN	O	O
by	NN	O	O
differential	NN	O	O
peanut	NN	O	O
lectin	NN	O	O
agglutination	NN	O	O
.	NN	O	O

These	NN	O	O
2	NN	O	O
cell	NN	O	O
subpopulations	NN	O	O
were	NN	O	O
analyzed	NN	O	O
for	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
content	NN	O	O
by	NN	O	O
using	NN	O	O
a	NN	O	O
whole	NN	O	O
cell	NN	O	O
assay	NN	O	O
,	NN	O	O
with	NN	O	O
(	NN	O	O
3H	NN	O	O
)	NN	O	O
-triamcinolone	NN	O	O
acetonide	NN	O	O
as	NN	O	O
tracer	NN	O	O
.	NN	O	O

The	NN	O	O
unagglutinated	NN	O	B-cell_type
thymocytes	NN	O	I-cell_type
(	NN	O	O
peanut	NN	O	O
negative	NN	O	O
)	NN	O	O
contained	NN	O	O
about	NN	O	O
2	NN	O	O
times	NN	O	O
more	NN	O	O
receptor	NN	O	O
sites	NN	O	O
per	NN	O	O
cell	NN	O	O
than	NN	O	O
agglutinated	NN	O	O
(	NN	O	O
peanut	NN	O	O
positive	NN	O	O
)	NN	O	O
ones	NN	O	O
(	NN	O	O
7650	NN	O	O
+/-	NN	O	O
1550	NN	O	O
S.D.	NN	O	O
verus	NN	O	O
3195	NN	O	O
+/-	NN	O	O
896	NN	O	O
S.D.	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
affinity	NN	O	O
for	NN	O	O
steroid	NN	O	O
was	NN	O	O
similar	NN	O	O
in	NN	O	O
both	NN	O	O
cell	NN	O	O
subsets	NN	O	O
,	NN	O	O
as	NN	O	O
was	NN	O	O
the	NN	O	O
stereospecificity	NN	O	O
for	NN	O	O
glucocorticoids	NN	O	O
,	NN	O	O
the	NN	O	O
time-course	NN	O	O
of	NN	O	O
steroid-receptor	NN	O	O
association	NN	O	O
,	NN	O	O
and	NN	O	O
cytoplasmic	NN	O	O
to	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
.	NN	O	O

Despite	NN	O	O
the	NN	O	O
greater	NN	O	O
number	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
sites	NN	O	O
,	NN	O	O
the	NN	O	O
peanut-negative	NN	O	B-cell_line
thymocyte	NN	O	I-cell_line
subpopulation	NN	O	I-cell_line
did	NN	O	O
not	NN	O	O
differ	NN	O	O
from	NN	O	O
the	NN	O	O
peanut-positive	NN	O	B-cell_line
one	NN	O	O
in	NN	O	O
its	NN	O	O
sensitivity	NN	O	O
to	NN	O	O
the	NN	O	O
inhibitory	NN	O	O
effects	NN	O	O
of	NN	O	O
triamcinolone	NN	O	O
acetonide	NN	O	O
,	NN	O	O
as	NN	O	O
determined	NN	O	O
by	NN	O	O
measurements	NN	O	O
of	NN	O	O
the	NN	O	O
incorporation	NN	O	O
of	NN	O	O
radiolabeled	NN	O	O
precursors	NN	O	O
of	NN	O	O
protein	NN	O	O
and	NN	O	O
DNA	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
the	NN	O	O
peanut-negative	NN	O	B-cell_line
subset	NN	O	I-cell_line
appeared	NN	O	O
more	NN	O	O
resistant	NN	O	O
in	NN	O	O
vitro	NN	O	O
to	NN	O	O
the	NN	O	O
steroid-induced	NN	O	O
cell	NN	O	O
lysis	NN	O	O
as	NN	O	O
compared	NN	O	O
to	NN	O	O
the	NN	O	O
peanut-positive	NN	O	B-cell_line
one	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
our	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
glucocorticoid	NN	O	O
receptor	NN	O	O
density	NN	O	O
and	NN	O	O
corticosensitivity	NN	O	O
are	NN	O	O
not	NN	O	O
directly	NN	O	O
correlated	NN	O	O
and	NN	O	O
that	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
sites	NN	O	O
may	NN	O	O
be	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
degree	NN	O	O
of	NN	O	O
immunologic	NN	O	O
maturation	NN	O	O

-DOCSTART-	O

Defective	NN	O	O
binding	NN	O	O
and	NN	O	O
function	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
end-organ	NN	O	O
resistance	NN	O	O
to	NN	O	O
1	NN	O	O
,	NN	O	O
25-dihydroxyvitamin	NN	O	O
D	NN	O	O
.	NN	O	O

Lectin-induced	NN	O	O
DNA	NN	O	O
synthesis	NN	O	O
by	NN	O	O
peripheral	NN	O	B-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
17	NN	O	O
normal	NN	O	O
donors	NN	O	O
was	NN	O	O
inhibited	NN	O	O
(	NN	O	O
40-60	NN	O	O
%	NN	O	O
)	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
at	NN	O	O
physiological	NN	O	O
concentrations	NN	O	O
(	NN	O	O
10	NN	O	O
(	NN	O	O
-10	NN	O	O
)	NN	O	O
-10	NN	O	O
(	NN	O	O
-9	NN	O	O
)	NN	O	O
M	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
lymphocytes	NN	O	B-cell_type
acquire	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
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
upon	NN	O	O
activation	NN	O	O
by	NN	O	O
the	NN	O	O
lectins	NN	O	B-protein
.	NN	O	O

This	NN	O	O
process	NN	O	O
precedes	NN	O	O
the	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
of	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
.	NN	O	O

We	NN	O	O
studied	NN	O	O
lymphocytes	NN	O	B-cell_type
from	NN	O	O
six	NN	O	O
patients	NN	O	O
from	NN	O	O
four	NN	O	O
different	NN	O	O
kindreds	NN	O	O
with	NN	O	O
the	NN	O	O
syndrome	NN	O	O
of	NN	O	O
hereditary	NN	O	O
end-organ	NN	O	O
resistance	NN	O	O
to	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D	NN	O	O
(	NN	O	O
the	NN	O	O
so-called	NN	O	O
vitamin	NN	O	O
D-dependent	NN	O	O
rickets	NN	O	O
type	NN	O	O
II	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
five	NN	O	O
patients	NN	O	O
(	NN	O	O
three	NN	O	O
kindreds	NN	O	O
)	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
did	NN	O	O
not	NN	O	O
acquire	NN	O	O
receptors	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
upon	NN	O	O
phytohemagglutinin-induced	NN	O	O
activation	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
in	NN	O	O
contrast	NN	O	O
to	NN	O	O
normal	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
the	NN	O	O
mitogenic	NN	O	O
stimulation	NN	O	O
of	NN	O	O
these	NN	O	O
patients	NN	O	O
'	NN	O	O
lymphocytes	NN	O	B-cell_type
by	NN	O	O
phytohemagglutinin	NN	O	B-protein
and	NN	O	O
concanavalin	NN	O	O
A	NN	O	O
was	NN	O	O
not	NN	O	O
inhibited	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

Activated	NN	O	O
lymphocytes	NN	O	B-cell_type
of	NN	O	O
the	NN	O	O
sixth	NN	O	O
patient	NN	O	O
from	NN	O	O
a	NN	O	O
fourth	NN	O	O
kindred	NN	O	O
exhibited	NN	O	O
normal	NN	O	O
binding	NN	O	O
of	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
but	NN	O	O
the	NN	O	O
hormone	NN	O	O
failed	NN	O	O
to	NN	O	O
inhibit	NN	O	O
the	NN	O	O
mitogenic	NN	O	O
stimulation	NN	O	O
.	NN	O	O

A	NN	O	O
similar	NN	O	O
pattern	NN	O	O
of	NN	O	O
the	NN	O	O
vitamin	NN	O	O
D	NN	O	O
effector	NN	O	O
system	NN	O	O
was	NN	O	O
previously	NN	O	O
observed	NN	O	O
in	NN	O	O
fibroblasts	NN	O	O
cultured	NN	O	O
from	NN	O	O
skin	NN	O	O
biopsies	NN	O	O
of	NN	O	O
the	NN	O	O
same	NN	O	O
group	NN	O	O
of	NN	O	O
patients	NN	O	O
.	NN	O	O

The	NN	O	O
conclusions	NN	O	O
from	NN	O	O
these	NN	O	O
findings	NN	O	O
are	NN	O	O
:	NN	O	O
(	NN	O	O
a	NN	O	O
)	NN	O	O
the	NN	O	O
inhibition	NN	O	O
of	NN	O	O
mitogenic	NN	O	O
stimulation	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
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
specific	NN	O	B-protein
functional	NN	O	I-protein
receptors	NN	O	I-protein
to	NN	O	O
the	NN	O	O
hormone	NN	O	O
;	NN	O	O
and	NN	O	O
(	NN	O	O
b	NN	O	O
)	NN	O	O
the	NN	O	O
receptors	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
in	NN	O	O
mononuclear	NN	O	B-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
probably	NN	O	O
controlled	NN	O	O
genetically	NN	O	O
by	NN	O	O
the	NN	O	O
same	NN	O	O
mechanisms	NN	O	O
as	NN	O	O
the	NN	O	O
effector	NN	O	O
system	NN	O	O
in	NN	O	O
well-characterized	NN	O	O
target	NN	O	O
organs	NN	O	O
of	NN	O	O
the	NN	O	O
hormone	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
intestine	NN	O	O
and	NN	O	O
kidney	NN	O	O
.	NN	O	O

-DOCSTART-	O

Glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
of	NN	O	O
mononuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
from	NN	O	O
myasthenia	NN	O	O
gravis	NN	O	O
patients	NN	O	O
.	NN	O	O

The	NN	O	O
present	NN	O	O
study	NN	O	O
was	NN	O	O
performed	NN	O	O
to	NN	O	O
analyse	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
GR	NN	O	B-protein
)	NN	O	O
binding	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
leukocytes	NN	O	I-cell_type
(	NN	O	O
MNL	NN	O	B-cell_type
)	NN	O	O
from	NN	O	O
39	NN	O	O
myasthenia	NN	O	O
gravis	NN	O	O
(	NN	O	O
MG	NN	O	O
)	NN	O	O
patients	NN	O	O
(	NN	O	O
unoperated	NN	O	O
patients	NN	O	O
(	NN	O	O
n	NN	O	O
=	NN	O	O
13	NN	O	O
)	NN	O	O
,	NN	O	O
thymectomized	NN	O	O
patients	NN	O	O
(	NN	O	O
n	NN	O	O
=	NN	O	O
14	NN	O	O
)	NN	O	O
and	NN	O	O
patients	NN	O	O
receiving	NN	O	O
glucocorticoids	NN	O	O
:	NN	O	O
thymectomized	NN	O	O
(	NN	O	O
n	NN	O	O
=	NN	O	O
11	NN	O	O
)	NN	O	O
and	NN	O	O
unoperated	NN	O	O
(	NN	O	O
n	NN	O	O
=	NN	O	O
6	NN	O	O
]	NN	O	O
.	NN	O	O

A	NN	O	O
whole	NN	O	O
cell	NN	O	O
binding	NN	O	O
assay	NN	O	O
with	NN	O	O
3	NN	O	O
(	NN	O	O
H	NN	O	O
)	NN	O	O
dexamethasone	NN	O	O
was	NN	O	O
used	NN	O	O
.	NN	O	O

GR	NN	O	B-protein
mean	NN	O	O
values	NN	O	O
were	NN	O	O
significantly	NN	O	O
higher	NN	O	O
in	NN	O	O
the	NN	O	O
MNL	NN	O	B-cell_type
of	NN	O	O
MG	NN	O	O
patients	NN	O	O
(	NN	O	O
thymectomized	NN	O	O
or	NN	O	O
not	NN	O	O
)	NN	O	O
not	NN	O	O
receiving	NN	O	O
glucocorticoid	NN	O	O
than	NN	O	O
in	NN	O	O
the	NN	O	O
MNL	NN	O	B-cell_type
of	NN	O	O
healthy	NN	O	O
donors	NN	O	O
.	NN	O	O

Affinity	NN	O	O
was	NN	O	O
within	NN	O	O
the	NN	O	O
normal	NN	O	O
range	NN	O	O
.	NN	O	O

Sex	NN	O	O
,	NN	O	O
age	NN	O	O
or	NN	O	O
clinical	NN	O	O
forms	NN	O	O
of	NN	O	O
illness	NN	O	O
did	NN	O	O
not	NN	O	O
influence	NN	O	O
the	NN	O	O
results	NN	O	O
.	NN	O	O

In	NN	O	O
patients	NN	O	O
receiving	NN	O	O
prednisone	NN	O	O
(	NN	O	O
Pd	NN	O	O
)	NN	O	O
the	NN	O	O
GR	NN	O	B-protein
values	NN	O	O
were	NN	O	O
significantly	NN	O	O
lower	NN	O	O
than	NN	O	O
in	NN	O	O
MG	NN	O	O
patients	NN	O	O
without	NN	O	O
Pd	NN	O	O
therapy	NN	O	O
,	NN	O	O
independent	NN	O	O
of	NN	O	O
Pd	NN	O	O
dose	NN	O	O
or	NN	O	O
time	NN	O	O
of	NN	O	O
administration	NN	O	O
.	NN	O	O

No	NN	O	O
differences	NN	O	O
in	NN	O	O
receptor	NN	O	O
binding	NN	O	O
between	NN	O	O
normal	NN	O	O
subjects	NN	O	O
and	NN	O	O
MG	NN	O	O
patients	NN	O	O
receiving	NN	O	O
Pd	NN	O	O
have	NN	O	O
been	NN	O	O
found	NN	O	O
.	NN	O	O

-DOCSTART-	O

Immunological	NN	O	O
interference	NN	O	O
of	NN	O	O
high	NN	O	O
dose	NN	O	O
corticosteroids	NN	O	O
.	NN	O	O

High-dose	NN	O	O
corticosteroids	NN	O	O
(	NN	O	O
HDC	NN	O	O
)	NN	O	O
will	NN	O	O
influence	NN	O	O
cellular	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
humoral	NN	O	O
participants	NN	O	O
of	NN	O	O
the	NN	O	O
immune	NN	O	O
response	NN	O	O
.	NN	O	O

The	NN	O	O
lymphoid	NN	O	O
tissue	NN	O	O
will	NN	O	O
decrease	NN	O	O
in	NN	O	O
size	NN	O	O
and	NN	O	O
weight	NN	O	O
after	NN	O	O
prolonged	NN	O	O
treatment	NN	O	O
with	NN	O	O
HDC	NN	O	O
.	NN	O	O

Lymphocyte	NN	O	O
functions	NN	O	O
will	NN	O	O
be	NN	O	O
impaired	NN	O	O
.	NN	O	O

Reduced	NN	O	O
synthesis	NN	O	O
of	NN	O	O
B-	NN	O	B-cell_type
as	NN	O	I-cell_type
well	NN	O	I-cell_type
as	NN	O	I-cell_type
T-lymphocytes	NN	O	I-cell_type
will	NN	O	O
be	NN	O	O
seen	NN	O	O
.	NN	O	O

The	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
on	NN	O	O
B-cell	NN	O	B-cell_type
function	NN	O	O
can	NN	O	O
be	NN	O	O
observed	NN	O	O
both	NN	O	O
as	NN	O	O
decreased	NN	O	O
serum	NN	O	O
levels	NN	O	O
of	NN	O	O
immunoglobulins	NN	O	B-protein
and	NN	O	O
as	NN	O	O
impaired	NN	O	O
binding	NN	O	O
of	NN	O	O
antibodies	NN	O	B-protein
and	NN	O	O
complement	NN	O	O
to	NN	O	O
the	NN	O	O
cellular	NN	O	O
surface	NN	O	O
.	NN	O	O

Reduced	NN	O	O
T-cell	NN	O	B-cell_type
function	NN	O	O
indicated	NN	O	O
by	NN	O	O
impaired	NN	O	O
stimulation	NN	O	O
by	NN	O	O
PHA	NN	O	B-protein
and	NN	O	O
porkweed	NN	O	B-protein
as	NN	O	O
well	NN	O	O
as	NN	O	O
by	NN	O	O
impaired	NN	O	O
lymphokinin	NN	O	O
effects	NN	O	O
on	NN	O	O
leukocyte	NN	O	B-cell_type
migration	NN	O	O
inhibition	NN	O	O
has	NN	O	O
been	NN	O	O
reported	NN	O	O
.	NN	O	O

Reduced	NN	O	O
lymphocyte	NN	O	O
adherence	NN	O	O
to	NN	O	O
antigen	NN	O	O
and	NN	O	O
suppressed	NN	O	O
lymphocyte	NN	O	O
reaction	NN	O	O
have	NN	O	O
also	NN	O	O
been	NN	O	O
observed	NN	O	O
.	NN	O	O

Humoral	NN	O	O
factors	NN	O	O
involved	NN	O	O
in	NN	O	O
chemotaxis	NN	O	O
,	NN	O	O
opsonisation	NN	O	O
,	NN	O	O
phagocytosis	NN	O	O
,	NN	O	O
vascular	NN	O	O
permeability	NN	O	O
leading	NN	O	O
to	NN	O	O
leakage	NN	O	O
of	NN	O	O
fluid	NN	O	O
and	NN	O	O
cells	NN	O	O
and	NN	O	O
factors	NN	O	O
involved	NN	O	O
in	NN	O	O
lysis	NN	O	O
of	NN	O	O
antigens	NN	O	O
are	NN	O	O
impaired	NN	O	O
.	NN	O	O

This	NN	O	O
can	NN	O	O
be	NN	O	O
explained	NN	O	O
partly	NN	O	O
by	NN	O	O
the	NN	O	O
observed	NN	O	O
reduced	NN	O	O
complement	NN	O	O
activation	NN	O	O
via	NN	O	O
the	NN	O	O
alternative	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
classical	NN	O	O
pathway	NN	O	O
in	NN	O	O
association	NN	O	O
with	NN	O	O
HDC	NN	O	O
therapy	NN	O	O
.	NN	O	O

Acute	NN	O	O
processes	NN	O	O
with	NN	O	O
increased	NN	O	O
vascular	NN	O	O
permeability	NN	O	O
and	NN	O	O
accumulation	NN	O	O
of	NN	O	O
leukocytes	NN	O	B-cell_type
as	NN	O	O
impairing	NN	O	O
factors	NN	O	O
could	NN	O	O
be	NN	O	O
influenced	NN	O	O
beneficially	NN	O	O
by	NN	O	O
HDC	NN	O	O
therapy	NN	O	O
.	NN	O	O

This	NN	O	O
positive	NN	O	O
effect	NN	O	O
can	NN	O	O
be	NN	O	O
seen	NN	O	O
in	NN	O	O
treatment	NN	O	O
of	NN	O	O
septic	NN	O	O
shock	NN	O	O
or	NN	O	O
rejection	NN	O	O
of	NN	O	O
a	NN	O	O
transplant	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
if	NN	O	O
sepsis	NN	O	O
or	NN	O	O
rejection	NN	O	O
is	NN	O	O
not	NN	O	O
rapidly	NN	O	O
reversed	NN	O	O
,	NN	O	O
complications	NN	O	O
such	NN	O	O
as	NN	O	O
multisystem	NN	O	O
organ	NN	O	O
failure	NN	O	O
and	NN	O	O
bacteremia	NN	O	O
are	NN	O	O
prone	NN	O	O
to	NN	O	O
appear	NN	O	O
.	NN	O	O

-DOCSTART-	O

Identification	NN	O	O
of	NN	O	O
human	NN	O	B-protein
leukemic	NN	O	I-protein
glucocorticoid	NN	O	I-protein
receptors	NN	O	I-protein
using	NN	O	O
affinity	NN	O	O
labeling	NN	O	O
and	NN	O	O
anti-human	NN	O	B-protein
glucocorticoid	NN	O	I-protein
receptor	NN	O	I-protein
antibodies	NN	O	I-protein
.	NN	O	O

Antisera	NN	O	O
raised	NN	O	O
against	NN	O	O
human	NN	O	B-protein
lymphoid	NN	O	I-protein
glucocorticoid	NN	O	I-protein
receptors	NN	O	I-protein
were	NN	O	O
used	NN	O	O
in	NN	O	O
combination	NN	O	O
with	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	O
receptor	NN	O	O
affinity	NN	O	O
label	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
dexamethasone	NN	O	O
21-mesylate	NN	O	O
[	NN	O	O
(	NN	O	O
3H	NN	O	O
]	NN	O	O
DM	NN	O	O
)	NN	O	O
to	NN	O	O
identify	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
of	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
B-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
and	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
leukemic	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
CEM-C7	NN	O	B-cell_line
.	NN	O	O

Antisera	NN	O	O
were	NN	O	O
obtained	NN	O	O
following	NN	O	O
immunization	NN	O	O
of	NN	O	O
New	NN	O	O
Zealand	NN	O	O
White	NN	O	O
rabbits	NN	O	O
with	NN	O	O
[	NN	O	B-protein
3H	NN	O	I-protein
]	NN	O	I-protein
triamcinolone	NN	O	I-protein
acetonide	NN	O	I-protein
[	NN	O	I-protein
(	NN	O	I-protein
3H	NN	O	I-protein
]	NN	O	I-protein
TA	NN	O	I-protein
)	NN	O	I-protein
-glucocorticoid	NN	O	I-protein
receptor	NN	O	I-protein
complexes	NN	O	I-protein
partially	NN	O	O
purified	NN	O	O
by	NN	O	O
two-stage	NN	O	O
DNA-cellulose	NN	O	O
chromatography	NN	O	O
.	NN	O	O

The	NN	O	O
presence	NN	O	O
of	NN	O	O
anti-human	NN	O	B-protein
glucocorticoid	NN	O	I-protein
receptor	NN	O	I-protein
antibodies	NN	O	I-protein
was	NN	O	O
verified	NN	O	O
by	NN	O	O
:	NN	O	O
(	NN	O	O
a	NN	O	O
)	NN	O	O
adsorption	NN	O	O
of	NN	O	O
[	NN	O	B-protein
3H	NN	O	I-protein
]	NN	O	I-protein
TA-receptor-antibody	NN	O	I-protein
complexes	NN	O	I-protein
to	NN	O	O
Protein	NN	O	B-protein
A	NN	O	I-protein
;	NN	O	O
(	NN	O	O
b	NN	O	O
)	NN	O	O
a	NN	O	O
shift	NN	O	O
to	NN	O	O
higher	NN	O	O
apparent	NN	O	O
molecular	NN	O	O
weight	NN	O	O
in	NN	O	O
the	NN	O	O
elution	NN	O	O
position	NN	O	O
from	NN	O	O
Sephacryl	NN	O	O
S300	NN	O	O
of	NN	O	O
[	NN	O	B-protein
3H	NN	O	I-protein
]	NN	O	I-protein
TA-receptor	NN	O	I-protein
complexes	NN	O	I-protein
incubated	NN	O	O
with	NN	O	O
immune	NN	O	O
serum	NN	O	O
;	NN	O	O
and	NN	O	O
(	NN	O	O
c	NN	O	O
)	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
immune	NN	O	O
serum	NN	O	O
to	NN	O	O
displace	NN	O	O
[	NN	O	B-protein
3H	NN	O	I-protein
]	NN	O	I-protein
TA-receptor	NN	O	I-protein
complexes	NN	O	I-protein
on	NN	O	O
sucrose	NN	O	O
gradients	NN	O	O
.	NN	O	O

These	NN	O	O
antibodies	NN	O	B-protein
also	NN	O	O
recognized	NN	O	O
rat	NN	O	O
liver	NN	O	O
and	NN	O	O
murine	NN	O	B-protein
S49	NN	O	I-protein
cell	NN	O	I-protein
glucocorticoid	NN	O	I-protein
receptors	NN	O	I-protein
.	NN	O	O

Sodium	NN	O	O
dodecyl	NN	O	O
sulfate-polyacrylamide	NN	O	O
gel	NN	O	O
electrophoresis	NN	O	O
of	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
DM-labeled	NN	O	O
IM-9	NN	O	O
cytosol	NN	O	O
identified	NN	O	O
a	NN	O	O
major	NN	O	O
competable	NN	O	O
band	NN	O	O
with	NN	O	O
a	NN	O	O
molecular	NN	O	O
weight	NN	O	O
of	NN	O	O
approximately	NN	O	O
90	NN	O	O
,	NN	O	O
000	NN	O	O
,	NN	O	O
three	NN	O	O
minor	NN	O	O
competable	NN	O	O
components	NN	O	O
with	NN	O	O
molecular	NN	O	O
weights	NN	O	O
of	NN	O	O
approximately	NN	O	O
78	NN	O	O
,	NN	O	O
000	NN	O	O
,	NN	O	O
approximately	NN	O	O
51	NN	O	O
,	NN	O	O
000	NN	O	O
,	NN	O	O
and	NN	O	O
approximately	NN	O	O
38	NN	O	O
,	NN	O	O
500	NN	O	O
,	NN	O	O
and	NN	O	O
at	NN	O	O
least	NN	O	O
21	NN	O	O
other	NN	O	O
noncompetable	NN	O	O
components	NN	O	O
.	NN	O	O

Following	NN	O	O
immunoprecipitation	NN	O	O
of	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
DM-labeled	NN	O	O
cytosol	NN	O	O
with	NN	O	O
immune	NN	O	O
serum	NN	O	O
,	NN	O	O
only	NN	O	O
the	NN	O	O
Mr	NN	O	O
90	NN	O	O
,	NN	O	O
000	NN	O	O
and	NN	O	O
78	NN	O	O
,	NN	O	O
000	NN	O	O
components	NN	O	O
were	NN	O	O
seen	NN	O	O
.	NN	O	O

Sodium	NN	O	O
dodecyl	NN	O	O
sulfate-polyacrylamide	NN	O	O
gel	NN	O	O
electrophoresis	NN	O	O
of	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
DM-labeled	NN	O	O
CEM-C7	NN	O	O
cytosol	NN	O	O
revealed	NN	O	O
a	NN	O	O
larger	NN	O	O
number	NN	O	O
of	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
DM-labeled	NN	O	O
components	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
after	NN	O	O
immunoprecipitation	NN	O	O
of	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
DM-labeled	NN	O	O
CEM-C7	NN	O	O
cytosol	NN	O	O
,	NN	O	O
a	NN	O	O
predominant	NN	O	O
competable	NN	O	O
component	NN	O	O
with	NN	O	O
a	NN	O	O
molecular	NN	O	O
weight	NN	O	O
of	NN	O	O
90	NN	O	O
,	NN	O	O
000	NN	O	O
was	NN	O	O
easily	NN	O	O
identified	NN	O	O
.	NN	O	O

This	NN	O	O
component	NN	O	O
was	NN	O	O
markedly	NN	O	O
diminished	NN	O	O
when	NN	O	O
cytosols	NN	O	O
from	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-cell_line
receptor-deficient	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
ICR-27	NN	O	B-cell_line
were	NN	O	O
used	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
combination	NN	O	O
of	NN	O	O
affinity	NN	O	O
labeling	NN	O	O
and	NN	O	O
anti-human	NN	O	B-protein
glucocorticoid	NN	O	I-protein
receptor	NN	O	I-protein
antibodies	NN	O	I-protein
is	NN	O	O
capable	NN	O	O
of	NN	O	O
providing	NN	O	O
direct	NN	O	O
physical	NN	O	O
identification	NN	O	O
of	NN	O	O
human	NN	O	B-protein
lymphoid	NN	O	I-protein
glucocorticoid	NN	O	I-protein
receptors	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Effect	NN	O	O
of	NN	O	O
thymosin	NN	O	O
on	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
activity	NN	O	O
and	NN	O	O
glucocorticoid	NN	O	O
sensitivity	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
thymocytes	NN	O	I-cell_type
.	NN	O	O

Incubation	NN	O	O
with	NN	O	O
thymosin	NN	O	O
fraction	NN	O	O
5	NN	O	O
,	NN	O	O
(	NN	O	O
TMS	NN	O	O
F5	NN	O	O
at	NN	O	O
300	NN	O	O
micrograms/ml	NN	O	O
)	NN	O	O
a	NN	O	O
partially	NN	O	O
purified	NN	O	O
thymic	NN	O	O
factor	NN	O	O
,	NN	O	O
reduced	NN	O	O
the	NN	O	O
steroid	NN	O	O
binding	NN	O	O
activity	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
infant	NN	O	I-cell_type
thymocytes	NN	O	I-cell_type
from	NN	O	O
9.6	NN	O	O
+/-	NN	O	O
2.1	NN	O	O
fmole/ml	NN	O	O
to	NN	O	O
5.0	NN	O	O
+/-	NN	O	O
2.0	NN	O	O
fmole/ml	NN	O	O
.	NN	O	O

The	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
activity	NN	O	O
in	NN	O	O
normal	NN	O	O
infant	NN	O	O
thymocytes	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
2	NN	O	O
,	NN	O	O
146	NN	O	O
+/-	NN	O	O
726	NN	O	O
(	NN	O	O
s.d.	NN	O	O
)	NN	O	O
sites	NN	O	O
per	NN	O	O
cell	NN	O	O
with	NN	O	O
dissociation	NN	O	O
constant	NN	O	O
of	NN	O	O
1.4	NN	O	O
+/-	NN	O	O
0.6	NN	O	O
X	NN	O	O
10	NN	O	O
(	NN	O	O
-8	NN	O	O
)	NN	O	O
M	NN	O	O
.	NN	O	O

TMS	NN	O	O
F5	NN	O	O
also	NN	O	O
increased	NN	O	O
the	NN	O	O
resistance	NN	O	O
of	NN	O	O
human	NN	O	O
thymocytes	NN	O	O
to	NN	O	O
the	NN	O	O
cytolytic	NN	O	O
effect	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
(	NN	O	O
2.5	NN	O	O
X	NN	O	O
10	NN	O	O
(	NN	O	O
-8	NN	O	O
)	NN	O	O
M	NN	O	O
)	NN	O	O
to	NN	O	O
168.6	NN	O	O
+/-	NN	O	O
30.2	NN	O	O
%	NN	O	O
of	NN	O	O
control	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

In	NN	O	O
animals	NN	O	O
,	NN	O	O
medullary	NN	O	B-cell_type
and	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
are	NN	O	O
more	NN	O	O
resistant	NN	O	O
to	NN	O	O
glucocorticoids	NN	O	O
than	NN	O	O
immature	NN	O	B-cell_type
thymic	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
results	NN	O	O
show	NN	O	O
that	NN	O	O
thymosin	NN	O	O
can	NN	O	O
induce	NN	O	O
changes	NN	O	O
consistent	NN	O	O
with	NN	O	O
differentiation	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
thymocytes	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
in	NN	O	O
vitro	NN	O	O
results	NN	O	O
are	NN	O	O
consistent	NN	O	O
with	NN	O	O
a	NN	O	O
physiological	NN	O	O
role	NN	O	O
of	NN	O	O
thymosin	NN	O	O
in	NN	O	O
intrathymic	NN	O	O
T	NN	O	O
cell	NN	O	O
maturation	NN	O	O
in	NN	O	O
man	NN	O	O
.	NN	O	O

Incubation	NN	O	O
of	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
malignant	NN	O	I-cell_line
thymus	NN	O	I-cell_line
derived	NN	O	I-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
(	NN	O	O
MOLT	NN	O	B-cell_line
3	NN	O	I-cell_line
)	NN	O	O
with	NN	O	O
TMS	NN	O	O
F5	NN	O	O
also	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
number	NN	O	O
of	NN	O	O
steroid	NN	O	O
binding	NN	O	O
sites	NN	O	O
to	NN	O	O
44.2	NN	O	O
+/-	NN	O	O
15.3	NN	O	O
%	NN	O	O
of	NN	O	O
control	NN	O	O
(	NN	O	O
P	NN	O	O
less	NN	O	O
than	NN	O	O
0.05	NN	O	O
)	NN	O	O
,	NN	O	O
but	NN	O	O
TMS	NN	O	O
F5	NN	O	O
did	NN	O	O
not	NN	O	O
significantly	NN	O	O
reduce	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	O
sensitivity	NN	O	O
of	NN	O	O
MOLT	NN	O	B-cell_line
3	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

Specific	NN	O	B-protein
high-affinity	NN	O	I-protein
receptors	NN	O	I-protein
for	NN	O	O
1	NN	O	O
,	NN	O	O
25-dihydroxyvitamin	NN	O	O
D3	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
presence	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
induction	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
following	NN	O	O
activation	NN	O	O
.	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
have	NN	O	O
high	NN	O	O
affinity	NN	O	O
binding	NN	O	O
sites	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
(	NN	O	O
Kd	NN	O	O
0.14	NN	O	O
nM	NN	O	O
,	NN	O	O
sedimentation	NN	O	O
coefficient	NN	O	O
3.7S	NN	O	O
)	NN	O	O
.	NN	O	O

Resting	NN	O	B-cell_type
human	NN	O	I-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
however	NN	O	O
,	NN	O	O
do	NN	O	O
not	NN	O	O
have	NN	O	O
a	NN	O	O
demonstrable	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

After	NN	O	O
activation	NN	O	O
with	NN	O	O
phytohemagglutinin	NN	O	B-protein
the	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
exhibit	NN	O	O
the	NN	O	O
receptor	NN	O	O
within	NN	O	O
24	NN	O	O
h	NN	O	O
,	NN	O	O
and	NN	O	O
this	NN	O	O
expression	NN	O	O
is	NN	O	O
blocked	NN	O	O
by	NN	O	O
cycloheximide	NN	O	O
.	NN	O	O

The	NN	O	O
receptor	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
has	NN	O	O
a	NN	O	O
sedimentation	NN	O	O
coefficient	NN	O	O
of	NN	O	O
3.7S	NN	O	O
and	NN	O	O
a	NN	O	O
high	NN	O	O
affinity	NN	O	O
(	NN	O	O
Kd	NN	O	O
0.10	NN	O	O
nM	NN	O	O
)	NN	O	O
for	NN	O	O
the	NN	O	O
ligand	NN	O	O
.	NN	O	O

-DOCSTART-	O

Effects	NN	O	O
of	NN	O	O
chronic	NN	O	O
glucocorticoid	NN	O	O
excess	NN	O	O
in	NN	O	O
man	NN	O	O
on	NN	O	O
insulin	NN	O	O
binding	NN	O	O
to	NN	O	O
circulating	NN	O	B-cell_type
cells	NN	O	I-cell_type
:	NN	O	O
differences	NN	O	O
between	NN	O	O
endogenous	NN	O	O
and	NN	O	O
exogenous	NN	O	O
hypercorticism	NN	O	O
.	NN	O	O

We	NN	O	O
measured	NN	O	O
[	NN	O	O
125I	NN	O	O
]	NN	O	O
insulin	NN	O	O
binding	NN	O	O
to	NN	O	O
circulating	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
or	NN	O	I-cell_type
erythrocytes	NN	O	I-cell_type
from	NN	O	O
16	NN	O	O
patients	NN	O	O
with	NN	O	O
chronic	NN	O	O
glucocorticoid	NN	O	O
excess	NN	O	O
,	NN	O	O
9	NN	O	O
chronically	NN	O	O
treated	NN	O	O
with	NN	O	O
prednisone	NN	O	O
and	NN	O	O
7	NN	O	O
with	NN	O	O
adrenocortical	NN	O	O
hyperfunction	NN	O	O
.	NN	O	O

With	NN	O	O
monocytes	NN	O	B-cell_type
,	NN	O	O
[	NN	O	O
125I	NN	O	O
]	NN	O	O
insulin	NN	O	O
binding	NN	O	O
was	NN	O	O
iincreased	NN	O	O
in	NN	O	O
all	NN	O	O
patients	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
binding	NN	O	O
data	NN	O	O
indicated	NN	O	O
that	NN	O	O
increased	NN	O	O
binding	NN	O	O
in	NN	O	O
patients	NN	O	O
treated	NN	O	O
with	NN	O	O
prednisone	NN	O	O
was	NN	O	O
due	NN	O	O
to	NN	O	O
an	NN	O	O
ncrease	NN	O	O
in	NN	O	O
receptor	NN	O	O
concentration	NN	O	O
,	NN	O	O
whereas	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
adrenocortical	NN	O	O
hyperfunction	NN	O	O
,	NN	O	O
it	NN	O	O
was	NN	O	O
due	NN	O	O
to	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
receptor	NN	O	O
affinity	NN	O	O
.	NN	O	O

With	NN	O	O
erythrocytes	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
adrenocortical	NN	O	O
hyperfunction	NN	O	O
there	NN	O	O
was	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
receptor	NN	O	O
affinity	NN	O	O
and	NN	O	O
a	NN	O	O
decrease	NN	O	O
in	NN	O	O
receptor	NN	O	O
concentration	NN	O	O
,	NN	O	O
so	NN	O	O
that	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
[	NN	O	O
125I	NN	O	O
]	NN	O	O
insulin	NN	O	O
was	NN	O	O
normal	NN	O	O
.	NN	O	O

The	NN	O	O
disparity	NN	O	O
of	NN	O	O
results	NN	O	O
between	NN	O	O
endogenous	NN	O	O
and	NN	O	O
exogenous	NN	O	O
hypercorticism	NN	O	O
,	NN	O	O
between	NN	O	O
the	NN	O	O
two	NN	O	O
cell	NN	O	O
types	NN	O	O
,	NN	O	O
and	NN	O	O
between	NN	O	O
the	NN	O	O
present	NN	O	O
studies	NN	O	O
and	NN	O	O
previous	NN	O	O
studies	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	O
excess	NN	O	O
on	NN	O	O
the	NN	O	O
insulin	NN	O	B-protein
receptor	NN	O	I-protein
are	NN	O	O
extremely	NN	O	O
complex	NN	O	O
and	NN	O	O
wide-ranging	NN	O	O
and	NN	O	O
that	NN	O	O
in	NN	O	O
this	NN	O	O
condition	NN	O	O
,	NN	O	O
extrapolations	NN	O	O
in	NN	O	O
humans	NN	O	O
from	NN	O	O
data	NN	O	O
with	NN	O	O
circulating	NN	O	B-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
liver	NN	O	O
and	NN	O	O
muscle	NN	O	O
may	NN	O	O
not	NN	O	O
be	NN	O	O
appropriate	NN	O	O
.	NN	O	O

-DOCSTART-	O

Reduced	NN	O	O
level	NN	O	O
of	NN	O	O
cellular	NN	O	B-protein
glucocorticoid	NN	O	I-protein
receptors	NN	O	I-protein
in	NN	O	O
patients	NN	O	O
with	NN	O	O
anorexia	NN	O	O
nervosa	NN	O	O
.	NN	O	O

Specific	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
were	NN	O	O
measured	NN	O	O
in	NN	O	O
circulating	NN	O	O
mononuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
from	NN	O	O
12	NN	O	O
patients	NN	O	O
with	NN	O	O
anorexia	NN	O	O
nervosa	NN	O	O
and	NN	O	O
21	NN	O	O
healthy	NN	O	O
control	NN	O	O
subjects	NN	O	O
.	NN	O	O

Cells	NN	O	O
from	NN	O	O
patients	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
contain	NN	O	O
a	NN	O	O
significantly	NN	O	O
(	NN	O	O
p	NN	O	O
less	NN	O	O
than	NN	O	O
0.01	NN	O	O
)	NN	O	O
lower	NN	O	O
level	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
3830	NN	O	O
+/-	NN	O	O
210	NN	O	O
sites/cell	NN	O	O
,	NN	O	O
mean	NN	O	O
+/-	NN	O	O
SE	NN	O	O
)	NN	O	O
than	NN	O	O
those	NN	O	O
from	NN	O	O
controls	NN	O	O
(	NN	O	O
4930	NN	O	O
+/-	NN	O	O
250	NN	O	O
sites/cell	NN	O	O
)	NN	O	O
.	NN	O	O

A	NN	O	O
partial	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
defect	NN	O	O
may	NN	O	O
well	NN	O	O
explain	NN	O	O
the	NN	O	O
abnormal	NN	O	O
cortisol	NN	O	O
metabolism	NN	O	O
and	NN	O	O
glucocorticoid	NN	O	O
resistance	NN	O	O
commonly	NN	O	O
found	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
anorexia	NN	O	O
nervosa	NN	O	O
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
presence	NN	O	O
of	NN	O	O
a	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
is	NN	O	O
well	NN	O	O
established	NN	O	O
,	NN	O	O
but	NN	O	O
factors	NN	O	O
affecting	NN	O	O
its	NN	O	O
regulation	NN	O	O
have	NN	O	O
not	NN	O	O
been	NN	O	O
described	NN	O	O
.	NN	O	O

Using	NN	O	O
a	NN	O	O
competitive	NN	O	O
binding	NN	O	O
whole	NN	O	O
cell	NN	O	O
assay	NN	O	O
,	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
[	NN	O	O
3H	NN	O	O
]	NN	O	O
-dexamethasone	NN	O	O
at	NN	O	O
24	NN	O	O
and	NN	O	O
37	NN	O	O
degrees	NN	O	O
C	NN	O	O
in	NN	O	O
untreated	NN	O	O
normal	NN	O	O
subjects	NN	O	O
and	NN	O	O
in	NN	O	O
healthy	NN	O	O
subjects	NN	O	O
taking	NN	O	O
various	NN	O	O
glucocorticoid	NN	O	O
preparations	NN	O	O
.	NN	O	O

At	NN	O	O
24	NN	O	O
degrees	NN	O	O
C	NN	O	O
normal	NN	O	B-cell_type
human	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
had	NN	O	O
6000	NN	O	O
binding	NN	O	O
sites/cell	NN	O	O
and	NN	O	O
a	NN	O	O
dissociation	NN	O	O
constant	NN	O	O
of	NN	O	O
4	NN	O	O
x	NN	O	O
10	NN	O	O
(	NN	O	O
-9	NN	O	O
)	NN	O	O
M	NN	O	O
.	NN	O	O

The	NN	O	O
administration	NN	O	O
of	NN	O	O
1	NN	O	O
mg	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
,	NN	O	O
5	NN	O	O
mg	NN	O	O
of	NN	O	O
prednisone	NN	O	O
,	NN	O	O
and	NN	O	O
37.5	NN	O	O
mg	NN	O	O
of	NN	O	O
cortisone	NN	O	O
acetate	NN	O	O
resulted	NN	O	O
in	NN	O	O
a	NN	O	O
30	NN	O	O
%	NN	O	O
decrease	NN	O	O
in	NN	O	O
binding	NN	O	O
sites	NN	O	O
after	NN	O	O
1	NN	O	O
week	NN	O	O
with	NN	O	O
no	NN	O	O
change	NN	O	O
in	NN	O	O
binding	NN	O	O
affinity	NN	O	O
.	NN	O	O

No	NN	O	O
changes	NN	O	O
in	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
binding	NN	O	O
sites	NN	O	O
was	NN	O	O
noted	NN	O	O
before	NN	O	O
1	NN	O	O
week	NN	O	O
and	NN	O	O
the	NN	O	O
diminished	NN	O	O
number	NN	O	O
persisted	NN	O	O
for	NN	O	O
1	NN	O	O
week	NN	O	O
after	NN	O	O
discontinuation	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	O
treatment	NN	O	O
.	NN	O	O

Lymphocytes	NN	O	B-cell_type
from	NN	O	O
hospitalized	NN	O	O
patients	NN	O	O
taking	NN	O	O
40-60	NN	O	O
mg	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
daily	NN	O	O
demonstrated	NN	O	O
the	NN	O	O
same	NN	O	O
change	NN	O	O
in	NN	O	O
number	NN	O	O
of	NN	O	O
binding	NN	O	O
sites	NN	O	O
that	NN	O	O
was	NN	O	O
seen	NN	O	O
in	NN	O	O
normal	NN	O	O
subjects	NN	O	O
taking	NN	O	O
1	NN	O	O
mg	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
.	NN	O	O

When	NN	O	O
binding	NN	O	O
assays	NN	O	O
were	NN	O	O
carried	NN	O	O
out	NN	O	O
at	NN	O	O
physiologic	NN	O	O
temperature	NN	O	O
there	NN	O	O
was	NN	O	O
the	NN	O	O
same	NN	O	O
decrease	NN	O	O
in	NN	O	O
number	NN	O	O
of	NN	O	O
binding	NN	O	O
sites	NN	O	O
after	NN	O	O
dexamethasone	NN	O	O
administration	NN	O	O
,	NN	O	O
and	NN	O	O
in	NN	O	O
addition	NN	O	O
,	NN	O	O
there	NN	O	O
was	NN	O	O
a	NN	O	O
two-fold	NN	O	O
increase	NN	O	O
in	NN	O	O
binding	NN	O	O
affinity	NN	O	O
.	NN	O	O

Glucocorticoid	NN	O	O
administration	NN	O	O
results	NN	O	O
in	NN	O	O
a	NN	O	O
time-dependent	NN	O	O
decrease	NN	O	O
in	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
lymphocyte	NN	O	O
glucocorticoid	NN	O	O
binding	NN	O	O
sites	NN	O	O
that	NN	O	O
is	NN	O	O
independent	NN	O	O
of	NN	O	O
the	NN	O	O
type	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	O
administered	NN	O	O
.	NN	O	O

This	NN	O	O
is	NN	O	O
the	NN	O	O
first	NN	O	O
in	NN	O	O
vivo	NN	O	O
demonstration	NN	O	O
that	NN	O	O
glucocorticoids	NN	O	O
modulate	NN	O	O
their	NN	O	O
own	NN	O	O
receptors	NN	O	O
in	NN	O	O
man	NN	O	O
.	NN	O	O

-DOCSTART-	O

Immunoglobulin	NN	O	O
localization	NN	O	O
in	NN	O	O
benign	NN	O	O
and	NN	O	O
malignant	NN	O	O
lesions	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	O
mammary	NN	O	O
gland	NN	O	O
.	NN	O	O

Using	NN	O	O
direct	NN	O	O
imunofluorescence	NN	O	O
,	NN	O	O
lesions	NN	O	O
from	NN	O	O
266	NN	O	O
human	NN	O	O
breast	NN	O	O
specimens	NN	O	O
were	NN	O	O
studied	NN	O	O
for	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
IgA	NN	O	O
,	NN	O	O
IgM	NN	O	O
,	NN	O	O
or	NN	O	O
IgG	NN	O	O
localization	NN	O	O
.	NN	O	O

The	NN	O	O
lesions	NN	O	O
included	NN	O	O
benign	NN	O	O
elements	NN	O	O
from	NN	O	O
66	NN	O	O
subcutaneous	NN	O	O
mastectomy	NN	O	O
specimens	NN	O	O
in	NN	O	O
which	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
simultaneous	NN	O	O
breast	NN	O	O
malignancy	NN	O	O
was	NN	O	O
documented	NN	O	O
,	NN	O	O
primary	NN	O	O
breast	NN	O	O
carcinomas	NN	O	O
from	NN	O	O
153	NN	O	O
mastectomy	NN	O	O
specimens	NN	O	O
,	NN	O	O
and	NN	O	O
47	NN	O	O
biopsies	NN	O	O
containing	NN	O	O
metastatic	NN	O	O
breast	NN	O	O
cancer	NN	O	O
.	NN	O	O

A	NN	O	O
statistically	NN	O	O
significant	NN	O	O
association	NN	O	O
of	NN	O	O
IgA	NN	O	B-protein
and	NN	O	O
IgM	NN	O	B-protein
with	NN	O	O
benign	NN	O	O
lesions	NN	O	O
was	NN	O	O
contrasted	NN	O	O
to	NN	O	O
the	NN	O	O
association	NN	O	O
of	NN	O	O
IgG	NN	O	B-protein
with	NN	O	O
malignant	NN	O	O
lesions	NN	O	O
.	NN	O	O

In	NN	O	O
both	NN	O	O
primary	NN	O	O
and	NN	O	O
metastatic	NN	O	O
lesions	NN	O	O
,	NN	O	O
IgG	NN	O	B-protein
localization	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
estrogen-receptor-poor	NN	O	O
primary	NN	O	O
cancers	NN	O	O
as	NN	O	O
compared	NN	O	O
with	NN	O	O
estrogen-receptor-rich	NN	O	O
primary	NN	O	O
cancers	NN	O	O
.	NN	O	O

Among	NN	O	O
primary	NN	O	O
breast	NN	O	O
cancer	NN	O	O
patients	NN	O	O
,	NN	O	O
IgG	NN	O	B-protein
localization	NN	O	O
in	NN	O	O
the	NN	O	O
tumor	NN	O	O
correlated	NN	O	O
with	NN	O	O
relative	NN	O	O
lymphopenia	NN	O	O
.	NN	O	O

A	NN	O	O
shorter	NN	O	O
disease-free	NN	O	O
interval	NN	O	O
was	NN	O	O
noted	NN	O	O
in	NN	O	O
association	NN	O	O
with	NN	O	O
IgG	NN	O	B-protein
localization	NN	O	O
among	NN	O	O
the	NN	O	O
metastatic	NN	O	O
breast	NN	O	O
lesions	NN	O	O
.	NN	O	O

No	NN	O	O
statistically	NN	O	O
significant	NN	O	O
association	NN	O	O
between	NN	O	O
stage	NN	O	O
of	NN	O	O
disease	NN	O	O
and	NN	O	O
immunoglobulin	NN	O	O
presence	NN	O	O
was	NN	O	O
demonstrable	NN	O	O
.	NN	O	O

Moderate-to-severe	NN	O	O
intraductal	NN	O	O
epithelial	NN	O	O
hyperplasias	NN	O	O
were	NN	O	O
more	NN	O	O
often	NN	O	O
associated	NN	O	O
with	NN	O	O
immunoglobulin	NN	O	B-protein
G	NN	O	I-protein
localization	NN	O	O
that	NN	O	O
were	NN	O	O
other	NN	O	O
benign	NN	O	O
lesions	NN	O	O

-DOCSTART-	O

Correlation	NN	O	O
of	NN	O	O
steroid	NN	O	B-protein
receptors	NN	O	I-protein
with	NN	O	O
histologic	NN	O	O
differentiation	NN	O	O
in	NN	O	O
mammary	NN	O	O
carcinoma	NN	O	O
.	NN	O	O

A	NN	O	O
Singapore	NN	O	O
experience	NN	O	O
.	NN	O	O

Cancer	NN	O	O
of	NN	O	O
the	NN	O	O
breast	NN	O	O
is	NN	O	O
the	NN	O	O
most	NN	O	O
common	NN	O	O
tumor	NN	O	O
in	NN	O	O
females	NN	O	O
in	NN	O	O
Singapore	NN	O	O
,	NN	O	O
with	NN	O	O
the	NN	O	O
rate	NN	O	O
of	NN	O	O
20.7	NN	O	O
per	NN	O	O
100	NN	O	O
,	NN	O	O
000	NN	O	O
per	NN	O	O
year	NN	O	O
(	NN	O	O
1977	NN	O	O
estimate	NN	O	O
)	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
predicted	NN	O	O
to	NN	O	O
increase	NN	O	O
to	NN	O	O
29.8	NN	O	O
per	NN	O	O
100	NN	O	O
,	NN	O	O
000	NN	O	O
women	NN	O	O
per	NN	O	O
year	NN	O	O
by	NN	O	O
1995	NN	O	O
.	NN	O	O

A	NN	O	O
detailed	NN	O	O
histopathologic	NN	O	O
review	NN	O	O
of	NN	O	O
50	NN	O	O
primary	NN	O	O
breast	NN	O	O
cancer	NN	O	O
tumors	NN	O	O
analyzed	NN	O	O
for	NN	O	O
estrogen	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
ER	NN	O	B-protein
)	NN	O	O
level	NN	O	O
was	NN	O	O
carried	NN	O	O
out	NN	O	O
and	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
morphologic	NN	O	O
features	NN	O	O
correlated	NN	O	O
with	NN	O	O
ER	NN	O	B-protein
results	NN	O	O
to	NN	O	O
identify	NN	O	O
any	NN	O	O
factors	NN	O	O
that	NN	O	O
will	NN	O	O
improve	NN	O	O
the	NN	O	O
management	NN	O	O
and	NN	O	O
prognosis	NN	O	O
for	NN	O	O
breast	NN	O	O
cancer	NN	O	O
.	NN	O	O

Cytosol	NN	O	O
was	NN	O	O
incubated	NN	O	O
with	NN	O	O
3H-estradiol	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
and	NN	O	O
absence	NN	O	O
of	NN	O	O
cold	NN	O	O
diethylstilbestrol	NN	O	O
,	NN	O	O
and	NN	O	O
bound	NN	O	O
and	NN	O	O
free	NN	O	O
hormone	NN	O	O
were	NN	O	O
separated	NN	O	O
by	NN	O	O
Dextran-coated	NN	O	O
charcoal	NN	O	O
method	NN	O	O
.	NN	O	O

Tumors	NN	O	O
binding	NN	O	O
more	NN	O	O
than	NN	O	O
5	NN	O	O
fmol/mg	NN	O	O
cytosol	NN	O	B-protein
protein	NN	O	I-protein
were	NN	O	O
classified	NN	O	O
as	NN	O	O
ER	NN	O	B-protein
-positive	NN	O	O
.	NN	O	O

Progesterone	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
PR	NN	O	B-protein
)	NN	O	O
level	NN	O	O
was	NN	O	O
analyzed	NN	O	O
in	NN	O	O
some	NN	O	O
specimens	NN	O	O
with	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
a	NN	O	O
similar	NN	O	O
method	NN	O	O
.	NN	O	O

Most	NN	O	O
of	NN	O	O
the	NN	O	O
patients	NN	O	O
were	NN	O	O
Chinese	NN	O	O
(	NN	O	O
90	NN	O	O
%	NN	O	O
)	NN	O	O
.	NN	O	O

Three	NN	O	O
patients	NN	O	O
were	NN	O	O
Malays	NN	O	O
,	NN	O	O
one	NN	O	O
was	NN	O	O
Indian	NN	O	O
,	NN	O	O
and	NN	O	O
one	NN	O	O
was	NN	O	O
European	NN	O	O
in	NN	O	O
this	NN	O	O
series	NN	O	O
.	NN	O	O

Results	NN	O	O
indicated	NN	O	O
that	NN	O	O
there	NN	O	O
was	NN	O	O
strong	NN	O	O
correlation	NN	O	O
between	NN	O	O
ER	NN	O	B-protein
level	NN	O	O
,	NN	O	O
age	NN	O	O
,	NN	O	O
and	NN	O	O
histologic	NN	O	O
grade	NN	O	O
of	NN	O	O
the	NN	O	O
tumors	NN	O	O
.	NN	O	O

No	NN	O	O
correlation	NN	O	O
existed	NN	O	O
between	NN	O	O
absence	NN	O	O
or	NN	O	O
presence	NN	O	O
of	NN	O	O
lymph	NN	O	O
node	NN	O	O
metastases	NN	O	O
and	NN	O	O
ER	NN	O	B-protein
.	NN	O	O

Although	NN	O	O
there	NN	O	O
was	NN	O	O
a	NN	O	O
trend	NN	O	O
for	NN	O	O
ER	NN	O	B-protein
-positive	NN	O	O
tumors	NN	O	O
to	NN	O	O
have	NN	O	O
a	NN	O	O
low-grade	NN	O	O
lymphocytic	NN	O	O
infiltration	NN	O	O
,	NN	O	O
the	NN	O	O
difference	NN	O	O
was	NN	O	O
not	NN	O	O
statistically	NN	O	O
significant	NN	O	O
.	NN	O	O

-DOCSTART-	O

Mononuclear	NN	O	B-cell_type
cells	NN	O	I-cell_type
infiltrating	NN	O	O
human	NN	O	O
mammary	NN	O	O
carcinomas	NN	O	O
:	NN	O	O
immunohistochemical	NN	O	O
analysis	NN	O	O
with	NN	O	O
monoclonal	NN	O	B-protein
antibodies	NN	O	I-protein
.	NN	O	O

Breast	NN	O	O
carcinomas	NN	O	O
were	NN	O	O
examined	NN	O	O
by	NN	O	O
the	NN	O	O
immunoperoxidase	NN	O	O
technique	NN	O	O
using	NN	O	O
antisera	NN	O	O
specific	NN	O	O
for	NN	O	O
lymphocyte	NN	O	B-cell_type
subsets	NN	O	I-cell_type
,	NN	O	O
monocytes	NN	O	B-cell_type
,	NN	O	O
NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
major	NN	O	B-protein
histocompatibility	NN	O	I-protein
antigens	NN	O	I-protein
(	NN	O	O
HLA-A	NN	O	B-protein
,	NN	O	O
-B	NN	O	B-protein
,	NN	O	O
-C	NN	O	O
;	NN	O	O
Ia-like	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Sixty-four	NN	O	O
per	NN	O	O
cent	NN	O	O
of	NN	O	O
the	NN	O	O
patients	NN	O	O
had	NN	O	O
a	NN	O	O
moderate	NN	O	O
or	NN	O	O
strong	NN	O	O
mononuclear	NN	O	O
cell	NN	O	O
infiltration	NN	O	O
,	NN	O	O
77	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
patients	NN	O	O
without	NN	O	O
mononuclear	NN	O	B-cell_type
cell	NN	O	I-cell_type
infiltration	NN	O	O
had	NN	O	O
receptors	NN	O	O
for	NN	O	O
estrogens	NN	O	O
as	NN	O	O
compared	NN	O	O
to	NN	O	O
51	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
patients	NN	O	O
with	NN	O	O
infiltration	NN	O	O
.	NN	O	O

The	NN	O	O
majority	NN	O	O
of	NN	O	O
the	NN	O	O
infiltrating	NN	O	B-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
;	NN	O	O
generally	NN	O	O
the	NN	O	O
OKT8	NN	O	B-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
predominant	NN	O	O
.	NN	O	O

The	NN	O	O
Leu	NN	O	O
3A/OKT8	NN	O	O
cell	NN	O	O
ratio	NN	O	O
was	NN	O	O
not	NN	O	O
related	NN	O	O
to	NN	O	O
histological	NN	O	O
type	NN	O	O
,	NN	O	O
tumor	NN	O	O
size	NN	O	O
,	NN	O	O
age	NN	O	O
of	NN	O	O
the	NN	O	O
patient	NN	O	O
or	NN	O	O
presence	NN	O	O
of	NN	O	O
metastases	NN	O	O
.	NN	O	O

Some	NN	O	O
of	NN	O	O
the	NN	O	O
T	NN	O	O
cells	NN	O	O
had	NN	O	O
the	NN	O	O
Ia	NN	O	B-protein
antigen	NN	O	I-protein
and	NN	O	O
were	NN	O	O
thus	NN	O	O
probably	NN	O	O
activated	NN	O	O
.	NN	O	O

The	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
either	NN	O	O
absent	NN	O	O
or	NN	O	O
less	NN	O	O
numerous	NN	O	O
than	NN	O	O
the	NN	O	O
T	NN	O	O
cells	NN	O	O
.	NN	O	O

There	NN	O	O
was	NN	O	O
no	NN	O	O
relation	NN	O	O
between	NN	O	O
their	NN	O	O
distribution	NN	O	O
and	NN	O	O
the	NN	O	O
various	NN	O	O
parameters	NN	O	O
studied	NN	O	O
.	NN	O	O

A	NN	O	O
few	NN	O	O
monocytes	NN	O	B-cell_type
were	NN	O	O
heterogeneous	NN	O	O
according	NN	O	O
to	NN	O	O
their	NN	O	O
markers	NN	O	O
(	NN	O	O
OKM	NN	O	B-protein
I	NN	O	I-protein
and	NN	O	O
acid	NN	O	O
phosphatase	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
6	NN	O	O
cases	NN	O	O
only	NN	O	O
there	NN	O	O
was	NN	O	O
a	NN	O	O
strong	NN	O	O
infiltration	NN	O	O
of	NN	O	O
mononuclear	NN	O	B-cell_type
cells	NN	O	I-cell_type
positive	NN	O	O
for	NN	O	O
acid	NN	O	B-protein
phosphatase	NN	O	I-protein
.	NN	O	O

The	NN	O	O
number	NN	O	O
of	NN	O	O
the	NN	O	O
natural	NN	O	B-cell_type
killer	NN	O	I-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
also	NN	O	O
low	NN	O	O
.	NN	O	O

Only	NN	O	O
a	NN	O	O
few	NN	O	O
mononuclear	NN	O	O
infiltrating	NN	O	O
cells	NN	O	O
had	NN	O	O
receptors	NN	O	O
for	NN	O	O
transferrin	NN	O	O
.	NN	O	O

There	NN	O	O
was	NN	O	O
a	NN	O	O
positive	NN	O	O
correlation	NN	O	O
between	NN	O	O
the	NN	O	O
inflammatory	NN	O	O
infiltration	NN	O	O
and	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
HLA	NN	O	B-protein
class-I	NN	O	I-protein
antigens	NN	O	I-protein
on	NN	O	O
tumor	NN	O	B-cell_type
cell	NN	O	I-cell_type
s	NN	O	O
.	NN	O	O

Some	NN	O	O
of	NN	O	O
the	NN	O	O
antisera	NN	O	O
specific	NN	O	O
for	NN	O	O
lymphocyte	NN	O	B-cell_type
subsets	NN	O	I-cell_type
also	NN	O	O
stained	NN	O	O
the	NN	O	O
breast	NN	O	B-cell_type
carcinoma	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
great	NN	O	O
variations	NN	O	O
in	NN	O	O
the	NN	O	O
subsets	NN	O	O
of	NN	O	O
mononuclear	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
breast	NN	O	O
carcinomas	NN	O	O
may	NN	O	O
correspond	NN	O	O
to	NN	O	O
various	NN	O	O
systems	NN	O	O
of	NN	O	O
defense	NN	O	O
against	NN	O	O
neoplasm	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
case	NN	O	O
of	NN	O	O
male	NN	O	O
pseudohermaphroditism	NN	O	O
with	NN	O	O
normal	NN	O	O
androgen	NN	O	B-protein
receptor	NN	O	I-protein
binding	NN	O	O
and	NN	O	O
47	NN	O	O
,	NN	O	O
XYY	NN	O	O
karyotype	NN	O	O
.	NN	O	O

A	NN	O	O
case	NN	O	O
of	NN	O	O
male	NN	O	O
pseudohermaphroditism	NN	O	O
with	NN	O	O
47	NN	O	O
,	NN	O	O
XYY	NN	O	O
karyotype	NN	O	O
in	NN	O	O
blood	NN	O	O
and	NN	O	O
cutaneous	NN	O	B-cell_type
fibroblasts	NN	O	I-cell_type
is	NN	O	O
described	NN	O	O
.	NN	O	O

The	NN	O	O
plasma	NN	O	O
testosterone	NN	O	O
response	NN	O	O
to	NN	O	O
HCG	NN	O	O
stimulation	NN	O	O
was	NN	O	O
slightly	NN	O	O
below	NN	O	O
the	NN	O	O
normal	NN	O	O
range	NN	O	O
on	NN	O	O
two	NN	O	O
occasions	NN	O	O
suggesting	NN	O	O
a	NN	O	O
deficit	NN	O	O
of	NN	O	O
gonadal	NN	O	O
function	NN	O	O
.	NN	O	O

A	NN	O	O
study	NN	O	O
of	NN	O	O
the	NN	O	O
receptors	NN	O	O
for	NN	O	O
dihydrotestosterone	NN	O	O
in	NN	O	O
fibroblasts	NN	O	B-cell_type
of	NN	O	O
genital	NN	O	O
and	NN	O	O
nongenital	NN	O	O
skin	NN	O	O
showed	NN	O	O
a	NN	O	O
normal	NN	O	O
concentration	NN	O	O
of	NN	O	O
receptors	NN	O	O
in	NN	O	O
genital	NN	O	O
skin	NN	O	O
;	NN	O	O
5-alpha-reductase	NN	O	B-protein
activity	NN	O	O
in	NN	O	O
fibroblasts	NN	O	B-cell_type
of	NN	O	O
the	NN	O	O
genital	NN	O	O
skin	NN	O	O
was	NN	O	O
low	NN	O	O
,	NN	O	O
but	NN	O	O
the	NN	O	O
plasma	NN	O	O
relationship	NN	O	O
testosterone/dihydrotestosterone	NN	O	O
under	NN	O	O
HCG	NN	O	O
stimulation	NN	O	O
was	NN	O	O
normal	NN	O	O
.	NN	O	O

The	NN	O	O
diagnostic	NN	O	O
possibility	NN	O	O
of	NN	O	O
a	NN	O	O
complete	NN	O	O
testicular	NN	O	O
feminization	NN	O	O
syndrome	NN	O	O
with	NN	O	O
normal	NN	O	O
receptors	NN	O	O
for	NN	O	O
dihydrotestosterone	NN	O	O
is	NN	O	O
commented	NN	O	O
on	NN	O	O
.	NN	O	O

-DOCSTART-	O

1	NN	O	O
,	NN	O	O
25-Dihydroxyvitamin	NN	O	O
D3	NN	O	O
inhibits	NN	O	O
antigen-induced	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
activation	NN	O	O
.	NN	O	O

The	NN	O	O
proliferative	NN	O	O
response	NN	O	O
of	NN	O	O
murine	NN	O	B-cell_type
spleen	NN	O	I-cell_type
and	NN	O	I-cell_type
thymus	NN	O	I-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
antigen	NN	O	B-protein
but	NN	O	O
not	NN	O	O
to	NN	O	O
lectin	NN	O	B-protein
was	NN	O	O
inhibited	NN	O	O
by	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
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
directly	NN	O	O
examine	NN	O	O
the	NN	O	O
effect	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
on	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
activation	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
other	NN	O	O
complicating	NN	O	O
interactions	NN	O	O
,	NN	O	O
we	NN	O	O
utilized	NN	O	O
a	NN	O	O
panel	NN	O	O
of	NN	O	O
cloned	NN	O	O
Ia-restricted	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
hybridomas	NN	O	I-cell_line
that	NN	O	O
secrete	NN	O	O
IL	NN	O	B-protein
2	NN	O	I-protein
on	NN	O	O
activation	NN	O	O
by	NN	O	O
cloned	NN	O	O
Ia-bearing	NN	O	B-cell_type
stimulator	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
TA3	NN	O	B-cell_type
)	NN	O	O
or	NN	O	O
when	NN	O	O
stimulated	NN	O	O
by	NN	O	O
mitogen	NN	O	O
.	NN	O	O

Physiologic	NN	O	O
concentrations	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
0.01	NN	O	O
to	NN	O	O
0.1	NN	O	O
nm	NN	O	O
)	NN	O	O
inhibited	NN	O	O
the	NN	O	O
antigen-induced	NN	O	O
secretion	NN	O	O
of	NN	O	O
IL	NN	O	O
2	NN	O	O
by	NN	O	O
several	NN	O	O
of	NN	O	O
these	NN	O	O
T	NN	O	B-cell_line
cell	NN	O	I-cell_line
hybridomas	NN	O	I-cell_line
.	NN	O	O

This	NN	O	O
inhibition	NN	O	O
was	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
concentration	NN	O	O
of	NN	O	O
the	NN	O	O
free	NN	O	O
hormone	NN	O	O
and	NN	O	O
could	NN	O	O
be	NN	O	O
overcome	NN	O	O
by	NN	O	O
increasing	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
Ia-bearing	NN	O	B-cell_line
stimulator	NN	O	I-cell_line
cells	NN	O	I-cell_line
used	NN	O	O
.	NN	O	O

Pretreatment	NN	O	O
of	NN	O	O
the	NN	O	O
T	NN	O	B-cell_line
hybridoma	NN	O	I-cell_line
but	NN	O	O
not	NN	O	O
the	NN	O	O
TA3	NN	O	B-cell_line
stimulator	NN	O	I-cell_line
cell	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
resulted	NN	O	O
in	NN	O	O
inhibition	NN	O	O
of	NN	O	O
activation	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
are	NN	O	O
consistent	NN	O	O
with	NN	O	O
the	NN	O	O
finding	NN	O	O
that	NN	O	O
specific	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
are	NN	O	O
present	NN	O	O
on	NN	O	O
the	NN	O	O
T	NN	O	O
cell	NN	O	O
hybridomas	NN	O	O
but	NN	O	O
are	NN	O	O
lacking	NN	O	O
in	NN	O	O
TA3	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

1	NN	O	O
,	NN	O	O
25-	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
failed	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
to	NN	O	O
inhibit	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
T	NN	O	B-cell_line
cell	NN	O	I-cell_line
hybridomas	NN	O	I-cell_line
by	NN	O	O
lectin	NN	O	B-protein
or	NN	O	O
by	NN	O	O
an	NN	O	O
anti-Thy-1	NN	O	B-protein
antibody	NN	O	I-protein
.	NN	O	O

These	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
1	NN	O	O
,	NN	O	O
25-	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
may	NN	O	O
be	NN	O	O
interfering	NN	O	O
with	NN	O	O
early	NN	O	O
events	NN	O	O
of	NN	O	O
antigen-induced	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
activation	NN	O	O
,	NN	O	O
perhaps	NN	O	O
by	NN	O	O
hindering	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
recognition	NN	O	O
of	NN	O	O
the	NN	O	O
relevant	NN	O	O
antigen	NN	O	O
on	NN	O	O
stimulator	NN	O	O
cell	NN	O	O
surfaces	NN	O	O
.	NN	O	O

This	NN	O	O
system	NN	O	O
should	NN	O	O
prove	NN	O	O
useful	NN	O	O
in	NN	O	O
studying	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
by	NN	O	O
which	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
acts	NN	O	O
to	NN	O	O
inhibit	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
and	NN	O	O
subsequent	NN	O	O
IL	NN	O	B-protein
2	NN	O	I-protein
production	NN	O	O
.	NN	O	O

-DOCSTART-	O

Glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
and	NN	O	O
steroid	NN	O	O
sensitivity	NN	O	O
in	NN	O	O
normal	NN	O	O
and	NN	O	O
neoplastic	NN	O	O
human	NN	O	O
lymphoid	NN	O	O
tissues	NN	O	O
:	NN	O	O
a	NN	O	O
review	NN	O	O
.	NN	O	O

The	NN	O	O
determination	NN	O	O
of	NN	O	O
estrogen	NN	O	B-protein
and	NN	O	I-protein
progesterone	NN	O	I-protein
receptors	NN	O	I-protein
in	NN	O	O
breast	NN	O	O
cancer	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
useful	NN	O	O
in	NN	O	O
predicting	NN	O	O
the	NN	O	O
response	NN	O	O
to	NN	O	O
endocrine	NN	O	O
therapy	NN	O	O
.	NN	O	O

Given	NN	O	O
their	NN	O	O
well-known	NN	O	O
inhibitory	NN	O	O
effects	NN	O	O
on	NN	O	O
lymphoid	NN	O	O
tissue	NN	O	O
,	NN	O	O
glucocorticoids	NN	O	O
have	NN	O	O
been	NN	O	O
used	NN	O	O
widely	NN	O	O
in	NN	O	O
the	NN	O	O
treatment	NN	O	O
of	NN	O	O
leukemia	NN	O	O
.	NN	O	O

Given	NN	O	O
these	NN	O	O
facts	NN	O	O
,	NN	O	O
over	NN	O	O
the	NN	O	O
last	NN	O	O
10	NN	O	O
years	NN	O	O
,	NN	O	O
several	NN	O	O
investigators	NN	O	O
have	NN	O	O
measured	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
normal	NN	O	O
and	NN	O	O
neoplastic	NN	O	O
lymphoid	NN	O	O
tissue	NN	O	O
to	NN	O	O
see	NN	O	O
whether	NN	O	O
their	NN	O	O
number	NN	O	O
correlated	NN	O	O
with	NN	O	O
glucocorticoid	NN	O	O
responsiveness	NN	O	O
in	NN	O	O
vitro	NN	O	O
or	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

No	NN	O	O
clear	NN	O	O
correlation	NN	O	O
could	NN	O	O
be	NN	O	O
established	NN	O	O
between	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
and	NN	O	O
the	NN	O	O
in	NN	O	O
vitro	NN	O	O
action	NN	O	O
of	NN	O	O
steroids	NN	O	O
in	NN	O	O
normal	NN	O	O
and	NN	O	O
neoplastic	NN	O	O
lymphoid	NN	O	O
tissue	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
attempts	NN	O	O
to	NN	O	O
correlate	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
levels	NN	O	O
in	NN	O	O
acute	NN	O	O
lymphocytic	NN	O	O
leukemia	NN	O	O
to	NN	O	O
in	NN	O	O
vivo	NN	O	O
steroid	NN	O	O
responsiveness	NN	O	O
and	NN	O	O
immunological	NN	O	O
type	NN	O	O
using	NN	O	O
the	NN	O	O
whole-cell-binding	NN	O	O
assay	NN	O	O
for	NN	O	O
receptor	NN	O	O
determination	NN	O	O
and	NN	O	O
selecting	NN	O	O
the	NN	O	O
patients	NN	O	O
according	NN	O	O
to	NN	O	O
age	NN	O	O
and	NN	O	O
immunological	NN	O	O
criteria	NN	O	O
have	NN	O	O
been	NN	O	O
more	NN	O	O
successful	NN	O	O
.	NN	O	O

-DOCSTART-	O

[	NN	O	O
Glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
normal	NN	O	B-cell_type
human	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
]	NN	O	O

Glucocorticoid	NN	O	B-cell_type
(	NN	O	I-cell_type
GC	NN	O	I-cell_type
)	NN	O	I-cell_type
receptors	NN	O	I-cell_type
were	NN	O	O
studied	NN	O	O
in	NN	O	O
intact	NN	O	O
lymphocytes	NN	O	B-cell_type
from	NN	O	O
11	NN	O	O
donors	NN	O	O
.	NN	O	O

GC	NN	O	O
binding	NN	O	O
parameters	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
highly	NN	O	O
reproducible	NN	O	O
in	NN	O	O
repeated	NN	O	O
experiments	NN	O	O
with	NN	O	O
lymphocytes	NN	O	B-cell_type
.	NN	O	O

It	NN	O	O
was	NN	O	O
shown	NN	O	O
that	NN	O	O
GC	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
donors	NN	O	O
'	NN	O	O
lymphocytes	NN	O	B-cell_type
could	NN	O	O
be	NN	O	O
distributed	NN	O	O
into	NN	O	O
two	NN	O	O
different	NN	O	O
classes	NN	O	O
similarly	NN	O	O
to	NN	O	O
the	NN	O	O
pattern	NN	O	O
seen	NN	O	O
in	NN	O	O
skin	NN	O	B-cell_type
fibroblasts	NN	O	I-cell_type
.	NN	O	O

Human	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
are	NN	O	O
an	NN	O	O
adequate	NN	O	O
object	NN	O	O
for	NN	O	O
studying	NN	O	O
genetically	NN	O	O
determined	NN	O	O
variability	NN	O	O
of	NN	O	O
GC	NN	O	B-protein
receptors	NN	O	I-protein
and	NN	O	O
its	NN	O	O
clinical	NN	O	O
importance	NN	O	O
.	NN	O	O

-DOCSTART-	O

Specific	NN	O	O
estrogen	NN	O	B-protein
binding	NN	O	I-protein
sites	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
lymphoid	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
thymic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
binding	NN	O	O
of	NN	O	O
estrogen	NN	O	O
in	NN	O	O
preparations	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	I-cell_type
as	NN	O	O
well	NN	O	O
as	NN	O	O
by	NN	O	O
splenic	NN	O	B-cell_type
and	NN	O	I-cell_type
thymic	NN	O	I-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
demonstrated	NN	O	O
by	NN	O	O
three	NN	O	O
different	NN	O	O
approaches	NN	O	O
(	NN	O	O
Dextran-coated	NN	O	O
charcoal	NN	O	O
method	NN	O	O
,	NN	O	O
whole	NN	O	O
cell	NN	O	O
assay	NN	O	O
,	NN	O	O
and	NN	O	O
gel	NN	O	O
filtration	NN	O	O
on	NN	O	O
a	NN	O	O
sepharose	NN	O	O
4B	NN	O	O
column	NN	O	O
)	NN	O	O
.	NN	O	O

Scatchard	NN	O	O
's	NN	O	O
analysis	NN	O	O
of	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
-moxestrol	NN	O	O
(	NN	O	O
R2858	NN	O	O
)	NN	O	O
and	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
-estradiol	NN	O	O
binding	NN	O	O
proves	NN	O	O
the	NN	O	O
existence	NN	O	O
of	NN	O	O
a	NN	O	O
single	NN	O	O
class	NN	O	O
of	NN	O	O
receptor	NN	O	B-protein
sites	NN	O	I-protein
having	NN	O	O
a	NN	O	O
dissociation	NN	O	O
constant	NN	O	O
of	NN	O	O
0.18-2.4	NN	O	O
X	NN	O	O
10	NN	O	O
(	NN	O	O
-9	NN	O	O
)	NN	O	O
M	NN	O	O
.	NN	O	O

Physicochemical	NN	O	O
properties	NN	O	O
of	NN	O	O
the	NN	O	O
binder	NN	O	O
,	NN	O	O
including	NN	O	O
binding	NN	O	O
capacity	NN	O	O
and	NN	O	O
steroid	NN	O	O
specificity	NN	O	O
,	NN	O	O
are	NN	O	O
quite	NN	O	O
similar	NN	O	O
to	NN	O	O
those	NN	O	O
reported	NN	O	O
for	NN	O	O
the	NN	O	O
thymus	NN	O	O
of	NN	O	O
small	NN	O	O
mammalian	NN	O	O
species	NN	O	O
or	NN	O	O
human	NN	O	O
thymoma	NN	O	O
.	NN	O	O

-DOCSTART-	O

Administration	NN	O	O
of	NN	O	O
fibroblast	NN	O	B-protein
interferon	NN	O	I-protein
to	NN	O	O
patients	NN	O	O
with	NN	O	O
advanced	NN	O	O
breast	NN	O	O
cancer	NN	O	O
:	NN	O	O
possible	NN	O	O
effects	NN	O	O
on	NN	O	O
skin	NN	O	O
metastasis	NN	O	O
and	NN	O	O
on	NN	O	O
hormone	NN	O	B-protein
receptors	NN	O	I-protein
.	NN	O	O

Eleven	NN	O	O
patients	NN	O	O
with	NN	O	O
metastasized	NN	O	O
breast	NN	O	O
cancer	NN	O	O
received	NN	O	O
8	NN	O	O
intramuscular	NN	O	O
injections	NN	O	O
of	NN	O	O
6	NN	O	O
x	NN	O	O
10	NN	O	O
(	NN	O	O
6	NN	O	O
)	NN	O	O
units	NN	O	O
of	NN	O	O
human	NN	O	B-protein
fibroblast	NN	O	I-protein
interferon	NN	O	I-protein
over	NN	O	O
a	NN	O	O
period	NN	O	O
of	NN	O	O
40	NN	O	O
days	NN	O	O
.	NN	O	O

The	NN	O	O
injections	NN	O	O
did	NN	O	O
not	NN	O	O
cause	NN	O	O
local	NN	O	O
irritation	NN	O	O
or	NN	O	O
inflammation	NN	O	O
.	NN	O	O

Fever	NN	O	O
occurred	NN	O	O
in	NN	O	O
only	NN	O	O
1	NN	O	O
of	NN	O	O
the	NN	O	O
11	NN	O	O
patients	NN	O	O
.	NN	O	O

Although	NN	O	O
several	NN	O	O
types	NN	O	O
of	NN	O	O
metastases	NN	O	O
were	NN	O	O
monitored	NN	O	O
,	NN	O	O
only	NN	O	O
skin	NN	O	O
nodules	NN	O	O
consistently	NN	O	O
(	NN	O	O
10	NN	O	O
out	NN	O	O
of	NN	O	O
11	NN	O	O
patients	NN	O	O
)	NN	O	O
exhibited	NN	O	O
changes	NN	O	O
that	NN	O	O
were	NN	O	O
suggestive	NN	O	O
of	NN	O	O
a	NN	O	O
therapeutic	NN	O	O
effect	NN	O	O
of	NN	O	O
the	NN	O	O
treatment	NN	O	O
regimen	NN	O	O
:	NN	O	O
either	NN	O	O
a	NN	O	O
simple	NN	O	O
decrease	NN	O	O
in	NN	O	O
size	NN	O	O
of	NN	O	O
some	NN	O	O
nodules	NN	O	O
or	NN	O	O
central	NN	O	O
necrosis	NN	O	O
accompanied	NN	O	O
by	NN	O	O
an	NN	O	O
inflammatory	NN	O	O
reaction	NN	O	O
.	NN	O	O

NK-activity	NN	O	O
of	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
leukocytes	NN	O	I-cell_type
was	NN	O	O
significantly	NN	O	O
increased	NN	O	O
after	NN	O	O
administration	NN	O	O
of	NN	O	O
the	NN	O	O
first	NN	O	O
dose	NN	O	O
;	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
subsequent	NN	O	O
injections	NN	O	O
was	NN	O	O
less	NN	O	O
clear	NN	O	O
.	NN	O	O

Receptors	NN	O	O
for	NN	O	O
estrogens	NN	O	O
and	NN	O	O
progestogens	NN	O	O
were	NN	O	O
increased	NN	O	O
in	NN	O	O
the	NN	O	O
tumor	NN	O	O
biopsies	NN	O	O
of	NN	O	O
2	NN	O	O
out	NN	O	O
of	NN	O	O
2	NN	O	O
and	NN	O	O
5	NN	O	O
out	NN	O	O
of	NN	O	O
6	NN	O	O
patients	NN	O	O
tested	NN	O	O
respectively	NN	O	O
.	NN	O	O

-DOCSTART-	O

Decreased	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
binding	NN	O	O
in	NN	O	O
adrenal	NN	O	O
insufficiency	NN	O	O
.	NN	O	O

To	NN	O	O
examine	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	O
deficiency	NN	O	O
on	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
,	NN	O	O
we	NN	O	O
examine	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
dexamethasone	NN	O	O
to	NN	O	O
lymphocytes	NN	O	B-cell_type
in	NN	O	O
normal	NN	O	O
subjects	NN	O	O
and	NN	O	O
patients	NN	O	O
with	NN	O	O
adrenal	NN	O	O
insufficiency	NN	O	O
before	NN	O	O
and	NN	O	O
after	NN	O	O
glucocorticoid	NN	O	O
replacement	NN	O	O
therapy	NN	O	O
.	NN	O	O

Using	NN	O	O
a	NN	O	O
whole	NN	O	O
cell	NN	O	O
competitive	NN	O	O
binding	NN	O	O
assay	NN	O	O
,	NN	O	O
normal	NN	O	B-cell_type
human	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
had	NN	O	O
5977	NN	O	O
+/-	NN	O	O
1487	NN	O	O
(	NN	O	O
mean	NN	O	O
+/-	NN	O	O
SD	NN	O	O
)	NN	O	O
binding	NN	O	O
sites/cell	NN	O	O
and	NN	O	O
a	NN	O	O
dissociation	NN	O	O
constant	NN	O	O
of	NN	O	O
10	NN	O	O
+/-	NN	O	O
2	NN	O	O
nM	NN	O	O
.	NN	O	O

Lymphocytes	NN	O	B-cell_type
from	NN	O	O
patients	NN	O	O
with	NN	O	O
untreated	NN	O	O
adrenal	NN	O	O
insufficiency	NN	O	O
had	NN	O	O
fewer	NN	O	O
binding	NN	O	B-protein
sites	NN	O	I-protein
(	NN	O	O
3364	NN	O	O
+/-322	NN	O	O
)	NN	O	O
and	NN	O	O
a	NN	O	O
2-fold	NN	O	O
increase	NN	O	O
in	NN	O	O
binding	NN	O	O
affinity	NN	O	O
(	NN	O	O
5.4	NN	O	O
+/-	NN	O	O
0.9	NN	O	O
mM	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
administration	NN	O	O
of	NN	O	O
conventional	NN	O	O
replacement	NN	O	O
doses	NN	O	O
of	NN	O	O
cortisone	NN	O	O
acetate	NN	O	O
for	NN	O	O
6	NN	O	O
months	NN	O	O
caused	NN	O	O
no	NN	O	O
change	NN	O	O
in	NN	O	O
receptor	NN	O	O
number	NN	O	O
,	NN	O	O
but	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
decrease	NN	O	O
in	NN	O	O
binding	NN	O	O
affinity	NN	O	O
toward	NN	O	O
normal	NN	O	O
.	NN	O	O

After	NN	O	O
long	NN	O	O
term	NN	O	O
glucocorticoid	NN	O	O
replacement	NN	O	O
therapy	NN	O	O
,	NN	O	O
binding	NN	O	O
parameters	NN	O	O
were	NN	O	O
similar	NN	O	O
to	NN	O	O
those	NN	O	O
in	NN	O	O
patients	NN	O	O
before	NN	O	O
treatment	NN	O	O
.	NN	O	O

The	NN	O	O
physiological	NN	O	O
implications	NN	O	O
of	NN	O	O
the	NN	O	O
decreased	NN	O	O
receptor	NN	O	O
number	NN	O	O
and	NN	O	O
increased	NN	O	O
binding	NN	O	O
affinity	NN	O	O
in	NN	O	O
adrenal	NN	O	O
insufficiency	NN	O	O
remain	NN	O	O
to	NN	O	O
be	NN	O	O
elucidated	NN	O	O
.	NN	O	O

-DOCSTART-	O

Glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
concentrations	NN	O	O
and	NN	O	O
terminal	NN	O	O
transferase	NN	O	B-protein
activity	NN	O	O
as	NN	O	O
indicators	NN	O	O
of	NN	O	O
prognosis	NN	O	O
in	NN	O	O
acute	NN	O	O
non-lymphocytic	NN	O	O
leukaemia	NN	O	O
.	NN	O	O

Activity	NN	O	O
of	NN	O	O
terminal	NN	O	B-protein
deoxynucleotidyl	NN	O	I-protein
transferase	NN	O	I-protein
(	NN	O	O
TdT	NN	O	B-protein
)	NN	O	O
,	NN	O	O
adenosine	NN	O	B-protein
deaminase	NN	O	I-protein
,	NN	O	O
and	NN	O	O
5'nucleotidase	NN	O	B-protein
and	NN	O	O
the	NN	O	O
cellular	NN	O	O
concentration	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
(	NN	O	I-protein
dexamethasone	NN	O	I-protein
)	NN	O	I-protein
receptor	NN	O	I-protein
were	NN	O	O
determined	NN	O	O
in	NN	O	O
25	NN	O	O
patients	NN	O	O
with	NN	O	O
acute	NN	O	O
non-lymphocytic	NN	O	O
leukaemia	NN	O	O
.	NN	O	O

All	NN	O	O
patients	NN	O	O
were	NN	O	O
treated	NN	O	O
according	NN	O	O
to	NN	O	O
a	NN	O	O
common	NN	O	O
protocol	NN	O	O
.	NN	O	O

Increased	NN	O	O
activity	NN	O	O
of	NN	O	O
TdT	NN	O	B-protein
(	NN	O	O
greater	NN	O	O
than	NN	O	O
0.1	NN	O	O
unit/microgram	NN	O	O
DNA	NN	O	O
)	NN	O	O
was	NN	O	O
found	NN	O	O
in	NN	O	O
11	NN	O	O
patients	NN	O	O
.	NN	O	O

This	NN	O	O
group	NN	O	O
of	NN	O	O
patients	NN	O	O
was	NN	O	O
shown	NN	O	O
to	NN	O	O
have	NN	O	O
higher	NN	O	O
remission	NN	O	O
and	NN	O	O
survival	NN	O	O
rates	NN	O	O
(	NN	O	O
p	NN	O	O
=	NN	O	O
0.06	NN	O	O
)	NN	O	O
compared	NN	O	O
with	NN	O	O
patients	NN	O	O
with	NN	O	O
low	NN	O	O
activity	NN	O	O
of	NN	O	O
TdT	NN	O	O
.	NN	O	O

The	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
concentration	NN	O	O
of	NN	O	O
the	NN	O	O
leukaemic	NN	O	B-cell_type
blast	NN	O	I-cell_type
cells	NN	O	I-cell_type
ranged	NN	O	O
from	NN	O	O
0	NN	O	O
to	NN	O	O
0.94	NN	O	O
fmol/microgram	NN	O	O
DNA	NN	O	O
.	NN	O	O

Thirteen	NN	O	O
patients	NN	O	O
had	NN	O	O
blast	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
a	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
concentration	NN	O	O
over	NN	O	O
0.22	NN	O	O
fmol/microgram	NN	O	O
DNA	NN	O	O
.	NN	O	O

These	NN	O	O
patients	NN	O	O
had	NN	O	O
significantly	NN	O	O
increased	NN	O	O
remission	NN	O	O
and	NN	O	O
survival	NN	O	O
rates	NN	O	O
(	NN	O	O
p	NN	O	O
=	NN	O	O
0.006	NN	O	O
)	NN	O	O
compared	NN	O	O
with	NN	O	O
those	NN	O	O
with	NN	O	O
a	NN	O	O
low	NN	O	O
receptor	NN	O	O
concentration	NN	O	O
.	NN	O	O

This	NN	O	O
finding	NN	O	O
can	NN	O	O
not	NN	O	O
be	NN	O	O
explained	NN	O	O
by	NN	O	O
a	NN	O	O
difference	NN	O	O
in	NN	O	O
sensitivity	NN	O	O
to	NN	O	O
glucocorticoids	NN	O	O
since	NN	O	O
these	NN	O	O
were	NN	O	O
not	NN	O	O
used	NN	O	O
as	NN	O	O
therapeutic	NN	O	O
agents	NN	O	O
.	NN	O	O

Adenosine	NN	O	B-protein
deaminase	NN	O	I-protein
and	NN	O	O
5'nucleotidase	NN	O	B-protein
activities	NN	O	O
both	NN	O	O
varied	NN	O	O
within	NN	O	O
two	NN	O	O
orders	NN	O	O
of	NN	O	O
magnitude	NN	O	O
.	NN	O	O

No	NN	O	O
correlation	NN	O	O
could	NN	O	O
be	NN	O	O
found	NN	O	O
between	NN	O	O
activities	NN	O	O
of	NN	O	O
these	NN	O	O
enzymes	NN	O	O
and	NN	O	O
remission	NN	O	O
or	NN	O	O
survival	NN	O	O
rate	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
show	NN	O	O
that	NN	O	O
measurements	NN	O	O
of	NN	O	O
TdT	NN	O	B-protein
activity	NN	O	O
and	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
concentration	NN	O	O
yield	NN	O	O
valuable	NN	O	O
prognostic	NN	O	O
information	NN	O	O
in	NN	O	O
acute	NN	O	O
non-lymphocytic	NN	O	O
leukaemia	NN	O	O

-DOCSTART-	O

[	NN	O	O
3H	NN	O	O
]	NN	O	O
cortivazol	NN	O	O
:	NN	O	O
a	NN	O	O
unique	NN	O	O
high	NN	O	O
affinity	NN	O	O
ligand	NN	O	O
for	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
.	NN	O	O

Cortivazol	NN	O	O
(	NN	O	O
CVZ	NN	O	O
)	NN	O	O
and	NN	O	O
deacylcortivazol	NN	O	O
(	NN	O	O
DAC	NN	O	O
)	NN	O	O
are	NN	O	O
pyrazolosteroids	NN	O	O
with	NN	O	O
potent	NN	O	O
glucocorticoid	NN	O	O
activity	NN	O	O
.	NN	O	O

In	NN	O	O
previous	NN	O	O
work	NN	O	O
we	NN	O	O
showed	NN	O	O
that	NN	O	O
DAC	NN	O	O
is	NN	O	O
40-fold	NN	O	O
more	NN	O	O
potent	NN	O	O
than	NN	O	O
dexamethasone	NN	O	O
(	NN	O	O
DEX	NN	O	O
)	NN	O	O
in	NN	O	O
lysing	NN	O	O
leukemic	NN	O	B-cell_type
lymphoblasts	NN	O	I-cell_type
.	NN	O	O

To	NN	O	O
assess	NN	O	O
the	NN	O	O
interaction	NN	O	O
between	NN	O	O
these	NN	O	O
atypical	NN	O	O
steroids	NN	O	O
and	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
,	NN	O	O
we	NN	O	O
examined	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
CVZ	NN	O	O
to	NN	O	O
cytosol	NN	O	O
from	NN	O	O
glucocorticoid-sensitive	NN	O	B-cell_line
and	NN	O	I-cell_line
-resistant	NN	O	I-cell_line
variants	NN	O	I-cell_line
of	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
leukemic	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
CEM	NN	O	B-cell_line
C7	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
glucocorticoid-sensitive	NN	O	B-cell_line
cells	NN	O	I-cell_line
[	NN	O	O
3H	NN	O	O
]	NN	O	O
CVZ	NN	O	O
causes	NN	O	O
a	NN	O	O
2-fold	NN	O	O
induction	NN	O	O
of	NN	O	O
glutamine	NN	O	B-protein
synthetase	NN	O	I-protein
and	NN	O	O
binds	NN	O	O
to	NN	O	O
a	NN	O	O
protein	NN	O	O
in	NN	O	O
the	NN	O	O
4.6	NN	O	O
S	NN	O	O
region	NN	O	O
of	NN	O	O
high	NN	O	O
salt	NN	O	O
sucrose	NN	O	O
gradients	NN	O	O
.	NN	O	O

On	NN	O	O
DEAE-cellulose	NN	O	O
chromatography	NN	O	O
,	NN	O	O
[	NN	O	B-protein
3H	NN	O	I-protein
]	NN	O	I-protein
CVZ-receptor	NN	O	I-protein
complexes	NN	O	I-protein
show	NN	O	O
a	NN	O	O
shift	NN	O	O
from	NN	O	O
high	NN	O	O
(	NN	O	O
0.25	NN	O	O
M	NN	O	O
KP	NN	O	O
)	NN	O	O
to	NN	O	O
low	NN	O	O
salt	NN	O	O
(	NN	O	O
0.09	NN	O	O
M	NN	O	O
KP	NN	O	O
)	NN	O	O
eluting	NN	O	O
forms	NN	O	O
upon	NN	O	O
activation	NN	O	O
.	NN	O	O

CVZ	NN	O	O
competes	NN	O	O
for	NN	O	O
a	NN	O	O
97	NN	O	O
,	NN	O	O
000-dalton	NN	O	O
protein	NN	O	O
labeled	NN	O	O
by	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
dexamethasone	NN	O	O
mesylate	NN	O	O
.	NN	O	O

Scatchard	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
CVZ	NN	O	O
in	NN	O	O
glucocorticoid-sensitive	NN	O	B-cell_line
cells	NN	O	I-cell_line
revealed	NN	O	O
a	NN	O	O
curvilinear	NN	O	O
plot	NN	O	O
which	NN	O	O
resolved	NN	O	O
into	NN	O	O
high	NN	O	O
(	NN	O	O
0.4	NN	O	O
nM	NN	O	O
)	NN	O	O
and	NN	O	O
low	NN	O	O
(	NN	O	O
11	NN	O	O
nM	NN	O	O
)	NN	O	O
affinity	NN	O	O
components	NN	O	O
.	NN	O	O

The	NN	O	O
receptor	NN	O	O
concentration	NN	O	O
of	NN	O	O
the	NN	O	O
low	NN	O	O
affinity	NN	O	O
site	NN	O	O
(	NN	O	O
0.30	NN	O	O
pmol/mg	NN	O	O
protein	NN	O	O
)	NN	O	O
was	NN	O	O
approximately	NN	O	O
twice	NN	O	O
that	NN	O	O
of	NN	O	O
the	NN	O	O
high	NN	O	B-protein
affinity	NN	O	I-protein
site	NN	O	I-protein
(	NN	O	O
0.14	NN	O	O
pmol/mg	NN	O	O
protein	NN	O	O
)	NN	O	O
.	NN	O	O

Dissociation	NN	O	O
experiments	NN	O	O
with	NN	O	O
dilution	NN	O	O
and/or	NN	O	O
excess	NN	O	O
unlabeled	NN	O	O
CVZ	NN	O	O
supported	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
independent	NN	O	O
sites	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
DEX	NN	O	O
to	NN	O	O
C7	NN	O	O
cytosol	NN	O	O
revealed	NN	O	O
a	NN	O	O
single	NN	O	O
class	NN	O	O
of	NN	O	O
binding	NN	O	O
sites	NN	O	O
(	NN	O	O
Kd	NN	O	O
=	NN	O	O
1.9	NN	O	O
nM	NN	O	O
;	NN	O	O
receptor	NN	O	O
concentration	NN	O	O
,	NN	O	O
0.46	NN	O	O
pmol/mg	NN	O	O
protein	NN	O	O
)	NN	O	O
.	NN	O	O

Examination	NN	O	O
of	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
CVZ	NN	O	O
using	NN	O	O
10	NN	O	O
(	NN	O	O
-5	NN	O	O
)	NN	O	O
M	NN	O	O
DEX	NN	O	O
as	NN	O	O
the	NN	O	O
competing	NN	O	O
ligand	NN	O	O
showed	NN	O	O
that	NN	O	O
DEX	NN	O	O
binds	NN	O	O
only	NN	O	O
to	NN	O	O
the	NN	O	O
low	NN	O	O
affinity	NN	O	O
site	NN	O	O
detected	NN	O	O
by	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
CVZ	NN	O	O
.	NN	O	O

In	NN	O	O
cytosol	NN	O	O
from	NN	O	O
a	NN	O	O
glucocorticoid-resistant	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
with	NN	O	O
virtually	NN	O	O
no	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
DEX	NN	O	O
binding	NN	O	O
,	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
CVZ	NN	O	O
detected	NN	O	O
a	NN	O	O
single	NN	O	O
high	NN	O	B-protein
affinity	NN	O	I-protein
binding	NN	O	I-protein
site	NN	O	I-protein
that	NN	O	O
was	NN	O	O
similar	NN	O	O
in	NN	O	O
dissociation	NN	O	O
constant	NN	O	O
(	NN	O	O
0.8	NN	O	O
nM	NN	O	O
)	NN	O	O
and	NN	O	O
receptor	NN	O	O
concentration	NN	O	O
(	NN	O	O
0.13	NN	O	O
pmol/mg	NN	O	O
protein	NN	O	O
)	NN	O	O
to	NN	O	O
the	NN	O	O
high	NN	O	B-protein
affinity	NN	O	I-protein
site	NN	O	I-protein
detected	NN	O	O
in	NN	O	O
the	NN	O	O
glucocorticoid-sensitive	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
C7	NN	O	B-cell_line
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
controlled	NN	O	O
pore	NN	O	O
glass	NN	O	O
bead	NN	O	O
assay	NN	O	O
for	NN	O	O
the	NN	O	O
measurement	NN	O	O
of	NN	O	O
cytoplasmic	NN	O	B-protein
and	NN	O	I-protein
nuclear	NN	O	I-protein
glucocorticoid	NN	O	I-protein
receptors	NN	O	I-protein
.	NN	O	O

An	NN	O	O
assay	NN	O	O
for	NN	O	O
the	NN	O	O
quantitation	NN	O	O
of	NN	O	O
cytoplasmic	NN	O	B-protein
and	NN	O	I-protein
nuclear	NN	O	I-protein
glucocorticoid	NN	O	I-protein
receptors	NN	O	I-protein
in	NN	O	O
lymphoid	NN	O	O
tissue	NN	O	O
has	NN	O	O
been	NN	O	O
developed	NN	O	O
using	NN	O	O
controlled	NN	O	O
pore	NN	O	O
glass	NN	O	O
(	NN	O	O
CPG	NN	O	O
)	NN	O	O
beads	NN	O	O
.	NN	O	O

Soluble	NN	O	O
receptor	NN	O	B-protein
--	NN	O	I-protein
3H-steroid	NN	O	I-protein
complex	NN	O	I-protein
(	NN	O	O
cytosol	NN	O	O
or	NN	O	O
nuclear	NN	O	O
extract	NN	O	O
)	NN	O	O
is	NN	O	O
adsorbed	NN	O	O
quantitatively	NN	O	O
within	NN	O	O
the	NN	O	O
crevasses	NN	O	O
of	NN	O	O
porous	NN	O	O
glass	NN	O	O
beads	NN	O	O
.	NN	O	O

Excess	NN	O	O
labeled	NN	O	O
steroid	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
most	NN	O	O
non-specifically	NN	O	O
bound	NN	O	O
steroid	NN	O	O
is	NN	O	O
easily	NN	O	O
washed	NN	O	O
away	NN	O	O
,	NN	O	O
leaving	NN	O	O
the	NN	O	O
hormone-receptor	NN	O	B-protein
complex	NN	O	I-protein
retained	NN	O	O
by	NN	O	O
the	NN	O	O
beads	NN	O	O
.	NN	O	O

Bound	NN	O	O
3H-steroid	NN	O	O
is	NN	O	O
eluted	NN	O	O
with	NN	O	O
ethanol	NN	O	O
and	NN	O	O
measured	NN	O	O
for	NN	O	O
radioactivity	NN	O	O
.	NN	O	O

This	NN	O	O
procedure	NN	O	O
which	NN	O	O
is	NN	O	O
simple	NN	O	O
,	NN	O	O
rapid	NN	O	O
,	NN	O	O
and	NN	O	O
highly	NN	O	O
reproducible	NN	O	O
is	NN	O	O
carried	NN	O	O
out	NN	O	O
using	NN	O	O
frozen	NN	O	O
samples	NN	O	O
(	NN	O	O
stable	NN	O	O
for	NN	O	O
many	NN	O	O
months	NN	O	O
)	NN	O	O
containing	NN	O	O
as	NN	O	O
few	NN	O	O
as	NN	O	O
1	NN	O	O
X	NN	O	O
10	NN	O	O
(	NN	O	O
7	NN	O	O
)	NN	O	O
cells	NN	O	O
.	NN	O	O

A	NN	O	O
comparison	NN	O	O
of	NN	O	O
the	NN	O	O
CPG	NN	O	O
assay	NN	O	O
to	NN	O	O
dextran	NN	O	O
coated	NN	O	O
charcoal	NN	O	O
and	NN	O	O
a	NN	O	O
whole	NN	O	O
cell	NN	O	O
assay	NN	O	O
demonstrates	NN	O	O
that	NN	O	O
CPG	NN	O	O
and	NN	O	O
dextran	NN	O	O
coated	NN	O	O
charcoal	NN	O	O
give	NN	O	O
equivalent	NN	O	O
measurements	NN	O	O
of	NN	O	O
cytosolic	NN	O	B-protein
receptor	NN	O	I-protein
concentration	NN	O	O
,	NN	O	O
while	NN	O	O
the	NN	O	O
CPG	NN	O	O
and	NN	O	O
whole	NN	O	O
cell	NN	O	O
assays	NN	O	O
provide	NN	O	O
equivalent	NN	O	O
values	NN	O	O
for	NN	O	O
total	NN	O	O
receptor	NN	O	O
content	NN	O	O
.	NN	O	O

-DOCSTART-	O

Plasmacytoid	NN	O	O
blast	NN	O	O
crisis	NN	O	O
in	NN	O	O
B-cell	NN	O	B-cell_type
chronic	NN	O	O
lymphocytic	NN	O	O
leukemia	NN	O	O
:	NN	O	O
effect	NN	O	O
of	NN	O	O
estradiol	NN	O	O
on	NN	O	O
growth	NN	O	O
and	NN	O	O
differentiation	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

Evolution	NN	O	O
of	NN	O	O
a	NN	O	O
case	NN	O	O
of	NN	O	O
chronic	NN	O	O
lymphocytic	NN	O	O
leukemia	NN	O	O
(	NN	O	O
CLL	NN	O	O
)	NN	O	O
into	NN	O	O
blast	NN	O	O
crisis	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
characterized	NN	O	O
by	NN	O	O
three	NN	O	O
unusual	NN	O	O
features	NN	O	O
(	NN	O	O
1	NN	O	O
)	NN	O	O
the	NN	O	O
phenotype	NN	O	O
of	NN	O	O
the	NN	O	O
emerging	NN	O	B-cell_type
blast	NN	O	I-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
that	NN	O	O
of	NN	O	O
pre-plasmacytoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
as	NN	O	O
shown	NN	O	O
by	NN	O	O
plasma	NN	O	O
cell	NN	O	O
morphology	NN	O	O
and	NN	O	O
an	NN	O	O
immunological	NN	O	O
phenotype	NN	O	O
corresponding	NN	O	O
partially	NN	O	O
with	NN	O	O
CLL-	NN	O	B-cell_type
or	NN	O	I-cell_type
intermediate	NN	O	I-cell_type
B-cells	NN	O	I-cell_type
,	NN	O	O
partially	NN	O	O
with	NN	O	O
plasma	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
terminal	NN	O	B-protein
transferase	NN	O	I-protein
-	NN	O	O
,	NN	O	O
common	NN	O	O
acute	NN	O	B-protein
lymphocytic	NN	O	I-protein
leukemia	NN	O	I-protein
antigen	NN	O	I-protein
-	NN	O	O
,	NN	O	O
Ia+	NN	O	B-protein
,	NN	O	O
surface	NN	O	B-protein
immunoglobulin	NN	O	I-protein
heavy	NN	O	I-protein
chains	NN	O	I-protein
-	NN	O	O
,	NN	O	O
surface	NN	O	B-protein
kappa	NN	O	I-protein
light	NN	O	I-protein
chains	NN	O	I-protein
+	NN	O	O
,	NN	O	O
intracytoplasmic	NN	O	B-protein
immunoglobulin	NN	O	I-protein
A+	NN	O	I-protein
and	NN	O	I-protein
G+	NN	O	I-protein
,	NN	O	O
BA-1+	NN	O	B-protein
,	NN	O	O
polyclonal	NN	O	O
gammaglobulin	NN	O	O
production	NN	O	O
)	NN	O	O
;	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
cytogenetic	NN	O	O
analysis	NN	O	O
of	NN	O	O
spontaneous	NN	O	O
metaphases	NN	O	O
revealed	NN	O	O
that	NN	O	O
in	NN	O	O
addition	NN	O	O
to	NN	O	O
the	NN	O	O
typical	NN	O	O
CLL	NN	O	O
abnormality	NN	O	O
,	NN	O	O
trisomy	NN	O	O
12	NN	O	O
,	NN	O	O
in	NN	O	O
all	NN	O	O
of	NN	O	O
the	NN	O	O
cells	NN	O	O
,	NN	O	O
an	NN	O	O
additional	NN	O	O
translocation	NN	O	O
between	NN	O	O
chromosomes	NN	O	B-DNA
14	NN	O	I-DNA
and	NN	O	I-DNA
17	NN	O	I-DNA
was	NN	O	O
present	NN	O	O
in	NN	O	O
40	NN	O	O
%	NN	O	O
with	NN	O	O
a	NN	O	O
presumptive	NN	O	O
breakpoint	NN	O	O
on	NN	O	O
chromosome	NN	O	B-DNA
14	NN	O	I-DNA
(	NN	O	O
q12-3	NN	O	O
)	NN	O	O
never	NN	O	O
described	NN	O	O
before	NN	O	O
(	NN	O	O
commonly	NN	O	O
q32	NN	O	O
)	NN	O	O
and	NN	O	O
(	NN	O	O
3	NN	O	O
)	NN	O	O
the	NN	O	O
progression	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
striking	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
expression	NN	O	O
by	NN	O	O
the	NN	O	O
transformed	NN	O	B-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
specific	NN	O	O
binding	NN	O	O
sites	NN	O	O
for	NN	O	O
estradiol	NN	O	O
(	NN	O	O
E2	NN	O	O
)	NN	O	O
due	NN	O	O
to	NN	O	O
an	NN	O	O
actual	NN	O	O
increase	NN	O	O
in	NN	O	O
total	NN	O	O
cellular	NN	O	B-protein
receptor	NN	O	I-protein
proteins	NN	O	I-protein
and	NN	O	O
not	NN	O	O
to	NN	O	O
a	NN	O	O
change	NN	O	O
in	NN	O	O
receptor	NN	O	O
affinity	NN	O	O
for	NN	O	O
E2	NN	O	O
.	NN	O	O

The	NN	O	O
functional	NN	O	O
status	NN	O	O
of	NN	O	O
the	NN	O	O
steroid	NN	O	B-protein
receptors	NN	O	I-protein
was	NN	O	O
confirmed	NN	O	O
by	NN	O	O
nuclear	NN	O	O
transfer	NN	O	O
of	NN	O	O
the	NN	O	O
cytoplasmic	NN	O	B-protein
hormone-receptor	NN	O	I-protein
complex	NN	O	I-protein
upon	NN	O	O
temperature	NN	O	O
activation	NN	O	O
.	NN	O	O

Since	NN	O	O
the	NN	O	O
rise	NN	O	O
in	NN	O	O
E2-receptor	NN	O	B-protein
display	NN	O	O
paralleled	NN	O	O
a	NN	O	O
large	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
proliferative	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
cells	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
a	NN	O	O
change	NN	O	O
in	NN	O	O
their	NN	O	O
maturation	NN	O	O
status	NN	O	O
the	NN	O	O
question	NN	O	O
was	NN	O	O
raised	NN	O	O
as	NN	O	O
to	NN	O	O
whether	NN	O	O
the	NN	O	O
E2-receptor	NN	O	B-protein
should	NN	O	O
be	NN	O	O
considered	NN	O	O
as	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
of	NN	O	O
cellular	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Exposure	NN	O	O
of	NN	O	O
the	NN	O	O
patient	NN	O	O
's	NN	O	O
blast	NN	O	B-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
E2	NN	O	O
in	NN	O	O
vitro	NN	O	O
resulted	NN	O	O
in	NN	O	O
cessation	NN	O	O
of	NN	O	O
cell	NN	O	O
growth	NN	O	O
following	NN	O	O
at	NN	O	O
least	NN	O	O
one	NN	O	O
mitosis	NN	O	O
after	NN	O	O
addition	NN	O	O
of	NN	O	O
the	NN	O	O
inducer	NN	O	O
as	NN	O	O
seen	NN	O	O
from	NN	O	O
the	NN	O	O
replacement	NN	O	O
of	NN	O	O
the	NN	O	O
large	NN	O	B-cell_type
blasts	NN	O	I-cell_type
by	NN	O	O
small	NN	O	O
CLL-like	NN	O	B-cell_type
cells	NN	O	I-cell_type
without	NN	O	O
definite	NN	O	O
signs	NN	O	O
of	NN	O	O
alteration	NN	O	O
of	NN	O	O
the	NN	O	O
differentiation	NN	O	O
status	NN	O	O
.	NN	O	O

This	NN	O	O
suggests	NN	O	O
the	NN	O	O
association	NN	O	O
of	NN	O	O
E2-receptor	NN	O	B-protein
expression	NN	O	O
with	NN	O	O
control	NN	O	O
of	NN	O	O
growth	NN	O	O
rather	NN	O	O
than	NN	O	O
cell	NN	O	O
maturation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Multiple	NN	O	O
forms	NN	O	O
and	NN	O	O
fragments	NN	O	O
of	NN	O	O
cytosolic	NN	O	B-protein
glucocorticoid	NN	O	I-protein
receptors	NN	O	I-protein
from	NN	O	O
human	NN	O	B-cell_type
leukemic	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
normal	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Therapy	NN	O	O
with	NN	O	O
glucocorticoids	NN	O	O
is	NN	O	O
generally	NN	O	O
more	NN	O	O
effective	NN	O	O
in	NN	O	O
acute	NN	O	O
lymphoblastic	NN	O	O
leukemia	NN	O	O
than	NN	O	O
in	NN	O	O
other	NN	O	O
types	NN	O	O
of	NN	O	O
human	NN	O	O
leukemia	NN	O	O
.	NN	O	O

Previous	NN	O	O
studies	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
have	NN	O	O
not	NN	O	O
revealed	NN	O	O
any	NN	O	O
consistent	NN	O	O
relationship	NN	O	O
between	NN	O	O
clinical	NN	O	O
responsiveness	NN	O	O
and	NN	O	O
the	NN	O	O
cellular	NN	O	O
or	NN	O	O
cytosolic	NN	O	O
concentration	NN	O	O
of	NN	O	O
glucocorticoid-binding	NN	O	B-protein
sites	NN	O	I-protein
.	NN	O	O

The	NN	O	O
objectives	NN	O	O
of	NN	O	O
this	NN	O	O
study	NN	O	O
were	NN	O	O
to	NN	O	O
determine	NN	O	O
whether	NN	O	O
there	NN	O	O
are	NN	O	O
intrinsic	NN	O	O
structural	NN	O	O
differences	NN	O	O
among	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
various	NN	O	O
types	NN	O	O
of	NN	O	O
leukemic	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
normal	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
to	NN	O	O
investigate	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
endogenous	NN	O	B-protein
peptidases	NN	O	I-protein
in	NN	O	O
receptor	NN	O	O
degradation	NN	O	O
.	NN	O	O

Cytosols	NN	O	O
were	NN	O	O
prepared	NN	O	O
from	NN	O	O
fresh	NN	O	B-cell_type
or	NN	O	I-cell_type
rapidly	NN	O	I-cell_type
frozen	NN	O	I-cell_type
leukocytes	NN	O	I-cell_type
from	NN	O	O
6	NN	O	O
healthy	NN	O	O
adults	NN	O	O
and	NN	O	O
35	NN	O	O
high-risk	NN	O	O
leukemia	NN	O	O
patients	NN	O	O
(	NN	O	O
median	NN	O	O
white	NN	O	O
blood	NN	O	O
cell	NN	O	O
count	NN	O	O
,	NN	O	O
150	NN	O	O
,	NN	O	O
000	NN	O	O
cells/microliter	NN	O	O
;	NN	O	O
median	NN	O	O
age	NN	O	O
,	NN	O	O
13	NN	O	O
years	NN	O	O
)	NN	O	O
.	NN	O	O

Receptors	NN	O	O
were	NN	O	O
labeled	NN	O	O
with	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
triamcinolone	NN	O	O
acetonide	NN	O	O
and	NN	O	O
quantitated	NN	O	O
by	NN	O	O
charcoal-dextran	NN	O	O
treatment	NN	O	O
or	NN	O	O
Sephadex	NN	O	O
LH-20	NN	O	O
chromatography	NN	O	O
.	NN	O	O

Mean	NN	O	O
and	NN	O	O
median	NN	O	O
cytosolic	NN	O	O
receptor	NN	O	O
concentrations	NN	O	O
in	NN	O	O
12	NN	O	O
acute	NN	O	O
lymphoblastic	NN	O	O
leukemia	NN	O	O
specimens	NN	O	O
lacking	NN	O	O
the	NN	O	O
standard	NN	O	O
B-cell	NN	O	O
or	NN	O	O
T-cell	NN	O	O
markers	NN	O	O
(	NN	O	O
``	NN	O	O
null	NN	O	O
cells	NN	O	O
''	NN	O	O
)	NN	O	O
were	NN	O	O
approximately	NN	O	O
4-fold	NN	O	O
higher	NN	O	O
than	NN	O	O
in	NN	O	O
23	NN	O	O
other	NN	O	O
leukemic	NN	O	B-cell_type
cell	NN	O	I-cell_type
specimens	NN	O	I-cell_type
.	NN	O	O

No	NN	O	O
other	NN	O	O
consistent	NN	O	O
differences	NN	O	O
in	NN	O	O
receptor	NN	O	O
content	NN	O	O
were	NN	O	O
observed	NN	O	O
.	NN	O	O

Agarose	NN	O	O
filtration	NN	O	O
and	NN	O	O
ultracentrifugation	NN	O	O
in	NN	O	O
hypotonic	NN	O	O
buffers	NN	O	O
containing	NN	O	O
20	NN	O	O
mM	NN	O	O
Na2MoO4	NN	O	O
revealed	NN	O	O
complexes	NN	O	O
of	NN	O	O
similar	NN	O	O
size	NN	O	O
and	NN	O	O
shape	NN	O	O
in	NN	O	O
all	NN	O	O
clinical	NN	O	O
specimens	NN	O	O
tested	NN	O	O
and	NN	O	O
two	NN	O	O
established	NN	O	O
leukemic	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

They	NN	O	O
had	NN	O	O
Stokes	NN	O	O
radii	NN	O	O
(	NN	O	O
Rs	NN	O	O
)	NN	O	O
of	NN	O	O
8.1	NN	O	O
+/-	NN	O	O
0.5	NN	O	O
(	NN	O	O
S.D.	NN	O	O
)	NN	O	O
nm	NN	O	O
(	NN	O	O
n	NN	O	O
=	NN	O	O
50	NN	O	O
)	NN	O	O
,	NN	O	O
sedimentation	NN	O	O
coefficients	NN	O	O
of	NN	O	O
9.5	NN	O	O
+/-	NN	O	O
0.3S	NN	O	O
(	NN	O	O
n	NN	O	O
=	NN	O	O
40	NN	O	O
)	NN	O	O
,	NN	O	O
molecular	NN	O	O
weights	NN	O	O
of	NN	O	O
approximately	NN	O	O
330	NN	O	O
,	NN	O	O
000	NN	O	O
,	NN	O	O
and	NN	O	O
axial	NN	O	O
ratios	NN	O	O
(	NN	O	O
a/b	NN	O	O
)	NN	O	O
of	NN	O	O
approximately	NN	O	O
12	NN	O	O
.	NN	O	O

In	NN	O	O
hypertonic	NN	O	O
,	NN	O	O
molybdate-free	NN	O	O
buffer	NN	O	O
,	NN	O	O
these	NN	O	O
oligomeric	NN	O	B-protein
complexes	NN	O	I-protein
were	NN	O	O
dissociated	NN	O	O
into	NN	O	O
subunits	NN	O	O
with	NN	O	O
Rs	NN	O	O
of	NN	O	O
5.9	NN	O	O
+/-	NN	O	O
0.3	NN	O	O
nm	NN	O	O
(	NN	O	O
n	NN	O	O
=	NN	O	O
12	NN	O	O
)	NN	O	O
and	NN	O	O
a/b	NN	O	O
of	NN	O	O
11	NN	O	O
to	NN	O	O
12	NN	O	O
,	NN	O	O
as	NN	O	O
observed	NN	O	O
previously	NN	O	O
for	NN	O	O
other	NN	O	O
receptors	NN	O	O
.	NN	O	O

Fragmentation	NN	O	O
of	NN	O	O
the	NN	O	O
oligomer	NN	O	B-protein
and	NN	O	O
the	NN	O	O
subunit	NN	O	B-protein
was	NN	O	O
evident	NN	O	O
in	NN	O	O
some	NN	O	O
cytosols	NN	O	O
.	NN	O	O

High	NN	O	O
activities	NN	O	O
of	NN	O	O
peptidases	NN	O	B-protein
of	NN	O	O
various	NN	O	O
specificities	NN	O	O
were	NN	O	O
detected	NN	O	O
in	NN	O	O
leukemic	NN	O	O
cell	NN	O	O
cytosols	NN	O	O
,	NN	O	O
as	NN	O	O
in	NN	O	O
other	NN	O	O
cytosols	NN	O	O
,	NN	O	O
by	NN	O	O
fluorometric	NN	O	O
assays	NN	O	O
with	NN	O	O
derivatives	NN	O	O
of	NN	O	O
7-amino-4-methylcoumarin	NN	O	O
.	NN	O	O

Receptor	NN	O	O
cleavage	NN	O	O
by	NN	O	O
these	NN	O	O
and	NN	O	O
other	NN	O	O
endogenous	NN	O	B-protein
enzymes	NN	O	I-protein
may	NN	O	O
account	NN	O	O
for	NN	O	O
previous	NN	O	O
observations	NN	O	O
of	NN	O	O
``	NN	O	O
abnormal	NN	O	O
''	NN	O	O
receptors	NN	O	O
in	NN	O	O
cytosols	NN	O	O
from	NN	O	O
some	NN	O	O
leukemic	NN	O	B-cell_type
specimens	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
intrinsic	NN	O	O
structural	NN	O	O
defects	NN	O	O
in	NN	O	O
the	NN	O	O
receptors	NN	O	O
are	NN	O	O
unlikely	NN	O	O
explanations	NN	O	O
for	NN	O	O
the	NN	O	O
unresponsiveness	NN	O	O
of	NN	O	O
some	NN	O	O
types	NN	O	O
of	NN	O	O
leukemia	NN	O	O
to	NN	O	O
steroid	NN	O	O
therapy	NN	O	O
.	NN	O	O

-DOCSTART-	O

Glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
number	NN	O	O
and	NN	O	O
intracellular	NN	O	O
water	NN	O	O
space	NN	O	O
.	NN	O	O

In	NN	O	O
order	NN	O	O
to	NN	O	O
elucidate	NN	O	O
the	NN	O	O
relationship	NN	O	O
between	NN	O	O
cell	NN	O	O
water	NN	O	O
content	NN	O	O
and	NN	O	O
number	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
,	NN	O	O
eleven	NN	O	O
normal	NN	O	O
and	NN	O	O
malignant	NN	O	B-cell_type
lymphoid	NN	O	I-cell_type
or	NN	O	I-cell_type
myelomonocytic	NN	O	I-cell_type
cell	NN	O	I-cell_type
types	NN	O	I-cell_type
originating	NN	O	O
from	NN	O	O
mouse	NN	O	O
,	NN	O	O
rat	NN	O	O
and	NN	O	O
man	NN	O	O
were	NN	O	O
investigated	NN	O	O
.	NN	O	O

The	NN	O	O
cellular	NN	O	O
water	NN	O	O
space	NN	O	O
was	NN	O	O
measured	NN	O	O
with	NN	O	O
3H2O	NN	O	O
,	NN	O	O
and	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
number	NN	O	O
was	NN	O	O
measured	NN	O	O
in	NN	O	O
a	NN	O	O
whole-cell	NN	O	O
binding	NN	O	O
assay	NN	O	O
with	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
dexamethasone	NN	O	O
at	NN	O	O
30	NN	O	O
and	NN	O	O
37	NN	O	O
degrees	NN	O	O
C	NN	O	O
.	NN	O	O

The	NN	O	O
intracellular	NN	O	O
water	NN	O	O
phase	NN	O	O
concentration	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
(	NN	O	O
around	NN	O	O
40	NN	O	O
nmol/l	NN	O	O
cell	NN	O	O
water	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
dependence	NN	O	O
of	NN	O	O
receptor	NN	O	O
affinity	NN	O	O
on	NN	O	O
temperature	NN	O	O
were	NN	O	O
similar	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
rodent	NN	O	I-cell_type
and	NN	O	I-cell_type
human	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

It	NN	O	O
is	NN	O	O
concluded	NN	O	O
that	NN	O	O
comparisons	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
levels	NN	O	O
are	NN	O	O
best	NN	O	O
made	NN	O	O
on	NN	O	O
the	NN	O	O
basis	NN	O	O
of	NN	O	O
intracellular	NN	O	B-protein
receptor	NN	O	I-protein
concentrations	NN	O	O
.	NN	O	O

-DOCSTART-	O

Covalent	NN	O	O
labeling	NN	O	O
of	NN	O	O
rat	NN	O	B-cell_type
thymocyte	NN	O	I-cell_type
and	NN	O	O
human	NN	O	B-protein
lymphoid	NN	O	I-protein
glucocorticoid	NN	O	I-protein
receptor	NN	O	I-protein
.	NN	O	O

Lymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
contain	NN	O	O
specific	NN	O	O
receptors	NN	O	B-protein
for	NN	O	O
glucocorticoids	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
used	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
dexamethasone-21-mesylate	NN	O	O
to	NN	O	O
label	NN	O	O
covalently	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
rat	NN	O	B-cell_type
thymic	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
in	NN	O	O
neoplastic	NN	O	B-cell_type
cells	NN	O	I-cell_type
obtained	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
acute	NN	O	O
lymphoblastic	NN	O	O
leukemia	NN	O	O
and	NN	O	O
malignant	NN	O	O
lymphoma	NN	O	O
.	NN	O	O

The	NN	O	O
covalently	NN	O	O
labeled	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
were	NN	O	O
identified	NN	O	O
by	NN	O	O
polyacrylamide	NN	O	O
gel	NN	O	O
electrophoresis	NN	O	O
(	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
0.1	NN	O	O
%	NN	O	O
sodium	NN	O	O
dodecyl	NN	O	O
sulfate	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
cytosolic	NN	O	O
fractions	NN	O	O
prepared	NN	O	O
from	NN	O	O
rat	NN	O	B-cell_type
thymic	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
-dexamethasone-21-mesylate	NN	O	O
labels	NN	O	O
a	NN	O	O
protein	NN	O	O
(	NN	O	O
Mr	NN	O	O
approximately	NN	O	O
equal	NN	O	O
to	NN	O	O
95	NN	O	O
,	NN	O	O
000	NN	O	O
)	NN	O	O
which	NN	O	O
was	NN	O	O
identified	NN	O	O
as	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
by	NN	O	O
the	NN	O	O
following	NN	O	O
criteria	NN	O	O
:	NN	O	O
(	NN	O	O
a	NN	O	O
)	NN	O	O
labeling	NN	O	O
of	NN	O	O
this	NN	O	O
moiety	NN	O	O
is	NN	O	O
inhibited	NN	O	O
by	NN	O	O
treatment	NN	O	O
with	NN	O	O
a	NN	O	O
100-fold	NN	O	O
molar	NN	O	O
excess	NN	O	O
of	NN	O	O
glucocorticoids	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
dexamethasone	NN	O	O
and	NN	O	O
triamcinolone	NN	O	O
acetonide	NN	O	O
;	NN	O	O
and	NN	O	O
(	NN	O	O
b	NN	O	O
)	NN	O	O
the	NN	O	O
covalently	NN	O	O
labeled	NN	O	O
Mr	NN	O	O
approximately	NN	O	O
equal	NN	O	O
to	NN	O	O
95	NN	O	O
,	NN	O	O
000	NN	O	O
protein	NN	O	O
is	NN	O	O
activated	NN	O	O
(	NN	O	O
by	NN	O	O
heating	NN	O	O
at	NN	O	O
20	NN	O	O
degrees	NN	O	O
for	NN	O	O
30	NN	O	O
min	NN	O	O
)	NN	O	O
to	NN	O	O
a	NN	O	O
form	NN	O	O
that	NN	O	O
binds	NN	O	O
to	NN	O	O
DNA-cellulose	NN	O	O
.	NN	O	O

When	NN	O	O
intact	NN	O	B-cell_type
thymocytes	NN	O	I-cell_type
are	NN	O	O
treated	NN	O	O
with	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
dexamethasone-21-mesylate	NN	O	O
,	NN	O	O
an	NN	O	O
Mr	NN	O	O
approximately	NN	O	O
equal	NN	O	O
to	NN	O	O
95	NN	O	O
,	NN	O	O
000	NN	O	O
moiety	NN	O	O
is	NN	O	O
also	NN	O	O
labeled	NN	O	O
covalently	NN	O	O
.	NN	O	O

Approximately	NN	O	O
35	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
can	NN	O	O
be	NN	O	O
labeled	NN	O	O
covalently	NN	O	O
when	NN	O	O
intact	NN	O	O
thymocytes	NN	O	B-cell_type
are	NN	O	O
treated	NN	O	O
with	NN	O	O
100	NN	O	O
nM	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
dexamethasone-21-mesylate	NN	O	O
for	NN	O	O
30	NN	O	O
min	NN	O	O
at	NN	O	O
4	NN	O	O
degrees	NN	O	O
.	NN	O	O

Neoplastic	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
acute	NN	O	O
lymphoblastic	NN	O	O
leukemia	NN	O	O
and	NN	O	O
malignant	NN	O	O
lymphoma	NN	O	O
were	NN	O	O
treated	NN	O	O
with	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
dexamethasone-21-mesylate	NN	O	O
.	NN	O	O

In	NN	O	O
all	NN	O	O
samples	NN	O	O
,	NN	O	O
an	NN	O	O
Mr	NN	O	O
approximately	NN	O	O
equal	NN	O	O
to	NN	O	O
95	NN	O	O
,	NN	O	O
000	NN	O	O
moiety	NN	O	O
was	NN	O	O
labeled	NN	O	O
covalently	NN	O	O
;	NN	O	O
labeling	NN	O	O
was	NN	O	O
inhibited	NN	O	O
by	NN	O	O
excess	NN	O	O
glucocorticoid	NN	O	O
.	NN	O	O

Smaller	NN	O	B-protein
moieties	NN	O	I-protein
were	NN	O	O
also	NN	O	O
identified	NN	O	O
by	NN	O	O
competition	NN	O	O
experiments	NN	O	O
;	NN	O	O
these	NN	O	O
may	NN	O	O
represent	NN	O	O
proteolytic	NN	O	B-protein
fragments	NN	O	I-protein
of	NN	O	O
the	NN	O	O
Mr	NN	O	O
approximately	NN	O	O
equal	NN	O	O
to	NN	O	O
95	NN	O	O
,	NN	O	O
000	NN	O	O
receptor	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
in	NN	O	O
rat	NN	O	B-cell_type
and	NN	O	I-cell_type
human	NN	O	I-cell_type
lymphoid	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
dexamethasone-21-mesylate	NN	O	O
can	NN	O	O
be	NN	O	O
used	NN	O	O
to	NN	O	O
label	NN	O	O
covalently	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
association	NN	O	O
of	NN	O	O
cytosol	NN	O	O
oestrogen	NN	O	B-protein
and	NN	O	I-protein
progesterone	NN	O	I-protein
receptors	NN	O	I-protein
with	NN	O	O
histological	NN	O	O
features	NN	O	O
of	NN	O	O
breast	NN	O	O
cancer	NN	O	O
and	NN	O	O
early	NN	O	O
recurrence	NN	O	O
of	NN	O	O
disease	NN	O	O
.	NN	O	O

Two	NN	O	O
hundred	NN	O	O
and	NN	O	O
eighty-eight	NN	O	O
primary	NN	O	O
breast	NN	O	O
tumours	NN	O	O
were	NN	O	O
examined	NN	O	O
for	NN	O	O
the	NN	O	O
presence	NN	O	O
or	NN	O	O
absence	NN	O	O
of	NN	O	O
oestrogen	NN	O	B-protein
(	NN	O	I-protein
REc	NN	O	I-protein
)	NN	O	I-protein
and	NN	O	I-protein
progesterone	NN	O	I-protein
(	NN	O	I-protein
RPc	NN	O	I-protein
)	NN	O	I-protein
receptors	NN	O	I-protein
.	NN	O	O

Analysis	NN	O	O
has	NN	O	O
shown	NN	O	O
a	NN	O	O
relative	NN	O	O
interdependence	NN	O	O
between	NN	O	O
the	NN	O	O
steroid	NN	O	B-protein
receptor	NN	O	I-protein
status	NN	O	O
of	NN	O	O
primary	NN	O	O
breast	NN	O	O
cancer	NN	O	O
and	NN	O	O
other	NN	O	O
prognostic	NN	O	O
variables	NN	O	O
such	NN	O	O
as	NN	O	O
histological	NN	O	O
grade	NN	O	O
,	NN	O	O
lymphocytic	NN	O	O
infiltration	NN	O	O
and	NN	O	O
tumour	NN	O	O
elastosis	NN	O	O
.	NN	O	O

There	NN	O	O
were	NN	O	O
significant	NN	O	O
associations	NN	O	O
between	NN	O	O
epithelial	NN	O	O
cellularity	NN	O	O
,	NN	O	O
stromal	NN	O	O
fibrosis	NN	O	O
and	NN	O	O
the	NN	O	O
value	NN	O	O
of	NN	O	O
REc	NN	O	B-protein
in	NN	O	O
those	NN	O	O
tumours	NN	O	O
in	NN	O	O
which	NN	O	O
the	NN	O	O
receptor	NN	O	O
was	NN	O	O
present	NN	O	O
.	NN	O	O

Cellularity	NN	O	O
and	NN	O	O
fibrosis	NN	O	O
were	NN	O	O
unrelated	NN	O	O
to	NN	O	O
the	NN	O	O
presence	NN	O	O
or	NN	O	O
absence	NN	O	O
of	NN	O	O
oestrogen	NN	O	B-protein
receptor	NN	O	I-protein
.	NN	O	O

By	NN	O	O
contrast	NN	O	O
,	NN	O	O
neither	NN	O	O
the	NN	O	O
presence	NN	O	O
or	NN	O	O
absence	NN	O	O
nor	NN	O	O
the	NN	O	O
value	NN	O	O
of	NN	O	O
RPc	NN	O	B-protein
could	NN	O	O
be	NN	O	O
related	NN	O	O
to	NN	O	O
cellularity	NN	O	O
or	NN	O	O
fibrosis	NN	O	O
.	NN	O	O

The	NN	O	O
value	NN	O	O
of	NN	O	O
REc	NN	O	B-protein
and	NN	O	O
RPc	NN	O	B-protein
analysis	NN	O	O
as	NN	O	O
an	NN	O	O
indicator	NN	O	O
of	NN	O	O
prognosis	NN	O	O
was	NN	O	O
examined	NN	O	O
in	NN	O	O
a	NN	O	O
sub-group	NN	O	O
of	NN	O	O
175	NN	O	O
patients	NN	O	O
receiving	NN	O	O
no	NN	O	O
additional	NN	O	O
treatment	NN	O	O
following	NN	O	O
mastectomy	NN	O	O
.	NN	O	O

Overall	NN	O	O
relapse-free	NN	O	O
survival	NN	O	O
(	NN	O	O
RFS	NN	O	O
)	NN	O	O
was	NN	O	O
no	NN	O	O
different	NN	O	O
for	NN	O	O
those	NN	O	O
patients	NN	O	O
with	NN	O	O
receptors	NN	O	O
compared	NN	O	O
to	NN	O	O
those	NN	O	O
without	NN	O	O
them	NN	O	O
(	NN	O	O
REc	NN	O	B-protein
P	NN	O	O
=	NN	O	O
0.11	NN	O	O
,	NN	O	O
RPc	NN	O	O
P	NN	O	O
=	NN	O	O
0.7	NN	O	O
)	NN	O	O
.	NN	O	O

There	NN	O	O
was	NN	O	O
no	NN	O	O
difference	NN	O	O
in	NN	O	O
RFS	NN	O	O
of	NN	O	O
receptor	NN	O	O
positive	NN	O	O
and	NN	O	O
negative	NN	O	O
tumours	NN	O	O
when	NN	O	O
the	NN	O	O
axillary	NN	O	O
node	NN	O	O
status	NN	O	O
was	NN	O	O
taken	NN	O	O
into	NN	O	O
account	NN	O	O
.	NN	O	O

-DOCSTART-	O

Clinical	NN	O	O
implications	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
human	NN	O	O
leukemia	NN	O	O
.	NN	O	O

Normal	NN	O	B-cell_type
lymphoid	NN	O	I-cell_type
cells	NN	O	I-cell_type
contain	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
.	NN	O	O

A	NN	O	O
variety	NN	O	O
of	NN	O	O
stimuli	NN	O	O
that	NN	O	O
activate	NN	O	O
these	NN	O	O
cells	NN	O	O
also	NN	O	O
induce	NN	O	O
increases	NN	O	O
in	NN	O	O
receptor	NN	O	O
concentration	NN	O	O
.	NN	O	O

Similar	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
can	NN	O	O
be	NN	O	O
detected	NN	O	O
in	NN	O	O
lymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
patients	NN	O	O
with	NN	O	O
acute	NN	O	O
lymphoblastic	NN	O	O
leukemia	NN	O	O
(	NN	O	O
ALL	NN	O	O
)	NN	O	O
.	NN	O	O

Absence	NN	O	O
of	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
usually	NN	O	O
found	NN	O	O
in	NN	O	O
treated	NN	O	O
patients	NN	O	O
)	NN	O	O
predicts	NN	O	O
lack	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	O
responsiveness	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
in	NN	O	O
our	NN	O	O
hands	NN	O	O
,	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
levels	NN	O	O
correlate	NN	O	O
with	NN	O	O
the	NN	O	O
duration	NN	O	O
of	NN	O	O
complete	NN	O	O
remission	NN	O	O
in	NN	O	O
ALL	NN	O	O
(	NN	O	O
though	NN	O	O
not	NN	O	O
in	NN	O	O
other	NN	O	O
forms	NN	O	O
of	NN	O	O
leukemia	NN	O	O
)	NN	O	O
.	NN	O	O

This	NN	O	O
association	NN	O	O
is	NN	O	O
independent	NN	O	O
of	NN	O	O
cell	NN	O	O
type	NN	O	O
,	NN	O	O
age	NN	O	O
,	NN	O	O
sex	NN	O	O
,	NN	O	O
or	NN	O	O
initial	NN	O	O
leukocyte	NN	O	B-cell_type
count	NN	O	O
.	NN	O	O

The	NN	O	O
level	NN	O	O
of	NN	O	O
receptor	NN	O	O
shows	NN	O	O
a	NN	O	O
negative	NN	O	O
correlation	NN	O	O
with	NN	O	O
increasing	NN	O	O
aggressiveness	NN	O	O
of	NN	O	O
the	NN	O	O
tumor	NN	O	O
(	NN	O	O
null-cell	NN	O	O
leukemia	NN	O	O
greater	NN	O	O
than	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
greater	NN	O	O
than	NN	O	O
Burkitt	NN	O	O
's	NN	O	O
lymphoma	NN	O	O
)	NN	O	O
.	NN	O	O

-DOCSTART-	O

Corticosteroid-mediated	NN	O	O
immunoregulation	NN	O	O
in	NN	O	O
man	NN	O	O
.	NN	O	O

Glucocorticoids	NN	O	O
have	NN	O	O
profound	NN	O	O
and	NN	O	O
complex	NN	O	O
effects	NN	O	O
on	NN	O	O
the	NN	O	O
human	NN	O	O
immune	NN	O	O
response	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
precise	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
the	NN	O	O
corticosteroid-induced	NN	O	O
immunoregulation	NN	O	O
in	NN	O	O
man	NN	O	O
have	NN	O	O
not	NN	O	O
been	NN	O	O
precisely	NN	O	O
defined	NN	O	O
.	NN	O	O

Intracytoplasmic	NN	O	B-protein
corticosteroid-specific	NN	O	I-protein
receptors	NN	O	I-protein
appear	NN	O	O
to	NN	O	O
be	NN	O	O
an	NN	O	O
important	NN	O	O
common	NN	O	O
pathway	NN	O	O
for	NN	O	O
steroid-induced	NN	O	O
changes	NN	O	O
,	NN	O	O
but	NN	O	O
variations	NN	O	O
of	NN	O	O
receptor	NN	O	O
parameters	NN	O	O
do	NN	O	O
not	NN	O	O
account	NN	O	O
for	NN	O	O
the	NN	O	O
multifaceted	NN	O	O
effects	NN	O	O
on	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
.	NN	O	O

Human	NN	O	B-cell_type
circulating	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
redistribute	NN	O	O
out	NN	O	O
of	NN	O	O
the	NN	O	O
intravascular	NN	O	O
compartment	NN	O	O
following	NN	O	O
treatment	NN	O	O
with	NN	O	O
corticosteroids	NN	O	O
.	NN	O	O

Although	NN	O	O
certain	NN	O	O
components	NN	O	O
at	NN	O	O
this	NN	O	O
redistribution	NN	O	O
phenomenon	NN	O	O
have	NN	O	O
been	NN	O	O
well-characterized	NN	O	O
,	NN	O	O
the	NN	O	O
importance	NN	O	O
of	NN	O	O
this	NN	O	O
compartmental	NN	O	O
cellular	NN	O	O
shift	NN	O	O
with	NN	O	O
respect	NN	O	O
to	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
corticosteroid-induced	NN	O	O
immunoregulation	NN	O	O
are	NN	O	O
less	NN	O	O
well-defined	NN	O	O
.	NN	O	O

Recent	NN	O	O
observations	NN	O	O
that	NN	O	O
activated	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
may	NN	O	O
be	NN	O	O
sensitive	NN	O	O
to	NN	O	O
the	NN	O	O
lytic	NN	O	O
effects	NN	O	O
of	NN	O	O
glucocorticoids	NN	O	O
suggest	NN	O	O
that	NN	O	O
under	NN	O	O
certain	NN	O	O
situations	NN	O	O
the	NN	O	O
elimination	NN	O	O
of	NN	O	O
selected	NN	O	O
subsets	NN	O	O
of	NN	O	O
cells	NN	O	O
may	NN	O	O
be	NN	O	O
a	NN	O	O
relevant	NN	O	O
mechanism	NN	O	O
of	NN	O	O
corticosteroid-mediated	NN	O	O
immunoregulation	NN	O	O
in	NN	O	O
man	NN	O	O
.	NN	O	O

Corticosteroid-mediated	NN	O	O
effects	NN	O	O
on	NN	O	O
monocyte	NN	O	B-cell_type
function	NN	O	O
may	NN	O	O
be	NN	O	O
an	NN	O	O
important	NN	O	O
mechanism	NN	O	O
of	NN	O	O
drug-induced	NN	O	B-cell_type
immunoregulation	NN	O	I-cell_type
in	NN	O	O
monocyte-dependent	NN	O	B-cell_type
responses	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
some	NN	O	O
experimental	NN	O	O
conditions	NN	O	O
,	NN	O	O
corticosteroids	NN	O	O
inhibit	NN	O	O
Interleukin	NN	O	B-protein
1	NN	O	I-protein
production	NN	O	O
by	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

The	NN	O	O
immunoregulatory	NN	O	O
effects	NN	O	O
of	NN	O	O
corticosteroids	NN	O	O
on	NN	O	O
lymphocyte	NN	O	B-cell_type
immune	NN	O	O
responses	NN	O	O
are	NN	O	O
complex	NN	O	O
.	NN	O	O

In	NN	O	O
vitro	NN	O	O
corticosteroids	NN	O	O
appear	NN	O	O
to	NN	O	O
selectively	NN	O	O
affect	NN	O	O
early	NN	O	O
immunoregulatory	NN	O	O
events	NN	O	O
as	NN	O	O
opposed	NN	O	O
to	NN	O	O
altering	NN	O	O
an	NN	O	O
established	NN	O	O
response	NN	O	O
.	NN	O	O

Multiple	NN	O	O
sites	NN	O	O
of	NN	O	O
steroid-induced	NN	O	O
modulations	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
B	NN	O	I-cell_type
cell	NN	O	I-cell_type
responses	NN	O	O
have	NN	O	O
been	NN	O	O
defined	NN	O	O
.	NN	O	O

-DOCSTART-	O

Human	NN	O	B-cell_line
lymphoid	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
and	NN	O	O
glucocorticoids	NN	O	O
:	NN	O	O
II	NN	O	O
.	NN	O	O

Whole	NN	O	O
cell	NN	O	O
and	NN	O	O
cytoplasmic	NN	O	O
binding	NN	O	O
properties	NN	O	O
of	NN	O	O
lymphoblastoid	NN	O	B-cell_line
,	NN	O	I-cell_line
leukaemia	NN	O	I-cell_line
and	NN	O	I-cell_line
lymphoma	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
glucocorticoid	NN	O	O
binding	NN	O	O
properties	NN	O	O
of	NN	O	O
18	NN	O	O
human	NN	O	B-cell_line
lymphoid	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
(	NN	O	O
HLCL	NN	O	B-cell_line
)	NN	O	O
have	NN	O	O
been	NN	O	O
investigated	NN	O	O
.	NN	O	O

The	NN	O	O
specificity	NN	O	O
of	NN	O	O
steroid	NN	O	O
binding	NN	O	O
was	NN	O	O
confirmed	NN	O	O
with	NN	O	O
various	NN	O	O
glucocorticoid	NN	O	O
agonists	NN	O	O
and	NN	O	O
antagonists	NN	O	O
.	NN	O	O

A	NN	O	O
gradation	NN	O	O
in	NN	O	O
whole	NN	O	O
cell	NN	O	O
and	NN	O	O
cytoplasmic	NN	O	O
glucocorticoid	NN	O	O
binding	NN	O	O
capacity	NN	O	O
was	NN	O	O
observed	NN	O	O
in	NN	O	O
the	NN	O	O
different	NN	O	O
cell	NN	O	O
line	NN	O	O
types	NN	O	O
:	NN	O	O
lymphoblastoid	NN	O	B-cell_line
greater	NN	O	O
than	NN	O	O
lymphoma	NN	O	B-cell_line
greater	NN	O	O
than	NN	O	O
leukaemia	NN	O	B-cell_line
.	NN	O	O

The	NN	O	O
cytoplasmic	NN	O	B-protein
receptors	NN	O	I-protein
of	NN	O	O
leukaemia	NN	O	B-cell_line
and	NN	O	I-cell_line
lymphoblastoid	NN	O	I-cell_line
lines	NN	O	I-cell_line
appeared	NN	O	O
to	NN	O	O
contain	NN	O	O
both	NN	O	O
proteinaceous	NN	O	O
and	NN	O	O
phospholipid	NN	O	O
components	NN	O	O
.	NN	O	O

Cytoplasmic	NN	O	B-protein
steroid-receptor	NN	O	I-protein
complexes	NN	O	I-protein
exhibited	NN	O	O
a	NN	O	O
wide	NN	O	O
range	NN	O	O
of	NN	O	O
sedimentation	NN	O	O
coefficients	NN	O	O
(	NN	O	O
8.5-11.3S	NN	O	O
)	NN	O	O
in	NN	O	O
low	NN	O	O
ionic	NN	O	O
strength	NN	O	O
buffer	NN	O	O
but	NN	O	O
there	NN	O	O
was	NN	O	O
no	NN	O	O
correlation	NN	O	O
with	NN	O	O
cell	NN	O	O
line	NN	O	O
type	NN	O	O
or	NN	O	O
glucocorticoid	NN	O	O
sensitivity	NN	O	O
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
these	NN	O	O
complexes	NN	O	O
by	NN	O	O
heat	NN	O	O
(	NN	O	O
37	NN	O	O
degrees	NN	O	O
C	NN	O	O
)	NN	O	O
or	NN	O	O
exposure	NN	O	O
to	NN	O	O
high	NN	O	O
ionic	NN	O	O
strength	NN	O	O
buffer	NN	O	O
(	NN	O	O
0.3	NN	O	O
M	NN	O	O
NaCl	NN	O	O
)	NN	O	O
induced	NN	O	O
nuclear	NN	O	O
binding	NN	O	O
of	NN	O	O
steroid	NN	O	O
but	NN	O	O
only	NN	O	O
complexes	NN	O	O
in	NN	O	O
high	NN	O	O
ionic	NN	O	O
strength	NN	O	O
buffer	NN	O	O
manifested	NN	O	O
changes	NN	O	O
in	NN	O	O
sedimentation	NN	O	O
coefficient	NN	O	O
.	NN	O	O

No	NN	O	O
correlation	NN	O	O
was	NN	O	O
observed	NN	O	O
between	NN	O	O
the	NN	O	O
level	NN	O	O
or	NN	O	O
nature	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	O
binding	NN	O	O
and	NN	O	O
the	NN	O	O
cytolethal	NN	O	O
or	NN	O	O
cytostatic	NN	O	O
responsiveness	NN	O	O
of	NN	O	O
HLCL	NN	O	B-cell_line
to	NN	O	O
glucocorticoid	NN	O	O
treatment	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

The	NN	O	O
resistance	NN	O	O
to	NN	O	O
cytolethal	NN	O	O
effects	NN	O	O
can	NN	O	O
not	NN	O	O
be	NN	O	O
ascribed	NN	O	O
to	NN	O	O
a	NN	O	O
failure	NN	O	O
of	NN	O	O
cells	NN	O	O
to	NN	O	O
take	NN	O	O
up	NN	O	O
and	NN	O	O
bind	NN	O	O
steroid	NN	O	O
or	NN	O	O
to	NN	O	O
significant	NN	O	O
differences	NN	O	O
in	NN	O	O
the	NN	O	O
molecular	NN	O	O
species	NN	O	O
of	NN	O	O
cytoplasmic	NN	O	B-protein
receptors	NN	O	I-protein
present	NN	O	O
.	NN	O	O

The	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
by	NN	O	O
which	NN	O	O
glucocorticoids	NN	O	O
achieve	NN	O	O
cytolethal	NN	O	O
responses	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

-DOCSTART-	O

CD28	NN	O	B-protein
-mediated	NN	O	O
activation	NN	O	O
in	NN	O	O
CD45RA+	NN	O	B-cell_type
and	NN	O	I-cell_type
CD45RO+	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
:	NN	O	O
enhanced	NN	O	O
levels	NN	O	O
of	NN	O	O
reactive	NN	O	O
oxygen	NN	O	O
intermediates	NN	O	O
and	NN	O	O
c-Rel	NN	O	B-protein
nuclear	NN	O	O
translocation	NN	O	O
in	NN	O	O
CD45RA+	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
have	NN	O	O
analyzed	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
complete	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
(	NN	O	O
anti-CD3	NN	O	B-protein
plus	NN	O	O
anti-CD28	NN	O	B-protein
)	NN	O	O
on	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
in	NN	O	O
CD45RA+	NN	O	B-cell_type
(	NN	O	I-cell_type
naive	NN	O	I-cell_type
)	NN	O	I-cell_type
and	NN	O	I-cell_type
CD45RO+	NN	O	I-cell_type
(	NN	O	I-cell_type
memory/effector	NN	O	I-cell_type
)	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Long	NN	O	O
exposure	NN	O	O
(	NN	O	O
24	NN	O	O
h	NN	O	O
)	NN	O	O
induced	NN	O	O
stronger	NN	O	O
NF-kappaB	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
in	NN	O	O
CD45RA+	NN	O	B-cell_type
cells	NN	O	I-cell_type
than	NN	O	O
in	NN	O	O
CD45RO+	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
c-Rel	NN	O	I-protein
protein	NN	O	I-protein
indicated	NN	O	O
that	NN	O	O
after	NN	O	O
anti-CD3+anti-CD28	NN	O	O
stimulation	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
c-Rel	NN	O	B-protein
was	NN	O	O
higher	NN	O	O
in	NN	O	O
CD45RA+	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
the	NN	O	O
cytoplasmic	NN	O	O
inhibitor	NN	O	O
IkappaBalpha	NN	O	B-protein
indicated	NN	O	O
that	NN	O	O
anti-CD3+anti-CD28	NN	O	O
stimulation	NN	O	O
induced	NN	O	O
a	NN	O	O
long-lasting	NN	O	O
degradation	NN	O	O
in	NN	O	O
CD45RA+	NN	O	B-cell_type
cells	NN	O	I-cell_type
but	NN	O	O
in	NN	O	O
CD45RO+	NN	O	B-cell_type
cells	NN	O	I-cell_type
the	NN	O	O
degradation	NN	O	O
process	NN	O	O
was	NN	O	O
more	NN	O	O
rapid	NN	O	O
.	NN	O	O

Because	NN	O	O
the	NN	O	O
CD28	NN	O	O
costimulus	NN	O	O
is	NN	O	O
known	NN	O	O
to	NN	O	O
induce	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
reactive	NN	O	O
oxygen	NN	O	O
intermediates	NN	O	O
(	NN	O	O
ROIs	NN	O	O
)	NN	O	O
,	NN	O	O
the	NN	O	O
intracellular	NN	O	O
ROI	NN	O	O
levels	NN	O	O
in	NN	O	O
CD45RA+	NN	O	B-cell_type
and	NN	O	I-cell_type
CD45RO+	NN	O	I-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
compared	NN	O	O
by	NN	O	O
flow	NN	O	O
cytometry	NN	O	O
.	NN	O	O

ROIs	NN	O	O
were	NN	O	O
produced	NN	O	O
in	NN	O	O
both	NN	O	O
cell	NN	O	O
types	NN	O	O
,	NN	O	O
but	NN	O	O
more	NN	O	O
strongly	NN	O	O
in	NN	O	O
CD45RA+	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
data	NN	O	O
presented	NN	O	O
in	NN	O	O
this	NN	O	O
study	NN	O	O
further	NN	O	O
emphasize	NN	O	O
the	NN	O	O
differences	NN	O	O
between	NN	O	O
CD45RA+	NN	O	B-cell_type
and	NN	O	I-cell_type
CD45RO+	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
in	NN	O	O
ROI-dependent	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
.	NN	O	O

-DOCSTART-	O

Ikaros	NN	O	B-protein
in	NN	O	O
hemopoietic	NN	O	B-cell_line
lineage	NN	O	I-cell_line
determination	NN	O	O
and	NN	O	O
homeostasis	NN	O	O
.	NN	O	O

Studies	NN	O	O
on	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
that	NN	O	O
control	NN	O	O
hemopoietic	NN	O	O
differentiation	NN	O	O
have	NN	O	O
focused	NN	O	O
on	NN	O	O
signaling	NN	O	O
cascades	NN	O	O
and	NN	O	O
nuclear	NN	O	O
effectors	NN	O	O
that	NN	O	O
drive	NN	O	O
this	NN	O	O
complex	NN	O	O
developmental	NN	O	O
system	NN	O	O
in	NN	O	O
a	NN	O	O
regulated	NN	O	O
fashion	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
review	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
Ikaros	NN	O	B-protein
,	NN	O	O
the	NN	O	O
founding	NN	O	O
member	NN	O	O
of	NN	O	O
a	NN	O	O
unique	NN	O	O
family	NN	O	O
of	NN	O	O
zinc	NN	O	B-protein
finger	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
in	NN	O	O
this	NN	O	O
developmental	NN	O	O
process	NN	O	O
.	NN	O	O

Studies	NN	O	O
on	NN	O	O
an	NN	O	O
Ikaros	NN	O	B-protein
null	NN	O	O
mutation	NN	O	O
have	NN	O	O
revealed	NN	O	O
an	NN	O	O
essential	NN	O	O
role	NN	O	O
for	NN	O	O
this	NN	O	O
factor	NN	O	O
in	NN	O	O
lymphoid	NN	O	B-cell_type
cell	NN	O	I-cell_type
fate	NN	O	O
determination	NN	O	O
and	NN	O	O
at	NN	O	O
subsequent	NN	O	O
branch	NN	O	O
points	NN	O	O
of	NN	O	O
the	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
differentiation	NN	O	O
pathway	NN	O	O
.	NN	O	O

Differences	NN	O	O
in	NN	O	O
the	NN	O	O
phenotypes	NN	O	O
of	NN	O	O
a	NN	O	O
null	NN	O	O
and	NN	O	O
a	NN	O	O
dominant	NN	O	O
negative	NN	O	O
(	NN	O	O
DN	NN	O	O
)	NN	O	O
Ikaros	NN	O	O
mutation	NN	O	O
provide	NN	O	O
insight	NN	O	O
into	NN	O	O
a	NN	O	O
regulatory	NN	O	O
network	NN	O	O
through	NN	O	O
which	NN	O	O
Ikaros	NN	O	B-protein
proteins	NN	O	I-protein
exert	NN	O	O
their	NN	O	O
effects	NN	O	O
in	NN	O	O
development	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
a	NN	O	O
comparative	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
hemopoietic	NN	O	B-cell_type
stem	NN	O	I-cell_type
cell	NN	O	I-cell_type
and	NN	O	O
precursor	NN	O	O
compartment	NN	O	O
resulting	NN	O	O
from	NN	O	O
the	NN	O	O
two	NN	O	O
Ikaros	NN	O	B-protein
mutations	NN	O	O
reveals	NN	O	O
a	NN	O	O
profound	NN	O	O
yet	NN	O	O
not	NN	O	O
absolute	NN	O	O
requirement	NN	O	O
for	NN	O	O
Ikaros	NN	O	B-protein
in	NN	O	O
the	NN	O	O
production	NN	O	O
and	NN	O	O
differentiation	NN	O	O
of	NN	O	O
these	NN	O	O
populations	NN	O	O
.	NN	O	O

-DOCSTART-	O

Overexpression	NN	O	O
of	NN	O	O
p65	NN	O	O
and	NN	O	O
c-Jun	NN	O	O
substitutes	NN	O	O
for	NN	O	O
B7-1	NN	O	O
costimulation	NN	O	O
by	NN	O	O
targeting	NN	O	O
the	NN	O	O
CD28RE	NN	O	B-DNA
within	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
role	NN	O	O
of	NN	O	O
Rel	NN	O	B-protein
and	NN	O	O
activation	NN	O	B-protein
protein-1	NN	O	I-protein
(	NN	O	O
AP-1	NN	O	B-protein
)	NN	O	O
in	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
in	NN	O	O
B7-1-	NN	O	B-cell_type
and	NN	O	I-cell_type
leukocyte	NN	O	I-cell_type
function-associated	NN	O	I-cell_type
Ag-3	NN	O	I-cell_type
(	NN	O	I-cell_type
LFA.	NN	O	I-cell_type
3	NN	O	I-cell_type
)	NN	O	I-cell_type
-costimulated	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
has	NN	O	O
been	NN	O	O
evaluated	NN	O	O
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
overexpression	NN	O	O
of	NN	O	O
c-Jun	NN	O	B-protein
but	NN	O	O
not	NN	O	O
c-Fos	NN	O	B-protein
increases	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
in	NN	O	O
both	NN	O	O
B7-1-	NN	O	B-cell_type
and	NN	O	I-cell_type
LFA-3-costimulated	NN	O	I-cell_type
Jurkat	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Cotransfection	NN	O	O
of	NN	O	O
both	NN	O	O
c-Jun	NN	O	O
and	NN	O	O
c-Fos	NN	O	O
substitutes	NN	O	O
for	NN	O	O
B7-1	NN	O	O
costimulation	NN	O	O
in	NN	O	O
driving	NN	O	O
an	NN	O	O
activation	NN	O	B-DNA
protein-1	NN	O	I-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
but	NN	O	O
not	NN	O	O
for	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Overexpression	NN	O	O
of	NN	O	O
Rel	NN	O	B-protein
proteins	NN	O	I-protein
demonstrated	NN	O	O
that	NN	O	O
p65-expressing	NN	O	B-cell_line
Jurkat	NN	O	I-cell_line
cells	NN	O	I-cell_line
transcribed	NN	O	O
equally	NN	O	O
well	NN	O	O
a	NN	O	O
nuclear	NN	O	B-DNA
factor	NN	O	I-DNA
kappabeta	NN	O	I-DNA
reporter	NN	O	I-DNA
construct	NN	O	I-DNA
when	NN	O	O
costimulated	NN	O	O
with	NN	O	O
B7-1	NN	O	B-protein
or	NN	O	O
LFA-3	NN	O	B-protein
,	NN	O	O
but	NN	O	O
transcription	NN	O	O
of	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
or	NN	O	O
CD28	NN	O	B-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	I-DNA
CD28RE	NN	O	I-DNA
)	NN	O	I-DNA
-driven	NN	O	I-DNA
reporters	NN	O	I-DNA
was	NN	O	O
superior	NN	O	O
in	NN	O	O
B7-1-costimulated	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Combined	NN	O	O
expression	NN	O	O
of	NN	O	O
c-Jun	NN	O	B-protein
and	NN	O	O
p65	NN	O	B-protein
induced	NN	O	O
vigorous	NN	O	O
transcription	NN	O	O
of	NN	O	O
IL-2	NN	O	B-DNA
promoter-	NN	O	I-DNA
and	NN	O	I-DNA
CD28RE-driven	NN	O	I-DNA
reporter	NN	O	I-DNA
constructs	NN	O	I-DNA
in	NN	O	O
both	NN	O	O
LFA-3-	NN	O	B-cell_type
and	NN	O	I-cell_type
B7-1-costimulated	NN	O	I-cell_type
Jurkat	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Mutating	NN	O	O
the	NN	O	O
CD28RE	NN	O	B-DNA
but	NN	O	O
not	NN	O	O
the	NN	O	O
upstream	NN	O	B-DNA
nuclear	NN	O	I-DNA
factor	NN	O	I-DNA
kappabeta-binding	NN	O	I-DNA
site	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
reduced	NN	O	O
B7-1	NN	O	B-protein
-driven	NN	O	O
transcription	NN	O	O
>	NN	O	O
90	NN	O	O
%	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
implicates	NN	O	O
a	NN	O	O
major	NN	O	O
role	NN	O	O
of	NN	O	O
the	NN	O	O
CD28RE	NN	O	B-DNA
in	NN	O	O
the	NN	O	O
integration	NN	O	O
of	NN	O	O
p65	NN	O	B-protein
/	NN	O	O
c-Jun	NN	O	B-protein
-mediated	NN	O	O
transcription	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

We	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
transition	NN	O	O
from	NN	O	O
an	NN	O	O
autocrine	NN	O	O
LFA-3	NN	O	B-protein
-driven	NN	O	O
immune	NN	O	O
response	NN	O	O
to	NN	O	O
a	NN	O	O
B7	NN	O	O
--	NN	O	O
induced	NN	O	O
paracrine	NN	O	O
immune	NN	O	O
response	NN	O	O
involves	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
c-Jun	NN	O	B-protein
and	NN	O	O
p65	NN	O	B-protein
,	NN	O	O
which	NN	O	O
target	NN	O	O
the	NN	O	O
CD28RE	NN	O	B-DNA
region	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Defects	NN	O	O
in	NN	O	O
actin-cap	NN	O	O
formation	NN	O	O
in	NN	O	O
Vav-deficient	NN	O	O
mice	NN	O	O
implicate	NN	O	O
an	NN	O	O
actin	NN	O	O
requirement	NN	O	O
for	NN	O	O
lymphocyte	NN	O	O
signal	NN	O	O
transduction	NN	O	O
.	NN	O	O

BACKGROUND	NN	O	O
:	NN	O	O
Antigen-receptor	NN	O	O
interactions	NN	O	O
on	NN	O	O
lymphocytes	NN	O	B-cell_type
result	NN	O	O
in	NN	O	O
local	NN	O	O
clustering	NN	O	O
of	NN	O	O
actin	NN	O	B-protein
,	NN	O	O
receptors	NN	O	O
and	NN	O	O
signaling	NN	O	O
molecules	NN	O	O
into	NN	O	O
an	NN	O	O
asymmetric	NN	O	O
membrane	NN	O	O
structure	NN	O	O
termed	NN	O	O
a	NN	O	O
cap	NN	O	O
.	NN	O	O

Although	NN	O	O
actin	NN	O	O
polymerization	NN	O	O
is	NN	O	O
known	NN	O	O
to	NN	O	O
be	NN	O	O
required	NN	O	O
,	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
underlying	NN	O	O
cap	NN	O	O
formation	NN	O	O
are	NN	O	O
unclear	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
studied	NN	O	O
the	NN	O	O
events	NN	O	O
underlying	NN	O	O
cap	NN	O	O
formation	NN	O	O
using	NN	O	O
mice	NN	O	O
bearing	NN	O	O
a	NN	O	O
null	NN	O	O
mutation	NN	O	O
in	NN	O	O
vav	NN	O	B-DNA
(	NN	O	O
vav	NN	O	B-DNA
-/-	NN	O	O
)	NN	O	O
,	NN	O	O
a	NN	O	O
gene	NN	O	O
that	NN	O	O
encodes	NN	O	O
a	NN	O	O
guanine-nucleotide	NN	O	B-protein
exchange	NN	O	I-protein
factor	NN	O	I-protein
for	NN	O	O
the	NN	O	O
GTPase	NN	O	B-protein
Rac	NN	O	I-protein
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
Lymphocytes	NN	O	B-cell_type
from	NN	O	O
vav	NN	O	B-DNA
-/-	NN	O	O
mice	NN	O	O
failed	NN	O	O
to	NN	O	O
form	NN	O	O
T-cell	NN	O	O
receptor	NN	O	O
caps	NN	O	O
following	NN	O	O
activation	NN	O	O
and	NN	O	O
had	NN	O	O
a	NN	O	O
defective	NN	O	O
actin	NN	O	O
cytoskeleton	NN	O	O
.	NN	O	O

The	NN	O	O
vav-/-	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
deficient	NN	O	O
in	NN	O	O
interleukin-2	NN	O	O
(	NN	O	O
IL-2	NN	O	O
)	NN	O	O
production	NN	O	O
and	NN	O	O
proliferation	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
peak	NN	O	O
of	NN	O	O
Ca2+	NN	O	O
mobilization	NN	O	O
was	NN	O	O
reduced	NN	O	O
although	NN	O	O
of	NN	O	O
normal	NN	O	O
duration	NN	O	O
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
Jun	NN	O	B-protein
N-terminal	NN	O	I-protein
kinase	NN	O	I-protein
or	NN	O	O
stress-activated	NN	O	B-protein
kinase	NN	O	I-protein
(	NN	O	O
JNK	NN	O	B-protein
or	NN	O	O
SAPK	NN	O	B-protein
)	NN	O	O
and	NN	O	O
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	O
MAPK	NN	O	B-protein
)	NN	O	O
and	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-ATc1	NN	O	B-DNA
and	NN	O	I-DNA
egr-1	NN	O	I-DNA
genes	NN	O	I-DNA
was	NN	O	O
normal	NN	O	O
.	NN	O	O

Despite	NN	O	O
the	NN	O	O
reduced	NN	O	O
Ca2+	NN	O	O
mobilization	NN	O	O
,	NN	O	O
translocation	NN	O	O
of	NN	O	O
cytoplasmic	NN	O	B-protein
NF-ATc	NN	O	I-protein
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
was	NN	O	O
normal	NN	O	O
,	NN	O	O
reflecting	NN	O	O
that	NN	O	O
the	NN	O	O
lower	NN	O	O
levels	NN	O	O
of	NN	O	O
Ca2+	NN	O	O
in	NN	O	O
vav-/-	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
still	NN	O	O
sufficient	NN	O	O
to	NN	O	O
activate	NN	O	O
calcineurin	NN	O	B-protein
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
lymphocytes	NN	O	B-cell_type
with	NN	O	O
cytochalasin	NN	O	O
D	NN	O	O
,	NN	O	O
which	NN	O	O
blocks	NN	O	O
actin	NN	O	O
polymerization	NN	O	O
,	NN	O	O
inhibited	NN	O	O
cap	NN	O	O
formation	NN	O	O
and	NN	O	O
produced	NN	O	O
defects	NN	O	O
in	NN	O	O
signaling	NN	O	O
and	NN	O	O
IL-2	NN	O	O
transcriptional	NN	O	O
induction	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
antigen-receptor	NN	O	O
signaling	NN	O	O
that	NN	O	O
were	NN	O	O
nearly	NN	O	O
identical	NN	O	O
to	NN	O	O
those	NN	O	O
seen	NN	O	O
in	NN	O	O
vav-/-	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
transfection	NN	O	O
studies	NN	O	O
,	NN	O	O
either	NN	O	O
constitutively	NN	O	O
active	NN	O	O
Vav	NN	O	B-protein
or	NN	O	O
Rac	NN	O	B-protein
could	NN	O	O
complement	NN	O	O
constitutively	NN	O	O
active	NN	O	O
calcineurin	NN	O	B-protein
to	NN	O	O
activate	NN	O	O
NF-AT-dependent	NN	O	O
transcription	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
:	NN	O	O
These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
Vav	NN	O	B-protein
is	NN	O	O
required	NN	O	O
for	NN	O	O
cap	NN	O	O
formation	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
the	NN	O	O
correlation	NN	O	O
between	NN	O	O
cap	NN	O	O
formation	NN	O	O
,	NN	O	O
IL-2	NN	O	O
production	NN	O	O
and	NN	O	O
proliferation	NN	O	O
supports	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
an	NN	O	O
actin-dependent	NN	O	O
pathway	NN	O	O
is	NN	O	O
a	NN	O	O
source	NN	O	O
of	NN	O	O
specialized	NN	O	O
growth	NN	O	O
regulatory	NN	O	O
signals	NN	O	O
.	NN	O	O

-DOCSTART-	O

CD14	NN	O	B-protein
-dependent	NN	O	O
activation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
endothelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
Bacteroides	NN	O	O
fragilis	NN	O	O
outer	NN	O	O
membrane	NN	O	O
.	NN	O	O

We	NN	O	O
studied	NN	O	O
the	NN	O	O
capacity	NN	O	O
of	NN	O	O
isolated	NN	O	O
Bacteriodes	NN	O	O
fragilis	NN	O	O
outer	NN	O	O
membrane	NN	O	O
,	NN	O	O
B.	NN	O	O
fragilis	NN	O	O
NCTC9343	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
;	NN	O	O
endotoxin	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
B.	NN	O	O
fragilis	NN	O	O
NCTC9343	NN	O	O
capsular	NN	O	O
polysaccharides	NN	O	O
to	NN	O	O
activate	NN	O	O
human	NN	O	O
umbilical	NN	O	O
vein	NN	O	O
endothelial	NN	O	O
cell	NN	O	O
(	NN	O	O
HUVEC	NN	O	O
)	NN	O	O
monolayers	NN	O	O
.	NN	O	O

To	NN	O	O
assess	NN	O	O
HUVEC	NN	O	O
activation	NN	O	O
,	NN	O	O
E-selectin	NN	O	B-protein
expression	NN	O	O
was	NN	O	O
measured	NN	O	O
by	NN	O	O
enzyme-linked	NN	O	O
immunosorbent	NN	O	O
assay	NN	O	O
(	NN	O	O
ELISA	NN	O	O
)	NN	O	O
,	NN	O	O
Northern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
for	NN	O	O
E-selectin-specific	NN	O	B-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
and	NN	O	O
electrophoretic	NN	O	O
gel	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
(	NN	O	O
EMSA	NN	O	O
)	NN	O	O
for	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
a	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
necessary	NN	O	O
for	NN	O	O
E-selectin	NN	O	B-DNA
gene	NN	O	I-DNA
activation	NN	O	O
.	NN	O	O

Exposure	NN	O	O
of	NN	O	O
HUVECs	NN	O	B-cell_type
to	NN	O	O
B.	NN	O	O
fragilis	NN	O	O
outer	NN	O	O
membrane	NN	O	O
fractions	NN	O	O
,	NN	O	O
separated	NN	O	O
from	NN	O	O
other	NN	O	O
components	NN	O	O
of	NN	O	O
the	NN	O	O
B.	NN	O	O
fragilis	NN	O	O
cell	NN	O	O
wall	NN	O	O
by	NN	O	O
isopycnic	NN	O	O
,	NN	O	O
sucrose	NN	O	O
gradient	NN	O	O
centrifugation	NN	O	O
,	NN	O	O
significantly	NN	O	O
increased	NN	O	O
surface	NN	O	O
expression	NN	O	O
of	NN	O	O
E-selectin	NN	O	B-protein
and	NN	O	O
induced	NN	O	O
functional	NN	O	O
endothelial	NN	O	B-cell_type
cell	NN	O	I-cell_type
-dependent	NN	O	O
leukocyte	NN	O	O
adhesion	NN	O	O
.	NN	O	O

B.	NN	O	O
fragilis	NN	O	O
outer	NN	O	O
membranes	NN	O	O
induced	NN	O	O
translocation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
to	NN	O	O
HUVEC	NN	O	O
nuclei	NN	O	O
and	NN	O	O
accumulation	NN	O	O
of	NN	O	O
E-selectin	NN	O	B-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
HUVEC	NN	O	O
cytoplasm	NN	O	O
.	NN	O	O

E-selectin	NN	O	O
expression	NN	O	O
induced	NN	O	O
by	NN	O	O
B.	NN	O	O
fragilis	NN	O	O
outer	NN	O	O
membranes	NN	O	O
was	NN	O	O
not	NN	O	O
blocked	NN	O	O
by	NN	O	O
polymixin	NN	O	O
B	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
E-selectin	NN	O	O
expression	NN	O	O
induced	NN	O	O
by	NN	O	O
outer	NN	O	O
membrane	NN	O	O
fractions	NN	O	O
purified	NN	O	O
from	NN	O	O
E.	NN	O	O
coli	NN	O	O
was	NN	O	O
competitively	NN	O	O
inhibited	NN	O	O
by	NN	O	O
polymixin	NN	O	O
B	NN	O	O
.	NN	O	O

Neither	NN	O	O
purified	NN	O	O
B.	NN	O	O
fragilis	NN	O	O
LPS	NN	O	O
,	NN	O	O
a	NN	O	O
prominent	NN	O	O
constituent	NN	O	O
of	NN	O	O
the	NN	O	O
outer	NN	O	O
membrane	NN	O	O
,	NN	O	O
nor	NN	O	O
purified	NN	O	O
B.	NN	O	O
fragilis	NN	O	O
capsular	NN	O	O
polysaccharides	NN	O	O
induced	NN	O	O
HUVEC	NN	O	O
activation	NN	O	O
.	NN	O	O

Two	NN	O	O
different	NN	O	O
monoclonal	NN	O	B-protein
antibodies	NN	O	I-protein
directed	NN	O	O
against	NN	O	O
human	NN	O	B-protein
CD14	NN	O	I-protein
completely	NN	O	O
inhibited	NN	O	O
B.	NN	O	O
fragilis	NN	O	O
outer	NN	O	O
membrane-induced	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
,	NN	O	O
E-selectin	NN	O	O
transcription	NN	O	O
,	NN	O	O
and	NN	O	O
E-selectin	NN	O	O
surface	NN	O	O
expression	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
the	NN	O	O
outer	NN	O	O
membrane	NN	O	O
component	NN	O	O
of	NN	O	O
the	NN	O	O
B.	NN	O	O
fragilis	NN	O	O
cell	NN	O	O
wall	NN	O	O
contains	NN	O	O
a	NN	O	O
proinflammatory	NN	O	O
factor	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
,	NN	O	O
that	NN	O	O
is	NN	O	O
not	NN	O	O
LPS	NN	O	O
,	NN	O	O
which	NN	O	O
induces	NN	O	O
human	NN	O	B-cell_type
endothelial	NN	O	I-cell_type
cell	NN	O	I-cell_type
activation	NN	O	O
by	NN	O	O
a	NN	O	O
soluble	NN	O	O
CD14	NN	O	B-protein
-dependent	NN	O	O
mechanism	NN	O	O
.	NN	O	O

-DOCSTART-	O

NF-kappaB	NN	O	B-protein
protects	NN	O	O
HIV-1-infected	NN	O	O
myeloid	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

HIV-1	NN	O	O
infection	NN	O	O
of	NN	O	O
primary	NN	O	B-cell_type
monocytic	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
myeloid	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
results	NN	O	O
in	NN	O	O
sustained	NN	O	O
NF-kappaB	NN	O	O
activation	NN	O	O
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
NF-kappaB	NN	O	O
induction	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
protecting	NN	O	O
cells	NN	O	O
from	NN	O	O
programmed	NN	O	O
cell	NN	O	O
death	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
sought	NN	O	O
to	NN	O	O
investigate	NN	O	O
whether	NN	O	O
constitutive	NN	O	O
NF-kappaB	NN	O	B-protein
activity	NN	O	O
in	NN	O	O
chronically	NN	O	O
HIV-1-infected	NN	O	O
promonocytic	NN	O	B-cell_line
U937	NN	O	I-cell_line
(	NN	O	I-cell_line
U9-IIIB	NN	O	I-cell_line
)	NN	O	I-cell_line
and	NN	O	I-cell_line
myeloblastic	NN	O	I-cell_line
PLB-985	NN	O	I-cell_line
(	NN	O	I-cell_line
PLB-IIIB	NN	O	I-cell_line
)	NN	O	I-cell_line
cells	NN	O	I-cell_line
affects	NN	O	O
apoptotic	NN	O	O
signaling	NN	O	O
.	NN	O	O

TNFalpha	NN	O	B-protein
and	NN	O	O
cycloheximide	NN	O	O
caused	NN	O	O
infected	NN	O	O
cells	NN	O	O
to	NN	O	O
undergo	NN	O	O
apoptosis	NN	O	O
more	NN	O	O
rapidly	NN	O	O
than	NN	O	O
parental	NN	O	B-cell_line
U937	NN	O	I-cell_line
and	NN	O	O
PLB-985	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
TNFalpha	NN	O	B-protein
-induced	NN	O	O
NF-kappaB	NN	O	O
activation	NN	O	O
using	NN	O	O
the	NN	O	O
antioxidant	NN	O	O
N-acetylcysteine	NN	O	O
(	NN	O	O
NAC	NN	O	O
)	NN	O	O
resulted	NN	O	O
in	NN	O	O
increased	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
both	NN	O	O
U937	NN	O	B-cell_line
and	NN	O	I-cell_line
U9-IIIB	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
while	NN	O	O
preactivation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
with	NN	O	O
the	NN	O	O
non-apoptotic	NN	O	B-protein
inducer	NN	O	I-protein
IL-1beta	NN	O	I-protein
caused	NN	O	O
a	NN	O	O
relative	NN	O	O
decrease	NN	O	O
in	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
constitutive	NN	O	O
NF-kappaB	NN	O	B-protein
activity	NN	O	O
in	NN	O	O
U9-IIIB	NN	O	B-cell_line
and	NN	O	I-cell_line
PLB-IIIB	NN	O	I-cell_line
cells	NN	O	I-cell_line
also	NN	O	O
induced	NN	O	O
apoptosis	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
NF-kappaB	NN	O	B-cell_type
protects	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
a	NN	O	O
persistent	NN	O	O
apoptotic	NN	O	O
signal	NN	O	O
.	NN	O	O

TNFalpha	NN	O	B-protein
plus	NN	O	O
NAC	NN	O	O
treatment	NN	O	O
resulted	NN	O	O
in	NN	O	O
a	NN	O	O
marked	NN	O	O
decrease	NN	O	O
in	NN	O	O
Bcl-2	NN	O	B-protein
protein	NN	O	I-protein
levels	NN	O	O
in	NN	O	O
HIV-1-infected	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
coupled	NN	O	O
with	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
Bax	NN	O	B-protein
protein	NN	O	I-protein
compared	NN	O	O
to	NN	O	O
uninfected	NN	O	O
cells	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
the	NN	O	O
difference	NN	O	O
in	NN	O	O
susceptibility	NN	O	O
to	NN	O	O
TNFalpha	NN	O	B-protein
-induced	NN	O	O
apoptosis	NN	O	O
may	NN	O	O
relate	NN	O	O
to	NN	O	O
the	NN	O	O
differences	NN	O	O
in	NN	O	O
relative	NN	O	O
levels	NN	O	O
of	NN	O	O
Bcl-2	NN	O	B-protein
and	NN	O	O
Bax	NN	O	B-protein
.	NN	O	O

The	NN	O	O
protective	NN	O	O
role	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
in	NN	O	O
blocking	NN	O	O
TNFalpha-	NN	O	O
and	NN	O	O
HIV-1-induced	NN	O	O
apoptosis	NN	O	O
was	NN	O	O
supported	NN	O	O
by	NN	O	O
studies	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
engineered	NN	O	O
to	NN	O	O
express	NN	O	O
IkappaB	NN	O	B-protein
alpha	NN	O	I-protein
repressor	NN	O	I-protein
mutants	NN	O	I-protein
(	NN	O	O
TD-IkappaB	NN	O	B-protein
)	NN	O	O
under	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
a	NN	O	O
tetracycline-responsive	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Cells	NN	O	O
underwent	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
TNFalpha	NN	O	B-protein
only	NN	O	O
when	NN	O	O
NF-kappaB	NN	O	O
activation	NN	O	O
was	NN	O	O
inhibited	NN	O	O
by	NN	O	O
TD-IkappaB	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

As	NN	O	O
was	NN	O	O
observed	NN	O	O
for	NN	O	O
the	NN	O	O
U9-IIIB	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
TNFalpha	NN	O	B-protein
treatment	NN	O	O
also	NN	O	O
induced	NN	O	O
a	NN	O	O
marked	NN	O	O
decrease	NN	O	O
in	NN	O	O
Bcl-2	NN	O	B-protein
protein	NN	O	I-protein
levels	NN	O	O
in	NN	O	O
TD-IkappaB	NN	O	B-cell_type
expressing	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
experiments	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
apoptotic	NN	O	O
signaling	NN	O	O
is	NN	O	O
perturbed	NN	O	O
in	NN	O	O
HIV-1-infected	NN	O	O
U9-IIIB	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
indicate	NN	O	O
that	NN	O	O
NF-kappaB	NN	O	O
activation	NN	O	O
may	NN	O	O
play	NN	O	O
an	NN	O	O
additional	NN	O	O
protective	NN	O	O
role	NN	O	O
against	NN	O	O
HIV-1-induced	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
myeloid	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Human	NN	O	B-cell_type
normal	NN	O	I-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
B-lymphocytes	NN	O	I-cell_type
are	NN	O	O
deficient	NN	O	O
in	NN	O	O
DNA-dependent	NN	O	O
protein	NN	O	O
kinase	NN	O	O
activity	NN	O	O
due	NN	O	O
to	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
variant	NN	O	O
form	NN	O	O
of	NN	O	O
the	NN	O	O
Ku86	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

The	NN	O	O
heterodimeric	NN	O	O
Ku	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
which	NN	O	O
comprises	NN	O	O
a	NN	O	O
86	NN	O	B-protein
kDa	NN	O	I-protein
(	NN	O	I-protein
Ku86	NN	O	I-protein
)	NN	O	I-protein
amd	NN	O	O
a	NN	O	O
70	NN	O	B-protein
kDa	NN	O	I-protein
(	NN	O	I-protein
Ku70	NN	O	I-protein
)	NN	O	I-protein
subunits	NN	O	I-protein
,	NN	O	O
is	NN	O	O
an	NN	O	O
abundant	NN	O	O
nuclear	NN	O	B-protein
DNA-binding	NN	O	I-protein
protein	NN	O	I-protein
which	NN	O	O
binds	NN	O	O
in	NN	O	O
vitro	NN	O	O
to	NN	O	O
DNA	NN	O	B-DNA
termini	NN	O	I-DNA
without	NN	O	O
sequence	NN	O	O
specificity	NN	O	O
.	NN	O	O

Ku	NN	O	B-protein
is	NN	O	O
the	NN	O	O
DNA-targeting	NN	O	B-protein
component	NN	O	I-protein
of	NN	O	O
the	NN	O	O
large	NN	O	O
catalytic	NN	O	B-protein
sub-unit	NN	O	I-protein
of	NN	O	O
the	NN	O	O
DNA-dependent	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
complex	NN	O	I-protein
(	NN	O	O
DNA-PK	NN	O	B-protein
[	NN	O	I-protein
CS	NN	O	I-protein
]	NN	O	I-protein
)	NN	O	O
,	NN	O	O
that	NN	O	O
plays	NN	O	O
a	NN	O	O
critical	NN	O	O
role	NN	O	O
in	NN	O	O
mammalian	NN	O	O
double-strand	NN	O	O
break	NN	O	O
repair	NN	O	O
and	NN	O	O
lymphoid	NN	O	O
V	NN	O	O
(	NN	O	O
D	NN	O	O
)	NN	O	O
J	NN	O	O
recombination	NN	O	O
.	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
,	NN	O	O
we	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
in	NN	O	O
addition	NN	O	O
to	NN	O	O
the	NN	O	O
major	NN	O	B-protein
Ku	NN	O	I-protein
x	NN	O	I-protein
DNA	NN	O	I-protein
complex	NN	O	I-protein
usually	NN	O	O
detected	NN	O	O
in	NN	O	O
cell	NN	O	O
line	NN	O	O
extracts	NN	O	O
,	NN	O	O
a	NN	O	O
second	NN	O	O
complex	NN	O	O
with	NN	O	O
faster	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
was	NN	O	O
observed	NN	O	O
in	NN	O	O
normal	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
(	NN	O	O
PBL	NN	O	B-cell_type
)	NN	O	O
extracts	NN	O	O
.	NN	O	O

The	NN	O	O
presence	NN	O	O
of	NN	O	O
this	NN	O	O
faster	NN	O	B-protein
migrating	NN	O	I-protein
complex	NN	O	I-protein
was	NN	O	O
restricted	NN	O	O
to	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
among	NN	O	O
the	NN	O	O
circulating	NN	O	B-cell_type
lymphocyte	NN	O	I-cell_type
population	NN	O	I-cell_type
.	NN	O	O

Western	NN	O	O
blot	NN	O	O
analysis	NN	O	O
revealed	NN	O	O
that	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
express	NN	O	O
a	NN	O	O
variant	NN	O	O
form	NN	O	O
of	NN	O	O
the	NN	O	O
Ku86	NN	O	B-protein
protein	NN	O	I-protein
with	NN	O	O
an	NN	O	O
apparent	NN	O	O
molecular	NN	O	O
weight	NN	O	O
of	NN	O	O
69	NN	O	O
kDa	NN	O	O
,	NN	O	O
and	NN	O	O
not	NN	O	O
the	NN	O	O
86	NN	O	B-protein
kDa-	NN	O	I-protein
full-length	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

Although	NN	O	O
the	NN	O	O
heterodimer	NN	O	B-protein
Ku70/variant-Ku86	NN	O	I-protein
binds	NN	O	O
to	NN	O	O
DNA-ends	NN	O	B-DNA
,	NN	O	O
this	NN	O	O
altered	NN	O	O
form	NN	O	O
of	NN	O	O
the	NN	O	O
Ku	NN	O	B-protein
heterodimer	NN	O	I-protein
has	NN	O	O
a	NN	O	O
decreased	NN	O	O
ability	NN	O	O
to	NN	O	O
recruit	NN	O	O
the	NN	O	O
catalytic	NN	O	B-protein
component	NN	O	I-protein
of	NN	O	O
the	NN	O	O
complex	NN	O	O
,	NN	O	O
DNA-PK	NN	O	B-protein
(	NN	O	I-protein
CS	NN	O	I-protein
)	NN	O	I-protein
,	NN	O	O
which	NN	O	O
contributes	NN	O	O
to	NN	O	O
an	NN	O	O
absence	NN	O	O
of	NN	O	O
detectable	NN	O	O
DNA-PK	NN	O	O
activity	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
data	NN	O	O
provide	NN	O	O
a	NN	O	O
molecular	NN	O	O
basis	NN	O	O
for	NN	O	O
the	NN	O	O
increased	NN	O	O
sensitivity	NN	O	O
of	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
ionizing	NN	O	O
radiation	NN	O	O
and	NN	O	O
identify	NN	O	O
a	NN	O	O
new	NN	O	O
mechanism	NN	O	O
of	NN	O	O
regulation	NN	O	O
of	NN	O	O
DNA-PK	NN	O	O
activity	NN	O	O
that	NN	O	O
operates	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

-DOCSTART-	O

Co-stimulation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
anti-CD3	NN	O	B-protein
monoclonal	NN	O	I-protein
antibodies	NN	O	I-protein
induces	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
CREB	NN	O	B-protein
.	NN	O	O

Phosphorylation	NN	O	O
of	NN	O	O
the	NN	O	O
cAMP-response	NN	O	B-protein
element	NN	O	I-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
CREB	NN	O	I-protein
within	NN	O	O
1	NN	O	O
h	NN	O	O
of	NN	O	O
CD2	NN	O	B-protein
but	NN	O	O
not	NN	O	O
CD3	NN	O	B-protein
cross-linking	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
PBMC	NN	O	I-cell_type
was	NN	O	O
recently	NN	O	O
demonstrated	NN	O	O
.	NN	O	O

The	NN	O	O
absence	NN	O	O
of	NN	O	O
P-CREB	NN	O	B-protein
following	NN	O	O
CD3	NN	O	O
cross-linking	NN	O	O
was	NN	O	O
unexpected	NN	O	O
,	NN	O	O
as	NN	O	O
other	NN	O	O
laboratories	NN	O	O
reported	NN	O	O
increased	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
CREB	NN	O	B-protein
following	NN	O	O
CD3	NN	O	O
cross-linking	NN	O	O
of	NN	O	O
the	NN	O	O
Jurkat	NN	O	B-cell_line
lymphocyte	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

Due	NN	O	O
to	NN	O	O
Jurkat	NN	O	B-cell_line
T-cells	NN	O	I-cell_line
being	NN	O	O
IL-2-independent	NN	O	O
,	NN	O	O
it	NN	O	O
was	NN	O	O
postulated	NN	O	O
that	NN	O	O
IL-2	NN	O	B-protein
might	NN	O	O
provide	NN	O	O
a	NN	O	O
necessary	NN	O	O
co-stimulus	NN	O	O
for	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
CREB	NN	O	B-protein
in	NN	O	O
primary	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
P-CREB	NN	O	B-protein
was	NN	O	O
evaluated	NN	O	O
following	NN	O	O
co-stimulation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
PBMC	NN	O	I-cell_type
through	NN	O	O
the	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	I-protein
CD2	NN	O	I-protein
or	NN	O	I-protein
CD3	NN	O	I-protein
receptors	NN	O	I-protein
.	NN	O	O

IL-2	NN	O	B-protein
did	NN	O	O
not	NN	O	O
further	NN	O	O
augment	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
CREB	NN	O	B-protein
following	NN	O	O
CD2	NN	O	O
cross-linking	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
while	NN	O	O
neither	NN	O	O
IL-2	NN	O	B-protein
nor	NN	O	O
CD3	NN	O	O
cross-linking	NN	O	O
alone	NN	O	O
induced	NN	O	O
P-CREB	NN	O	B-protein
,	NN	O	O
a	NN	O	O
4.5-fold	NN	O	O
increase	NN	O	O
in	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
CREB	NN	O	B-protein
within	NN	O	O
1	NN	O	O
h	NN	O	O
of	NN	O	O
IL-2/CD3	NN	O	O
co-stimulation	NN	O	O
was	NN	O	O
observed	NN	O	O
.	NN	O	O

Phosphorylation	NN	O	O
was	NN	O	O
not	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
cAMP	NN	O	O
,	NN	O	O
and	NN	O	O
inhibition	NN	O	O
of	NN	O	O
PKA	NN	O	O
signaling	NN	O	O
had	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
P-CREB	NN	O	B-protein
.	NN	O	O

Consistent	NN	O	O
with	NN	O	O
signal	NN	O	O
transduction	NN	O	O
through	NN	O	O
p56lck	NN	O	B-protein
or	NN	O	O
p59fyn	NN	O	B-protein
,	NN	O	O
inhibition	NN	O	O
of	NN	O	O
PTK	NN	O	O
signaling	NN	O	O
reduced	NN	O	O
phosphorylation	NN	O	O
50	NN	O	O
%	NN	O	O
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
inhibiting	NN	O	O
PKC	NN	O	O
signaling	NN	O	O
with	NN	O	O
calphostin	NN	O	O
C	NN	O	O
further	NN	O	O
increased	NN	O	O
P-CREB	NN	O	O
levels	NN	O	O
3-fold	NN	O	O
over	NN	O	O
that	NN	O	O
observed	NN	O	O
in	NN	O	O
IL-2/CD3	NN	O	B-cell_type
co-stimulated	NN	O	I-cell_type
cells	NN	O	I-cell_type
not	NN	O	O
pretreated	NN	O	O
with	NN	O	O
a	NN	O	O
PKC	NN	O	O
inhibitor	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
previous	NN	O	O
studies	NN	O	O
performed	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
exogenous	NN	O	B-protein
IL-2	NN	O	I-protein
,	NN	O	O
no	NN	O	O
increase	NN	O	O
in	NN	O	O
binding	NN	O	O
of	NN	O	O
CREB	NN	O	B-protein
to	NN	O	O
a	NN	O	O
32P-labeled	NN	O	O
oligonucleotide	NN	O	O
probe	NN	O	O
was	NN	O	O
observed	NN	O	O
by	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
CD3	NN	O	B-protein
signaling	NN	O	O
pathways	NN	O	O
provide	NN	O	O
a	NN	O	O
necessary	NN	O	O
and	NN	O	O
co-operative	NN	O	O
stimulus	NN	O	O
promoting	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
CREB	NN	O	B-protein
following	NN	O	O
receptor	NN	O	O
cross-linking	NN	O	O
.	NN	O	O

-DOCSTART-	O

HIV-1	NN	O	O
infection	NN	O	O
induces	NN	O	O
a	NN	O	O
selective	NN	O	O
reduction	NN	O	O
in	NN	O	O
STAT5	NN	O	O
protein	NN	O	O
expression	NN	O	O
.	NN	O	O

HIV-1	NN	O	O
infection	NN	O	O
is	NN	O	O
accompanied	NN	O	O
by	NN	O	O
qualitative	NN	O	O
and	NN	O	O
quantitative	NN	O	O
defects	NN	O	O
in	NN	O	O
CD4+	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Loss	NN	O	O
of	NN	O	O
immune	NN	O	O
function	NN	O	O
in	NN	O	O
HIV	NN	O	O
patients	NN	O	O
is	NN	O	O
usually	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
profound	NN	O	O
dysregulation	NN	O	O
of	NN	O	O
cytokine	NN	O	B-protein
production	NN	O	O
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
whether	NN	O	O
cytokine	NN	O	B-protein
signaling	NN	O	O
defects	NN	O	O
occur	NN	O	O
during	NN	O	O
HIV	NN	O	O
infection	NN	O	O
,	NN	O	O
PHA	NN	O	B-cell_type
blasts	NN	O	I-cell_type
from	NN	O	O
healthy	NN	O	O
human	NN	O	O
donors	NN	O	O
were	NN	O	O
infected	NN	O	O
with	NN	O	O
two	NN	O	O
strains	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
and	NN	O	O
screened	NN	O	O
for	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
STAT	NN	O	B-protein
proteins	NN	O	I-protein
used	NN	O	O
in	NN	O	O
cytokine	NN	O	B-protein
signaling	NN	O	O
.	NN	O	O

A	NN	O	O
selective	NN	O	O
decrease	NN	O	O
in	NN	O	O
STAT5B	NN	O	B-protein
was	NN	O	O
seen	NN	O	O
8	NN	O	O
days	NN	O	O
after	NN	O	O
infection	NN	O	O
with	NN	O	O
the	NN	O	O
BZ167	NN	O	O
dual-tropic	NN	O	O
HIV	NN	O	O
isolate	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
with	NN	O	O
the	NN	O	O
Ba-L	NN	O	O
,	NN	O	O
M-tropic	NN	O	O
strain	NN	O	O
.	NN	O	O

Based	NN	O	O
on	NN	O	O
these	NN	O	O
findings	NN	O	O
,	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
HIV-infected	NN	O	O
patients	NN	O	O
in	NN	O	O
different	NN	O	O
stages	NN	O	O
of	NN	O	O
disease	NN	O	O
were	NN	O	O
also	NN	O	O
tested	NN	O	O
for	NN	O	O
STAT	NN	O	O
expression	NN	O	O
;	NN	O	O
decreases	NN	O	O
in	NN	O	O
STAT5A	NN	O	B-protein
,	NN	O	O
STAT5B	NN	O	B-protein
,	NN	O	O
and	NN	O	O
STAT1alpha	NN	O	B-protein
were	NN	O	O
observed	NN	O	O
in	NN	O	O
all	NN	O	O
patients	NN	O	O
.	NN	O	O

The	NN	O	O
reduction	NN	O	O
in	NN	O	O
STATs	NN	O	B-protein
seen	NN	O	O
in	NN	O	O
vivo	NN	O	O
and	NN	O	O
in	NN	O	O
vitro	NN	O	O
after	NN	O	O
HIV	NN	O	O
infection	NN	O	O
may	NN	O	O
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
loss	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
function	NN	O	O
in	NN	O	O
HIV	NN	O	O
disease	NN	O	O
.	NN	O	O

-DOCSTART-	O

Transcription	NN	O	B-protein
factors	NN	O	I-protein
that	NN	O	O
regulate	NN	O	O
monocyte	NN	O	B-cell_type
/	NN	O	O
macrophage	NN	O	B-cell_type
differentiation	NN	O	O
.	NN	O	O

Although	NN	O	O
all	NN	O	O
the	NN	O	O
cells	NN	O	O
in	NN	O	O
an	NN	O	O
organism	NN	O	O
contain	NN	O	O
the	NN	O	O
same	NN	O	O
genetic	NN	O	O
information	NN	O	O
,	NN	O	O
differences	NN	O	O
in	NN	O	O
the	NN	O	O
cell	NN	O	O
phenotype	NN	O	O
arise	NN	O	O
from	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
lineage-specific	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

During	NN	O	O
myelopoiesis	NN	O	O
,	NN	O	O
external	NN	O	O
differentiating	NN	O	O
signals	NN	O	O
regulate	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
set	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

The	NN	O	O
combined	NN	O	O
action	NN	O	O
of	NN	O	O
these	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
subsequently	NN	O	O
determines	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
myeloid-specific	NN	O	B-DNA
genes	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
generation	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
macrophages	NN	O	B-cell_type
.	NN	O	O

In	NN	O	O
particular	NN	O	O
,	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
PU.1	NN	O	I-protein
has	NN	O	O
a	NN	O	O
critical	NN	O	O
role	NN	O	O
in	NN	O	O
this	NN	O	O
process	NN	O	O
.	NN	O	O

We	NN	O	O
review	NN	O	O
the	NN	O	O
contribution	NN	O	O
of	NN	O	O
several	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
to	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
macrophage	NN	O	B-cell_type
development	NN	O	O

-DOCSTART-	O

Transcription	NN	O	B-protein
factor	NN	O	I-protein
LKLF	NN	O	I-protein
is	NN	O	O
sufficient	NN	O	O
to	NN	O	O
program	NN	O	O
T	NN	O	O
cell	NN	O	O
quiescence	NN	O	O
via	NN	O	O
a	NN	O	O
c-Myc	NN	O	O
--	NN	O	O
dependent	NN	O	O
pathway	NN	O	O
.	NN	O	O

T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
circulate	NN	O	O
in	NN	O	O
a	NN	O	O
quiescent	NN	O	O
state	NN	O	O
until	NN	O	O
they	NN	O	O
encounter	NN	O	O
cognate	NN	O	B-protein
antigen	NN	O	I-protein
bound	NN	O	O
to	NN	O	O
the	NN	O	O
surface	NN	O	O
of	NN	O	O
an	NN	O	O
antigen-presenting	NN	O	B-cell_type
cell	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
molecular	NN	O	O
pathways	NN	O	O
that	NN	O	O
regulate	NN	O	O
T	NN	O	O
cell	NN	O	O
quiescence	NN	O	O
remain	NN	O	O
largely	NN	O	O
unknown	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
forced	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
lung	NN	O	B-protein
Kruppel-like	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
LKLF	NN	O	B-protein
)	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
is	NN	O	O
sufficient	NN	O	O
to	NN	O	O
program	NN	O	O
a	NN	O	O
quiescent	NN	O	O
phenotype	NN	O	O
characterized	NN	O	O
by	NN	O	O
decreased	NN	O	O
proliferation	NN	O	O
,	NN	O	O
reduced	NN	O	O
cell	NN	O	O
size	NN	O	O
and	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
and	NN	O	O
decreased	NN	O	O
surface	NN	O	O
expression	NN	O	O
of	NN	O	O
activation	NN	O	O
markers	NN	O	O
.	NN	O	O

Conversely	NN	O	O
,	NN	O	O
LKLF-deficient	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
produced	NN	O	O
by	NN	O	O
gene	NN	O	O
targeting	NN	O	O
showed	NN	O	O
increased	NN	O	O
proliferation	NN	O	O
,	NN	O	O
increased	NN	O	O
cell	NN	O	O
size	NN	O	O
and	NN	O	O
enhanced	NN	O	O
expression	NN	O	O
of	NN	O	O
surface	NN	O	O
activation	NN	O	O
markers	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

LKLF	NN	O	B-protein
appeared	NN	O	O
to	NN	O	O
function	NN	O	O
,	NN	O	O
at	NN	O	O
least	NN	O	O
in	NN	O	O
part	NN	O	O
,	NN	O	O
by	NN	O	O
decreasing	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
proto-oncogene	NN	O	B-DNA
encoding	NN	O	O
c-Myc	NN	O	B-protein
.	NN	O	O

Forced	NN	O	O
expression	NN	O	O
of	NN	O	O
LKLF	NN	O	B-protein
was	NN	O	O
associated	NN	O	O
with	NN	O	O
markedly	NN	O	O
decreased	NN	O	O
c-Myc	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
many	NN	O	O
effects	NN	O	O
of	NN	O	O
LKLF	NN	O	B-protein
expression	NN	O	O
were	NN	O	O
mimicked	NN	O	O
by	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
dominant-negative	NN	O	B-protein
MadMyc	NN	O	I-protein
protein	NN	O	I-protein
and	NN	O	O
rescued	NN	O	O
by	NN	O	O
overexpression	NN	O	O
of	NN	O	O
c-Myc	NN	O	B-protein
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
LKLF	NN	O	B-protein
is	NN	O	O
both	NN	O	O
necessary	NN	O	O
and	NN	O	O
sufficient	NN	O	O
to	NN	O	O
program	NN	O	O
quiescence	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
functions	NN	O	O
,	NN	O	O
in	NN	O	O
part	NN	O	O
,	NN	O	O
by	NN	O	O
negatively	NN	O	O
regulating	NN	O	O
a	NN	O	O
c-Myc	NN	O	O
--	NN	O	O
dependent	NN	O	O
pathway	NN	O	O
.	NN	O	O

-DOCSTART-	O

HTLV-1	NN	O	B-protein
p12	NN	O	I-protein
(	NN	O	I-protein
I	NN	O	I-protein
)	NN	O	I-protein
protein	NN	O	I-protein
enhances	NN	O	O
STAT5	NN	O	O
activation	NN	O	O
and	NN	O	O
decreases	NN	O	O
the	NN	O	O
interleukin-2	NN	O	B-protein
requirement	NN	O	O
for	NN	O	O
proliferation	NN	O	O
of	NN	O	O
primary	NN	O	B-cell_type
human	NN	O	I-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
p12	NN	O	B-protein
(	NN	O	I-protein
I	NN	O	I-protein
)	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
encoded	NN	O	O
by	NN	O	O
the	NN	O	O
pX	NN	O	B-DNA
open	NN	O	I-DNA
reading	NN	O	I-DNA
frame	NN	O	I-DNA
I	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
human	NN	O	O
T-lymphotropic	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
(	NN	O	O
HTLV-1	NN	O	O
)	NN	O	O
,	NN	O	O
is	NN	O	O
a	NN	O	O
hydrophobic	NN	O	B-protein
protein	NN	O	I-protein
that	NN	O	O
localizes	NN	O	O
to	NN	O	O
the	NN	O	O
endoplasmic	NN	O	O
reticulum	NN	O	O
and	NN	O	O
the	NN	O	O
Golgi	NN	O	O
.	NN	O	O

Although	NN	O	O
p12	NN	O	B-protein
(	NN	O	I-protein
I	NN	O	I-protein
)	NN	O	I-protein
contains	NN	O	O
4	NN	O	O
minimal	NN	O	O
proline-rich	NN	O	O
,	NN	O	O
src	NN	O	B-protein
homology	NN	O	I-protein
3-binding	NN	O	I-protein
motifs	NN	O	I-protein
(	NN	O	O
PXXP	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
characteristic	NN	O	O
commonly	NN	O	O
found	NN	O	O
in	NN	O	O
proteins	NN	O	O
involved	NN	O	O
in	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
,	NN	O	O
it	NN	O	O
has	NN	O	O
not	NN	O	O
been	NN	O	O
known	NN	O	O
whether	NN	O	O
p12	NN	O	B-protein
(	NN	O	I-protein
I	NN	O	I-protein
)	NN	O	I-protein
has	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
modulating	NN	O	O
intracellular	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
.	NN	O	O

This	NN	O	O
study	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
p12	NN	O	B-protein
(	NN	O	I-protein
I	NN	O	I-protein
)	NN	O	I-protein
binds	NN	O	O
to	NN	O	O
the	NN	O	O
cytoplasmic	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
the	NN	O	O
interleukin-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
chain	NN	O	I-protein
that	NN	O	O
is	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
recruitment	NN	O	O
of	NN	O	O
the	NN	O	O
Jak1	NN	O	B-protein
and	NN	O	I-protein
Jak3	NN	O	I-protein
kinases	NN	O	I-protein
.	NN	O	O

As	NN	O	O
a	NN	O	O
result	NN	O	O
of	NN	O	O
this	NN	O	O
interaction	NN	O	O
,	NN	O	O
p12	NN	O	B-protein
(	NN	O	I-protein
I	NN	O	I-protein
)	NN	O	I-protein
increases	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
5	NN	O	I-protein
(	NN	O	O
STAT5	NN	O	B-protein
)	NN	O	O
DNA	NN	O	O
binding	NN	O	O
and	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
and	NN	O	O
this	NN	O	O
effect	NN	O	O
depends	NN	O	O
on	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
both	NN	O	O
IL-2R	NN	O	B-protein
beta	NN	O	I-protein
and	NN	O	I-protein
gamma	NN	O	I-protein
(	NN	O	I-protein
c	NN	O	I-protein
)	NN	O	I-protein
chains	NN	O	I-protein
and	NN	O	O
Jak3	NN	O	B-protein
.	NN	O	O

Transduction	NN	O	O
of	NN	O	O
primary	NN	O	B-cell_line
human	NN	O	I-cell_line
peripheral	NN	O	I-cell_line
blood	NN	O	I-cell_line
mononuclear	NN	O	I-cell_line
cells	NN	O	I-cell_line
(	NN	O	O
PBMCs	NN	O	B-cell_type
)	NN	O	O
with	NN	O	O
a	NN	O	O
human	NN	O	B-DNA
immunodeficiency	NN	O	I-DNA
virus	NN	O	I-DNA
type	NN	O	I-DNA
1-based	NN	O	I-DNA
retroviral	NN	O	I-DNA
vector	NN	O	I-DNA
expressing	NN	O	O
p12	NN	O	B-protein
(	NN	O	I-protein
I	NN	O	I-protein
)	NN	O	I-protein
also	NN	O	O
resulted	NN	O	O
in	NN	O	O
increased	NN	O	O
STAT5	NN	O	B-protein
phosphorylation	NN	O	O
and	NN	O	O
DNA	NN	O	O
binding	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
p12	NN	O	B-protein
(	NN	O	I-protein
I	NN	O	I-protein
)	NN	O	I-protein
could	NN	O	O
increase	NN	O	O
proliferation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
PBMCs	NN	O	I-cell_type
only	NN	O	O
after	NN	O	O
stimulation	NN	O	O
of	NN	O	O
T-cell	NN	O	B-protein
receptors	NN	O	I-protein
by	NN	O	O
treatment	NN	O	O
of	NN	O	O
cells	NN	O	O
with	NN	O	O
low	NN	O	O
concentrations	NN	O	O
of	NN	O	O
alphaCD3	NN	O	B-protein
and	NN	O	I-protein
alphaCD28	NN	O	I-protein
antibodies	NN	O	I-protein
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
the	NN	O	O
proliferative	NN	O	O
advantage	NN	O	O
of	NN	O	O
p12	NN	O	B-cell_type
(	NN	O	I-cell_type
I	NN	O	I-cell_type
)	NN	O	I-cell_type
-transduced	NN	O	I-cell_type
PBMCs	NN	O	I-cell_type
was	NN	O	O
evident	NN	O	O
mainly	NN	O	O
at	NN	O	O
low	NN	O	O
concentrations	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
.	NN	O	O

Together	NN	O	O
,	NN	O	O
these	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
p12	NN	O	B-protein
(	NN	O	I-protein
I	NN	O	I-protein
)	NN	O	I-protein
may	NN	O	O
confer	NN	O	O
a	NN	O	O
proliferative	NN	O	O
advantage	NN	O	O
on	NN	O	O
HTLV-1-infected	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
suboptimal	NN	O	O
antigen	NN	O	O
stimulation	NN	O	O
and	NN	O	O
that	NN	O	O
this	NN	O	O
event	NN	O	O
may	NN	O	O
account	NN	O	O
for	NN	O	O
the	NN	O	O
clonal	NN	O	O
proliferation	NN	O	O
of	NN	O	O
infected	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

(	NN	O	O
Blood.	NN	O	O
2001	NN	O	O
;	NN	O	O
98	NN	O	O
:	NN	O	O
823-829	NN	O	O
)	NN	O	O

-DOCSTART-	O

Single	NN	O	O
dose	NN	O	O
intranasal	NN	O	O
administration	NN	O	O
of	NN	O	O
retinal	NN	O	B-protein
autoantigen	NN	O	I-protein
generates	NN	O	O
a	NN	O	O
rapid	NN	O	O
accumulation	NN	O	O
and	NN	O	O
cell	NN	O	O
activation	NN	O	O
in	NN	O	O
draining	NN	O	O
lymph	NN	O	O
node	NN	O	O
and	NN	O	O
spleen	NN	O	O
:	NN	O	O
implications	NN	O	O
for	NN	O	O
tolerance	NN	O	O
therapy	NN	O	O
.	NN	O	O

BACKGROUND/AIMS	NN	O	O
:	NN	O	O
A	NN	O	O
single	NN	O	O
intranasal	NN	O	O
delivery	NN	O	O
of	NN	O	O
retinal	NN	O	B-protein
autoantigen	NN	O	I-protein
suppresses	NN	O	O
effectively	NN	O	O
experimental	NN	O	O
autoimmune	NN	O	O
uveoretinitis	NN	O	O
(	NN	O	O
EAU	NN	O	O
)	NN	O	O
.	NN	O	O

To	NN	O	O
further	NN	O	O
unravel	NN	O	O
underlying	NN	O	O
mechanisms	NN	O	O
the	NN	O	O
authors	NN	O	O
wished	NN	O	O
to	NN	O	O
determine	NN	O	O
,	NN	O	O
firstly	NN	O	O
,	NN	O	O
the	NN	O	O
kinetics	NN	O	O
of	NN	O	O
antigen	NN	O	O
delivery	NN	O	O
and	NN	O	O
,	NN	O	O
secondly	NN	O	O
,	NN	O	O
the	NN	O	O
early	NN	O	O
cellular	NN	O	O
responses	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
initial	NN	O	O
stages	NN	O	O
of	NN	O	O
nasal	NN	O	O
mucosal	NN	O	O
tolerance	NN	O	O
induction	NN	O	O
.	NN	O	O

METHODS	NN	O	O
:	NN	O	O
Flow	NN	O	O
cytometry	NN	O	O
,	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
assays	NN	O	O
,	NN	O	O
and	NN	O	O
microscopy	NN	O	O
were	NN	O	O
used	NN	O	O
to	NN	O	O
track	NN	O	O
antigen	NN	O	B-protein
following	NN	O	O
a	NN	O	O
single	NN	O	O
,	NN	O	O
intranasal	NN	O	O
dose	NN	O	O
of	NN	O	O
Alexa-488	NN	O	B-protein
labelled	NN	O	I-protein
retinal	NN	O	I-protein
antigen	NN	O	I-protein
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
A	NN	O	O
rapid	NN	O	O
accumulation	NN	O	O
of	NN	O	O
antigen	NN	O	B-protein
within	NN	O	O
both	NN	O	O
superficial	NN	O	O
cervical	NN	O	O
lymph	NN	O	O
nodes	NN	O	O
(	NN	O	O
SCLN	NN	O	O
)	NN	O	O
and	NN	O	O
spleen	NN	O	O
was	NN	O	O
observed	NN	O	O
after	NN	O	O
30	NN	O	O
minutes	NN	O	O
.	NN	O	O

Significant	NN	O	O
proliferative	NN	O	O
responses	NN	O	O
to	NN	O	O
IRBP	NN	O	O
were	NN	O	O
elicited	NN	O	O
by	NN	O	O
48	NN	O	O
hours	NN	O	O
indicating	NN	O	O
that	NN	O	O
systemic	NN	O	O
priming	NN	O	O
of	NN	O	O
naive	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
retinal	NN	O	B-protein
antigen	NN	O	I-protein
had	NN	O	O
occurred	NN	O	O
.	NN	O	O

Cell	NN	O	O
activation	NN	O	O
was	NN	O	O
further	NN	O	O
confirmed	NN	O	O
by	NN	O	O
immunoprecipitation	NN	O	O
studies	NN	O	O
,	NN	O	O
which	NN	O	O
demonstrated	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
STAT4	NN	O	B-protein
but	NN	O	O
not	NN	O	O
STAT6	NN	O	B-protein
in	NN	O	O
both	NN	O	O
lymph	NN	O	O
nodes	NN	O	O
and	NN	O	O
spleen	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
at	NN	O	O
24	NN	O	O
hours	NN	O	O
,	NN	O	O
STAT4	NN	O	O
heterodimerisation	NN	O	O
with	NN	O	O
STAT	NN	O	B-protein
3	NN	O	I-protein
was	NN	O	O
only	NN	O	O
observed	NN	O	O
in	NN	O	O
spleen	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
:	NN	O	O
The	NN	O	O
results	NN	O	O
provide	NN	O	O
novel	NN	O	O
evidence	NN	O	O
that	NN	O	O
following	NN	O	O
a	NN	O	O
single	NN	O	O
intranasal	NN	O	O
application	NN	O	O
rapid	NN	O	O
transfer	NN	O	O
of	NN	O	O
antigen	NN	O	B-protein
occurs	NN	O	O
.	NN	O	O

Resulting	NN	O	O
T	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
develops	NN	O	O
consequent	NN	O	O
to	NN	O	O
differential	NN	O	O
cell	NN	O	O
signalling	NN	O	O
in	NN	O	O
SCLN	NN	O	O
and	NN	O	O
spleen	NN	O	O
.	NN	O	O

Further	NN	O	O
understanding	NN	O	O
of	NN	O	O
these	NN	O	O
underlying	NN	O	O
cellular	NN	O	O
mechanisms	NN	O	O
,	NN	O	O
in	NN	O	O
particular	NN	O	O
as	NN	O	O
is	NN	O	O
inferred	NN	O	O
by	NN	O	O
the	NN	O	O
results	NN	O	O
the	NN	O	O
contribution	NN	O	O
of	NN	O	O
local	NN	O	O
versus	NN	O	O
systemic	NN	O	O
tolerance	NN	O	O
induction	NN	O	O
,	NN	O	O
may	NN	O	O
assist	NN	O	O
in	NN	O	O
strategies	NN	O	O
to	NN	O	O
clinically	NN	O	O
apply	NN	O	O
mucosal	NN	O	O
tolerance	NN	O	O
therapy	NN	O	O
successfully	NN	O	O
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
interleukin	NN	O	B-protein
(	NN	O	I-protein
IL	NN	O	I-protein
)	NN	O	I-protein
-18	NN	O	I-protein
receptor	NN	O	I-protein
alpha	NN	O	I-protein
chain	NN	O	I-protein
expression	NN	O	O
on	NN	O	O
CD4	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
during	NN	O	O
T	NN	O	O
helper	NN	O	O
(	NN	O	O
Th	NN	O	O
)	NN	O	O
1/Th2	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Critical	NN	O	O
downregulatory	NN	O	O
role	NN	O	O
of	NN	O	O
IL-4	NN	O	B-protein
.	NN	O	O

Interleukin	NN	O	B-protein
(	NN	O	I-protein
IL	NN	O	I-protein
)	NN	O	I-protein
-18	NN	O	I-protein
has	NN	O	O
been	NN	O	O
well	NN	O	O
characterized	NN	O	O
as	NN	O	O
a	NN	O	O
costimulatory	NN	O	B-protein
factor	NN	O	I-protein
for	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
IL-12-mediated	NN	O	B-protein
interferon	NN	O	I-protein
(	NN	O	I-protein
IFN	NN	O	I-protein
)	NN	O	I-protein
-gamma	NN	O	I-protein
production	NN	O	O
by	NN	O	O
T	NN	O	B-cell_type
helper	NN	O	I-cell_type
(	NN	O	I-cell_type
Th	NN	O	I-cell_type
)	NN	O	I-cell_type
1	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
but	NN	O	O
also	NN	O	O
can	NN	O	O
induce	NN	O	O
IL-4	NN	O	O
production	NN	O	O
and	NN	O	O
thus	NN	O	O
facilitate	NN	O	O
the	NN	O	O
differentiation	NN	O	O
of	NN	O	O
Th2	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

To	NN	O	O
determine	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
by	NN	O	O
which	NN	O	O
IL-18	NN	O	B-protein
might	NN	O	O
regulate	NN	O	O
these	NN	O	O
diametrically	NN	O	O
distinct	NN	O	O
immune	NN	O	O
responses	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
analyzed	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
IL-18	NN	O	B-protein
receptor	NN	O	I-protein
alpha	NN	O	I-protein
chain	NN	O	I-protein
(	NN	O	O
IL-18Ralpha	NN	O	B-protein
)	NN	O	O
expression	NN	O	O
.	NN	O	O

The	NN	O	O
majority	NN	O	O
of	NN	O	O
peripheral	NN	O	B-cell_type
CD4	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
constitutively	NN	O	O
expressed	NN	O	O
the	NN	O	O
IL-18Ralpha	NN	O	B-protein
.	NN	O	O

Upon	NN	O	O
antigen	NN	O	O
stimulation	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
IL-12	NN	O	B-protein
,	NN	O	O
marked	NN	O	O
enhancement	NN	O	O
of	NN	O	O
IL-18Ralpha	NN	O	B-protein
expression	NN	O	O
was	NN	O	O
observed	NN	O	O
.	NN	O	O

IL-12	NN	O	B-protein
-mediated	NN	O	O
upregulation	NN	O	O
of	NN	O	O
IL-18Ralpha	NN	O	B-protein
required	NN	O	O
IFN-gamma	NN	O	B-protein
.	NN	O	O

Activated	NN	O	B-cell_type
CD4	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
that	NN	O	O
expressed	NN	O	O
low	NN	O	O
levels	NN	O	O
of	NN	O	O
IL-18Ralpha	NN	O	B-protein
could	NN	O	O
produce	NN	O	O
IFN-gamma	NN	O	B-protein
when	NN	O	O
stimulated	NN	O	O
with	NN	O	O
the	NN	O	O
combination	NN	O	O
of	NN	O	O
IL-12	NN	O	B-protein
and	NN	O	O
IL-18	NN	O	B-protein
,	NN	O	O
while	NN	O	O
CD4	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
which	NN	O	O
expressed	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
IL-18Ralpha	NN	O	B-protein
could	NN	O	O
respond	NN	O	O
to	NN	O	O
IL-18	NN	O	B-protein
alone	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
T	NN	O	O
cell	NN	O	O
stimulation	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
IL-4	NN	O	B-protein
resulted	NN	O	O
in	NN	O	O
a	NN	O	O
downregulation	NN	O	O
of	NN	O	O
IL-18Ralpha	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

Both	NN	O	O
IL-4	NN	O	B-cell_type
(	NN	O	I-cell_type
-/	NN	O	I-cell_type
)	NN	O	I-cell_type
-	NN	O	I-cell_type
and	NN	O	I-cell_type
signal	NN	O	I-cell_type
transducer	NN	O	I-cell_type
and	NN	O	I-cell_type
activator	NN	O	I-cell_type
of	NN	O	I-cell_type
transcription	NN	O	I-cell_type
(	NN	O	I-cell_type
Stat	NN	O	I-cell_type
)	NN	O	I-cell_type
6	NN	O	I-cell_type
(	NN	O	I-cell_type
-/	NN	O	I-cell_type
)	NN	O	I-cell_type
-	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
expressed	NN	O	O
higher	NN	O	O
levels	NN	O	O
of	NN	O	O
IL-18Ralpha	NN	O	B-protein
after	NN	O	O
TCR	NN	O	O
stimulation	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
Stat6	NN	O	O
(	NN	O	O
-/	NN	O	O
)	NN	O	O
-	NN	O	O
mice	NN	O	O
produced	NN	O	O
more	NN	O	O
IFN-gamma	NN	O	B-protein
in	NN	O	O
response	NN	O	O
to	NN	O	O
IL-18	NN	O	B-protein
than	NN	O	O
wild-type	NN	O	O
controls	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
positive/negative	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
IL-18Ralpha	NN	O	B-protein
by	NN	O	O
the	NN	O	O
major	NN	O	O
inductive	NN	O	O
cytokines	NN	O	B-protein
(	NN	O	O
IL-12	NN	O	B-protein
and	NN	O	O
IL-4	NN	O	B-protein
)	NN	O	O
determines	NN	O	O
the	NN	O	O
capacity	NN	O	O
of	NN	O	O
IL-18	NN	O	B-protein
to	NN	O	O
polarize	NN	O	O
an	NN	O	O
immune	NN	O	O
response	NN	O	O
.	NN	O	O

-DOCSTART-	O

Glucocorticoid-regulated	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

Glucocorticoids	NN	O	O
are	NN	O	O
the	NN	O	O
most	NN	O	O
effective	NN	O	O
antiinflammatory	NN	O	O
drugs	NN	O	O
used	NN	O	O
in	NN	O	O
the	NN	O	O
treatment	NN	O	O
of	NN	O	O
asthma	NN	O	O
.	NN	O	O

They	NN	O	O
act	NN	O	O
by	NN	O	O
binding	NN	O	O
to	NN	O	O
a	NN	O	O
specific	NN	O	O
receptor	NN	O	O
(	NN	O	O
GR	NN	O	B-protein
)	NN	O	O
that	NN	O	O
,	NN	O	O
upon	NN	O	O
activation	NN	O	O
,	NN	O	O
translocates	NN	O	O
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
and	NN	O	O
either	NN	O	O
increases	NN	O	O
(	NN	O	O
transactivates	NN	O	O
)	NN	O	O
or	NN	O	O
decreases	NN	O	O
(	NN	O	O
transrepresses	NN	O	O
)	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
pro-inflammatory	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
such	NN	O	O
as	NN	O	O
activator	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	I-protein
AP	NN	O	I-protein
)	NN	O	I-protein
-1	NN	O	I-protein
,	NN	O	O
signal	NN	O	B-protein
transducers	NN	O	I-protein
and	NN	O	I-protein
activators	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
(	NN	O	O
STATs	NN	O	B-protein
)	NN	O	O
,	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
(	NN	O	O
NFAT	NN	O	B-protein
)	NN	O	O
and	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	I-protein
NF	NN	O	I-protein
)	NN	O	I-protein
-kappa	NN	O	I-protein
B	NN	O	I-protein
is	NN	O	O
thought	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
major	NN	O	O
action	NN	O	O
of	NN	O	O
glucocorticoids	NN	O	O
.	NN	O	O

Acetylation	NN	O	O
of	NN	O	O
histones	NN	O	B-protein
allows	NN	O	O
unwinding	NN	O	O
of	NN	O	O
the	NN	O	O
local	NN	O	B-DNA
DNA	NN	O	I-DNA
structure	NN	O	I-DNA
and	NN	O	O
enables	NN	O	O
RNA	NN	O	B-protein
polymerase	NN	O	I-protein
II	NN	O	I-protein
to	NN	O	O
enhance	NN	O	O
gene	NN	O	O
transcription	NN	O	O
.	NN	O	O

Histone	NN	O	O
acetylation	NN	O	O
is	NN	O	O
regulated	NN	O	O
by	NN	O	O
a	NN	O	O
balance	NN	O	O
between	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
histone	NN	O	B-protein
acetyltransferases	NN	O	I-protein
(	NN	O	O
HATs	NN	O	B-protein
)	NN	O	O
and	NN	O	O
histone	NN	O	B-protein
deacetylases	NN	O	I-protein
(	NN	O	O
HDACs	NN	O	B-protein
)	NN	O	O
.	NN	O	O

GR	NN	O	B-protein
acts	NN	O	O
as	NN	O	O
a	NN	O	O
direct	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
NF-kappa	NN	O	O
B-induced	NN	O	O
HAT	NN	O	O
activity	NN	O	O
and	NN	O	O
also	NN	O	O
by	NN	O	O
recruiting	NN	O	O
HDAC2	NN	O	B-protein
to	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B/HAT	NN	O	I-protein
complex	NN	O	I-protein
.	NN	O	O

A	NN	O	O
sub-group	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
glucocorticoid-insensitive	NN	O	O
asthma	NN	O	O
have	NN	O	O
an	NN	O	O
inability	NN	O	O
to	NN	O	O
induce	NN	O	O
histone	NN	O	O
acetylation	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
dexamethasone	NN	O	O
suggesting	NN	O	O
reduced	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
GR-specific	NN	O	B-protein
HAT	NN	O	I-protein
.	NN	O	O

This	NN	O	O
suggests	NN	O	O
that	NN	O	O
pharmacological	NN	O	O
manipulation	NN	O	O
of	NN	O	O
specific	NN	O	O
histone	NN	O	O
acetylation	NN	O	O
status	NN	O	O
is	NN	O	O
a	NN	O	O
potentially	NN	O	O
useful	NN	O	O
approach	NN	O	O
for	NN	O	O
the	NN	O	O
treatment	NN	O	O
of	NN	O	O
inflammatory	NN	O	O
diseases	NN	O	O
.	NN	O	O

Identification	NN	O	O
of	NN	O	O
the	NN	O	O
precise	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
activated	NN	O	B-protein
GR	NN	O	I-protein
recruits	NN	O	O
HDAC2	NN	O	B-protein
may	NN	O	O
reveal	NN	O	O
new	NN	O	O
targets	NN	O	O
for	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
drugs	NN	O	O
that	NN	O	O
may	NN	O	O
dissociate	NN	O	O
the	NN	O	O
antiinflammatory	NN	O	O
actions	NN	O	O
of	NN	O	O
glucocorticoids	NN	O	O
from	NN	O	O
their	NN	O	O
side	NN	O	O
effects	NN	O	O
that	NN	O	O
are	NN	O	O
largely	NN	O	O
due	NN	O	O
to	NN	O	O
gene	NN	O	O
induction	NN	O	O
.	NN	O	O

Copyright	NN	O	O
Academic	NN	O	O
Press	NN	O	O
.	NN	O	O

-DOCSTART-	O

Differential	NN	O	O
ultraviolet-B-induced	NN	O	O
immunomodulation	NN	O	O
in	NN	O	O
XPA	NN	O	O
,	NN	O	O
XPC	NN	O	O
,	NN	O	O
and	NN	O	O
CSB	NN	O	O
DNA	NN	O	O
repair-deficient	NN	O	O
mice	NN	O	O
.	NN	O	O

Ultraviolet	NN	O	O
B	NN	O	O
irradiation	NN	O	O
has	NN	O	O
serious	NN	O	O
consequences	NN	O	O
for	NN	O	O
cellular	NN	O	O
immunity	NN	O	O
and	NN	O	O
can	NN	O	O
suppress	NN	O	O
the	NN	O	O
rejection	NN	O	O
of	NN	O	O
skin	NN	O	O
tumors	NN	O	O
and	NN	O	O
the	NN	O	O
resistance	NN	O	O
to	NN	O	O
infectious	NN	O	O
diseases	NN	O	O
.	NN	O	O

DNA	NN	O	O
damage	NN	O	O
plays	NN	O	O
a	NN	O	O
crucial	NN	O	O
role	NN	O	O
in	NN	O	O
these	NN	O	O
immunomodulatory	NN	O	O
effects	NN	O	O
of	NN	O	O
ultraviolet	NN	O	O
B	NN	O	O
,	NN	O	O
as	NN	O	O
impaired	NN	O	O
repair	NN	O	O
of	NN	O	O
ultraviolet-B-induced	NN	O	O
DNA	NN	O	O
damage	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
cause	NN	O	O
suppression	NN	O	O
of	NN	O	O
cellular	NN	O	O
immunity	NN	O	O
.	NN	O	O

Ultraviolet-B-induced	NN	O	O
DNA	NN	O	O
damage	NN	O	O
is	NN	O	O
repaired	NN	O	O
by	NN	O	O
the	NN	O	O
nucleotide	NN	O	O
excision	NN	O	O
repair	NN	O	O
mechanism	NN	O	O
very	NN	O	O
efficiently	NN	O	O
.	NN	O	O

Nucleotide	NN	O	O
excision	NN	O	O
repair	NN	O	O
comprises	NN	O	O
two	NN	O	O
subpathways	NN	O	O
:	NN	O	O
transcription-coupled	NN	O	O
and	NN	O	O
global	NN	O	O
genome	NN	O	O
repair	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
the	NN	O	O
immunologic	NN	O	O
consequences	NN	O	O
of	NN	O	O
specific	NN	O	O
nucleotide	NN	O	O
excision	NN	O	O
repair	NN	O	O
defects	NN	O	O
in	NN	O	O
three	NN	O	O
mouse	NN	O	O
models	NN	O	O
,	NN	O	O
XPA	NN	O	O
,	NN	O	O
XPC	NN	O	O
,	NN	O	O
and	NN	O	O
CSB	NN	O	O
mutant	NN	O	O
mice	NN	O	O
,	NN	O	O
were	NN	O	O
investigated	NN	O	O
.	NN	O	O

XPA	NN	O	O
mice	NN	O	O
carry	NN	O	O
a	NN	O	O
total	NN	O	O
nucleotide	NN	O	O
excision	NN	O	O
repair	NN	O	O
defect	NN	O	O
,	NN	O	O
whereas	NN	O	O
XPC	NN	O	O
and	NN	O	O
CSB	NN	O	O
mice	NN	O	O
only	NN	O	O
lack	NN	O	O
global	NN	O	O
genome	NN	O	O
and	NN	O	O
transcription-coupled	NN	O	O
nucleotide	NN	O	O
excision	NN	O	O
repair	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

Our	NN	O	O
data	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
cellular	NN	O	O
immune	NN	O	O
parameters	NN	O	O
in	NN	O	O
XPA	NN	O	O
,	NN	O	O
XPC	NN	O	O
,	NN	O	O
and	NN	O	O
CSB	NN	O	O
mice	NN	O	O
are	NN	O	O
normal	NN	O	O
compared	NN	O	O
with	NN	O	O
their	NN	O	O
wild-type	NN	O	O
(	NN	O	O
control	NN	O	O
)	NN	O	O
littermates	NN	O	O
.	NN	O	O

This	NN	O	O
may	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
reported	NN	O	O
altered	NN	O	O
cellular	NN	O	O
responses	NN	O	O
in	NN	O	O
xeroderma	NN	O	O
pigmentosum	NN	O	O
patients	NN	O	O
are	NN	O	O
not	NN	O	O
constitutive	NN	O	O
but	NN	O	O
could	NN	O	O
be	NN	O	O
due	NN	O	O
to	NN	O	O
external	NN	O	O
factors	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
ultraviolet	NN	O	O
B	NN	O	O
.	NN	O	O

Upon	NN	O	O
exposure	NN	O	O
to	NN	O	O
ultraviolet	NN	O	O
B	NN	O	O
,	NN	O	O
only	NN	O	O
XPA	NN	O	O
mice	NN	O	O
are	NN	O	O
very	NN	O	O
sensitive	NN	O	O
to	NN	O	O
ultraviolet-B-induced	NN	O	O
inhibition	NN	O	O
of	NN	O	O
Th1-mediated	NN	O	O
contact	NN	O	O
hypersensitivity	NN	O	O
responses	NN	O	O
and	NN	O	O
interferon-gamma	NN	O	B-protein
production	NN	O	O
in	NN	O	O
skin	NN	O	O
draining	NN	O	O
lymph	NN	O	O
nodes	NN	O	O
.	NN	O	O

Lipopolysaccharide-stimulated	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
and	NN	O	O
interleukin-10	NN	O	B-protein
production	NN	O	O
are	NN	O	O
significantly	NN	O	O
augmented	NN	O	O
in	NN	O	O
both	NN	O	O
XPA	NN	O	O
and	NN	O	O
CSB	NN	O	O
mice	NN	O	O
after	NN	O	O
ultraviolet	NN	O	O
B	NN	O	O
exposure	NN	O	O
.	NN	O	O

Lymph	NN	O	O
node	NN	O	O
cell	NN	O	O
numbers	NN	O	O
were	NN	O	O
increased	NN	O	O
very	NN	O	O
significantly	NN	O	O
in	NN	O	O
XPA	NN	O	O
,	NN	O	O
mildly	NN	O	O
increased	NN	O	O
in	NN	O	O
CSB	NN	O	O
,	NN	O	O
and	NN	O	O
not	NN	O	O
in	NN	O	O
XPC	NN	O	O
mice	NN	O	O
.	NN	O	O

In	NN	O	O
general	NN	O	O
XPC	NN	O	O
mice	NN	O	O
do	NN	O	O
not	NN	O	O
exhibit	NN	O	O
any	NN	O	O
indication	NN	O	O
of	NN	O	O
enhanced	NN	O	O
ultraviolet	NN	O	O
B	NN	O	O
susceptibility	NN	O	O
with	NN	O	O
regard	NN	O	O
to	NN	O	O
the	NN	O	O
immune	NN	O	O
parameters	NN	O	O
analyzed	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
both	NN	O	O
global	NN	O	O
genome	NN	O	O
repair	NN	O	O
and	NN	O	O
transcription-coupled	NN	O	O
repair	NN	O	O
are	NN	O	O
needed	NN	O	O
to	NN	O	O
prevent	NN	O	O
immunomodulation	NN	O	O
by	NN	O	O
ultraviolet	NN	O	O
B	NN	O	O
,	NN	O	O
whereas	NN	O	O
transcription-coupled	NN	O	O
repair	NN	O	O
is	NN	O	O
the	NN	O	O
major	NN	O	O
DNA	NN	O	O
repair	NN	O	O
subpathway	NN	O	O
of	NN	O	O
nucleotide	NN	O	O
excision	NN	O	O
repair	NN	O	O
that	NN	O	O
prevents	NN	O	O
the	NN	O	O
acute	NN	O	O
ultraviolet-B-induced	NN	O	O
effects	NN	O	O
such	NN	O	O
as	NN	O	O
erythema	NN	O	O
.	NN	O	O

-DOCSTART-	O

Biomechanical	NN	O	O
strain	NN	O	O
induces	NN	O	O
class	NN	O	O
a	NN	O	O
scavenger	NN	O	B-protein
receptor	NN	O	I-protein
expression	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocyte/macrophages	NN	O	I-cell_type
and	NN	O	O
THP-1	NN	O	B-cell_type
cells	NN	O	I-cell_type
:	NN	O	O
a	NN	O	O
potential	NN	O	O
mechanism	NN	O	O
of	NN	O	O
increased	NN	O	O
atherosclerosis	NN	O	O
in	NN	O	O
hypertension	NN	O	O
.	NN	O	O

BACKGROUND	NN	O	O
:	NN	O	O
Although	NN	O	O
hypertension	NN	O	O
is	NN	O	O
an	NN	O	O
important	NN	O	O
risk	NN	O	O
factor	NN	O	O
for	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
atherosclerosis	NN	O	O
,	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
for	NN	O	O
this	NN	O	O
interaction	NN	O	O
are	NN	O	O
incompletely	NN	O	O
described	NN	O	O
.	NN	O	O

Previous	NN	O	O
studies	NN	O	O
have	NN	O	O
suggested	NN	O	O
that	NN	O	O
biomechanical	NN	O	O
strain	NN	O	O
regulates	NN	O	O
macrophage	NN	O	O
phenotype	NN	O	O
.	NN	O	O

We	NN	O	O
tested	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
biomechanical	NN	O	O
strain	NN	O	O
can	NN	O	O
induce	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
class	NN	O	B-protein
A	NN	O	I-protein
scavenger	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
SRA	NN	O	B-protein
)	NN	O	O
,	NN	O	O
an	NN	O	O
important	NN	O	O
lipoprotein	NN	O	B-protein
receptor	NN	O	I-protein
in	NN	O	O
atherogenesis	NN	O	O
.	NN	O	O

METHODS	NN	O	O
AND	NN	O	O
RESULTS	NN	O	O
:	NN	O	O
Human	NN	O	B-cell_type
monocyte/macrophages	NN	O	I-cell_type
or	NN	O	O
THP-1	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
cultured	NN	O	O
in	NN	O	O
a	NN	O	O
device	NN	O	O
that	NN	O	O
imposes	NN	O	O
uniform	NN	O	O
biaxial	NN	O	O
cyclic	NN	O	O
1-Hz	NN	O	O
strains	NN	O	O
of	NN	O	O
0	NN	O	O
%	NN	O	O
,	NN	O	O
1	NN	O	O
%	NN	O	O
,	NN	O	O
2	NN	O	O
%	NN	O	O
,	NN	O	O
or	NN	O	O
3	NN	O	O
%	NN	O	O
,	NN	O	O
and	NN	O	O
SRA	NN	O	B-protein
expression	NN	O	O
was	NN	O	O
analyzed	NN	O	O
.	NN	O	O

Mechanical	NN	O	O
strains	NN	O	O
induced	NN	O	O
SRA	NN	O	B-RNA
mRNA	NN	O	I-RNA
(	NN	O	O
3.5+/-0.6-fold	NN	O	O
at	NN	O	O
3	NN	O	O
%	NN	O	O
strain	NN	O	O
for	NN	O	O
48	NN	O	O
hours	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
SRA	NN	O	B-protein
protein	NN	O	I-protein
in	NN	O	O
THP-1	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
an	NN	O	O
amplitude-dependent	NN	O	O
manner	NN	O	O
.	NN	O	O

This	NN	O	O
induction	NN	O	O
was	NN	O	O
accompanied	NN	O	O
by	NN	O	O
augmented	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
class	NN	O	B-protein
B	NN	O	I-protein
scavenger	NN	O	I-protein
receptor	NN	O	I-protein
CD36	NN	O	I-protein
(	NN	O	O
2.8+/-0.3-fold	NN	O	O
,	NN	O	O
P	NN	O	O
<	NN	O	O
0.001	NN	O	O
)	NN	O	O
but	NN	O	O
not	NN	O	O
by	NN	O	O
increased	NN	O	O
peroxisome	NN	O	B-protein
proliferator-activated	NN	O	I-protein
receptor-gamma	NN	O	I-protein
expression	NN	O	O
.	NN	O	O

To	NN	O	O
evaluate	NN	O	O
this	NN	O	O
effect	NN	O	O
in	NN	O	O
vivo	NN	O	O
,	NN	O	O
apolipoprotein	NN	O	O
E	NN	O	O
(	NN	O	O
-/-	NN	O	O
)	NN	O	O
mice	NN	O	O
were	NN	O	O
randomly	NN	O	O
assigned	NN	O	O
to	NN	O	O
receive	NN	O	O
standard	NN	O	O
chow	NN	O	O
,	NN	O	O
a	NN	O	O
high-cholesterol	NN	O	O
diet	NN	O	O
,	NN	O	O
or	NN	O	O
a	NN	O	O
high-cholesterol	NN	O	O
diet	NN	O	O
with	NN	O	O
hypertension	NN	O	O
induced	NN	O	O
by	NN	O	O
angiotensin	NN	O	O
II	NN	O	O
infusion	NN	O	O
for	NN	O	O
8	NN	O	O
weeks	NN	O	O
.	NN	O	O

Immunohistochemistry	NN	O	O
revealed	NN	O	O
that	NN	O	O
among	NN	O	O
macrophages	NN	O	B-cell_type
in	NN	O	O
atherosclerotic	NN	O	O
lesions	NN	O	O
of	NN	O	O
the	NN	O	O
aorta	NN	O	O
,	NN	O	O
the	NN	O	O
proportion	NN	O	O
of	NN	O	O
macrophages	NN	O	B-cell_type
with	NN	O	O
SRA	NN	O	B-protein
expression	NN	O	O
was	NN	O	O
highest	NN	O	O
in	NN	O	O
hypertensive	NN	O	O
animals	NN	O	O
on	NN	O	O
a	NN	O	O
high-cholesterol	NN	O	O
diet	NN	O	O
(	NN	O	O
43.9+/-0.7	NN	O	O
%	NN	O	O
,	NN	O	O
versus	NN	O	O
12.0+/-2.0	NN	O	O
%	NN	O	O
for	NN	O	O
normotensive	NN	O	O
animals	NN	O	O
on	NN	O	O
a	NN	O	O
high-cholesterol	NN	O	O
diet	NN	O	O
and	NN	O	O
4.7+/-4.7	NN	O	O
%	NN	O	O
for	NN	O	O
animals	NN	O	O
on	NN	O	O
standard	NN	O	O
chow	NN	O	O
;	NN	O	O
P	NN	O	O
<	NN	O	O
0.001	NN	O	O
)	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
:	NN	O	O
Biomechanical	NN	O	O
strain	NN	O	O
induces	NN	O	O
SRA	NN	O	B-protein
expression	NN	O	O
by	NN	O	O
monocyte/macrophages	NN	O	B-cell_type
,	NN	O	O
suggesting	NN	O	O
a	NN	O	O
novel	NN	O	O
mechanism	NN	O	O
for	NN	O	O
promotion	NN	O	O
of	NN	O	O
atherosclerosis	NN	O	O
in	NN	O	O
hypertensive	NN	O	O
patients	NN	O	O
.	NN	O	O

-DOCSTART-	O

High	NN	O	O
glucose	NN	O	O
induces	NN	O	O
MCP-1	NN	O	O
expression	NN	O	O
partly	NN	O	O
via	NN	O	O
tyrosine	NN	O	O
kinase-AP-1	NN	O	O
pathway	NN	O	O
in	NN	O	O
peritoneal	NN	O	B-cell_type
mesothelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

BACKGROUND	NN	O	O
:	NN	O	O
High	NN	O	O
glucose	NN	O	O
in	NN	O	O
peritoneal	NN	O	O
dialysis	NN	O	O
solutions	NN	O	O
has	NN	O	O
been	NN	O	O
implicated	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
peritoneal	NN	O	O
fibrosis	NN	O	O
in	NN	O	O
chronic	NN	O	O
ambulatory	NN	O	O
peritoneal	NN	O	O
dialysis	NN	O	O
(	NN	O	O
CAPD	NN	O	O
)	NN	O	O
patients	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
are	NN	O	O
not	NN	O	O
very	NN	O	O
clear	NN	O	O
.	NN	O	O

Peritoneal	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
seem	NN	O	O
to	NN	O	O
participate	NN	O	O
in	NN	O	O
the	NN	O	O
process	NN	O	O
of	NN	O	O
peritoneal	NN	O	O
fibrosis	NN	O	O
and	NN	O	O
monocyte	NN	O	B-protein
chemoattractant	NN	O	I-protein
protein-1	NN	O	I-protein
(	NN	O	O
MCP-1	NN	O	B-protein
)	NN	O	O
plays	NN	O	O
a	NN	O	O
key	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
recruitment	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
toward	NN	O	O
the	NN	O	O
peritoneal	NN	O	O
cavity	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
about	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
high	NN	O	O
glucose	NN	O	O
on	NN	O	O
MCP-1	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
its	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathway	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
peritoneal	NN	O	I-cell_type
mesothelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

METHODS	NN	O	O
:	NN	O	O
Mesothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
cultured	NN	O	O
with	NN	O	O
glucose	NN	O	O
(	NN	O	O
5	NN	O	O
to	NN	O	O
100	NN	O	O
mmol/L	NN	O	O
)	NN	O	O
or	NN	O	O
mannitol	NN	O	O
chronically	NN	O	O
for	NN	O	O
up	NN	O	O
to	NN	O	O
seven	NN	O	O
days	NN	O	O
.	NN	O	O

MCP-1	NN	O	B-protein
expression	NN	O	O
of	NN	O	O
mRNA	NN	O	O
and	NN	O	O
protein	NN	O	O
was	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
enzyme-linked	NN	O	O
immunosorbent	NN	O	O
assay	NN	O	O
(	NN	O	O
ELISA	NN	O	O
)	NN	O	O
.	NN	O	O

Chemotactic	NN	O	O
activity	NN	O	O
of	NN	O	O
high-glucose-conditioned	NN	O	O
culture	NN	O	O
supernatant	NN	O	O
was	NN	O	O
measured	NN	O	O
by	NN	O	O
chemotactic	NN	O	O
assay	NN	O	O
.	NN	O	O

To	NN	O	O
examine	NN	O	O
the	NN	O	O
roles	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
activator	NN	O	B-protein
protein-1	NN	O	I-protein
(	NN	O	O
AP-1	NN	O	B-protein
)	NN	O	O
and	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
(	NN	O	O
NF-kappaB	NN	O	B-protein
)	NN	O	O
,	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
(	NN	O	O
EMSA	NN	O	O
)	NN	O	O
was	NN	O	O
performed	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
Glucose	NN	O	O
induced	NN	O	O
MCP-1	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
in	NN	O	O
a	NN	O	O
time-	NN	O	O
and	NN	O	O
dose-dependent	NN	O	O
manner	NN	O	O
.	NN	O	O

MCP-1	NN	O	B-protein
protein	NN	O	I-protein
in	NN	O	O
cell	NN	O	O
culture	NN	O	O
supernant	NN	O	O
was	NN	O	O
also	NN	O	O
increased	NN	O	O
.	NN	O	O

Equivalent	NN	O	O
concentrations	NN	O	O
of	NN	O	O
mannitol	NN	O	O
had	NN	O	O
no	NN	O	O
significant	NN	O	O
effect	NN	O	O
.	NN	O	O

High-glucose-conditioned	NN	O	O
supernatant	NN	O	O
possessed	NN	O	O
an	NN	O	O
increased	NN	O	O
chemotactic	NN	O	O
activity	NN	O	O
for	NN	O	O
monocytes	NN	O	B-cell_type
,	NN	O	O
which	NN	O	O
was	NN	O	O
neutralized	NN	O	O
by	NN	O	O
anti-MCP-1	NN	O	B-protein
antibody	NN	O	I-protein
.	NN	O	O

EMSA	NN	O	O
revealed	NN	O	O
that	NN	O	O
glucose	NN	O	O
increased	NN	O	O
the	NN	O	O
AP-1	NN	O	B-protein
binding	NN	O	O
activity	NN	O	O
in	NN	O	O
a	NN	O	O
time-	NN	O	O
and	NN	O	O
dose-dependent	NN	O	O
manner	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
NF-kappaB	NN	O	B-protein
.	NN	O	O

Curcumin	NN	O	O
,	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
,	NN	O	O
dose-dependently	NN	O	O
suppressed	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
MCP-1	NN	O	B-RNA
mRNA	NN	O	I-RNA
by	NN	O	O
high	NN	O	O
glucose	NN	O	O
.	NN	O	O

Tyrosine	NN	O	O
kinase	NN	O	O
inhibitors	NN	O	O
such	NN	O	O
as	NN	O	O
genistein	NN	O	O
(	NN	O	O
12.5	NN	O	O
to	NN	O	O
50	NN	O	O
micromol/L	NN	O	O
)	NN	O	O
and	NN	O	O
herbimycin	NN	O	O
A	NN	O	O
(	NN	O	O
0.1	NN	O	O
to	NN	O	O
1	NN	O	O
micromol/L	NN	O	O
)	NN	O	O
inhibited	NN	O	O
the	NN	O	O
high-glucose-induced	NN	O	O
MCP-1	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
in	NN	O	O
a	NN	O	O
dose-dependent	NN	O	O
manner	NN	O	O
,	NN	O	O
and	NN	O	O
also	NN	O	O
suppressed	NN	O	O
the	NN	O	O
high-glucose-induced	NN	O	O
AP-1	NN	O	B-protein
binding	NN	O	O
activity	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
:	NN	O	O
:	NN	O	O
High	NN	O	O
glucose	NN	O	O
induced	NN	O	O
mesothelial	NN	O	O
MCP-1	NN	O	B-protein
expression	NN	O	O
partly	NN	O	O
via	NN	O	O
the	NN	O	O
tyrosine	NN	O	O
kinase-	NN	O	O
AP-1	NN	O	B-protein
pathway	NN	O	O
.	NN	O	O

-DOCSTART-	O

Signal	NN	O	O
thresholds	NN	O	O
and	NN	O	O
modular	NN	O	O
synergy	NN	O	O
during	NN	O	O
expression	NN	O	O
of	NN	O	O
costimulatory	NN	O	O
molecules	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
analyzed	NN	O	O
intracellular	NN	O	O
pathways	NN	O	O
modulating	NN	O	O
surface	NN	O	O
densities	NN	O	O
of	NN	O	O
CD80	NN	O	B-protein
and	NN	O	O
CD86	NN	O	B-protein
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
activated	NN	O	O
through	NN	O	O
ligation	NN	O	O
of	NN	O	O
the	NN	O	O
Ag	NN	O	B-protein
receptor	NN	O	I-protein
,	NN	O	O
and	NN	O	O
the	NN	O	O
adhesion	NN	O	B-protein
molecule	NN	O	I-protein
CD54	NN	O	B-protein
.	NN	O	O

Whereas	NN	O	O
B	NN	O	B-protein
cell	NN	O	I-protein
Ag	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
BCR	NN	O	B-protein
)	NN	O	O
cross-linking	NN	O	O
alone	NN	O	O
stimulated	NN	O	O
increased	NN	O	O
expression	NN	O	O
of	NN	O	O
CD86	NN	O	B-protein
,	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
CD80	NN	O	B-protein
required	NN	O	O
dual	NN	O	O
stimulation	NN	O	O
with	NN	O	O
anti-IgM	NN	O	B-protein
and	NN	O	O
anti-CD54	NN	O	B-protein
.	NN	O	O

The	NN	O	O
principal	NN	O	O
downstream	NN	O	O
component	NN	O	O
contributed	NN	O	O
by	NN	O	O
BCR	NN	O	B-protein
signaling	NN	O	O
,	NN	O	O
toward	NN	O	O
both	NN	O	O
CD80	NN	O	B-protein
and	NN	O	O
CD86	NN	O	B-protein
induction	NN	O	O
,	NN	O	O
was	NN	O	O
the	NN	O	O
elevated	NN	O	O
concentration	NN	O	O
of	NN	O	O
free	NN	O	O
cytoplasmic	NN	O	O
Ca	NN	O	O
(	NN	O	O
2+	NN	O	O
)	NN	O	O
,	NN	O	O
recruited	NN	O	O
by	NN	O	O
way	NN	O	O
of	NN	O	O
capacitative	NN	O	O
influx	NN	O	O
.	NN	O	O

This	NN	O	O
alone	NN	O	O
was	NN	O	O
sufficient	NN	O	O
to	NN	O	O
generate	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
CD86	NN	O	B-protein
levels	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
CD80	NN	O	B-protein
enhancement	NN	O	O
required	NN	O	O
the	NN	O	O
concerted	NN	O	O
action	NN	O	O
of	NN	O	O
both	NN	O	O
intracellular	NN	O	O
Ca	NN	O	O
(	NN	O	O
2+	NN	O	O
)	NN	O	O
concentration	NN	O	O
and	NN	O	O
CD54-initiated	NN	O	O
pathways	NN	O	O
.	NN	O	O

The	NN	O	O
nexus	NN	O	O
between	NN	O	O
anti-IgM	NN	O	O
and	NN	O	O
anti-CD54	NN	O	O
stimulation	NN	O	O
,	NN	O	O
in	NN	O	O
the	NN	O	O
context	NN	O	O
of	NN	O	O
CD80	NN	O	B-protein
regulation	NN	O	O
,	NN	O	O
was	NN	O	O
identified	NN	O	O
to	NN	O	O
involve	NN	O	O
a	NN	O	O
self-propagating	NN	O	O
process	NN	O	O
of	NN	O	O
sequential	NN	O	O
synergy	NN	O	O
.	NN	O	O

The	NN	O	O
first	NN	O	O
step	NN	O	O
involved	NN	O	O
amplified	NN	O	O
accumulation	NN	O	O
of	NN	O	O
intracellular	NN	O	O
cAMP	NN	O	O
,	NN	O	O
as	NN	O	O
a	NN	O	O
result	NN	O	O
of	NN	O	O
cross-talk	NN	O	O
between	NN	O	O
BCR	NN	O	B-protein
-mobilized	NN	O	O
Ca	NN	O	O
(	NN	O	O
2+	NN	O	O
)	NN	O	O
and	NN	O	O
CD54-derived	NN	O	O
signals	NN	O	O
.	NN	O	O

This	NN	O	O
then	NN	O	O
facilitated	NN	O	O
a	NN	O	O
second	NN	O	O
synergistic	NN	O	O
interaction	NN	O	O
between	NN	O	O
Ca	NN	O	O
(	NN	O	O
2+	NN	O	O
)	NN	O	O
and	NN	O	O
cAMP	NN	O	O
,	NN	O	O
culminating	NN	O	O
in	NN	O	O
CD80	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

Our	NN	O	O
findings	NN	O	O
of	NN	O	O
distinct	NN	O	O
signal	NN	O	O
transducer	NN	O	O
requirements	NN	O	O
,	NN	O	O
with	NN	O	O
the	NN	O	O
added	NN	O	O
consequences	NN	O	O
of	NN	O	O
cross-talk	NN	O	O
,	NN	O	O
offers	NN	O	O
an	NN	O	O
explanation	NN	O	O
for	NN	O	O
variable	NN	O	O
modulation	NN	O	O
of	NN	O	O
costimulatory	NN	O	O
molecule	NN	O	O
expression	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
diverse	NN	O	O
physiological	NN	O	O
stimuli	NN	O	O
.	NN	O	O

Importantly	NN	O	O
,	NN	O	O
these	NN	O	O
results	NN	O	O
also	NN	O	O
reveal	NN	O	O
how	NN	O	O
concentration	NN	O	O
threshold	NN	O	O
barriers	NN	O	O
for	NN	O	O
recruitment	NN	O	O
of	NN	O	O
individual	NN	O	O
second	NN	O	O
messengers	NN	O	O
can	NN	O	O
be	NN	O	O
overcome	NN	O	O
by	NN	O	O
constructive	NN	O	O
convergence	NN	O	O
of	NN	O	O
signaling	NN	O	O
modules	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
role	NN	O	O
of	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
in	NN	O	O
neoplastic	NN	O	O
transformation	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
review	NN	O	O
,	NN	O	O
we	NN	O	O
focus	NN	O	O
on	NN	O	O
new	NN	O	O
data	NN	O	O
from	NN	O	O
basic	NN	O	O
,	NN	O	O
translational	NN	O	O
and	NN	O	O
clinical	NN	O	O
research	NN	O	O
relating	NN	O	O
to	NN	O	O
the	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
(	NN	O	O
EBV	NN	O	O
)	NN	O	O
.	NN	O	O

Beside	NN	O	O
its	NN	O	O
well-known	NN	O	O
tropism	NN	O	O
for	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
epithelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
EBV	NN	O	O
also	NN	O	O
infects	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
granulocytes	NN	O	B-cell_type
.	NN	O	O

After	NN	O	O
primary	NN	O	O
infection	NN	O	O
,	NN	O	O
EBV	NN	O	O
persists	NN	O	O
throughout	NN	O	O
the	NN	O	O
life	NN	O	O
span	NN	O	O
in	NN	O	O
resting	NN	O	B-cell_type
memory	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
from	NN	O	O
where	NN	O	O
it	NN	O	O
is	NN	O	O
reactivated	NN	O	O
upon	NN	O	O
breakdown	NN	O	O
of	NN	O	O
cellular	NN	O	O
immunity	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
process	NN	O	O
of	NN	O	O
neoplastic	NN	O	O
transformation	NN	O	O
,	NN	O	O
the	NN	O	O
EBV-encoded	NN	O	B-DNA
latent	NN	O	I-DNA
membrane	NN	O	I-DNA
protein	NN	O	I-DNA
1	NN	O	I-DNA
(	NN	O	I-DNA
LMP1	NN	O	I-DNA
)	NN	O	I-DNA
oncogene	NN	O	I-DNA
represents	NN	O	O
the	NN	O	O
major	NN	O	O
driving	NN	O	O
force	NN	O	O
.	NN	O	O

LMP1	NN	O	B-protein
acts	NN	O	O
like	NN	O	O
a	NN	O	O
constitutively	NN	O	B-protein
activated	NN	O	I-protein
receptor	NN	O	I-protein
of	NN	O	O
the	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
receptor	NN	O	I-protein
family	NN	O	I-protein
and	NN	O	O
allows	NN	O	O
the	NN	O	O
amplification	NN	O	O
or	NN	O	O
bypassing	NN	O	O
of	NN	O	O
physiological	NN	O	O
regulatory	NN	O	O
signals	NN	O	O
through	NN	O	O
direct	NN	O	O
and	NN	O	O
indirect	NN	O	O
interactions	NN	O	O
with	NN	O	O
proteins	NN	O	O
of	NN	O	O
the	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
receptor-associated	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	I-protein
TRAF	NN	O	I-protein
)	NN	O	I-protein
family	NN	O	I-protein
.	NN	O	O

TRAF2	NN	O	B-protein
-mediated	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
,	NN	O	O
AP-1	NN	O	B-protein
induction	NN	O	O
and	NN	O	O
JAK3	NN	O	B-protein
/	NN	O	O
STAT	NN	O	B-protein
activation	NN	O	O
may	NN	O	O
result	NN	O	O
in	NN	O	O
sustained	NN	O	O
proliferation	NN	O	O
leading	NN	O	O
to	NN	O	O
lymphoma	NN	O	B-cell_line
.	NN	O	O

The	NN	O	O
ability	NN	O	O
of	NN	O	O
LMP1	NN	O	B-protein
to	NN	O	O
suppress	NN	O	O
germinal	NN	O	O
center	NN	O	O
formation	NN	O	O
and	NN	O	O
its	NN	O	O
capacity	NN	O	O
to	NN	O	O
mediate	NN	O	O
its	NN	O	O
own	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
shed	NN	O	O
new	NN	O	O
light	NN	O	O
on	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
EBV-associated	NN	O	O
latency	NN	O	O
type	NN	O	O
II	NN	O	O
lymphoproliferations	NN	O	O
like	NN	O	O
Hodgkin	NN	O	O
's	NN	O	O
disease	NN	O	O
and	NN	O	O
angioimmunoblastic	NN	O	O
lymphadenopathy	NN	O	O
.	NN	O	O

The	NN	O	O
carboxy	NN	O	B-protein
terminus	NN	O	I-protein
of	NN	O	O
LMP1	NN	O	B-protein
is	NN	O	O
also	NN	O	O
a	NN	O	O
reliable	NN	O	O
marker	NN	O	O
for	NN	O	O
individual	NN	O	O
EBV	NN	O	O
strain	NN	O	O
identification	NN	O	O
and	NN	O	O
thus	NN	O	O
offers	NN	O	O
new	NN	O	O
possibilities	NN	O	O
in	NN	O	O
tracing	NN	O	O
the	NN	O	O
molecular	NN	O	O
events	NN	O	O
leading	NN	O	O
to	NN	O	O
posttransplant	NN	O	O
lymphoproliferative	NN	O	O
disorders	NN	O	O
(	NN	O	O
PTLDs	NN	O	O
)	NN	O	O
.	NN	O	O

Cytotoxic	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
directed	NN	O	O
against	NN	O	O
well-characterized	NN	O	O
epitopes	NN	O	O
of	NN	O	O
EBV	NN	O	B-DNA
latency	NN	O	I-DNA
genes	NN	O	I-DNA
represent	NN	O	O
an	NN	O	O
already	NN	O	O
successful	NN	O	O
and	NN	O	O
promising	NN	O	O
therapeutic	NN	O	O
approach	NN	O	O
to	NN	O	O
EBV-associated	NN	O	B-cell_line
lymphomas	NN	O	I-cell_line
,	NN	O	O
in	NN	O	O
particular	NN	O	O
PTLDs	NN	O	O

-DOCSTART-	O

Interferon-alpha	NN	O	B-protein
drives	NN	O	O
T	NN	O	O
cell-mediated	NN	O	O
immunopathology	NN	O	O
in	NN	O	O
the	NN	O	O
intestine	NN	O	O
.	NN	O	O

The	NN	O	O
ability	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
-alpha	NN	O	I-protein
to	NN	O	O
induce	NN	O	O
autoimmunity	NN	O	O
and	NN	O	O
exacerbate	NN	O	O
Th1	NN	O	O
diseases	NN	O	O
is	NN	O	O
well	NN	O	O
known	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
recently	NN	O	O
described	NN	O	O
enhanced	NN	O	O
expression	NN	O	O
of	NN	O	O
IFN-alpha	NN	O	B-protein
in	NN	O	O
the	NN	O	O
mucosa	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
celiac	NN	O	O
disease	NN	O	O
(	NN	O	O
CD	NN	O	O
)	NN	O	O
,	NN	O	O
a	NN	O	O
gluten-sensitive	NN	O	O
Th1-mediated	NN	O	O
enteropathy	NN	O	O
,	NN	O	O
characterized	NN	O	O
by	NN	O	O
villous	NN	O	O
atrophy	NN	O	O
and	NN	O	O
crypt	NN	O	O
cell	NN	O	O
hyperplasia	NN	O	O
.	NN	O	O

Previous	NN	O	O
studies	NN	O	O
from	NN	O	O
this	NN	O	O
laboratory	NN	O	O
have	NN	O	O
shown	NN	O	O
that	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
activation	NN	O	O
in	NN	O	O
explant	NN	O	O
cultures	NN	O	O
of	NN	O	O
human	NN	O	O
fetal	NN	O	O
gut	NN	O	O
can	NN	O	O
also	NN	O	O
result	NN	O	O
in	NN	O	O
villous	NN	O	O
atrophy	NN	O	O
and	NN	O	O
crypt	NN	O	O
cell	NN	O	O
hyperplasia	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
,	NN	O	O
therefore	NN	O	O
,	NN	O	O
examined	NN	O	O
changes	NN	O	O
that	NN	O	O
take	NN	O	O
place	NN	O	O
in	NN	O	O
explant	NN	O	O
cultures	NN	O	O
of	NN	O	O
human	NN	O	O
fetal	NN	O	O
gut	NN	O	O
after	NN	O	O
activation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
anti-CD3	NN	O	B-protein
and/or	NN	O	O
IFN-alpha	NN	O	B-protein
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	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
anti-CD3	NN	O	B-protein
alone	NN	O	O
elicits	NN	O	O
a	NN	O	O
small	NN	O	O
IFN-gamma	NN	O	B-protein
and	NN	O	O
TNF-alpha	NN	O	B-protein
response	NN	O	O
with	NN	O	O
no	NN	O	O
tissue	NN	O	O
injury	NN	O	O
.	NN	O	O

Similarly	NN	O	O
,	NN	O	O
no	NN	O	O
changes	NN	O	O
are	NN	O	O
seen	NN	O	O
in	NN	O	O
explants	NN	O	O
cultured	NN	O	O
with	NN	O	O
IFN-alpha	NN	O	B-protein
alone	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
addition	NN	O	O
of	NN	O	O
IFN-alpha	NN	O	B-protein
with	NN	O	O
anti-CD3	NN	O	B-protein
results	NN	O	O
in	NN	O	O
enhanced	NN	O	O
Th1	NN	O	O
response	NN	O	O
and	NN	O	O
crypt	NN	O	O
cell	NN	O	O
hyperplasia	NN	O	O
.	NN	O	O

This	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
enhanced	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
STAT1	NN	O	B-protein
,	NN	O	O
STAT3	NN	O	B-protein
,	NN	O	O
and	NN	O	O
Fyn	NN	O	B-protein
,	NN	O	I-protein
a	NN	O	I-protein
Src	NN	O	I-protein
homology	NN	O	I-protein
tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
,	NN	O	O
which	NN	O	O
interacts	NN	O	O
with	NN	O	O
both	NN	O	O
TCR	NN	O	O
and	NN	O	O
IFN-alpha	NN	O	O
signal	NN	O	O
components	NN	O	O
.	NN	O	O

Together	NN	O	O
these	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
IFN-alpha	NN	O	B-protein
can	NN	O	O
facilitate	NN	O	O
activation	NN	O	O
of	NN	O	O
Th1-reactive	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
gut	NN	O	O
and	NN	O	O
drive	NN	O	O
immunopathology	NN	O	O
.	NN	O	O

-DOCSTART-	O

Suppression	NN	O	O
of	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
production	NN	O	O
by	NN	O	O
cAMP	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
:	NN	O	O
dissociation	NN	O	O
with	NN	O	O
mRNA	NN	O	O
level	NN	O	O
and	NN	O	O
independent	NN	O	O
of	NN	O	O
interleukin-10	NN	O	B-protein
.	NN	O	O

BACKGROUND	NN	O	O
:	NN	O	O
Elevation	NN	O	O
of	NN	O	O
cellular	NN	O	O
cAMP	NN	O	O
inhibits	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
-stimulated	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
production	NN	O	O
and	NN	O	O
increases	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
interleukin	NN	O	B-protein
(	NN	O	I-protein
IL	NN	O	I-protein
)	NN	O	I-protein
-10	NN	O	I-protein
in	NN	O	O
mononuclear	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

TNF-alpha	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
obligates	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappaB	NN	O	I-protein
(	NN	O	O
NF-kappaB	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Exogenous	NN	O	O
IL-10	NN	O	B-protein
inhibits	NN	O	O
NF-kappaB	NN	O	B-protein
in	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
thus	NN	O	O
attenuates	NN	O	O
TNF-alpha	NN	O	B-protein
production	NN	O	O
.	NN	O	O

We	NN	O	O
examined	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
endogenous	NN	O	O
IL-10	NN	O	B-protein
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
and	NN	O	O
TNF-alpha	NN	O	B-protein
production	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
by	NN	O	O
cAMP	NN	O	O
.	NN	O	O

METHODS	NN	O	O
:	NN	O	O
Human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
were	NN	O	O
stimulated	NN	O	O
with	NN	O	O
Escherichia	NN	O	O
coli	NN	O	O
LPS	NN	O	O
(	NN	O	O
100	NN	O	O
ng/ml	NN	O	O
)	NN	O	O
with	NN	O	O
and	NN	O	O
without	NN	O	O
forskolin	NN	O	O
(	NN	O	O
FSK	NN	O	O
,	NN	O	O
50	NN	O	O
microM	NN	O	O
)	NN	O	O
or	NN	O	O
dibutyryl	NN	O	O
cyclic	NN	O	O
AMP	NN	O	O
(	NN	O	O
dbcAMP	NN	O	O
,	NN	O	O
100	NN	O	O
microM	NN	O	O
)	NN	O	O
.	NN	O	O

Cytokine	NN	O	B-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
and	NN	O	O
IL-10	NN	O	B-protein
)	NN	O	O
release	NN	O	O
was	NN	O	O
measured	NN	O	O
by	NN	O	O
immunoassay	NN	O	O
.	NN	O	O

TNF-alpha	NN	O	B-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
measured	NN	O	O
by	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
and	NN	O	O
NF-kappaB	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
was	NN	O	O
assessed	NN	O	O
by	NN	O	O
gel	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
cAMP-elevating	NN	O	O
agents	NN	O	O
inhibited	NN	O	O
LPS-stimulated	NN	O	O
TNF-alpha	NN	O	B-protein
release	NN	O	O
(	NN	O	O
0.77	NN	O	O
+/-	NN	O	O
0.13	NN	O	O
ng/10	NN	O	O
(	NN	O	O
6	NN	O	O
)	NN	O	O
cells	NN	O	O
in	NN	O	O
LPS	NN	O	O
+	NN	O	O
dbcAMP	NN	O	O
and	NN	O	O
0.68	NN	O	O
+/-	NN	O	O
0.19	NN	O	O
ng/10	NN	O	O
(	NN	O	O
6	NN	O	O
)	NN	O	O
cells	NN	O	O
in	NN	O	O
LPS	NN	O	O
+	NN	O	O
FSK	NN	O	O
,	NN	O	O
both	NN	O	O
P	NN	O	O
<	NN	O	O
0.05	NN	O	O
vs	NN	O	O
1.61	NN	O	O
+/-	NN	O	O
0.34	NN	O	O
ng/10	NN	O	O
(	NN	O	O
6	NN	O	O
)	NN	O	O
cells	NN	O	O
in	NN	O	O
LPS	NN	O	O
alone	NN	O	O
)	NN	O	O
.	NN	O	O

Conversely	NN	O	O
,	NN	O	O
cAMP	NN	O	O
enhanced	NN	O	O
LPS-stimulated	NN	O	O
IL-10	NN	O	B-protein
release	NN	O	O
(	NN	O	O
100	NN	O	O
+/-	NN	O	O
21.5	NN	O	O
pg/10	NN	O	O
(	NN	O	O
6	NN	O	O
)	NN	O	O
cells	NN	O	O
in	NN	O	O
LPS	NN	O	O
+	NN	O	O
dbcAMP	NN	O	O
and	NN	O	O
110	NN	O	O
+/-	NN	O	O
25.2	NN	O	O
pg/10	NN	O	O
(	NN	O	O
6	NN	O	O
)	NN	O	O
cells	NN	O	O
in	NN	O	O
LPS	NN	O	O
+	NN	O	O
FSK	NN	O	O
,	NN	O	O
both	NN	O	O
P	NN	O	O
<	NN	O	O
0.05	NN	O	O
vs	NN	O	O
53.3	NN	O	O
+/-	NN	O	O
12.8	NN	O	O
pg/10	NN	O	O
(	NN	O	O
6	NN	O	O
)	NN	O	O
cells	NN	O	O
in	NN	O	O
LPS	NN	O	O
alone	NN	O	O
)	NN	O	O
.	NN	O	O

Neither	NN	O	O
TNF-alpha	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
nor	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
stimulated	NN	O	O
by	NN	O	O
LPS	NN	O	O
was	NN	O	O
inhibited	NN	O	O
by	NN	O	O
the	NN	O	O
cAMP-elevating	NN	O	O
agents	NN	O	O
.	NN	O	O

Neutralization	NN	O	O
of	NN	O	O
IL-10	NN	O	B-protein
with	NN	O	O
a	NN	O	O
specific	NN	O	O
antibody	NN	O	B-protein
did	NN	O	O
not	NN	O	O
attenuate	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
cAMP-elevating	NN	O	O
agents	NN	O	O
on	NN	O	O
TNF-alpha	NN	O	B-protein
production	NN	O	O
.	NN	O	O

CONCLUSION	NN	O	O
:	NN	O	O
The	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
cAMP	NN	O	O
inhibits	NN	O	O
LPS-stimulated	NN	O	O
TNF-alpha	NN	O	B-protein
production	NN	O	O
through	NN	O	O
a	NN	O	O
posttranscriptional	NN	O	O
mechanism	NN	O	O
that	NN	O	O
is	NN	O	O
independent	NN	O	O
of	NN	O	O
endogenous	NN	O	O
IL-10	NN	O	B-protein
.	NN	O	O

Copyright	NN	O	O
2001	NN	O	O
Academic	NN	O	O
Press	NN	O	O
.	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
of	NN	O	O
the	NN	O	O
p21	NN	O	B-DNA
(	NN	O	I-DNA
CIP1/WAF1	NN	O	I-DNA
)	NN	O	I-DNA
promoter	NN	O	I-DNA
by	NN	O	O
bone	NN	O	B-protein
morphogenetic	NN	O	I-protein
protein-2	NN	O	I-protein
in	NN	O	O
mouse	NN	O	O
B	NN	O	B-cell_line
lineage	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

BMPs	NN	O	B-protein
exert	NN	O	O
a	NN	O	O
negative	NN	O	O
growth	NN	O	O
effect	NN	O	O
on	NN	O	O
various	NN	O	O
types	NN	O	O
of	NN	O	O
cells	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
BMP-2	NN	O	B-protein
inhibited	NN	O	O
the	NN	O	O
growth	NN	O	O
of	NN	O	O
HS-72	NN	O	B-cell_line
mouse	NN	O	I-cell_line
hybridoma	NN	O	I-cell_line
cells	NN	O	I-cell_line
by	NN	O	O
inducing	NN	O	O
p21	NN	O	O
(	NN	O	O
CIP1/WAF1	NN	O	O
)	NN	O	O
expression	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
BMP-2	NN	O	B-protein
activated	NN	O	O
the	NN	O	O
mouse	NN	O	O
p21	NN	O	B-DNA
(	NN	O	I-DNA
CIP1/WAF1	NN	O	I-DNA
)	NN	O	I-DNA
promoter	NN	O	I-DNA
in	NN	O	O
HS-72	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
and	NN	O	O
that	NN	O	O
a	NN	O	O
29-base	NN	O	B-DNA
pair	NN	O	I-DNA
(	NN	O	I-DNA
b	NN	O	I-DNA
)	NN	O	I-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
promoter	NN	O	O
(	NN	O	O
-1928/-1900	NN	O	O
relative	NN	O	O
to	NN	O	O
the	NN	O	O
TATA	NN	O	B-DNA
box	NN	O	I-DNA
)	NN	O	O
,	NN	O	O
conserved	NN	O	O
between	NN	O	O
mice	NN	O	O
and	NN	O	O
humans	NN	O	O
,	NN	O	O
was	NN	O	O
responsive	NN	O	O
to	NN	O	O
BMP-2	NN	O	B-protein
as	NN	O	O
well	NN	O	O
as	NN	O	O
expression	NN	O	O
of	NN	O	O
Smad1	NN	O	B-protein
,	NN	O	O
Smad4	NN	O	B-protein
,	NN	O	O
and	NN	O	O
constitutively	NN	O	O
active	NN	O	O
mutants	NN	O	O
of	NN	O	O
BMP	NN	O	B-protein
type	NN	O	I-protein
I	NN	O	I-protein
receptors	NN	O	I-protein
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
an	NN	O	O
oligonucleotide	NN	O	O
containing	NN	O	O
the	NN	O	O
29-b	NN	O	B-DNA
region	NN	O	I-DNA
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
associated	NN	O	O
with	NN	O	O
Smad4	NN	O	B-protein
and	NN	O	O
phosphorylated	NN	O	B-protein
Smad1	NN	O	I-protein
in	NN	O	O
the	NN	O	O
nuclear	NN	O	O
extract	NN	O	O
of	NN	O	O
BMP-2	NN	O	B-protein
-stimulated	NN	O	O
HS-72	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
BMP-2	NN	O	B-protein
might	NN	O	O
activate	NN	O	O
p21	NN	O	O
(	NN	O	O
CIP1/WAF1	NN	O	O
)	NN	O	O
transcription	NN	O	O
by	NN	O	O
inducing	NN	O	O
a	NN	O	O
binding	NN	O	O
of	NN	O	O
Smad4	NN	O	B-protein
and	NN	O	O
Smad1	NN	O	B-protein
to	NN	O	O
the	NN	O	O
29-b	NN	O	B-DNA
region	NN	O	I-DNA
in	NN	O	O
HS-72	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

Dendritic	NN	O	B-cell_type
cell	NN	O	I-cell_type
development	NN	O	O
from	NN	O	O
common	NN	O	O
myeloid	NN	O	B-cell_type
progenitors	NN	O	I-cell_type
.	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
are	NN	O	O
professional	NN	O	O
antigen-presenting	NN	O	B-cell_type
cells	NN	O	I-cell_type
which	NN	O	O
both	NN	O	O
initiate	NN	O	O
adaptive	NN	O	O
immune	NN	O	O
responses	NN	O	O
and	NN	O	O
control	NN	O	O
tolerance	NN	O	O
to	NN	O	O
self-antigens	NN	O	B-protein
.	NN	O	O

It	NN	O	O
has	NN	O	O
been	NN	O	O
suggested	NN	O	O
that	NN	O	O
these	NN	O	O
different	NN	O	O
effects	NN	O	O
on	NN	O	O
responder	NN	O	O
cells	NN	O	O
depend	NN	O	O
on	NN	O	O
subsets	NN	O	O
of	NN	O	O
DCs	NN	O	B-cell_type
arising	NN	O	O
from	NN	O	O
either	NN	O	O
myeloid	NN	O	O
or	NN	O	O
lymphoid	NN	O	O
hematopoietic	NN	O	O
origins	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
model	NN	O	O
,	NN	O	O
CD8	NN	O	B-cell_type
alpha+	NN	O	I-cell_type
Mac-1-	NN	O	I-cell_type
DCs	NN	O	I-cell_type
are	NN	O	O
supposed	NN	O	O
to	NN	O	O
be	NN	O	O
of	NN	O	O
lymphoid	NN	O	O
while	NN	O	O
CD8	NN	O	B-cell_type
alpha-	NN	O	I-cell_type
Mac-1+	NN	O	I-cell_type
DCs	NN	O	I-cell_type
are	NN	O	O
supposed	NN	O	O
to	NN	O	O
be	NN	O	O
of	NN	O	O
myeloid	NN	O	O
origin	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
summarize	NN	O	O
our	NN	O	O
findings	NN	O	O
that	NN	O	O
both	NN	O	O
CD8	NN	O	B-cell_type
alpha+	NN	O	I-cell_type
and	NN	O	I-cell_type
CD8	NN	O	I-cell_type
alpha-	NN	O	I-cell_type
DCs	NN	O	I-cell_type
can	NN	O	O
arise	NN	O	O
from	NN	O	O
clonogenic	NN	O	B-cell_type
common	NN	O	I-cell_type
myeloid	NN	O	I-cell_type
progenitors	NN	O	I-cell_type
(	NN	O	O
CMPs	NN	O	B-cell_type
)	NN	O	O
in	NN	O	O
both	NN	O	O
thymus	NN	O	O
and	NN	O	O
spleen	NN	O	O
.	NN	O	O

Therefore	NN	O	O
CD8	NN	O	B-protein
alpha	NN	O	I-protein
expression	NN	O	O
DCs	NN	O	B-cell_type
does	NN	O	O
not	NN	O	O
indicate	NN	O	O
a	NN	O	O
lymphoid	NN	O	O
origin	NN	O	O
and	NN	O	O
differences	NN	O	O
among	NN	O	O
CD8	NN	O	B-cell_type
alpha+	NN	O	I-cell_type
and	NN	O	I-cell_type
CD8	NN	O	I-cell_type
alpha-	NN	O	I-cell_type
DCs	NN	O	I-cell_type
might	NN	O	O
rather	NN	O	O
reflect	NN	O	O
maturation	NN	O	O
status	NN	O	O
than	NN	O	O
ontogeny	NN	O	O
.	NN	O	O

On	NN	O	O
the	NN	O	O
basis	NN	O	O
of	NN	O	O
transplantation	NN	O	O
studies	NN	O	O
,	NN	O	O
it	NN	O	O
seems	NN	O	O
likely	NN	O	O
that	NN	O	O
most	NN	O	O
of	NN	O	O
the	NN	O	O
DCs	NN	O	B-cell_type
in	NN	O	O
secondary	NN	O	O
lymphoid	NN	O	O
organs	NN	O	O
and	NN	O	O
a	NN	O	O
substantial	NN	O	O
fraction	NN	O	O
of	NN	O	O
thymic	NN	O	B-cell_type
DCs	NN	O	I-cell_type
are	NN	O	O
myeloid-derived	NN	O	O
.	NN	O	O

-DOCSTART-	O

Macrophage	NN	O	O
stimulation	NN	O	O
with	NN	O	O
Murabutide	NN	O	O
,	NN	O	O
an	NN	O	O
HIV-suppressive	NN	O	O
muramyl	NN	O	O
peptide	NN	O	O
derivative	NN	O	O
,	NN	O	O
selectively	NN	O	O
activates	NN	O	O
extracellular	NN	O	B-protein
signal-regulated	NN	O	I-protein
kinases	NN	O	I-protein
1	NN	O	I-protein
and	NN	O	I-protein
2	NN	O	I-protein
,	NN	O	O
C/EBPbeta	NN	O	B-protein
and	NN	O	O
STAT1	NN	O	B-protein
:	NN	O	O
role	NN	O	O
of	NN	O	O
CD14	NN	O	B-protein
and	NN	O	O
Toll-like	NN	O	B-protein
receptors	NN	O	I-protein
2	NN	O	I-protein
and	NN	O	I-protein
4	NN	O	I-protein
.	NN	O	O

The	NN	O	O
smallest	NN	O	O
unit	NN	O	O
of	NN	O	O
bacterial	NN	O	O
peptidoglycans	NN	O	O
known	NN	O	O
to	NN	O	O
be	NN	O	O
endowed	NN	O	O
with	NN	O	O
biological	NN	O	O
activities	NN	O	O
is	NN	O	O
muramyl	NN	O	O
dipeptide	NN	O	O
(	NN	O	O
MDP	NN	O	O
)	NN	O	O
.	NN	O	O

A	NN	O	O
clinically	NN	O	O
acceptable	NN	O	O
synthetic	NN	O	O
derivative	NN	O	O
of	NN	O	O
MDP	NN	O	O
,	NN	O	O
namely	NN	O	O
murabutide	NN	O	O
(	NN	O	O
MB	NN	O	O
)	NN	O	O
,	NN	O	O
has	NN	O	O
been	NN	O	O
found	NN	O	O
to	NN	O	O
present	NN	O	O
interesting	NN	O	O
pharmacological	NN	O	O
properties	NN	O	O
and	NN	O	O
to	NN	O	O
suppress	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
in	NN	O	O
monocyte-derived	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
(	NN	O	O
MDM	NN	O	B-cell_type
)	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
addressed	NN	O	O
the	NN	O	O
signaling	NN	O	O
events	NN	O	O
activated	NN	O	O
in	NN	O	O
MDM	NN	O	B-cell_type
following	NN	O	O
stimulation	NN	O	O
with	NN	O	O
either	NN	O	O
MB	NN	O	O
or	NN	O	O
the	NN	O	O
potent	NN	O	O
immunostimulant	NN	O	O
LPS	NN	O	O
.	NN	O	O

We	NN	O	O
also	NN	O	O
examined	NN	O	O
whether	NN	O	O
signaling	NN	O	O
by	NN	O	O
muramyl	NN	O	O
peptides	NN	O	O
involves	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
cell	NN	O	B-protein
surface	NN	O	I-protein
receptors	NN	O	I-protein
,	NN	O	O
including	NN	O	O
CD14	NN	O	B-protein
and	NN	O	O
Toll-like	NN	O	B-protein
receptor	NN	O	I-protein
2	NN	O	I-protein
(	NN	O	O
TLR2	NN	O	B-protein
)	NN	O	O
or	NN	O	O
TLR4	NN	O	B-protein
that	NN	O	O
are	NN	O	O
known	NN	O	O
to	NN	O	O
be	NN	O	O
signal-transducing	NN	O	B-protein
receptors	NN	O	I-protein
for	NN	O	O
other	NN	O	O
bacterial	NN	O	O
cell	NN	O	O
wall	NN	O	O
components	NN	O	O
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
,	NN	O	O
unlike	NN	O	O
LPS	NN	O	O
,	NN	O	O
the	NN	O	O
safe	NN	O	O
immunomodulator	NN	O	O
MB	NN	O	O
selectively	NN	O	O
activates	NN	O	O
extracellular	NN	O	B-protein
signal-regulated	NN	O	I-protein
kinases	NN	O	I-protein
(	NN	O	I-protein
Erk	NN	O	I-protein
)	NN	O	I-protein
1/2	NN	O	I-protein
,	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
detectable	NN	O	O
Jun	NN	O	B-protein
N-terminal	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	O
JNK	NN	O	B-protein
)	NN	O	O
or	NN	O	O
p38	NN	O	B-protein
mitogen-activated	NN	O	I-protein
kinase	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
STAT1	NN	O	B-protein
activation	NN	O	O
but	NN	O	O
weak	NN	O	O
or	NN	O	O
no	NN	O	O
activation	NN	O	O
of	NN	O	O
STAT3	NN	O	B-protein
or	NN	O	O
STAT5	NN	O	B-protein
respectively	NN	O	O
,	NN	O	O
could	NN	O	O
be	NN	O	O
detected	NN	O	O
in	NN	O	O
MB-stimulated	NN	O	B-cell_type
MDM	NN	O	I-cell_type
.	NN	O	O

Using	NN	O	O
MonoMac6	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
we	NN	O	O
observed	NN	O	O
high	NN	O	O
C/EBPbeta	NN	O	B-protein
and	NN	O	O
AP-1	NN	O	B-protein
but	NN	O	O
weaker	NN	O	O
and	NN	O	O
transient	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
by	NN	O	O
MB	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
the	NN	O	O
truncated	NN	O	O
form	NN	O	O
of	NN	O	O
C/EBPbeta	NN	O	B-protein
,	NN	O	O
known	NN	O	O
to	NN	O	O
repress	NN	O	O
HIV-1	NN	O	O
transcription	NN	O	O
,	NN	O	O
was	NN	O	O
detected	NN	O	O
in	NN	O	O
extracts	NN	O	O
from	NN	O	O
MB-treated	NN	O	B-cell_line
THP-1	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Surprisingly	NN	O	O
,	NN	O	O
neither	NN	O	O
MB	NN	O	O
nor	NN	O	O
MDP	NN	O	O
were	NN	O	O
able	NN	O	O
to	NN	O	O
transduce	NN	O	O
signals	NN	O	O
via	NN	O	O
CD14	NN	O	B-protein
and	NN	O	O
TLR2	NN	O	B-protein
or	NN	O	I-protein
4	NN	O	I-protein
.	NN	O	O

These	NN	O	O
findings	NN	O	O
present	NN	O	O
major	NN	O	O
differences	NN	O	O
in	NN	O	O
the	NN	O	O
early	NN	O	O
cell	NN	O	O
activation	NN	O	O
process	NN	O	O
between	NN	O	O
LPS	NN	O	O
and	NN	O	O
muramyl	NN	O	O
peptides	NN	O	O
,	NN	O	O
and	NN	O	O
strongly	NN	O	O
argue	NN	O	O
for	NN	O	O
the	NN	O	O
implication	NN	O	O
of	NN	O	O
co-receptors	NN	O	B-protein
other	NN	O	O
than	NN	O	O
TLR2	NN	O	B-protein
and	NN	O	O
TLR4	NN	O	B-protein
in	NN	O	O
mediating	NN	O	O
the	NN	O	O
signaling	NN	O	O
events	NN	O	O
induced	NN	O	O
by	NN	O	O
defined	NN	O	O
subunits	NN	O	O
of	NN	O	O
bacterial	NN	O	O
peptidoglycans	NN	O	O
.	NN	O	O

-DOCSTART-	O

Nuclear	NN	O	B-protein
peroxisome	NN	O	I-protein
proliferator-activated	NN	O	I-protein
receptors	NN	O	I-protein
alpha	NN	O	I-protein
and	NN	O	I-protein
gamma	NN	O	I-protein
have	NN	O	O
opposing	NN	O	O
effects	NN	O	O
on	NN	O	O
monocyte	NN	O	O
chemotaxis	NN	O	O
in	NN	O	O
endometriosis	NN	O	O
.	NN	O	O

The	NN	O	O
peroxisome	NN	O	B-protein
proliferator-activated	NN	O	I-protein
receptors	NN	O	I-protein
(	NN	O	I-protein
PPARs	NN	O	I-protein
)	NN	O	I-protein
alpha	NN	O	I-protein
and	NN	O	I-protein
gamma	NN	O	I-protein
are	NN	O	O
nuclear	NN	O	B-protein
receptors	NN	O	I-protein
that	NN	O	O
play	NN	O	O
important	NN	O	O
roles	NN	O	O
in	NN	O	O
inflammatory	NN	O	O
diseases	NN	O	O
like	NN	O	O
ulcerative	NN	O	O
colitis	NN	O	O
and	NN	O	O
arthritis	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
examined	NN	O	O
the	NN	O	O
possible	NN	O	O
role	NN	O	O
of	NN	O	O
PPARs	NN	O	B-protein
in	NN	O	O
macrophage	NN	O	O
attraction	NN	O	O
into	NN	O	O
the	NN	O	O
peritoneal	NN	O	O
cavity	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
endometriosis	NN	O	O
.	NN	O	O

We	NN	O	O
identified	NN	O	O
PPAR-alpha	NN	O	B-RNA
and	NN	O	I-RNA
-gamma	NN	O	I-RNA
messenger	NN	O	O
RNA	NN	O	O
by	NN	O	O
RT-PCR	NN	O	O
and	NN	O	O
protein	NN	O	O
by	NN	O	O
immunoblotting	NN	O	O
of	NN	O	O
lysates	NN	O	O
of	NN	O	O
peritoneal	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
and	NN	O	O
monocytic	NN	O	B-cell_line
U937	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Using	NN	O	O
immunocytochemistry	NN	O	O
,	NN	O	O
we	NN	O	O
localized	NN	O	O
PPAR-alpha	NN	O	B-protein
and	NN	O	I-protein
-gamma	NN	O	I-protein
within	NN	O	O
the	NN	O	O
nuclei	NN	O	O
of	NN	O	O
both	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

Monocyte	NN	O	O
chemotactic	NN	O	O
activity	NN	O	O
of	NN	O	O
peritoneal	NN	O	O
fluid	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
endometriosis	NN	O	O
was	NN	O	O
quantified	NN	O	O
in	NN	O	O
Boyden	NN	O	O
chambers	NN	O	O
.	NN	O	O

Migration	NN	O	O
of	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
was	NN	O	O
increased	NN	O	O
by	NN	O	O
WY	NN	O	O
14643	NN	O	O
and	NN	O	O
reduced	NN	O	O
by	NN	O	O
rosiglitazone	NN	O	O
.	NN	O	O

Peritoneal	NN	O	O
fluid	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
endometriosis	NN	O	O
activated	NN	O	O
U937	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
PPAR-alpha/GAL4	NN	O	B-DNA
luciferase	NN	O	I-DNA
reporter	NN	O	I-DNA
.	NN	O	O

By	NN	O	O
contrast	NN	O	O
,	NN	O	O
peritoneal	NN	O	O
fluid	NN	O	O
did	NN	O	O
not	NN	O	O
cause	NN	O	O
significant	NN	O	O
activation	NN	O	O
of	NN	O	O
PPAR-gamma/GAL4	NN	O	B-DNA
constructs	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
U937	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
PPAR	NN	O	B-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
luciferase	NN	O	I-DNA
reporter	NN	O	I-DNA
showed	NN	O	O
disease	NN	O	O
stage-dependent	NN	O	O
up-regulation	NN	O	O
when	NN	O	O
treated	NN	O	O
with	NN	O	O
peritoneal	NN	O	O
fluid	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
endometriosis	NN	O	O
.	NN	O	O

Treatment	NN	O	O
with	NN	O	O
peritoneal	NN	O	O
fluid	NN	O	O
from	NN	O	O
healthy	NN	O	O
controls	NN	O	O
down-regulated	NN	O	O
PPAR	NN	O	O
response	NN	O	O
element	NN	O	O
transactivation	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
peritoneal	NN	O	O
fluid	NN	O	O
of	NN	O	O
endometriosis	NN	O	O
patients	NN	O	O
contains	NN	O	O
activators	NN	O	O
of	NN	O	O
PPAR-alpha	NN	O	B-protein
that	NN	O	O
stimulate	NN	O	O
macrophage	NN	O	O
chemotaxis	NN	O	O
.	NN	O	O

Inhibitors	NN	O	O
of	NN	O	O
PPAR-alpha	NN	O	B-protein
or	NN	O	O
activators	NN	O	O
of	NN	O	O
PPAR-gamma	NN	O	B-protein
could	NN	O	O
be	NN	O	O
developed	NN	O	O
for	NN	O	O
the	NN	O	O
treatment	NN	O	O
of	NN	O	O
inflammation	NN	O	O
associated	NN	O	O
with	NN	O	O
endometriosis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Long-term-impaired	NN	O	O
expression	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
and	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	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
of	NN	O	O
trauma	NN	O	O
patients	NN	O	O
.	NN	O	O

Nuclear	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	I-protein
NF	NN	O	I-protein
)	NN	O	I-protein
-kappa	NN	O	I-protein
B	NN	O	I-protein
expression	NN	O	O
and	NN	O	O
dimer	NN	O	O
characteristics	NN	O	O
were	NN	O	O
studied	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
PBMCs	NN	O	B-cell_type
)	NN	O	O
of	NN	O	O
major-trauma	NN	O	O
patients	NN	O	O
and	NN	O	O
healthy	NN	O	O
controls	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
PBMCs	NN	O	B-cell_type
on	NN	O	O
days	NN	O	O
1	NN	O	O
,	NN	O	O
3	NN	O	O
,	NN	O	O
5	NN	O	O
,	NN	O	O
and	NN	O	O
10	NN	O	O
after	NN	O	O
trauma	NN	O	O
revealed	NN	O	O
that	NN	O	O
expression	NN	O	O
of	NN	O	O
both	NN	O	O
p65p50	NN	O	B-protein
heterodimers	NN	O	I-protein
and	NN	O	O
p50p50	NN	O	B-protein
homodimers	NN	O	I-protein
was	NN	O	O
significantly	NN	O	O
reduced	NN	O	O
compared	NN	O	O
with	NN	O	O
that	NN	O	O
in	NN	O	O
controls	NN	O	O
.	NN	O	O

In	NN	O	O
vitro	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
stimulation	NN	O	O
of	NN	O	O
PBMCs	NN	O	B-cell_type
induced	NN	O	O
NF-kappa	NN	O	O
B	NN	O	O
translocation	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
throughout	NN	O	O
the	NN	O	O
survey	NN	O	O
,	NN	O	O
p65p50	NN	O	O
activation	NN	O	O
remained	NN	O	O
significantly	NN	O	O
lower	NN	O	O
in	NN	O	O
trauma	NN	O	O
patients	NN	O	O
than	NN	O	O
in	NN	O	O
controls	NN	O	O
.	NN	O	O

After	NN	O	O
LPS	NN	O	O
stimulation	NN	O	O
in	NN	O	O
vitro	NN	O	O
,	NN	O	O
the	NN	O	O
p65p50/p50p50	NN	O	O
ratio	NN	O	O
was	NN	O	O
significantly	NN	O	O
lower	NN	O	O
in	NN	O	O
PBMCs	NN	O	B-cell_type
from	NN	O	O
trauma	NN	O	O
patients	NN	O	O
than	NN	O	O
from	NN	O	O
healthy	NN	O	O
controls	NN	O	O
.	NN	O	O

The	NN	O	O
ex	NN	O	O
vivo	NN	O	O
expression	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
was	NN	O	O
higher	NN	O	O
in	NN	O	O
PBMCs	NN	O	B-cell_type
of	NN	O	O
controls	NN	O	O
than	NN	O	O
of	NN	O	O
trauma	NN	O	O
patients	NN	O	O
.	NN	O	O

LPS	NN	O	O
did	NN	O	O
not	NN	O	O
induce	NN	O	O
I	NN	O	O
kappa	NN	O	O
B	NN	O	O
expression	NN	O	O
in	NN	O	O
PBMCs	NN	O	B-cell_type
from	NN	O	O
trauma	NN	O	O
patients	NN	O	O
,	NN	O	O
but	NN	O	O
strong	NN	O	O
induction	NN	O	O
was	NN	O	O
obtained	NN	O	O
with	NN	O	O
staphylococci	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
this	NN	O	O
defect	NN	O	O
is	NN	O	O
not	NN	O	O
universal	NN	O	O
and	NN	O	O
depends	NN	O	O
on	NN	O	O
the	NN	O	O
nature	NN	O	O
of	NN	O	O
the	NN	O	O
activating	NN	O	O
signal	NN	O	O
.	NN	O	O

Although	NN	O	O
no	NN	O	O
direct	NN	O	O
correlation	NN	O	O
was	NN	O	O
found	NN	O	O
between	NN	O	O
levels	NN	O	O
of	NN	O	O
interleukin-10	NN	O	B-protein
or	NN	O	O
transforming	NN	O	B-protein
growth	NN	O	I-protein
factor-beta	NN	O	I-protein
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
these	NN	O	O
immunosuppressive	NN	O	B-protein
cytokines	NN	O	I-protein
were	NN	O	O
significantly	NN	O	O
elevated	NN	O	O
in	NN	O	O
trauma	NN	O	O
patients	NN	O	O
by	NN	O	O
10	NN	O	O
days	NN	O	O
after	NN	O	O
admission	NN	O	O
.	NN	O	O

The	NN	O	O
long-term	NN	O	O
low-basal	NN	O	O
and	NN	O	O
LPS-induced	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
recalled	NN	O	O
long-term	NN	O	O
immunoparalysis	NN	O	O
observed	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
severe	NN	O	O
inflammatory	NN	O	O
stress	NN	O	O
such	NN	O	O
as	NN	O	O
trauma	NN	O	O
.	NN	O	O

-DOCSTART-	O

Hepatic	NN	O	O
ischemia-reperfusion	NN	O	O
injury	NN	O	O
.	NN	O	O

BACKGROUND	NN	O	O
:	NN	O	O
The	NN	O	O
morbidity	NN	O	O
associated	NN	O	O
with	NN	O	O
liver	NN	O	O
transplantation	NN	O	O
and	NN	O	O
major	NN	O	O
hepatic	NN	O	O
resections	NN	O	O
is	NN	O	O
partly	NN	O	O
a	NN	O	O
result	NN	O	O
of	NN	O	O
ischemia-reperfusion	NN	O	O
injury	NN	O	O
.	NN	O	O

DATA	NN	O	O
SOURCES	NN	O	O
:	NN	O	O
The	NN	O	O
entire	NN	O	O
world	NN	O	O
literature	NN	O	O
on	NN	O	O
the	NN	O	O
subject	NN	O	O
was	NN	O	O
searched	NN	O	O
via	NN	O	O
Medline	NN	O	O
.	NN	O	O

Keywords	NN	O	O
included	NN	O	O
reperfusion	NN	O	O
injury	NN	O	O
,	NN	O	O
transplantation	NN	O	O
,	NN	O	O
liver	NN	O	O
resection	NN	O	O
,	NN	O	O
nitric	NN	O	O
oxide	NN	O	O
,	NN	O	O
endothelin	NN	O	O
,	NN	O	O
cytokines	NN	O	B-protein
,	NN	O	O
Kupffer	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
ischemic/ischaemic	NN	O	O
preconditioning	NN	O	O
,	NN	O	O
and	NN	O	O
nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
.	NN	O	O

CONCLUSIONS	NN	O	O
:	NN	O	O
An	NN	O	O
imbalance	NN	O	O
between	NN	O	O
endothelin	NN	O	O
and	NN	O	O
nitric	NN	O	O
oxide	NN	O	O
levels	NN	O	O
results	NN	O	O
in	NN	O	O
failure	NN	O	O
of	NN	O	O
the	NN	O	O
hepatic	NN	O	O
microcirculation	NN	O	O
at	NN	O	O
the	NN	O	O
onset	NN	O	O
of	NN	O	O
reperfusion	NN	O	O
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
in	NN	O	O
the	NN	O	O
liver	NN	O	O
promotes	NN	O	O
proinflammatory	NN	O	B-protein
cytokine	NN	O	I-protein
and	NN	O	O
adhesion	NN	O	B-protein
molecule	NN	O	I-protein
synthesis	NN	O	O
.	NN	O	O

These	NN	O	O
result	NN	O	O
in	NN	O	O
oxygen-derived	NN	O	O
free	NN	O	O
radical	NN	O	O
production	NN	O	O
and	NN	O	O
neutrophil	NN	O	O
recruitment	NN	O	O
,	NN	O	O
further	NN	O	O
contributing	NN	O	O
to	NN	O	O
cellular	NN	O	O
injury	NN	O	O
.	NN	O	O

Various	NN	O	O
therapeutic	NN	O	O
modalities	NN	O	O
acting	NN	O	O
on	NN	O	O
the	NN	O	O
above	NN	O	O
mediators	NN	O	O
have	NN	O	O
been	NN	O	O
successfully	NN	O	O
used	NN	O	O
to	NN	O	O
attenuate	NN	O	O
reperfusion	NN	O	O
injury	NN	O	O
in	NN	O	O
animal	NN	O	O
models	NN	O	O
of	NN	O	O
hepatic	NN	O	O
transplantation	NN	O	O
and	NN	O	O
resection	NN	O	O
.	NN	O	O

Application	NN	O	O
of	NN	O	O
the	NN	O	O
knowledge	NN	O	O
gained	NN	O	O
from	NN	O	O
animal	NN	O	O
models	NN	O	O
of	NN	O	O
hepatic	NN	O	O
ischemia-reperfusion	NN	O	O
to	NN	O	O
the	NN	O	O
clinical	NN	O	O
setting	NN	O	O
will	NN	O	O
improve	NN	O	O
the	NN	O	O
outcome	NN	O	O
of	NN	O	O
hepatic	NN	O	O
surgery	NN	O	O
.	NN	O	O

-DOCSTART-	O

Down-regulation	NN	O	O
of	NN	O	O
IL-12	NN	O	B-DNA
p40	NN	O	I-DNA
gene	NN	O	I-DNA
in	NN	O	O
Plasmodium	NN	O	O
berghei-infected	NN	O	O
mice	NN	O	O
.	NN	O	O

We	NN	O	O
analyzed	NN	O	O
the	NN	O	O
mechanism	NN	O	O
that	NN	O	O
causes	NN	O	O
suppression	NN	O	O
of	NN	O	O
IL-12	NN	O	B-DNA
p40	NN	O	I-DNA
gene	NN	O	I-DNA
induction	NN	O	O
during	NN	O	O
Plasmodium	NN	O	O
berghei	NN	O	O
infection	NN	O	O
.	NN	O	O

Although	NN	O	O
IL-12	NN	O	B-protein
together	NN	O	O
with	NN	O	O
IFN-gamma	NN	O	B-protein
plays	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
protection	NN	O	O
against	NN	O	O
pathogenic	NN	O	O
infection	NN	O	O
,	NN	O	O
the	NN	O	O
IL-12	NN	O	B-protein
p70	NN	O	I-protein
protein	NN	O	I-protein
production	NN	O	O
of	NN	O	O
infected	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
is	NN	O	O
lower	NN	O	O
than	NN	O	O
that	NN	O	O
by	NN	O	O
the	NN	O	O
uninfected	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
showed	NN	O	O
in	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
that	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
IL-12	NN	O	B-DNA
p40	NN	O	I-DNA
gene	NN	O	I-DNA
but	NN	O	O
not	NN	O	O
IL-12	NN	O	B-DNA
p35	NN	O	I-DNA
gene	NN	O	I-DNA
in	NN	O	O
macrophages	NN	O	B-cell_type
of	NN	O	O
P.	NN	O	O
berghei-infected	NN	O	O
mice	NN	O	O
was	NN	O	O
profoundly	NN	O	O
inhibited	NN	O	O
.	NN	O	O

The	NN	O	O
inhibition	NN	O	O
was	NN	O	O
induced	NN	O	O
by	NN	O	O
interaction	NN	O	O
with	NN	O	O
macrophages	NN	O	B-cell_type
that	NN	O	O
had	NN	O	O
contacted	NN	O	O
with	NN	O	O
P.	NN	O	B-cell_type
berghei-infected	NN	O	I-cell_type
erythrocytes	NN	O	I-cell_type
and	NN	O	O
was	NN	O	O
mediated	NN	O	O
by	NN	O	O
a	NN	O	O
soluble	NN	O	O
factor	NN	O	O
,	NN	O	O
IL-10	NN	O	B-protein
.	NN	O	O

There	NN	O	O
was	NN	O	O
comparable	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
in	NN	O	O
uninfected	NN	O	B-cell_type
and	NN	O	I-cell_type
infected	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
induction	NN	O	O
of	NN	O	O
IFN-regulatory	NN	O	B-DNA
factor-1	NN	O	I-DNA
gene	NN	O	I-DNA
was	NN	O	O
comparable	NN	O	O
in	NN	O	O
transcription	NN	O	O
level	NN	O	O
in	NN	O	O
uninfected	NN	O	B-cell_type
and	NN	O	I-cell_type
infected	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
while	NN	O	O
the	NN	O	O
unidentified	NN	O	O
complex	NN	O	O
formation	NN	O	O
of	NN	O	O
IFN-regulatory	NN	O	B-protein
factor-1	NN	O	I-protein
was	NN	O	O
observed	NN	O	O
in	NN	O	O
infected	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
the	NN	O	O
inhibition	NN	O	O
of	NN	O	O
the	NN	O	O
IL-12	NN	O	B-DNA
p40	NN	O	I-DNA
gene	NN	O	I-DNA
induction	NN	O	O
appeared	NN	O	O
to	NN	O	O
be	NN	O	O
regulated	NN	O	O
at	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
level	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
Friend	NN	O	B-protein
of	NN	O	I-protein
GATA	NN	O	I-protein
proteins	NN	O	I-protein
U-shaped	NN	O	O
,	NN	O	O
FOG-1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
FOG-2	NN	O	B-protein
function	NN	O	O
as	NN	O	O
negative	NN	O	O
regulators	NN	O	O
of	NN	O	O
blood	NN	O	O
,	NN	O	O
heart	NN	O	O
,	NN	O	O
and	NN	O	O
eye	NN	O	O
development	NN	O	O
in	NN	O	O
Drosophila	NN	O	O
.	NN	O	O

Friend	NN	O	B-protein
of	NN	O	I-protein
GATA	NN	O	I-protein
(	NN	O	I-protein
FOG	NN	O	I-protein
)	NN	O	I-protein
proteins	NN	O	I-protein
regulate	NN	O	O
GATA	NN	O	O
factor-activated	NN	O	O
gene	NN	O	O
transcription	NN	O	O
.	NN	O	O

During	NN	O	O
vertebrate	NN	O	O
hematopoiesis	NN	O	O
,	NN	O	O
FOG	NN	O	B-protein
and	NN	O	I-protein
GATA	NN	O	I-protein
proteins	NN	O	I-protein
cooperate	NN	O	O
to	NN	O	O
promote	NN	O	O
erythrocyte	NN	O	O
and	NN	O	O
megakaryocyte	NN	O	O
differentiation	NN	O	O
.	NN	O	O

The	NN	O	O
Drosophila	NN	O	B-protein
FOG	NN	O	I-protein
homologue	NN	O	I-protein
U-shaped	NN	O	I-protein
(	NN	O	O
Ush	NN	O	B-protein
)	NN	O	O
is	NN	O	O
expressed	NN	O	O
similarly	NN	O	O
in	NN	O	O
the	NN	O	O
blood	NN	O	O
cell	NN	O	O
anlage	NN	O	O
during	NN	O	O
embryogenesis	NN	O	O
.	NN	O	O

During	NN	O	O
hematopoiesis	NN	O	O
,	NN	O	O
the	NN	O	O
acute	NN	O	O
myeloid	NN	O	O
leukemia	NN	O	O
1	NN	O	O
homologue	NN	O	O
Lozenge	NN	O	B-cell_type
and	NN	O	I-cell_type
Glial	NN	O	I-cell_type
cells	NN	O	I-cell_type
missing	NN	O	O
are	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
crystal	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
plasmatocytes	NN	O	B-cell_type
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
additional	NN	O	O
factors	NN	O	O
have	NN	O	O
been	NN	O	O
predicted	NN	O	O
to	NN	O	O
control	NN	O	O
crystal	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
Ush	NN	O	B-protein
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
hemocyte	NN	O	B-cell_type
precursors	NN	O	I-cell_type
and	NN	O	O
plasmatocytes	NN	O	B-cell_type
throughout	NN	O	O
embryogenesis	NN	O	O
and	NN	O	O
larval	NN	O	O
development	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
GATA	NN	O	B-protein
factor	NN	O	I-protein
Serpent	NN	O	I-protein
is	NN	O	O
essential	NN	O	O
for	NN	O	O
Ush	NN	O	B-protein
embryonic	NN	O	O
expression	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
loss	NN	O	O
of	NN	O	O
ush	NN	O	O
function	NN	O	O
results	NN	O	O
in	NN	O	O
an	NN	O	O
overproduction	NN	O	O
of	NN	O	O
crystal	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
whereas	NN	O	O
forced	NN	O	O
expression	NN	O	O
of	NN	O	O
Ush	NN	O	B-protein
reduces	NN	O	O
this	NN	O	O
cell	NN	O	O
population	NN	O	O
.	NN	O	O

Murine	NN	O	B-protein
FOG-1	NN	O	I-protein
and	NN	O	I-protein
FOG-2	NN	O	I-protein
also	NN	O	O
can	NN	O	O
repress	NN	O	O
crystal	NN	O	O
cell	NN	O	O
production	NN	O	O
,	NN	O	O
but	NN	O	O
a	NN	O	O
mutant	NN	O	O
version	NN	O	O
of	NN	O	O
FOG-2	NN	O	B-protein
lacking	NN	O	O
a	NN	O	O
conserved	NN	O	O
motif	NN	O	O
that	NN	O	O
binds	NN	O	O
the	NN	O	O
corepressor	NN	O	B-protein
C-terminal	NN	O	I-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
fails	NN	O	O
to	NN	O	O
affect	NN	O	O
the	NN	O	O
cell	NN	O	O
lineage	NN	O	O
.	NN	O	O

The	NN	O	O
GATA	NN	O	B-protein
factor	NN	O	I-protein
Pannier	NN	O	I-protein
(	NN	O	O
Pnr	NN	O	B-protein
)	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
eye	NN	O	O
and	NN	O	O
heart	NN	O	O
development	NN	O	O
in	NN	O	O
Drosophila	NN	O	O
.	NN	O	O

When	NN	O	O
Ush	NN	O	B-protein
,	NN	O	O
FOG-1	NN	O	B-protein
,	NN	O	O
FOG-2	NN	O	B-protein
,	NN	O	O
or	NN	O	O
mutant	NN	O	B-protein
FOG-2	NN	O	I-protein
is	NN	O	O
coexpressed	NN	O	O
with	NN	O	O
Pnr	NN	O	B-protein
during	NN	O	O
these	NN	O	O
developmental	NN	O	O
processes	NN	O	O
,	NN	O	O
severe	NN	O	O
eye	NN	O	O
and	NN	O	O
heart	NN	O	O
phenotypes	NN	O	O
result	NN	O	O
,	NN	O	O
consistent	NN	O	O
with	NN	O	O
a	NN	O	O
conserved	NN	O	O
negative	NN	O	O
regulation	NN	O	O
of	NN	O	O
Pnr	NN	O	B-protein
function	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
fly	NN	O	O
and	NN	O	O
mouse	NN	O	O
FOG	NN	O	B-protein
proteins	NN	O	I-protein
function	NN	O	O
similarly	NN	O	O
in	NN	O	O
three	NN	O	O
distinct	NN	O	O
cellular	NN	O	O
contexts	NN	O	O
in	NN	O	O
Drosophila	NN	O	O
,	NN	O	O
but	NN	O	O
may	NN	O	O
use	NN	O	O
different	NN	O	O
mechanisms	NN	O	O
to	NN	O	O
regulate	NN	O	O
genetic	NN	O	O
events	NN	O	O
in	NN	O	O
blood	NN	O	O
vs	NN	O	O
.	NN	O	O
cardial	NN	O	B-cell_line
or	NN	O	I-cell_line
eye	NN	O	I-cell_line
cell	NN	O	I-cell_line
lineages	NN	O	I-cell_line

-DOCSTART-	O

The	NN	O	O
RING	NN	O	B-protein
finger	NN	O	I-protein
protein	NN	O	I-protein
Siah-1	NN	O	B-protein
regulates	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
the	NN	O	O
transcriptional	NN	O	B-protein
coactivator	NN	O	I-protein
OBF-1	NN	O	B-protein
.	NN	O	O

The	NN	O	O
transcriptional	NN	O	B-protein
coactivator	NN	O	I-protein
OBF-1	NN	O	B-protein
,	NN	O	O
which	NN	O	O
interacts	NN	O	O
with	NN	O	O
Oct-1	NN	O	B-protein
and	NN	O	O
Oct-2	NN	O	B-protein
and	NN	O	O
the	NN	O	O
octamer	NN	O	B-DNA
site	NN	O	I-DNA
DNA	NN	O	I-DNA
,	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
critical	NN	O	O
for	NN	O	O
development	NN	O	O
of	NN	O	O
a	NN	O	O
normal	NN	O	O
immune	NN	O	O
response	NN	O	O
and	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
germinal	NN	O	O
centers	NN	O	O
in	NN	O	O
secondary	NN	O	O
lymphoid	NN	O	O
organs	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
have	NN	O	O
identified	NN	O	O
the	NN	O	O
RING	NN	O	B-protein
finger	NN	O	I-protein
protein	NN	O	I-protein
Siah-1	NN	O	B-protein
as	NN	O	O
a	NN	O	O
protein	NN	O	O
interacting	NN	O	O
specifically	NN	O	O
with	NN	O	O
OBF-1	NN	O	B-protein
.	NN	O	O

This	NN	O	O
interaction	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
C-terminal	NN	O	B-protein
part	NN	O	O
of	NN	O	O
Siah-1	NN	O	B-protein
and	NN	O	O
by	NN	O	O
residues	NN	O	O
in	NN	O	O
the	NN	O	O
N-terminus	NN	O	B-protein
of	NN	O	O
OBF-1	NN	O	B-protein
,	NN	O	O
partly	NN	O	O
distinct	NN	O	O
from	NN	O	O
the	NN	O	O
residues	NN	O	O
required	NN	O	O
for	NN	O	O
formation	NN	O	O
of	NN	O	O
a	NN	O	O
complex	NN	O	O
with	NN	O	O
the	NN	O	O
Oct	NN	O	B-protein
POU	NN	O	I-protein
domains	NN	O	I-protein
and	NN	O	O
the	NN	O	O
DNA	NN	O	O
.	NN	O	O

Interaction	NN	O	O
between	NN	O	O
Siah-1	NN	O	B-protein
and	NN	O	O
OBF-1	NN	O	B-protein
leads	NN	O	O
to	NN	O	O
downregulation	NN	O	O
of	NN	O	O
OBF-1	NN	O	O
protein	NN	O	O
level	NN	O	O
but	NN	O	O
not	NN	O	O
mRNA	NN	O	O
,	NN	O	O
and	NN	O	O
to	NN	O	O
a	NN	O	O
corresponding	NN	O	O
reduction	NN	O	O
in	NN	O	O
octamer	NN	O	O
site-dependent	NN	O	O
transcription	NN	O	O
activation	NN	O	O
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
the	NN	O	O
ubiquitin-proteasome	NN	O	O
pathway	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
leads	NN	O	O
to	NN	O	O
elevated	NN	O	O
levels	NN	O	O
of	NN	O	O
OBF-1	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
in	NN	O	O
immunized	NN	O	O
mice	NN	O	O
,	NN	O	O
OBF-1	NN	O	B-protein
protein	NN	O	I-protein
amounts	NN	O	O
are	NN	O	O
dramatically	NN	O	O
increased	NN	O	O
in	NN	O	O
primary	NN	O	B-cell_type
activated	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
without	NN	O	O
concomitant	NN	O	O
increase	NN	O	O
in	NN	O	O
OBF-1	NN	O	B-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
Siah-1	NN	O	B-protein
is	NN	O	O
part	NN	O	O
of	NN	O	O
a	NN	O	O
novel	NN	O	O
regulatory	NN	O	O
loop	NN	O	O
controlling	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
OBF-1	NN	O	B-protein
protein	NN	O	I-protein
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
beta-catenin	NN	O	O
--	NN	O	O
TCF-1	NN	O	O
pathway	NN	O	O
ensures	NN	O	O
CD4	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
CD8	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
thymocyte	NN	O	I-cell_type
survival	NN	O	O
.	NN	O	O

The	NN	O	O
association	NN	O	O
of	NN	O	O
trans-acting	NN	O	B-protein
T	NN	O	I-protein
cell	NN	O	I-protein
factors	NN	O	I-protein
(	NN	O	O
TCFs	NN	O	B-protein
)	NN	O	O
or	NN	O	O
lymphoid	NN	O	B-protein
enhancer	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
with	NN	O	O
their	NN	O	O
coactivator	NN	O	O
beta-catenin	NN	O	O
mediates	NN	O	O
transient	NN	O	O
transcriptional	NN	O	O
responses	NN	O	O
to	NN	O	O
extracellular	NN	O	O
Wnt	NN	O	O
signals	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
here	NN	O	O
that	NN	O	O
T	NN	O	O
cell	NN	O	O
maturation	NN	O	O
depends	NN	O	O
on	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
the	NN	O	O
beta-catenin	NN	O	B-protein
--	NN	O	I-protein
binding	NN	O	I-protein
domain	NN	O	I-protein
in	NN	O	O
TCF-1	NN	O	B-protein
.	NN	O	O

This	NN	O	O
domain	NN	O	O
is	NN	O	O
necessary	NN	O	O
to	NN	O	O
mediate	NN	O	O
the	NN	O	O
survival	NN	O	O
of	NN	O	O
immature	NN	O	O
CD4	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
CD8	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
double-positive	NN	O	I-cell_type
(	NN	O	I-cell_type
DP	NN	O	I-cell_type
)	NN	O	I-cell_type
thymocytes	NN	O	I-cell_type
.	NN	O	O

Accelerated	NN	O	O
spontaneous	NN	O	O
thymocyte	NN	O	O
death	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
TCF-1	NN	O	B-protein
correlates	NN	O	O
with	NN	O	O
aberrantly	NN	O	O
low	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
anti-apoptotic	NN	O	B-protein
protein	NN	O	I-protein
Bcl-x	NN	O	B-protein
(	NN	O	I-protein
L	NN	O	I-protein
)	NN	O	I-protein
.	NN	O	O

Increasing	NN	O	O
anti-apoptotic	NN	O	O
effectors	NN	O	O
in	NN	O	O
thymocytes	NN	O	B-cell_type
by	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
a	NN	O	O
Bcl-2	NN	O	B-DNA
transgene	NN	O	I-DNA
rescued	NN	O	O
TCF-1-deficient	NN	O	B-cell_type
DP	NN	O	I-cell_type
thymocytes	NN	O	I-cell_type
from	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
TCF-1	NN	O	B-protein
,	NN	O	O
upon	NN	O	O
association	NN	O	O
with	NN	O	O
beta-catenin	NN	O	O
,	NN	O	O
transiently	NN	O	O
ensures	NN	O	O
the	NN	O	O
survival	NN	O	O
of	NN	O	O
immature	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
which	NN	O	O
enables	NN	O	O
them	NN	O	O
to	NN	O	O
generate	NN	O	O
and	NN	O	O
edit	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	I-protein
TCR	NN	O	I-protein
)	NN	O	I-protein
alpha	NN	O	I-protein
chains	NN	O	I-protein
and	NN	O	O
attempt	NN	O	O
TCR	NN	O	B-protein
-mediated	NN	O	O
positive	NN	O	O
selection	NN	O	O
.	NN	O	O

-DOCSTART-	O

TRAIL/Apo2L	NN	O	B-protein
ligand	NN	O	I-protein
selectively	NN	O	O
induces	NN	O	O
apoptosis	NN	O	O
and	NN	O	O
overcomes	NN	O	O
drug	NN	O	O
resistance	NN	O	O
in	NN	O	O
multiple	NN	O	O
myeloma	NN	O	O
:	NN	O	O
therapeutic	NN	O	O
applications	NN	O	O
.	NN	O	O

Multiple	NN	O	O
myeloma	NN	O	O
(	NN	O	O
MM	NN	O	O
)	NN	O	O
remains	NN	O	O
incurable	NN	O	O
and	NN	O	O
novel	NN	O	O
treatments	NN	O	O
are	NN	O	O
urgently	NN	O	O
needed	NN	O	O
.	NN	O	O

Preclinical	NN	O	O
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
in	NN	O	O
vivo	NN	O	O
evaluations	NN	O	O
were	NN	O	O
performed	NN	O	O
to	NN	O	O
assess	NN	O	O
the	NN	O	O
potential	NN	O	O
therapeutic	NN	O	O
applications	NN	O	O
of	NN	O	O
human	NN	O	O
recombinant	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
-related	NN	O	I-protein
apoptosis-inducing	NN	O	I-protein
ligand/Apo2	NN	O	I-protein
ligand	NN	O	I-protein
(	NN	O	O
TRAIL/Apo2L	NN	O	B-protein
)	NN	O	O
in	NN	O	O
MM	NN	O	O
.	NN	O	O

TRAIL/Apo2L	NN	O	B-protein
potently	NN	O	O
induced	NN	O	O
apoptosis	NN	O	O
of	NN	O	O
MM	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
patients	NN	O	O
and	NN	O	O
the	NN	O	O
majority	NN	O	O
of	NN	O	O
MM	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
including	NN	O	O
cells	NN	O	O
sensitive	NN	O	O
or	NN	O	O
resistant	NN	O	O
to	NN	O	O
dexamethasone	NN	O	O
(	NN	O	O
Dex	NN	O	O
)	NN	O	O
,	NN	O	O
doxorubicin	NN	O	O
(	NN	O	O
Dox	NN	O	O
)	NN	O	O
,	NN	O	O
melphalan	NN	O	O
,	NN	O	O
and	NN	O	O
mitoxantrone	NN	O	O
.	NN	O	O

TRAIL/Apo2L	NN	O	B-protein
also	NN	O	O
overcame	NN	O	O
the	NN	O	O
survival	NN	O	O
effect	NN	O	O
of	NN	O	O
interleukin	NN	O	B-protein
6	NN	O	I-protein
on	NN	O	O
MM	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
did	NN	O	O
not	NN	O	O
affect	NN	O	O
the	NN	O	O
survival	NN	O	O
of	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
and	NN	O	I-cell_type
bone	NN	O	I-cell_type
marrow	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
purified	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
healthy	NN	O	O
donors	NN	O	O
.	NN	O	O

The	NN	O	O
status	NN	O	O
of	NN	O	O
the	NN	O	O
TRAIL	NN	O	B-protein
receptors	NN	O	I-protein
(	NN	O	O
assessed	NN	O	O
by	NN	O	O
immunoblotting	NN	O	O
and	NN	O	O
flow	NN	O	O
cytometry	NN	O	O
)	NN	O	O
could	NN	O	O
not	NN	O	O
predict	NN	O	O
TRAIL	NN	O	O
sensitivity	NN	O	O
of	NN	O	O
MM	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
anti-MM	NN	O	O
activity	NN	O	O
of	NN	O	O
TRAIL/Apo2L	NN	O	B-protein
was	NN	O	O
confirmed	NN	O	O
in	NN	O	O
nu/xid/bg	NN	O	O
mice	NN	O	O
xenografted	NN	O	O
with	NN	O	O
human	NN	O	B-cell_type
MM	NN	O	I-cell_type
cells	NN	O	I-cell_type
;	NN	O	O
TRAIL	NN	O	B-protein
(	NN	O	O
500	NN	O	O
microg	NN	O	O
intraperitoneally	NN	O	O
daily	NN	O	O
for	NN	O	O
14	NN	O	O
days	NN	O	O
)	NN	O	O
was	NN	O	O
well	NN	O	O
tolerated	NN	O	O
and	NN	O	O
significantly	NN	O	O
suppressed	NN	O	O
the	NN	O	O
growth	NN	O	O
of	NN	O	O
plasmacytomas	NN	O	B-cell_type
.	NN	O	O

Dox	NN	O	O
up-regulated	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
TRAIL	NN	O	B-protein
receptor	NN	O	I-protein
death	NN	O	B-protein
receptor	NN	O	I-protein
5	NN	O	I-protein
(	NN	O	O
DR5	NN	O	B-protein
)	NN	O	O
and	NN	O	O
synergistically	NN	O	O
enhanced	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
TRAIL	NN	O	B-protein
not	NN	O	O
only	NN	O	O
against	NN	O	O
MM	NN	O	B-cell_type
cells	NN	O	I-cell_type
sensitive	NN	O	O
to	NN	O	O
,	NN	O	O
but	NN	O	O
also	NN	O	O
against	NN	O	O
those	NN	O	O
resistant	NN	O	O
to	NN	O	O
,	NN	O	O
Dex-	NN	O	O
or	NN	O	O
Dox-induced	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

Nuclear	NN	O	O
factor	NN	O	O
(	NN	O	O
NF	NN	O	O
)	NN	O	O
-kappaB	NN	O	O
inhibitors	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
SN50	NN	O	O
(	NN	O	O
a	NN	O	O
cell-permeable	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
and	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
)	NN	O	O
or	NN	O	O
the	NN	O	O
proteasome	NN	O	O
inhibitor	NN	O	O
PS-341	NN	O	O
,	NN	O	O
enhanced	NN	O	O
the	NN	O	O
proapoptotic	NN	O	O
activity	NN	O	O
of	NN	O	O
TRAIL/Apo2L	NN	O	B-protein
against	NN	O	O
TRAIL-sensitive	NN	O	B-cell_type
MM	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
whereas	NN	O	O
SN50	NN	O	O
reversed	NN	O	O
the	NN	O	O
TRAIL	NN	O	O
resistance	NN	O	O
of	NN	O	O
ARH-77	NN	O	B-cell_type
and	NN	O	I-cell_type
IM-9	NN	O	I-cell_type
MM	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Importantly	NN	O	O
,	NN	O	O
normal	NN	O	B-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
were	NN	O	O
not	NN	O	O
sensitized	NN	O	O
to	NN	O	O
TRAIL	NN	O	B-protein
by	NN	O	O
either	NN	O	O
Dox	NN	O	O
,	NN	O	O
SN50	NN	O	O
,	NN	O	O
or	NN	O	O
PS-341	NN	O	O
.	NN	O	O

These	NN	O	O
preclinical	NN	O	O
studies	NN	O	O
suggest	NN	O	O
that	NN	O	O
TRAIL/Apo2L	NN	O	B-protein
can	NN	O	O
overcome	NN	O	O
conventional	NN	O	O
drug	NN	O	O
resistance	NN	O	O
and	NN	O	O
provide	NN	O	O
the	NN	O	O
basis	NN	O	O
for	NN	O	O
clinical	NN	O	O
trials	NN	O	O
of	NN	O	O
TRAIL	NN	O	B-protein
-based	NN	O	O
treatment	NN	O	O
regimens	NN	O	O
to	NN	O	O
improve	NN	O	O
outcome	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
MM	NN	O	O
.	NN	O	O

(	NN	O	O
Blood.	NN	O	O
2001	NN	O	O
;	NN	O	O
98	NN	O	O
:	NN	O	O
795-804	NN	O	O
)	NN	O	O

-DOCSTART-	O

CD28	NN	O	B-protein
costimulation	NN	O	O
is	NN	O	O
required	NN	O	O
not	NN	O	O
only	NN	O	O
to	NN	O	O
induce	NN	O	O
IL-12	NN	O	B-protein
receptor	NN	O	I-protein
but	NN	O	O
also	NN	O	O
to	NN	O	O
render	NN	O	O
janus	NN	O	B-protein
kinases/STAT4	NN	O	I-protein
responsive	NN	O	O
to	NN	O	O
IL-12	NN	O	B-protein
stimulation	NN	O	O
in	NN	O	O
TCR-triggered	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
activation	NN	O	O
of	NN	O	O
resting	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
for	NN	O	O
the	NN	O	O
acquisition	NN	O	O
of	NN	O	O
various	NN	O	O
functions	NN	O	O
depends	NN	O	O
on	NN	O	O
whether	NN	O	O
CD28	NN	O	O
costimulatory	NN	O	O
signals	NN	O	O
are	NN	O	O
provided	NN	O	O
upon	NN	O	O
T	NN	O	O
cell	NN	O	O
receptor	NN	O	O
stimulation	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
how	NN	O	O
CD28	NN	O	O
costimulation	NN	O	O
functions	NN	O	O
to	NN	O	O
allow	NN	O	O
TCR	NN	O	B-protein
-triggered	NN	O	O
resting	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
acquire	NN	O	O
IL-12	NN	O	B-protein
responsiveness	NN	O	O
.	NN	O	O

When	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
stimulated	NN	O	O
with	NN	O	O
low	NN	O	O
doses	NN	O	O
of	NN	O	O
anti-CD3	NN	O	B-protein
mAb	NN	O	I-protein
,	NN	O	O
CD28	NN	O	O
costimulation	NN	O	O
was	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
optimal	NN	O	O
levels	NN	O	O
of	NN	O	O
IL-12	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
IL-12R	NN	O	B-protein
)	NN	O	O
expression	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
stimulation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
high	NN	O	O
doses	NN	O	O
of	NN	O	O
anti-CD3	NN	O	B-protein
alone	NN	O	O
induced	NN	O	O
comparable	NN	O	O
levels	NN	O	O
of	NN	O	O
IL-12R	NN	O	B-protein
expression	NN	O	O
to	NN	O	O
those	NN	O	O
induced	NN	O	O
upon	NN	O	O
CD28	NN	O	O
costimulation	NN	O	O
.	NN	O	O

Nevertheless	NN	O	O
,	NN	O	O
there	NN	O	O
was	NN	O	O
a	NN	O	O
substantial	NN	O	O
difference	NN	O	O
in	NN	O	O
IL-12	NN	O	B-protein
responsiveness	NN	O	O
between	NN	O	O
these	NN	O	O
two	NN	O	O
groups	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
:	NN	O	O
compared	NN	O	O
to	NN	O	O
anti-CD28-costimulated	NN	O	B-cell_type
T	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
that	NN	O	O
were	NN	O	O
not	NN	O	O
costimulated	NN	O	O
with	NN	O	O
anti-CD28	NN	O	B-protein
exhibited	NN	O	O
decreased	NN	O	O
levels	NN	O	O
of	NN	O	O
Janus	NN	O	B-protein
kinases	NN	O	I-protein
(	NN	O	O
JAK	NN	O	B-protein
)	NN	O	O
JAK2/TYK2	NN	O	B-protein
and	NN	O	O
STAT4	NN	O	B-protein
phosphorylation	NN	O	O
and	NN	O	O
IFN-y	NN	O	O
production	NN	O	O
following	NN	O	O
IL-12	NN	O	B-protein
stimulation	NN	O	O
.	NN	O	O

Importantly	NN	O	O
,	NN	O	O
STAT6	NN	O	O
phosphorylation	NN	O	O
following	NN	O	O
IL-4	NN	O	O
stimulation	NN	O	O
was	NN	O	O
not	NN	O	O
decreased	NN	O	O
in	NN	O	O
anti-CD28-uncostimulated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
resutls	NN	O	O
indicate	NN	O	O
that	NN	O	O
CD28	NN	O	O
costimulation	NN	O	O
not	NN	O	O
only	NN	O	O
contributes	NN	O	O
to	NN	O	O
up-regulating	NN	O	O
IL-12R	NN	O	B-protein
expression	NN	O	O
but	NN	O	O
is	NN	O	O
also	NN	O	O
required	NN	O	O
to	NN	O	O
render	NN	O	O
JAKs/STAT4	NN	O	B-protein
responsive	NN	O	O
to	NN	O	O
IL-12	NN	O	B-protein
stimulation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Constitutively	NN	O	O
activated	NN	O	O
Akt-1	NN	O	B-protein
is	NN	O	O
vital	NN	O	O
for	NN	O	O
the	NN	O	O
survival	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
monocyte-differentiated	NN	O	I-cell_type
macrophages	NN	O	I-cell_type
.	NN	O	O

Role	NN	O	O
of	NN	O	O
Mcl-1	NN	O	B-protein
,	NN	O	O
independent	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	I-protein
NF	NN	O	I-protein
)	NN	O	I-protein
-kappaB	NN	O	I-protein
,	NN	O	O
Bad	NN	O	B-protein
,	NN	O	O
or	NN	O	O
caspase	NN	O	O
activation	NN	O	O
.	NN	O	O

Recent	NN	O	O
data	NN	O	O
from	NN	O	O
mice	NN	O	O
deficient	NN	O	O
for	NN	O	O
phosphatase	NN	O	B-protein
and	NN	O	O
tensin	NN	O	B-DNA
homologue	NN	O	I-DNA
deleted	NN	O	O
from	NN	O	O
chromosome	NN	O	B-DNA
10	NN	O	I-DNA
or	NN	O	O
src	NN	O	B-protein
homology	NN	O	I-protein
2	NN	O	I-protein
domain-containing	NN	O	I-protein
5	NN	O	I-protein
'	NN	O	I-protein
inositol	NN	O	I-protein
phosphatase	NN	O	I-protein
,	NN	O	O
phosphatases	NN	O	B-protein
that	NN	O	O
negatively	NN	O	O
regulate	NN	O	O
the	NN	O	O
phosphatidylinositol	NN	O	B-protein
3-kinase	NN	O	I-protein
(	NN	O	O
PI3K	NN	O	B-protein
)	NN	O	O
pathway	NN	O	O
,	NN	O	O
revealed	NN	O	O
an	NN	O	O
increased	NN	O	O
number	NN	O	O
of	NN	O	O
macrophages	NN	O	B-cell_type
in	NN	O	O
these	NN	O	O
animals	NN	O	O
,	NN	O	O
suggesting	NN	O	O
an	NN	O	O
essential	NN	O	O
role	NN	O	O
for	NN	O	O
the	NN	O	O
PI3K	NN	O	B-protein
pathway	NN	O	O
for	NN	O	O
macro-phage	NN	O	B-cell_type
survival	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
focused	NN	O	O
on	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
the	NN	O	O
PI3K-regulated	NN	O	B-protein
serine/threonine	NN	O	I-protein
kinase	NN	O	I-protein
Akt-1	NN	O	B-protein
in	NN	O	O
modulating	NN	O	O
macrophage	NN	O	B-cell_type
survival	NN	O	O
.	NN	O	O

Akt-1	NN	O	B-protein
was	NN	O	O
constitutively	NN	O	O
activated	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
and	NN	O	O
addition	NN	O	O
of	NN	O	O
the	NN	O	O
PI3K	NN	O	B-protein
inhibitor	NN	O	O
,	NN	O	O
LY294002	NN	O	O
,	NN	O	O
suppressed	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
Akt-1	NN	O	B-protein
and	NN	O	O
induced	NN	O	O
cell	NN	O	O
death	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
suppression	NN	O	O
of	NN	O	O
Akt-1	NN	O	B-protein
by	NN	O	O
inhibition	NN	O	O
of	NN	O	O
PI3K	NN	O	B-protein
or	NN	O	O
a	NN	O	O
dominant	NN	O	B-protein
negative	NN	O	I-protein
(	NN	O	I-protein
DN	NN	O	I-protein
)	NN	O	I-protein
Akt-1	NN	O	I-protein
resulted	NN	O	O
in	NN	O	O
loss	NN	O	O
of	NN	O	O
mitochondrial	NN	O	O
transmembrane	NN	O	O
potential	NN	O	O
,	NN	O	O
activation	NN	O	O
of	NN	O	O
caspases-9	NN	O	B-protein
and	NN	O	I-protein
-3	NN	O	I-protein
,	NN	O	O
and	NN	O	O
DNA	NN	O	O
fragmentation	NN	O	O
.	NN	O	O

The	NN	O	O
effects	NN	O	O
of	NN	O	O
PI3K	NN	O	B-protein
inhibition	NN	O	O
were	NN	O	O
reversed	NN	O	O
by	NN	O	O
the	NN	O	O
ectopic	NN	O	O
expression	NN	O	O
of	NN	O	O
constitutively	NN	O	O
activated	NN	O	O
Akt-1	NN	O	B-protein
or	NN	O	O
Bcl-x	NN	O	B-protein
(	NN	O	I-protein
L	NN	O	I-protein
)	NN	O	I-protein
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
PI3K	NN	O	B-protein
/Akt-1	NN	O	O
pathway	NN	O	O
either	NN	O	O
by	NN	O	O
LY294002	NN	O	O
or	NN	O	O
DN	NN	O	B-protein
Akt-1	NN	O	I-protein
had	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
constitutive	NN	O	O
or	NN	O	O
inducible	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
-kappaB	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
.	NN	O	O

However	NN	O	O
,	NN	O	O
after	NN	O	O
inhibition	NN	O	O
of	NN	O	O
the	NN	O	O
PI3K	NN	O	B-protein
/	NN	O	O
Akt-1	NN	O	B-protein
pathway	NN	O	O
,	NN	O	O
a	NN	O	O
marked	NN	O	O
decrease	NN	O	O
in	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
antiapoptotic	NN	O	B-protein
molecule	NN	O	I-protein
Mcl-1	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
other	NN	O	O
Bcl-2	NN	O	B-protein
family	NN	O	I-protein
members	NN	O	I-protein
was	NN	O	O
observed	NN	O	O
,	NN	O	O
and	NN	O	O
Mcl-1	NN	O	B-protein
rescued	NN	O	O
macrophages	NN	O	B-cell_type
from	NN	O	O
LY294002-induced	NN	O	O
cell	NN	O	O
death	NN	O	O
.	NN	O	O

Further	NN	O	O
,	NN	O	O
inhibition	NN	O	O
of	NN	O	O
Mcl-1	NN	O	B-protein
by	NN	O	O
antisense	NN	O	O
oligonucleotides	NN	O	O
,	NN	O	O
also	NN	O	O
resulted	NN	O	O
in	NN	O	O
macrophage	NN	O	B-cell_type
apoptosis	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
our	NN	O	O
findings	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
constitutive	NN	O	O
activation	NN	O	O
of	NN	O	O
Akt-1	NN	O	B-protein
regulates	NN	O	O
macrophage	NN	O	B-cell_type
survival	NN	O	O
through	NN	O	O
Mcl-1	NN	O	B-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
independent	NN	O	O
of	NN	O	O
caspases	NN	O	B-protein
,	NN	O	O
NF-kappaB	NN	O	B-protein
,	NN	O	O
or	NN	O	O
Bad	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

GRbeta	NN	O	O
expression	NN	O	O
in	NN	O	O
nasal	NN	O	B-cell_type
polyp	NN	O	I-cell_type
inflammatory	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
its	NN	O	O
relationship	NN	O	O
to	NN	O	O
the	NN	O	O
anti-inflammatory	NN	O	O
effects	NN	O	O
of	NN	O	O
intranasal	NN	O	O
fluticasone	NN	O	O
.	NN	O	O

BACKGROUND	NN	O	O
:	NN	O	O
Nasal	NN	O	O
polyposis	NN	O	O
disease	NN	O	O
is	NN	O	O
an	NN	O	O
inflammatory	NN	O	O
disorder	NN	O	O
with	NN	O	O
intense	NN	O	O
eosinophilic	NN	O	O
infiltration	NN	O	O
of	NN	O	O
respiratory	NN	O	O
mucosa	NN	O	O
that	NN	O	O
is	NN	O	O
often	NN	O	O
difficult	NN	O	O
to	NN	O	O
control	NN	O	O
with	NN	O	O
topical	NN	O	O
steroids	NN	O	O
.	NN	O	O

Recent	NN	O	O
evidence	NN	O	O
suggests	NN	O	O
that	NN	O	O
overexpression	NN	O	O
of	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
splice	NN	O	I-protein
variant	NN	O	I-protein
GRbeta	NN	O	B-protein
in	NN	O	O
inflammatory	NN	O	B-cell_type
cells	NN	O	I-cell_type
might	NN	O	O
contribute	NN	O	O
to	NN	O	O
steroid	NN	O	O
insensitivity	NN	O	O
in	NN	O	O
diseases	NN	O	O
such	NN	O	O
as	NN	O	O
asthma	NN	O	O
.	NN	O	O

OBJECTIVE	NN	O	O
:	NN	O	O
The	NN	O	O
purposes	NN	O	O
of	NN	O	O
this	NN	O	O
investigation	NN	O	O
were	NN	O	O
to	NN	O	O
determine	NN	O	O
whether	NN	O	O
nasal	NN	O	B-cell_type
polyp	NN	O	I-cell_type
(	NN	O	I-cell_type
NP	NN	O	I-cell_type
)	NN	O	I-cell_type
inflammatory	NN	O	I-cell_type
cells	NN	O	I-cell_type
overexpress	NN	O	O
GRbeta	NN	O	B-protein
and	NN	O	O
to	NN	O	O
examine	NN	O	O
whether	NN	O	O
GRbeta	NN	O	B-protein
overexpression	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
insensitivity	NN	O	O
to	NN	O	O
the	NN	O	O
potent	NN	O	O
topical	NN	O	O
steroid	NN	O	O
fluticasone	NN	O	O
propionate	NN	O	O
(	NN	O	O
FP	NN	O	O
)	NN	O	O
.	NN	O	O

METHODS	NN	O	O
:	NN	O	O
Biopsies	NN	O	O
were	NN	O	O
obtained	NN	O	O
from	NN	O	O
10	NN	O	O
subjects	NN	O	O
with	NN	O	O
NPs	NN	O	O
before	NN	O	O
and	NN	O	O
4	NN	O	O
weeks	NN	O	O
after	NN	O	O
treatment	NN	O	O
with	NN	O	O
intranasal	NN	O	O
FP	NN	O	O
.	NN	O	O

Middle	NN	O	O
turbinates	NN	O	O
biopsies	NN	O	O
from	NN	O	O
6	NN	O	O
healthy	NN	O	O
,	NN	O	O
nonallergic	NN	O	O
subjects	NN	O	O
served	NN	O	O
as	NN	O	O
normal	NN	O	O
controls	NN	O	O
.	NN	O	O

Biopsies	NN	O	O
were	NN	O	O
immunostained	NN	O	O
for	NN	O	O
inflammatory	NN	O	O
cell	NN	O	O
markers	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
GRbeta	NN	O	B-protein
and	NN	O	O
probed	NN	O	O
for	NN	O	O
various	NN	O	O
cytokine	NN	O	B-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

The	NN	O	O
anti-inflammatory	NN	O	O
response	NN	O	O
to	NN	O	O
FP	NN	O	O
was	NN	O	O
examined	NN	O	O
in	NN	O	O
relation	NN	O	O
to	NN	O	O
pretreatment	NN	O	O
levels	NN	O	O
of	NN	O	O
GRbeta	NN	O	O
expression	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
The	NN	O	O
total	NN	O	O
numbers	NN	O	O
of	NN	O	O
inflammatory	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
increased	NN	O	O
in	NN	O	O
NPs	NN	O	B-cell_type
.	NN	O	O

The	NN	O	O
percentage	NN	O	O
of	NN	O	O
inflammatory	NN	O	B-cell_type
cells	NN	O	I-cell_type
expressing	NN	O	O
GRbeta	NN	O	B-protein
was	NN	O	O
also	NN	O	O
increased	NN	O	O
(	NN	O	O
40.5	NN	O	O
%	NN	O	O
+/-	NN	O	O
19.2	NN	O	O
%	NN	O	O
vs	NN	O	O
16.1	NN	O	O
%	NN	O	O
+/-	NN	O	O
4.0	NN	O	O
%	NN	O	O
,	NN	O	O
P	NN	O	O
=.009	NN	O	O
)	NN	O	O
.	NN	O	O

GRbeta	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
NPs	NN	O	B-cell_type
was	NN	O	O
almost	NN	O	O
exclusive	NN	O	O
to	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
eosinophils	NN	O	B-cell_type
,	NN	O	O
and	NN	O	O
macrophages	NN	O	B-cell_type
.	NN	O	O

An	NN	O	O
inverse	NN	O	O
correlation	NN	O	O
was	NN	O	O
observed	NN	O	O
between	NN	O	O
the	NN	O	O
baseline	NN	O	O
inflammatory	NN	O	O
cell	NN	O	O
GRbeta	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
the	NN	O	O
reduction	NN	O	O
after	NN	O	O
FP	NN	O	O
treatment	NN	O	O
in	NN	O	O
EG2-positive	NN	O	B-cell_type
eosinophils	NN	O	I-cell_type
,	NN	O	O
CD4-positive	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
endothelial	NN	O	O
VCAM-1	NN	O	B-protein
expression	NN	O	O
,	NN	O	O
and	NN	O	O
IL-4	NN	O	B-cell_type
mRNA-positive	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

NPs	NN	O	B-cell_type
that	NN	O	O
were	NN	O	O
``	NN	O	O
FP-insensitive	NN	O	O
''	NN	O	O
in	NN	O	O
terms	NN	O	O
of	NN	O	O
suppression	NN	O	O
of	NN	O	O
eosinophil	NN	O	B-cell_type
numbers	NN	O	O
(	NN	O	O
major	NN	O	B-protein
basic	NN	O	I-protein
protein	NN	O	I-protein
-positive	NN	O	O
)	NN	O	O
had	NN	O	O
a	NN	O	O
significantly	NN	O	O
greater	NN	O	O
percentage	NN	O	O
of	NN	O	O
GRbeta-positive	NN	O	B-cell_type
inflammatory	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
a	NN	O	O
higher	NN	O	O
ratio	NN	O	O
of	NN	O	O
GRbeta-positive/GRalpha-positive	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
increased	NN	O	O
numbers	NN	O	O
of	NN	O	O
GRbeta-positive	NN	O	B-cell_type
eosinophils	NN	O	I-cell_type
and	NN	O	O
macrophages	NN	O	B-cell_type
in	NN	O	O
comparison	NN	O	O
with	NN	O	O
those	NN	O	O
that	NN	O	O
were	NN	O	O
``	NN	O	O
FP-sensitive.	NN	O	O
''	NN	O	O

``	NN	O	B-cell_type
FP-insensitive	NN	O	I-cell_type
''	NN	O	I-cell_type
NPs	NN	O	I-cell_type
also	NN	O	O
demonstrated	NN	O	O
a	NN	O	O
higher	NN	O	O
percentage	NN	O	O
of	NN	O	O
IL-5-positive	NN	O	B-cell_type
inflammatory	NN	O	I-cell_type
cells	NN	O	I-cell_type
expressing	NN	O	O
GRbeta	NN	O	B-protein
before	NN	O	O
and	NN	O	O
after	NN	O	O
FP	NN	O	O
treatment	NN	O	O
.	NN	O	O

CONCLUSION	NN	O	O
:	NN	O	O
GRbeta	NN	O	B-protein
expression	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
marker	NN	O	O
of	NN	O	O
steroid	NN	O	O
insensitivity	NN	O	O
in	NN	O	O
NPs	NN	O	B-cell_type
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
GRbeta	NN	O	B-protein
by	NN	O	O
NP	NN	O	B-cell_type
inflammatory	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
particularly	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
eosinophils	NN	O	B-cell_type
,	NN	O	O
might	NN	O	O
render	NN	O	O
them	NN	O	O
resistant	NN	O	O
to	NN	O	O
suppression	NN	O	O
by	NN	O	O
topical	NN	O	O
steroids	NN	O	O
and	NN	O	O
thereby	NN	O	O
contribute	NN	O	O
to	NN	O	O
persistent	NN	O	O
NP	NN	O	B-cell_type
inflammation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
galectin-10	NN	O	B-protein
(	NN	O	O
eosinophil	NN	O	B-protein
Charcot-Leyden	NN	O	I-protein
crystal	NN	O	I-protein
protein	NN	O	I-protein
)	NN	O	O
:	NN	O	O
a	NN	O	O
GC	NN	O	B-DNA
box	NN	O	I-DNA
(	NN	O	O
-44	NN	O	O
to	NN	O	O
-50	NN	O	O
)	NN	O	O
controls	NN	O	O
butyric	NN	O	O
acid	NN	O	O
induction	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

Galectin-10	NN	O	B-protein
(	NN	O	O
gal-10	NN	O	B-protein
,	NN	O	O
also	NN	O	O
known	NN	O	O
as	NN	O	O
Charcot-Leyden	NN	O	B-protein
crystal	NN	O	I-protein
protein	NN	O	I-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
galectin	NN	O	B-protein
family	NN	O	I-protein
of	NN	O	O
beta-galactoside	NN	O	B-protein
binding	NN	O	I-protein
proteins	NN	O	I-protein
that	NN	O	O
is	NN	O	O
expressed	NN	O	O
uniquely	NN	O	O
in	NN	O	O
eosinophilic	NN	O	B-cell_type
and	NN	O	I-cell_type
basophilic	NN	O	I-cell_type
leukocytes	NN	O	I-cell_type
.	NN	O	O

To	NN	O	O
gain	NN	O	O
a	NN	O	O
better	NN	O	O
understanding	NN	O	O
of	NN	O	O
galectin	NN	O	O
gene	NN	O	O
expression	NN	O	O
,	NN	O	O
we	NN	O	O
present	NN	O	O
an	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
encoding	NN	O	O
gal-10	NN	O	B-protein
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
the	NN	O	O
minimal	NN	O	B-DNA
promoter	NN	O	I-DNA
revealed	NN	O	O
nine	NN	O	O
consensus-binding	NN	O	B-DNA
sites	NN	O	I-DNA
for	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
including	NN	O	O
several	NN	O	O
that	NN	O	O
are	NN	O	O
also	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
minimal	NN	O	B-DNA
promoters	NN	O	I-DNA
of	NN	O	O
galectins	NN	O	B-DNA
-1	NN	O	I-DNA
,	NN	O	I-DNA
-2	NN	O	I-DNA
,	NN	O	I-DNA
and	NN	O	I-DNA
-3	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
decrease	NN	O	O
in	NN	O	O
gal-10	NN	O	O
promoter	NN	O	O
activity	NN	O	O
after	NN	O	O
disruption	NN	O	O
of	NN	O	O
either	NN	O	O
the	NN	O	O
GC	NN	O	B-DNA
box	NN	O	I-DNA
(	NN	O	O
-44	NN	O	O
to	NN	O	O
-50	NN	O	O
)	NN	O	O
or	NN	O	O
the	NN	O	O
Oct	NN	O	B-DNA
site	NN	O	I-DNA
(	NN	O	O
-255	NN	O	O
to	NN	O	O
-261	NN	O	O
)	NN	O	O
suggests	NN	O	O
that	NN	O	O
these	NN	O	O
sites	NN	O	O
,	NN	O	O
along	NN	O	O
with	NN	O	O
the	NN	O	O
previously	NN	O	O
characterized	NN	O	O
GATA	NN	O	B-DNA
and	NN	O	I-DNA
EoTF	NN	O	I-DNA
sites	NN	O	I-DNA
,	NN	O	O
are	NN	O	O
necessary	NN	O	O
for	NN	O	O
full	NN	O	O
promoter	NN	O	O
activity	NN	O	O
.	NN	O	O

By	NN	O	O
supershift	NN	O	O
analysis	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
binding	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
Sp1	NN	O	B-protein
and	NN	O	O
Oct1	NN	O	B-protein
to	NN	O	O
the	NN	O	O
consensus	NN	O	B-DNA
GC	NN	O	I-DNA
box	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
Oct	NN	O	B-DNA
site	NN	O	I-DNA
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

Similar	NN	O	O
to	NN	O	O
gal-1	NN	O	B-DNA
,	NN	O	O
gal-10	NN	O	O
expression	NN	O	O
is	NN	O	O
induced	NN	O	O
by	NN	O	O
butyric	NN	O	O
acid	NN	O	O
,	NN	O	O
an	NN	O	O
effect	NN	O	O
that	NN	O	O
is	NN	O	O
lost	NN	O	O
upon	NN	O	O
ablation	NN	O	O
of	NN	O	O
the	NN	O	O
GC	NN	O	B-DNA
box	NN	O	I-DNA
.	NN	O	O

Additionally	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
AML3	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
consensus	NN	O	B-DNA
AML	NN	O	I-DNA
site	NN	O	I-DNA
and	NN	O	O
YY1	NN	O	B-protein
binding	NN	O	I-protein
to	NN	O	O
the	NN	O	O
Inr	NN	O	B-DNA
sequence	NN	O	I-DNA
,	NN	O	O
both	NN	O	O
elements	NN	O	O
functioning	NN	O	O
as	NN	O	O
silencers	NN	O	O
in	NN	O	O
the	NN	O	O
gal-10	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Tumour-stromal	NN	O	O
interactions	NN	O	O
.	NN	O	O

Role	NN	O	O
of	NN	O	O
the	NN	O	O
stroma	NN	O	O
in	NN	O	O
mammary	NN	O	O
development	NN	O	O
.	NN	O	O

Mammary	NN	O	O
development	NN	O	O
depends	NN	O	O
on	NN	O	O
branching	NN	O	O
morphogenesis	NN	O	O
,	NN	O	O
namely	NN	O	O
the	NN	O	O
bifurcation	NN	O	O
and	NN	O	O
extension	NN	O	O
of	NN	O	O
ductal	NN	O	O
growth	NN	O	O
points	NN	O	O
(	NN	O	O
end	NN	O	O
buds	NN	O	O
)	NN	O	O
and	NN	O	O
secretory	NN	O	O
lobules	NN	O	O
into	NN	O	O
a	NN	O	O
more	NN	O	O
or	NN	O	O
less	NN	O	O
fatty	NN	O	O
stroma	NN	O	O
.	NN	O	O

Because	NN	O	O
breast	NN	O	O
carcinomas	NN	O	O
are	NN	O	O
overwhelmingly	NN	O	O
ductal	NN	O	O
in	NN	O	O
origin	NN	O	O
,	NN	O	O
this	NN	O	O
review	NN	O	O
focuses	NN	O	O
on	NN	O	O
stromal	NN	O	O
influences	NN	O	O
guiding	NN	O	O
postnatal	NN	O	O
ductal	NN	O	O
development	NN	O	O
and	NN	O	O
there	NN	O	O
is	NN	O	O
only	NN	O	O
the	NN	O	O
briefest	NN	O	O
account	NN	O	O
of	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
embryonic	NN	O	O
stroma	NN	O	O
(	NN	O	O
mesenchyme	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
stroma	NN	O	O
as	NN	O	O
the	NN	O	O
necessary	NN	O	O
target	NN	O	O
for	NN	O	O
endocrine	NN	O	O
mammogens	NN	O	O
and	NN	O	O
the	NN	O	O
source	NN	O	O
of	NN	O	O
stimulatory	NN	O	O
growth	NN	O	O
factors	NN	O	O
is	NN	O	O
described	NN	O	O
and	NN	O	O
the	NN	O	O
importance	NN	O	O
of	NN	O	O
mammary	NN	O	O
epithelium-induced	NN	O	O
modifications	NN	O	O
of	NN	O	O
the	NN	O	O
periductal	NN	O	O
stroma	NN	O	O
is	NN	O	O
emphasized	NN	O	O
.	NN	O	O

Evidence	NN	O	O
is	NN	O	O
presented	NN	O	O
that	NN	O	O
if	NN	O	O
they	NN	O	O
are	NN	O	O
to	NN	O	O
grow	NN	O	O
,	NN	O	O
end	NN	O	O
buds	NN	O	O
must	NN	O	O
condition	NN	O	O
proximal	NN	O	O
fatty	NN	O	O
stroma	NN	O	O
by	NN	O	O
recruiting	NN	O	O
white	NN	O	B-cell_type
blood	NN	O	I-cell_type
cells	NN	O	I-cell_type
as	NN	O	O
well	NN	O	O
as	NN	O	O
inducing	NN	O	O
stromal	NN	O	O
cell	NN	O	O
division	NN	O	O
and	NN	O	O
,	NN	O	O
possibly	NN	O	O
,	NN	O	O
estrogen	NN	O	B-protein
receptors	NN	O	I-protein
.	NN	O	O

The	NN	O	O
induction	NN	O	O
of	NN	O	O
a	NN	O	O
fibrous	NN	O	O
stromal	NN	O	O
tunic	NN	O	O
around	NN	O	O
the	NN	O	O
end	NN	O	O
bud	NN	O	O
is	NN	O	O
described	NN	O	O
and	NN	O	O
its	NN	O	O
likely	NN	O	O
role	NN	O	O
as	NN	O	O
a	NN	O	O
complex	NN	O	O
ductal	NN	O	O
morphogen	NN	O	O
is	NN	O	O
discussed	NN	O	O
;	NN	O	O
a	NN	O	O
possible	NN	O	O
role	NN	O	O
in	NN	O	O
growth	NN	O	O
inhibition	NN	O	O
is	NN	O	O
also	NN	O	O
considered	NN	O	O
.	NN	O	O

Although	NN	O	O
the	NN	O	O
signals	NN	O	O
governing	NN	O	O
fibrotic	NN	O	O
induction	NN	O	O
,	NN	O	O
ductal	NN	O	O
morphogenesis	NN	O	O
,	NN	O	O
and	NN	O	O
growth	NN	O	O
inhibition	NN	O	O
are	NN	O	O
unknown	NN	O	O
,	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
transforming	NN	O	O
growth	NN	O	B-protein
factor-beta	NN	O	I-protein
is	NN	O	O
highly	NN	O	O
likely	NN	O	O
and	NN	O	O
is	NN	O	O
discussed	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
a	NN	O	O
need	NN	O	O
for	NN	O	O
new	NN	O	O
conceptual	NN	O	O
and	NN	O	O
experimental	NN	O	O
approaches	NN	O	O
to	NN	O	O
understanding	NN	O	O
stromal-epithelial	NN	O	O
signaling	NN	O	O
is	NN	O	O
discussed	NN	O	O
.	NN	O	O

-DOCSTART-	O

Adipophilin	NN	O	B-protein
is	NN	O	O
a	NN	O	O
sensitive	NN	O	O
marker	NN	O	O
for	NN	O	O
lipid	NN	O	O
loading	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
blood	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

Adipophilin	NN	O	B-protein
,	NN	O	O
a	NN	O	O
marker	NN	O	O
of	NN	O	O
lipid	NN	O	O
accumulation	NN	O	O
initially	NN	O	O
described	NN	O	O
in	NN	O	O
adipocytes	NN	O	B-cell_type
,	NN	O	O
was	NN	O	O
recently	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
induced	NN	O	O
in	NN	O	O
macrophage	NN	O	B-cell_type
foam	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
even	NN	O	O
freshly	NN	O	B-cell_type
isolated	NN	O	I-cell_type
blood	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
express	NN	O	O
adipophilin	NN	O	B-protein
and	NN	O	O
that	NN	O	O
the	NN	O	O
amount	NN	O	O
of	NN	O	O
adipophilin	NN	O	B-protein
protein	NN	O	I-protein
is	NN	O	O
variable	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
from	NN	O	O
different	NN	O	O
healthy	NN	O	O
individuals	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
physiological	NN	O	O
expression	NN	O	O
of	NN	O	O
adipophilin	NN	O	B-protein
does	NN	O	O
not	NN	O	O
correlate	NN	O	O
with	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
free	NN	O	O
fatty	NN	O	O
acids	NN	O	O
,	NN	O	O
cholesterylesters	NN	O	O
or	NN	O	O
free	NN	O	O
cholesterol	NN	O	O
.	NN	O	O

Enzymatically	NN	O	B-protein
modified	NN	O	I-protein
low-density	NN	O	I-protein
lipoprotein	NN	O	I-protein
(	NN	O	O
E-LDL	NN	O	B-protein
)	NN	O	O
induces	NN	O	O
rapid	NN	O	O
foam	NN	O	O
cell	NN	O	O
formation	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
upregulates	NN	O	O
adipophilin	NN	O	B-protein
mRNA	NN	O	O
and	NN	O	O
protein	NN	O	O
within	NN	O	O
2	NN	O	O
h	NN	O	O
of	NN	O	O
incubation	NN	O	O
.	NN	O	O

This	NN	O	O
rapid	NN	O	O
induction	NN	O	O
of	NN	O	O
adipophilin	NN	O	B-protein
is	NN	O	O
accompanied	NN	O	O
by	NN	O	O
a	NN	O	O
significant	NN	O	O
increase	NN	O	O
of	NN	O	O
free	NN	O	O
fatty	NN	O	O
acids	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
incubated	NN	O	O
with	NN	O	O
E-LDL	NN	O	B-protein
.	NN	O	O

Adipophilin	NN	O	B-protein
facilitates	NN	O	O
the	NN	O	O
uptake	NN	O	O
of	NN	O	O
free	NN	O	O
fatty	NN	O	O
acids	NN	O	O
,	NN	O	O
and	NN	O	O
here	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
free	NN	O	O
fatty	NN	O	O
acids	NN	O	O
increase	NN	O	O
is	NN	O	O
related	NN	O	O
to	NN	O	O
the	NN	O	O
early	NN	O	O
upregulation	NN	O	O
of	NN	O	O
adipophilin	NN	O	O
expression	NN	O	O
in	NN	O	O
blood	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

Fatty	NN	O	O
acids	NN	O	O
are	NN	O	O
ligands	NN	O	O
for	NN	O	O
peroxisome	NN	O	B-protein
proliferator-activated	NN	O	I-protein
receptor-gamma	NN	O	I-protein
(	NN	O	O
PPARgamma	NN	O	B-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
upregulation	NN	O	O
of	NN	O	O
adipophilin	NN	O	B-RNA
mRNA	NN	O	I-RNA
by	NN	O	O
PPARgamma	NN	O	B-protein
agonists	NN	O	O
like	NN	O	O
15d-PGJ	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
and	NN	O	O
ciglitazone	NN	O	O
indicates	NN	O	O
that	NN	O	O
PPARgamma	NN	O	B-protein
may	NN	O	O
mediate	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
adipophilin	NN	O	O
expression	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
blood	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Constitutive	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
melanoma	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
results	NN	O	O
from	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
class	NN	O	B-DNA
II	NN	O	I-DNA
transactivator	NN	O	I-DNA
abnormally	NN	O	O
initiated	NN	O	O
from	NN	O	O
its	NN	O	O
B	NN	O	B-DNA
cell-specific	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
melanoma	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
two	NN	O	O
different	NN	O	O
patterns	NN	O	O
of	NN	O	O
MHC	NN	O	O
class	NN	O	O
II	NN	O	O
expression	NN	O	O
have	NN	O	O
been	NN	O	O
described	NN	O	O
,	NN	O	O
either	NN	O	O
an	NN	O	O
IFN	NN	O	O
gamma-inducible	NN	O	O
expression	NN	O	O
of	NN	O	O
HLA-DR	NN	O	B-DNA
and	NN	O	O
HLA-DP	NN	O	B-DNA
,	NN	O	O
with	NN	O	O
a	NN	O	O
faint	NN	O	O
or	NN	O	O
null	NN	O	O
expression	NN	O	O
of	NN	O	O
HLA-DQ	NN	O	B-DNA
,	NN	O	O
resembling	NN	O	O
that	NN	O	O
described	NN	O	O
for	NN	O	O
melanocytes	NN	O	B-cell_type
,	NN	O	O
or	NN	O	O
a	NN	O	O
constitutive	NN	O	O
expression	NN	O	O
,	NN	O	O
i.e.	NN	O	O
,	NN	O	O
IFN-gamma	NN	O	O
independent	NN	O	O
,	NN	O	O
of	NN	O	O
all	NN	O	O
three	NN	O	O
HLA-D	NN	O	B-DNA
isotypes	NN	O	I-DNA
.	NN	O	O

As	NN	O	O
this	NN	O	O
latter	NN	O	O
phenotype	NN	O	O
has	NN	O	O
been	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
more	NN	O	O
rapid	NN	O	O
progression	NN	O	O
of	NN	O	O
melanoma	NN	O	O
tumors	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
analyzed	NN	O	O
in	NN	O	O
different	NN	O	O
melanoma	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
the	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
leading	NN	O	O
to	NN	O	O
this	NN	O	O
abnormal	NN	O	O
pattern	NN	O	O
of	NN	O	O
MHC	NN	O	O
class	NN	O	O
II	NN	O	O
expression	NN	O	O
.	NN	O	O

In	NN	O	O
agreement	NN	O	O
with	NN	O	O
the	NN	O	O
evidence	NN	O	O
of	NN	O	O
a	NN	O	O
coordinate	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
HLA-D	NN	O	B-DNA
genes	NN	O	I-DNA
in	NN	O	O
these	NN	O	O
cell	NN	O	O
lines	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
shown	NN	O	O
the	NN	O	O
constitutive	NN	O	O
expression	NN	O	O
of	NN	O	O
CIITA	NN	O	B-RNA
(	NN	O	I-RNA
class	NN	O	I-RNA
II	NN	O	I-RNA
transactivator	NN	O	I-RNA
)	NN	O	I-RNA
transcripts	NN	O	I-RNA
,	NN	O	O
CIITA	NN	O	B-protein
being	NN	O	O
known	NN	O	O
as	NN	O	O
the	NN	O	O
master	NN	O	O
switch	NN	O	O
of	NN	O	O
MHC	NN	O	O
class	NN	O	O
II	NN	O	O
expression	NN	O	O
.	NN	O	O

Unexpectedly	NN	O	O
,	NN	O	O
these	NN	O	O
transcripts	NN	O	O
initiate	NN	O	O
from	NN	O	O
promoter	NN	O	B-DNA
III	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
CIITA	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	O
a	NN	O	O
promoter	NN	O	O
that	NN	O	O
is	NN	O	O
mainly	NN	O	O
used	NN	O	O
constitutively	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

This	NN	O	O
expression	NN	O	O
was	NN	O	O
further	NN	O	O
shown	NN	O	O
to	NN	O	O
occur	NN	O	O
through	NN	O	O
factor	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
acting	NN	O	O
on	NN	O	O
the	NN	O	O
enhancer	NN	O	O
located	NN	O	O
upstream	NN	O	O
of	NN	O	O
CIITA	NN	O	B-DNA
promoter	NN	O	I-DNA
III	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
was	NN	O	O
previously	NN	O	O
described	NN	O	O
in	NN	O	O
epithelioid	NN	O	B-cell_type
cells	NN	O	I-cell_type
as	NN	O	O
an	NN	O	O
IFN-gamma-response	NN	O	O
sequence	NN	O	O
.	NN	O	O

The	NN	O	O
hypothesis	NN	O	O
of	NN	O	O
a	NN	O	O
general	NN	O	O
abnormality	NN	O	O
of	NN	O	O
the	NN	O	O
IFN-gamma	NN	O	O
transduction	NN	O	O
pathway	NN	O	O
was	NN	O	O
dismissed	NN	O	O
.	NN	O	O

Constitutive	NN	O	O
transcription	NN	O	O
of	NN	O	O
CIITA	NN	O	B-DNA
from	NN	O	O
promoter	NN	O	B-DNA
III	NN	O	I-DNA
having	NN	O	O
been	NN	O	O
observed	NN	O	O
in	NN	O	O
unrelated	NN	O	O
melanoma	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
we	NN	O	O
propose	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
this	NN	O	O
phenomenon	NN	O	O
might	NN	O	O
not	NN	O	O
be	NN	O	O
a	NN	O	O
random	NN	O	O
event	NN	O	O
,	NN	O	O
but	NN	O	O
could	NN	O	O
be	NN	O	O
linked	NN	O	O
to	NN	O	O
the	NN	O	O
neoplasic	NN	O	O
state	NN	O	O
of	NN	O	O
the	NN	O	O
melanoma	NN	O	B-cell_type
cells	NN	O	I-cell_type

-DOCSTART-	O

Caspase	NN	O	B-protein
-mediated	NN	O	O
calcineurin	NN	O	O
activation	NN	O	O
contributes	NN	O	O
to	NN	O	O
IL-2	NN	O	B-protein
release	NN	O	O
during	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
.	NN	O	O

Calcineurin	NN	O	B-protein
,	NN	O	O
a	NN	O	O
Ca	NN	O	B-protein
(	NN	O	I-protein
2+	NN	O	I-protein
)	NN	O	I-protein
/calmodulin-dependent	NN	O	I-protein
Ser/Thr	NN	O	I-protein
phosphatase	NN	O	I-protein
(	NN	O	I-protein
protein	NN	O	I-protein
phosphatase	NN	O	I-protein
2B	NN	O	I-protein
)	NN	O	I-protein
,	NN	O	O
plays	NN	O	O
a	NN	O	O
critical	NN	O	O
role	NN	O	O
in	NN	O	O
IL-2	NN	O	O
production	NN	O	O
during	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
.	NN	O	O

It	NN	O	O
has	NN	O	O
been	NN	O	O
previously	NN	O	O
reported	NN	O	O
that	NN	O	O
IL-2	NN	O	B-protein
release	NN	O	O
in	NN	O	O
activated	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
requires	NN	O	O
caspase-like	NN	O	O
activity	NN	O	O
(	NN	O	O
Posmantur	NN	O	O
et	NN	O	O
al.	NN	O	O
(	NN	O	O
1998	NN	O	O
)	NN	O	O
Exp.	NN	O	O
Cell.	NN	O	O
Res.	NN	O	O
244	NN	O	O
,	NN	O	O
302-309	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
here	NN	O	O
that	NN	O	O
the	NN	O	O
60-kDa	NN	O	B-protein
catalytic	NN	O	I-protein
subunit	NN	O	I-protein
of	NN	O	O
calcineurin	NN	O	B-protein
A	NN	O	I-protein
(	NN	O	O
Cn	NN	O	B-protein
A	NN	O	I-protein
)	NN	O	O
was	NN	O	O
partially	NN	O	O
cleaved	NN	O	O
to	NN	O	O
a	NN	O	O
45-kDa	NN	O	B-protein
form	NN	O	O
in	NN	O	O
phytohemagglutinin	NN	O	O
A	NN	O	O
(	NN	O	O
PHA	NN	O	O
)	NN	O	O
or	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
+	NN	O	O
ionomycin	NN	O	O
(	NN	O	O
P	NN	O	O
+	NN	O	O
I	NN	O	O
)	NN	O	O
-activated	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
parallel	NN	O	O
,	NN	O	O
proteolytic	NN	O	O
activation	NN	O	O
of	NN	O	O
upstream	NN	O	O
caspases	NN	O	B-protein
(	NN	O	O
caspase-8	NN	O	O
and	NN	O	O
-9	NN	O	O
)	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
effector	NN	O	B-protein
caspase-3	NN	O	I-protein
was	NN	O	O
also	NN	O	O
observed	NN	O	O
.	NN	O	O

Cn	NN	O	B-protein
A	NN	O	I-protein
cleavage	NN	O	O
was	NN	O	O
caspase	NN	O	O
mediated	NN	O	O
,	NN	O	O
since	NN	O	O
it	NN	O	O
was	NN	O	O
inhibitable	NN	O	O
by	NN	O	O
pan-caspase	NN	O	O
inhibitor	NN	O	O
Cbz-Asp-CH	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
OC	NN	O	O
(	NN	O	O
O	NN	O	O
)	NN	O	O
-2	NN	O	O
,	NN	O	O
6-dichlorobenzene	NN	O	O
(	NN	O	O
Z-D-DCB	NN	O	O
)	NN	O	O
.	NN	O	O

Cn	NN	O	B-protein
A	NN	O	I-protein
cleavage	NN	O	O
was	NN	O	O
also	NN	O	O
observed	NN	O	O
when	NN	O	O
purified	NN	O	O
calcineurin	NN	O	B-protein
was	NN	O	O
digested	NN	O	O
in	NN	O	O
vitro	NN	O	O
with	NN	O	O
caspase-3	NN	O	B-protein
.	NN	O	O

Truncated	NN	O	B-protein
Cn	NN	O	I-protein
A	NN	O	I-protein
was	NN	O	O
associated	NN	O	O
with	NN	O	O
enhanced	NN	O	O
phosphatase	NN	O	O
activity	NN	O	O
and	NN	O	O
reduced	NN	O	O
calmodulin	NN	O	O
sensitivity	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
in	NN	O	O
PHA	NN	O	O
or	NN	O	O
P	NN	O	O
+	NN	O	O
I-activated	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
dephosphorylation	NN	O	O
of	NN	O	O
calcineurin	NN	O	B-protein
substrate	NN	O	O
NFATc	NN	O	B-protein
(	NN	O	O
a	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
known	NN	O	O
to	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
transactivation	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
)	NN	O	O
,	NN	O	O
was	NN	O	O
also	NN	O	O
suppressed	NN	O	O
by	NN	O	O
Z-D-DCB	NN	O	O
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
our	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
caspase-mediated	NN	O	O
cleavage	NN	O	O
of	NN	O	O
Cn	NN	O	B-protein
A	NN	O	I-protein
contributes	NN	O	O
to	NN	O	O
IL-2	NN	O	O
production	NN	O	O
during	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
.	NN	O	O

Copyright	NN	O	O
2001	NN	O	O
Academic	NN	O	O
Press	NN	O	O
.	NN	O	O

-DOCSTART-	O

Maturation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
dendritic	NN	O	I-cell_type
cells	NN	O	I-cell_type
as	NN	O	O
sulfasalazine	NN	O	O
target	NN	O	O
.	NN	O	O

AIM	NN	O	O
:	NN	O	O
Sulfasalazine	NN	O	O
,	NN	O	O
a	NN	O	O
nonsteroidal	NN	O	O
anti-inflammatory	NN	O	O
drug	NN	O	O
,	NN	O	O
is	NN	O	O
effective	NN	O	O
in	NN	O	O
treating	NN	O	O
some	NN	O	O
autoimmune	NN	O	O
diseases	NN	O	O
,	NN	O	O
but	NN	O	O
its	NN	O	O
mechanism	NN	O	O
of	NN	O	O
action	NN	O	O
is	NN	O	O
unclear	NN	O	O
.	NN	O	O

To	NN	O	O
determine	NN	O	O
whether	NN	O	O
dendritic	NN	O	B-cell_type
cells	NN	O	I-cell_type
could	NN	O	O
be	NN	O	O
a	NN	O	O
possible	NN	O	O
target	NN	O	O
of	NN	O	O
the	NN	O	O
drug	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
sulfasalazine	NN	O	O
and	NN	O	O
its	NN	O	O
metabolites	NN	O	O
,	NN	O	O
aminosalicylate	NN	O	O
and	NN	O	O
sulfapyridine	NN	O	O
,	NN	O	O
on	NN	O	O
in	NN	O	O
vitro	NN	O	O
maturation	NN	O	O
(	NN	O	O
terminal	NN	O	O
differentiation	NN	O	O
)	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
myeloid	NN	O	I-cell_type
dendritic	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

METHODS	NN	O	O
:	NN	O	O
We	NN	O	O
prepared	NN	O	O
immature	NN	O	B-cell_type
dendritic	NN	O	I-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
incubating	NN	O	O
CD14-positive	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
granulocyte-	NN	O	B-protein
macrophage	NN	O	I-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
and	NN	O	O
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

The	NN	O	O
cells	NN	O	O
were	NN	O	O
matured	NN	O	O
by	NN	O	O
addition	NN	O	O
of	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	I-protein
TNF	NN	O	I-protein
)	NN	O	I-protein
-a	NN	O	I-protein
,	NN	O	O
IL-1	NN	O	B-protein
beta	NN	O	I-protein
,	NN	O	O
and	NN	O	O
prostaglandin	NN	O	B-protein
E2	NN	O	I-protein
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
sulfasalazine	NN	O	O
or	NN	O	O
its	NN	O	O
metabolites	NN	O	O
--	NN	O	O
aminosalicylate	NN	O	O
and	NN	O	O
sulfapyridine	NN	O	O
,	NN	O	O
or	NN	O	O
their	NN	O	O
combinations	NN	O	O
.	NN	O	O

We	NN	O	O
quantified	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
drugs	NN	O	O
on	NN	O	O
the	NN	O	O
dendritic	NN	O	O
cell	NN	O	O
characteristics	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
stimulation	NN	O	O
of	NN	O	O
autologous	NN	O	O
and	NN	O	O
allogeneic	NN	O	O
pan-T	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
,	NN	O	O
surface	NN	O	O
marker	NN	O	O
phenotype	NN	O	O
,	NN	O	O
IL-12	NN	O	O
p40	NN	O	O
subunit	NN	O	O
secretion	NN	O	O
,	NN	O	O
and	NN	O	O
activation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	I-protein
NF	NN	O	I-protein
)	NN	O	I-protein
-kappa	NN	O	I-protein
B	NN	O	I-protein
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
Dendritic	NN	O	B-cell_type
cells	NN	O	I-cell_type
treated	NN	O	O
with	NN	O	O
sulfasalazine	NN	O	O
(	NN	O	O
1.25	NN	O	O
micromol/L	NN	O	O
or	NN	O	O
2.5	NN	O	O
micromol/L	NN	O	O
)	NN	O	O
could	NN	O	O
not	NN	O	O
stimulate	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
p	NN	O	O
<	NN	O	O
0.028	NN	O	O
,	NN	O	O
two-sided	NN	O	O
paired	NN	O	O
t-test	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
distinction	NN	O	O
to	NN	O	O
drug-free	NN	O	O
maturing	NN	O	O
dendritic	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
2.5	NN	O	O
micromol/L	NN	O	O
sulfasalazine	NN	O	O
upregulated	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
CD14	NN	O	B-protein
and	NN	O	O
CD68	NN	O	B-protein
and	NN	O	O
downregulated	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
CD40	NN	O	B-protein
,	NN	O	O
CD80	NN	O	B-protein
,	NN	O	O
and	NN	O	O
CD83	NN	O	B-protein
(	NN	O	O
for	NN	O	O
all	NN	O	O
CD	NN	O	O
markers	NN	O	O
,	NN	O	O
p	NN	O	O
<	NN	O	O
0.03	NN	O	O
for	NN	O	O
difference	NN	O	O
between	NN	O	O
measurements	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
and	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
sulfasalazine	NN	O	O
)	NN	O	O
.	NN	O	O

From	NN	O	O
concentration-dependent	NN	O	O
changes	NN	O	O
in	NN	O	O
CD83	NN	O	O
expression	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
an	NN	O	O
apparent	NN	O	O
ID50	NN	O	O
>	NN	O	O
>	NN	O	O
1.5	NN	O	O
micromol/L	NN	O	O
sulfasalazine	NN	O	O
.	NN	O	O

The	NN	O	O
apparent	NN	O	O
ID50	NN	O	O
value	NN	O	O
for	NN	O	O
aminosalicylate-inhibited	NN	O	O
maturation	NN	O	O
was	NN	O	O
4	NN	O	O
micromol/L	NN	O	O
.	NN	O	O

Sulfapyridine	NN	O	O
had	NN	O	O
no	NN	O	O
effect	NN	O	O
.	NN	O	O

At	NN	O	O
1.25	NN	O	O
micromol/L	NN	O	O
,	NN	O	O
sulfasalazine	NN	O	O
largely	NN	O	O
inhibited	NN	O	O
NF-kB	NN	O	O
activation	NN	O	O
in	NN	O	O
dendritic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

CONCLUSION	NN	O	O
:	NN	O	O
Maturing	NN	O	B-cell_type
human	NN	O	I-cell_type
dendritic	NN	O	I-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
hundred-fold	NN	O	O
more	NN	O	O
sensitive	NN	O	O
to	NN	O	O
sulfasalazine	NN	O	O
than	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
the	NN	O	O
most	NN	O	O
sensitive	NN	O	O
human	NN	O	O
cells	NN	O	O
described	NN	O	O
so	NN	O	O
far	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
dendritic	NN	O	O
cell	NN	O	O
maturation	NN	O	O
is	NN	O	O
an	NN	O	O
important	NN	O	O
target	NN	O	O
of	NN	O	O
sulfasalazine	NN	O	O
.	NN	O	O

Because	NN	O	O
of	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
dendritic	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
(	NN	O	O
auto	NN	O	O
)	NN	O	O
immunity	NN	O	O
,	NN	O	O
inhibition	NN	O	O
of	NN	O	O
their	NN	O	O
maturation	NN	O	O
might	NN	O	O
provide	NN	O	O
a	NN	O	O
target	NN	O	O
for	NN	O	O
further	NN	O	O
optimization	NN	O	O
of	NN	O	O
sulfasalazine	NN	O	O
therapy	NN	O	O
.	NN	O	O

-DOCSTART-	O

Defective	NN	O	O
function	NN	O	O
of	NN	O	O
the	NN	O	O
proteasome	NN	O	B-protein
in	NN	O	O
autoimmunity	NN	O	O
:	NN	O	O
involvement	NN	O	O
of	NN	O	O
impaired	NN	O	O
NF-kappaB	NN	O	O
activation	NN	O	O
.	NN	O	O

Type	NN	O	O
1	NN	O	O
diabetes	NN	O	O
(	NN	O	O
also	NN	O	O
known	NN	O	O
as	NN	O	O
insulin-dependent	NN	O	O
diabetes	NN	O	O
mellitus	NN	O	O
or	NN	O	O
juvenile-onset	NN	O	O
diabetes	NN	O	O
)	NN	O	O
is	NN	O	O
usually	NN	O	O
caused	NN	O	O
by	NN	O	O
T	NN	O	O
cell-mediated	NN	O	O
autoimmunity	NN	O	O
,	NN	O	O
with	NN	O	O
a	NN	O	O
prediabetic	NN	O	O
state	NN	O	O
characterized	NN	O	O
by	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
autoantibodies	NN	O	B-protein
specific	NN	O	O
for	NN	O	O
proteins	NN	O	O
expressed	NN	O	O
by	NN	O	O
pancreatic	NN	O	B-cell_type
beta	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
nonobese	NN	O	O
patient	NN	O	O
with	NN	O	O
diabetes	NN	O	O
(	NN	O	O
NOD	NN	O	O
)	NN	O	O
mouse	NN	O	O
is	NN	O	O
a	NN	O	O
spontaneous	NN	O	O
model	NN	O	O
of	NN	O	O
type	NN	O	O
1	NN	O	O
diabetes	NN	O	O
with	NN	O	O
a	NN	O	O
strong	NN	O	O
genetic	NN	O	O
component	NN	O	O
that	NN	O	O
maps	NN	O	O
to	NN	O	O
the	NN	O	O
major	NN	O	B-DNA
histocompatibility	NN	O	I-DNA
complex	NN	O	I-DNA
(	NN	O	I-DNA
MHC	NN	O	I-DNA
)	NN	O	I-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
genome	NN	O	B-DNA
.	NN	O	O

A	NN	O	O
specific	NN	O	O
proteasome	NN	O	B-protein
defect	NN	O	O
has	NN	O	O
been	NN	O	O
identified	NN	O	O
in	NN	O	O
NOD	NN	O	O
mouse	NN	O	O
in	NN	O	O
select	NN	O	O
lymphocytic	NN	O	B-cell_line
and	NN	O	I-cell_line
monocytic	NN	O	I-cell_line
lineages	NN	O	I-cell_line
that	NN	O	O
results	NN	O	O
from	NN	O	O
down-regulation	NN	O	O
of	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
proteasome	NN	O	B-protein
subunit	NN	O	I-protein
LMP2	NN	O	I-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
encoded	NN	O	O
by	NN	O	O
a	NN	O	O
gene	NN	O	O
in	NN	O	O
the	NN	O	O
MHC	NN	O	B-DNA
genomic	NN	O	I-DNA
region	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
defect	NN	O	O
prevents	NN	O	O
the	NN	O	O
proteolytic	NN	O	O
processing	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
production	NN	O	O
and	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
nuclear	NN	O	I-protein
factor-kappaB	NN	O	I-protein
(	NN	O	O
NF-kappaB	NN	O	B-protein
)	NN	O	O
,	NN	O	O
which	NN	O	O
plays	NN	O	O
important	NN	O	O
roles	NN	O	O
in	NN	O	O
immune	NN	O	O
and	NN	O	O
inflammatory	NN	O	O
responses	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
increases	NN	O	O
the	NN	O	O
susceptibility	NN	O	O
of	NN	O	O
the	NN	O	O
affected	NN	O	O
cells	NN	O	O
to	NN	O	O
apoptosis	NN	O	O
induced	NN	O	O
by	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
.	NN	O	O

The	NN	O	O
novel	NN	O	O
role	NN	O	O
of	NN	O	O
the	NN	O	O
proteasome	NN	O	B-protein
in	NN	O	O
dysfunction	NN	O	O
in	NN	O	O
autoimmunity	NN	O	O
is	NN	O	O
presented	NN	O	O
and	NN	O	O
documented	NN	O	O
to	NN	O	O
be	NN	O	O
both	NN	O	O
tissue	NN	O	O
and	NN	O	O
developmental	NN	O	O
stage	NN	O	O
specific	NN	O	O
.	NN	O	O

We	NN	O	O
propose	NN	O	O
a	NN	O	O
role	NN	O	O
of	NN	O	O
the	NN	O	O
proteasome	NN	O	B-protein
as	NN	O	O
a	NN	O	O
step	NN	O	O
in	NN	O	O
disease	NN	O	O
pathogenesis	NN	O	O
and	NN	O	O
tissue	NN	O	O
targeting	NN	O	O
.	NN	O	O

-DOCSTART-	O

Down-regulation	NN	O	O
of	NN	O	O
TDT	NN	O	O
transcription	NN	O	O
in	NN	O	O
CD4	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
CD8	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
thymocytes	NN	O	I-cell_type
by	NN	O	O
Ikaros	NN	O	B-protein
proteins	NN	O	I-protein
in	NN	O	O
direct	NN	O	O
competition	NN	O	O
with	NN	O	O
an	NN	O	O
Ets	NN	O	B-protein
activator	NN	O	I-protein
.	NN	O	O

Ikaros	NN	O	B-protein
is	NN	O	O
a	NN	O	O
unique	NN	O	O
regulator	NN	O	O
of	NN	O	O
lymphopoiesis	NN	O	O
that	NN	O	O
associates	NN	O	O
with	NN	O	O
pericentromeric	NN	O	B-DNA
heterochromatin	NN	O	I-DNA
and	NN	O	O
has	NN	O	O
been	NN	O	O
implicated	NN	O	O
in	NN	O	O
heritable	NN	O	O
gene	NN	O	O
inactivation	NN	O	O
.	NN	O	O

Binding	NN	O	O
and	NN	O	O
competition	NN	O	O
experiments	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
Ikaros	NN	O	B-protein
dimers	NN	O	I-protein
compete	NN	O	O
with	NN	O	O
an	NN	O	O
Ets	NN	O	B-protein
activator	NN	O	I-protein
for	NN	O	O
occupancy	NN	O	O
of	NN	O	O
the	NN	O	O
lymphocyte-specific	NN	O	B-DNA
TdT	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Mutations	NN	O	O
that	NN	O	O
selectively	NN	O	O
disrupt	NN	O	O
Ikaros	NN	O	O
binding	NN	O	O
to	NN	O	O
an	NN	O	O
integrated	NN	O	B-DNA
TdT	NN	O	I-DNA
promoter	NN	O	I-DNA
had	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
promoter	NN	O	O
function	NN	O	O
in	NN	O	O
a	NN	O	O
CD4	NN	O	B-cell_line
(	NN	O	I-cell_line
+	NN	O	I-cell_line
)	NN	O	I-cell_line
CD8	NN	O	I-cell_line
(	NN	O	I-cell_line
+	NN	O	I-cell_line
)	NN	O	I-cell_line
thymocyte	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

However	NN	O	O
,	NN	O	O
these	NN	O	O
mutations	NN	O	O
abolished	NN	O	O
down-regulation	NN	O	O
on	NN	O	O
differentiation	NN	O	O
,	NN	O	O
providing	NN	O	O
evidence	NN	O	O
that	NN	O	O
Ikaros	NN	O	B-protein
plays	NN	O	O
a	NN	O	O
direct	NN	O	O
role	NN	O	O
in	NN	O	O
repression	NN	O	O
.	NN	O	O

Reduced	NN	O	O
access	NN	O	O
to	NN	O	O
restriction	NN	O	O
enzyme	NN	O	O
cleavage	NN	O	O
suggested	NN	O	O
that	NN	O	O
chromatin	NN	O	O
alterations	NN	O	O
accompany	NN	O	O
down-regulation	NN	O	O
.	NN	O	O

The	NN	O	O
Ikaros-dependent	NN	O	O
down-regulation	NN	O	O
event	NN	O	O
and	NN	O	O
the	NN	O	O
observed	NN	O	O
chromatin	NN	O	O
alterations	NN	O	O
appear	NN	O	O
to	NN	O	O
precede	NN	O	O
pericentromeric	NN	O	O
repositioning	NN	O	O
.	NN	O	O

Current	NN	O	O
models	NN	O	O
propose	NN	O	O
that	NN	O	O
the	NN	O	O
functions	NN	O	O
of	NN	O	O
Ikaros	NN	O	B-protein
should	NN	O	O
be	NN	O	O
disrupted	NN	O	O
by	NN	O	O
a	NN	O	O
small	NN	O	O
isoform	NN	O	B-protein
that	NN	O	O
retains	NN	O	O
the	NN	O	O
dimerization	NN	O	B-protein
domain	NN	O	I-protein
and	NN	O	O
lacks	NN	O	O
the	NN	O	O
DNA-binding	NN	O	B-protein
domain	NN	O	I-protein
.	NN	O	O

Surprisingly	NN	O	O
,	NN	O	O
in	NN	O	O
the	NN	O	O
CD4	NN	O	B-cell_line
(	NN	O	I-cell_line
+	NN	O	I-cell_line
)	NN	O	I-cell_line
CD8	NN	O	I-cell_line
(	NN	O	I-cell_line
+	NN	O	I-cell_line
)	NN	O	I-cell_line
thymocyte	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
overexpression	NN	O	O
of	NN	O	O
a	NN	O	O
small	NN	O	B-protein
Ikaros	NN	O	I-protein
isoform	NN	O	I-protein
had	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
differentiation	NN	O	O
or	NN	O	O
on	NN	O	O
the	NN	O	O
pericentromeric	NN	O	O
targeting	NN	O	O
and	NN	O	O
DNA-binding	NN	O	O
properties	NN	O	O
of	NN	O	O
Ikaros	NN	O	B-protein
.	NN	O	O

Rather	NN	O	O
,	NN	O	O
the	NN	O	O
small	NN	O	O
isoform	NN	O	B-protein
assembled	NN	O	O
into	NN	O	O
multimeric	NN	O	B-protein
complexes	NN	O	I-protein
with	NN	O	O
DNA-bound	NN	O	B-protein
Ikaros	NN	O	I-protein
at	NN	O	O
the	NN	O	O
pericentromeric	NN	O	O
foci	NN	O	O
.	NN	O	O

The	NN	O	O
capacity	NN	O	O
for	NN	O	O
in	NN	O	O
vivo	NN	O	O
multimer	NN	O	O
formation	NN	O	O
suggests	NN	O	O
that	NN	O	O
interactions	NN	O	O
between	NN	O	O
Ikaros	NN	O	B-protein
dimers	NN	O	I-protein
bound	NN	O	O
to	NN	O	O
the	NN	O	O
TdT	NN	O	B-DNA
promoter	NN	O	I-DNA
and	NN	O	O
those	NN	O	O
bound	NN	O	O
to	NN	O	O
pericentromeric	NN	O	B-DNA
repeat	NN	O	I-DNA
sequences	NN	O	I-DNA
may	NN	O	O
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
pericentromeric	NN	O	O
repositioning	NN	O	O
of	NN	O	O
the	NN	O	O
inactive	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Type	NN	O	B-protein
I	NN	O	I-protein
interferons	NN	O	I-protein
and	NN	O	O
IL-12	NN	O	B-protein
:	NN	O	O
convergence	NN	O	O
and	NN	O	O
cross-regulation	NN	O	O
among	NN	O	O
mediators	NN	O	O
of	NN	O	O
cellular	NN	O	O
immunity	NN	O	O
.	NN	O	O

Therapeutic	NN	O	O
use	NN	O	O
of	NN	O	O
type	NN	O	B-protein
I	NN	O	I-protein
IFN	NN	O	I-protein
(	NN	O	O
IFN-alpha/beta	NN	O	B-protein
)	NN	O	O
has	NN	O	O
become	NN	O	O
common	NN	O	O
.	NN	O	O

Many	NN	O	O
of	NN	O	O
the	NN	O	O
diverse	NN	O	O
diseases	NN	O	O
targeted	NN	O	O
are	NN	O	O
marked	NN	O	O
by	NN	O	O
pathogenetic	NN	O	O
abnormalities	NN	O	O
in	NN	O	O
cell-mediated	NN	O	O
immunity	NN	O	O
(	NN	O	O
CMI	NN	O	O
)	NN	O	O
,	NN	O	O
these	NN	O	O
cellular	NN	O	O
immune	NN	O	O
responses	NN	O	O
either	NN	O	O
causing	NN	O	O
injury	NN	O	O
to	NN	O	O
the	NN	O	O
host	NN	O	O
,	NN	O	O
lacking	NN	O	O
sufficient	NN	O	O
vigor	NN	O	O
for	NN	O	O
virus	NN	O	O
or	NN	O	O
tumor	NN	O	O
clearance	NN	O	O
,	NN	O	O
or	NN	O	O
both	NN	O	O
.	NN	O	O

In	NN	O	O
general	NN	O	O
,	NN	O	O
therapeutic	NN	O	O
efficacy	NN	O	O
is	NN	O	O
limited	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
thus	NN	O	O
notable	NN	O	O
that	NN	O	O
the	NN	O	O
pleiotropic	NN	O	O
effects	NN	O	O
of	NN	O	O
type	NN	O	B-protein
I	NN	O	I-protein
IFN	NN	O	I-protein
on	NN	O	O
CMI	NN	O	O
remain	NN	O	O
poorly	NN	O	O
understood	NN	O	O
.	NN	O	O

We	NN	O	O
characterized	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
type	NN	O	B-protein
I	NN	O	I-protein
IFN	NN	O	I-protein
on	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
IL-12	NN	O	B-protein
,	NN	O	O
the	NN	O	O
central	NN	O	B-protein
immunoregulatory	NN	O	I-protein
cytokine	NN	O	I-protein
of	NN	O	O
the	NN	O	O
CD4	NN	O	B-protein
(	NN	O	I-protein
+	NN	O	I-protein
)	NN	O	I-protein
T	NN	O	I-protein
cell	NN	O	I-protein
arm	NN	O	I-protein
of	NN	O	O
CMI	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
type	NN	O	B-protein
I	NN	O	I-protein
IFN	NN	O	I-protein
are	NN	O	O
potent	NN	O	O
inhibitors	NN	O	O
of	NN	O	O
IL-12	NN	O	O
production	NN	O	O
by	NN	O	O
human	NN	O	B-cell_type
monocytes/macrophages	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
underlying	NN	O	O
mechanism	NN	O	O
involves	NN	O	O
transcriptional	NN	O	O
inhibition	NN	O	O
of	NN	O	O
the	NN	O	O
IL-12p40	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	O
marked	NN	O	O
by	NN	O	O
down-regulation	NN	O	O
of	NN	O	O
PU.1	NN	O	O
binding	NN	O	O
activity	NN	O	O
at	NN	O	O
the	NN	O	O
upstream	NN	O	B-DNA
Ets	NN	O	I-DNA
site	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
IL-12p40	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Type	NN	O	B-protein
I	NN	O	I-protein
IFN	NN	O	I-protein
have	NN	O	O
previously	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
able	NN	O	O
to	NN	O	O
substitute	NN	O	O
for	NN	O	O
IL-12	NN	O	B-protein
in	NN	O	O
driving	NN	O	O
IFN-gamma	NN	O	O
production	NN	O	O
from	NN	O	O
T	NN	O	B-cell_type
and	NN	O	I-cell_type
NK	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
ability	NN	O	O
of	NN	O	O
IFN-alpha/beta	NN	O	B-protein
to	NN	O	O
suppress	NN	O	O
IL-12	NN	O	O
production	NN	O	O
while	NN	O	O
up-regulating	NN	O	O
IFN-gamma	NN	O	O
production	NN	O	O
suggests	NN	O	O
a	NN	O	O
possible	NN	O	O
mechanistic	NN	O	O
basis	NN	O	O
for	NN	O	O
the	NN	O	O
difficulties	NN	O	O
of	NN	O	O
employing	NN	O	O
these	NN	O	O
cytokines	NN	O	B-protein
in	NN	O	O
diseases	NN	O	O
involving	NN	O	O
abnormalities	NN	O	O
of	NN	O	O
CMI	NN	O	O
.	NN	O	O

-DOCSTART-	O

Nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
suppressive	NN	O	O
and	NN	O	O
inhibitor-kappaB	NN	O	B-protein
stimulatory	NN	O	O
effects	NN	O	O
of	NN	O	O
troglitazone	NN	O	O
in	NN	O	O
obese	NN	O	O
patients	NN	O	O
with	NN	O	O
type	NN	O	O
2	NN	O	O
diabetes	NN	O	O
:	NN	O	O
evidence	NN	O	O
of	NN	O	O
an	NN	O	O
antiinflammatory	NN	O	O
action	NN	O	O
?	NN	O	O

It	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
recently	NN	O	O
that	NN	O	O
troglitazone	NN	O	O
exerts	NN	O	O
an	NN	O	O
anti-inflammatory	NN	O	O
effect	NN	O	O
,	NN	O	O
in	NN	O	O
vitro	NN	O	O
,	NN	O	O
and	NN	O	O
in	NN	O	O
experimental	NN	O	O
animals	NN	O	O
.	NN	O	O

To	NN	O	O
test	NN	O	O
these	NN	O	O
properties	NN	O	O
in	NN	O	O
humans	NN	O	O
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
troglitazone	NN	O	O
on	NN	O	O
the	NN	O	O
proinflammatory	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
nuclear	NN	O	I-protein
factor-kappaB	NN	O	I-protein
and	NN	O	O
its	NN	O	O
inhibitory	NN	O	B-protein
protein	NN	O	I-protein
IkappaB	NN	O	I-protein
in	NN	O	O
mononuclear	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
MNC	NN	O	B-cell_type
)	NN	O	O
and	NN	O	O
plasma	NN	O	B-protein
soluble	NN	O	I-protein
intracellular	NN	O	I-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
,	NN	O	O
monocyte	NN	O	B-protein
chemoattractant	NN	O	I-protein
protein-1	NN	O	I-protein
,	NN	O	O
plasminogen	NN	O	B-protein
activator	NN	O	I-protein
inhibitor-1	NN	O	I-protein
,	NN	O	O
and	NN	O	O
C-reactive	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

We	NN	O	O
also	NN	O	O
examined	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
troglitazone	NN	O	O
on	NN	O	O
reactive	NN	O	O
oxygen	NN	O	O
species	NN	O	O
generation	NN	O	O
,	NN	O	O
p47	NN	O	O
(	NN	O	O
phox	NN	O	O
)	NN	O	O
subunit	NN	O	O
expression	NN	O	O
,	NN	O	O
9-hydroxyoctadecadienoic	NN	O	O
acid	NN	O	O
(	NN	O	O
9-HODE	NN	O	O
)	NN	O	O
,	NN	O	O
13-HODE	NN	O	O
,	NN	O	O
o-tyrosine	NN	O	O
,	NN	O	O
and	NN	O	O
m-tyrosine	NN	O	O
in	NN	O	O
obese	NN	O	O
patients	NN	O	O
with	NN	O	O
type	NN	O	O
2	NN	O	O
diabetes	NN	O	O
.	NN	O	O

Seven	NN	O	O
obese	NN	O	O
patients	NN	O	O
with	NN	O	O
type	NN	O	O
2	NN	O	O
diabetes	NN	O	O
were	NN	O	O
treated	NN	O	O
with	NN	O	O
troglitazone	NN	O	O
(	NN	O	O
400	NN	O	O
mg/day	NN	O	O
)	NN	O	O
for	NN	O	O
4	NN	O	O
weeks	NN	O	O
.	NN	O	O

Blood	NN	O	O
samples	NN	O	O
were	NN	O	O
obtained	NN	O	O
at	NN	O	O
weekly	NN	O	O
intervals	NN	O	O
.	NN	O	O

Nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
binding	NN	O	O
activity	NN	O	O
in	NN	O	O
MNC	NN	O	B-cell_type
nuclear	NN	O	O
extracts	NN	O	O
was	NN	O	O
significantly	NN	O	O
inhibited	NN	O	O
after	NN	O	O
troglitazone	NN	O	O
treatment	NN	O	O
at	NN	O	O
week	NN	O	O
1	NN	O	O
and	NN	O	O
continued	NN	O	O
to	NN	O	O
be	NN	O	O
inhibited	NN	O	O
up	NN	O	O
to	NN	O	O
week	NN	O	O
4	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
IkappaB	NN	O	O
protein	NN	O	O
levels	NN	O	O
increased	NN	O	O
significantly	NN	O	O
after	NN	O	O
troglitazone	NN	O	O
treatment	NN	O	O
at	NN	O	O
week	NN	O	O
1	NN	O	O
,	NN	O	O
and	NN	O	O
this	NN	O	O
increase	NN	O	O
persisted	NN	O	O
throughout	NN	O	O
the	NN	O	O
study	NN	O	O
.	NN	O	O

Plasma	NN	O	B-protein
monocyte	NN	O	I-protein
chemoattractant	NN	O	I-protein
protein-1	NN	O	I-protein
and	NN	O	O
soluble	NN	O	B-protein
intracellular	NN	O	I-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
concentrations	NN	O	O
did	NN	O	O
not	NN	O	O
decrease	NN	O	O
significantly	NN	O	O
after	NN	O	O
troglitazone	NN	O	O
treatment	NN	O	O
,	NN	O	O
although	NN	O	O
there	NN	O	O
was	NN	O	O
a	NN	O	O
trend	NN	O	O
toward	NN	O	O
inhibition	NN	O	O
.	NN	O	O

Reactive	NN	O	O
oxygen	NN	O	O
species	NN	O	O
generation	NN	O	O
by	NN	O	O
polymorphonuclear	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
MNC	NN	O	B-cell_type
,	NN	O	O
p47	NN	O	B-protein
(	NN	O	I-protein
phox	NN	O	I-protein
)	NN	O	I-protein
subunit	NN	O	I-protein
protein	NN	O	I-protein
quantities	NN	O	O
,	NN	O	O
plasminogen	NN	O	B-protein
activator	NN	O	I-protein
inhibitor-1	NN	O	I-protein
,	NN	O	O
and	NN	O	O
C-reactive	NN	O	B-protein
protein	NN	O	I-protein
levels	NN	O	O
decreased	NN	O	O
significantly	NN	O	O
after	NN	O	O
troglitazone	NN	O	O
intake	NN	O	O
.	NN	O	O

13-HODE/linoleic	NN	O	O
acid	NN	O	O
and	NN	O	O
9-HODE/linoleic	NN	O	O
acid	NN	O	O
ratios	NN	O	O
also	NN	O	O
decreased	NN	O	O
after	NN	O	O
troglitazone	NN	O	O
intake	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
o-tyrosine/phenylalanine	NN	O	O
and	NN	O	O
m-tyrosine/phenylalanine	NN	O	O
ratios	NN	O	O
did	NN	O	O
not	NN	O	O
change	NN	O	O
significantly	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
show	NN	O	O
that	NN	O	O
troglitazone	NN	O	O
has	NN	O	O
profound	NN	O	O
antiinflammatory	NN	O	O
effects	NN	O	O
in	NN	O	O
addition	NN	O	O
to	NN	O	O
antioxidant	NN	O	O
effects	NN	O	O
in	NN	O	O
obese	NN	O	O
type	NN	O	O
2	NN	O	O
diabetics	NN	O	O
;	NN	O	O
these	NN	O	O
effects	NN	O	O
may	NN	O	O
be	NN	O	O
relevant	NN	O	O
to	NN	O	O
the	NN	O	O
recently	NN	O	O
described	NN	O	O
beneficial	NN	O	O
antiatherosclerotic	NN	O	O
effects	NN	O	O
of	NN	O	O
troglitazone	NN	O	O
at	NN	O	O
the	NN	O	O
vascular	NN	O	O
level	NN	O	O
.	NN	O	O

-DOCSTART-	O

OX40	NN	O	O
stimulation	NN	O	O
by	NN	O	O
gp34/OX40	NN	O	B-protein
ligand	NN	O	I-protein
enhances	NN	O	O
productive	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
infection	NN	O	O
.	NN	O	O

OX40	NN	O	B-protein
is	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	I-protein
TNF	NN	O	I-protein
)	NN	O	I-protein
receptor	NN	O	I-protein
superfamily	NN	O	I-protein
and	NN	O	O
known	NN	O	O
to	NN	O	O
be	NN	O	O
an	NN	O	O
important	NN	O	O
costimulatory	NN	O	B-protein
molecule	NN	O	I-protein
expressed	NN	O	O
on	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
costimulation	NN	O	O
of	NN	O	O
OX40	NN	O	B-protein
in	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
(	NN	O	O
HIV-1	NN	O	O
)	NN	O	O
infection	NN	O	O
by	NN	O	O
its	NN	O	O
natural	NN	O	O
ligand	NN	O	O
,	NN	O	O
gp34	NN	O	B-protein
,	NN	O	O
the	NN	O	O
OX40-transfected	NN	O	B-cell_line
ACH-2	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	I-cell_line
ACH-2/OX40	NN	O	I-cell_line
,	NN	O	O
chronically	NN	O	O
infected	NN	O	O
with	NN	O	O
HIV-1	NN	O	O
,	NN	O	O
was	NN	O	O
cocultured	NN	O	O
with	NN	O	O
paraformaldehyde	NN	O	O
(	NN	O	O
PFA	NN	O	O
)	NN	O	O
-fixed	NN	O	O
gp34-transfected	NN	O	B-cell_line
mouse	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	I-cell_line
SV-T2/gp34	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
results	NN	O	O
showed	NN	O	O
that	NN	O	O
HIV-1	NN	O	O
production	NN	O	O
was	NN	O	O
strongly	NN	O	O
induced	NN	O	O
.	NN	O	O

This	NN	O	O
was	NN	O	O
followed	NN	O	O
by	NN	O	O
apparent	NN	O	O
apoptosis	NN	O	O
,	NN	O	O
and	NN	O	O
both	NN	O	O
processes	NN	O	O
were	NN	O	O
specifically	NN	O	O
inhibited	NN	O	O
by	NN	O	O
the	NN	O	O
gp34	NN	O	B-protein
-specific	NN	O	O
neutralizing	NN	O	B-protein
monoclonal	NN	O	I-protein
antibody	NN	O	I-protein
5A8	NN	O	I-protein
.	NN	O	O

Endogenous	NN	O	O
TNF	NN	O	B-protein
alpha	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
and	NN	O	O
TNF-beta	NN	O	B-protein
production	NN	O	O
were	NN	O	O
not	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
enhanced	NN	O	O
HIV-1	NN	O	O
production	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
enhanced	NN	O	O
HIV-1	NN	O	O
transcription	NN	O	O
in	NN	O	O
gp34	NN	O	B-protein
-stimulated	NN	O	O
ACH-2/OX40	NN	O	B-cell_line
cells	NN	O	I-cell_line
was	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
the	NN	O	O
OX40-gp34	NN	O	O
interaction	NN	O	O
activated	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
consisting	NN	O	O
of	NN	O	O
p50	NN	O	B-protein
and	NN	O	I-protein
p65	NN	O	I-protein
subunits	NN	O	I-protein
.	NN	O	O

When	NN	O	O
primary	NN	O	O
activated	NN	O	O
CD4	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
acutely	NN	O	O
infected	NN	O	O
with	NN	O	O
HIV-1	NN	O	O
(	NN	O	O
NL4-3	NN	O	O
)	NN	O	O
(	NN	O	O
CXCR4-using	NN	O	B-cell_line
T-cell-line-tropic	NN	O	I-cell_line
)	NN	O	O
were	NN	O	O
cocultured	NN	O	O
with	NN	O	O
PFA-fixed	NN	O	O
gp34	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
human	NN	O	I-cell_type
T-cell	NN	O	I-cell_type
leukemia	NN	O	O
virus	NN	O	O
type	NN	O	O
1-bearing	NN	O	O
MT-2	NN	O	B-cell_line
cells	NN	O	I-cell_line
or	NN	O	O
SV-T2/gp34	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
HIV-1	NN	O	O
production	NN	O	O
was	NN	O	O
also	NN	O	O
markedly	NN	O	O
enhanced	NN	O	O
.	NN	O	O

The	NN	O	O
enhancement	NN	O	O
was	NN	O	O
again	NN	O	O
significantly	NN	O	O
inhibited	NN	O	O
by	NN	O	O
5A8	NN	O	B-protein
.	NN	O	O

The	NN	O	O
present	NN	O	O
study	NN	O	O
first	NN	O	O
shows	NN	O	O
that	NN	O	O
OX40-gp34	NN	O	O
interaction	NN	O	O
stimulates	NN	O	O
HIV-1	NN	O	O
expression	NN	O	O
and	NN	O	O
suggests	NN	O	O
that	NN	O	O
OX40	NN	O	O
triggering	NN	O	O
by	NN	O	O
gp34	NN	O	B-protein
may	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
enhancing	NN	O	O
HIV-1	NN	O	O
production	NN	O	O
in	NN	O	O
both	NN	O	O
acutely	NN	O	O
and	NN	O	O
latently	NN	O	O
infected	NN	O	O
CD4	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

-DOCSTART-	O

Troglitazone	NN	O	O
,	NN	O	O
a	NN	O	O
PPARgamma	NN	O	O
ligand	NN	O	O
,	NN	O	O
inhibits	NN	O	O
osteopontin	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes/macrophage	NN	O	I-cell_type
THP-1	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Peroxizome	NN	O	B-protein
proliferator-activated	NN	O	I-protein
receptor-gamma	NN	O	I-protein
(	NN	O	O
PPARgamma	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
receptor	NN	O	I-protein
family	NN	O	I-protein
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
that	NN	O	O
regulate	NN	O	O
adipocyte	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Recent	NN	O	O
studies	NN	O	O
indicate	NN	O	O
that	NN	O	O
liganded	NN	O	B-protein
PPARgamma	NN	O	I-protein
not	NN	O	O
only	NN	O	O
promotes	NN	O	O
differentiation	NN	O	O
but	NN	O	O
also	NN	O	O
inhibits	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
macrophages	NN	O	B-cell_type
.	NN	O	O

Osteopontin	NN	O	B-protein
,	NN	O	O
a	NN	O	O
component	NN	O	O
of	NN	O	O
extracellular	NN	O	O
matrix	NN	O	O
,	NN	O	O
is	NN	O	O
synthesized	NN	O	O
by	NN	O	O
macrophages	NN	O	B-cell_type
in	NN	O	O
atherosclerotic	NN	O	O
plaques	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
examined	NN	O	O
whether	NN	O	O
PPARgamma	NN	O	B-protein
ligand	NN	O	O
regulates	NN	O	O
osteopontin	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
THP-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
a	NN	O	O
cell	NN	O	O
line	NN	O	O
derived	NN	O	O
from	NN	O	O
human	NN	O	B-cell_type
monocytic	NN	O	I-cell_type
leukemia	NN	O	I-cell_type
cells	NN	O	I-cell_type
which	NN	O	O
can	NN	O	O
differentiate	NN	O	O
to	NN	O	O
macrophage	NN	O	B-cell_type
upon	NN	O	O
stimulation	NN	O	O
with	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
PMA	NN	O	O
.	NN	O	O

Northern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
showed	NN	O	O
that	NN	O	O
osteopontin	NN	O	O
expression	NN	O	O
is	NN	O	O
markedly	NN	O	O
induced	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
PMA	NN	O	O
.	NN	O	O

Troglitazone	NN	O	O
,	NN	O	O
a	NN	O	O
PPARgamma	NN	O	B-protein
ligand	NN	O	O
,	NN	O	O
dramatically	NN	O	O
attenuated	NN	O	O
the	NN	O	O
PMA-induced	NN	O	O
osteopontin	NN	O	O
expression	NN	O	O
.	NN	O	O

Transient	NN	O	O
transfection	NN	O	O
assays	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
osteopontin	NN	O	I-DNA
promoter/luciferase	NN	O	I-DNA
construct	NN	O	I-DNA
which	NN	O	O
contains	NN	O	O
a	NN	O	O
5'-flanking	NN	O	B-DNA
region	NN	O	I-DNA
between	NN	O	O
-1500	NN	O	O
and	NN	O	O
+87	NN	O	O
relative	NN	O	O
to	NN	O	O
the	NN	O	O
transcription	NN	O	B-DNA
start	NN	O	I-DNA
site	NN	O	I-DNA
demonstrate	NN	O	O
that	NN	O	O
either	NN	O	O
treatment	NN	O	O
with	NN	O	O
troglitazone	NN	O	O
or	NN	O	O
cotransfection	NN	O	O
of	NN	O	O
PPARgamma	NN	O	B-DNA
expression	NN	O	I-DNA
vector	NN	O	I-DNA
inhibits	NN	O	O
osteopontin	NN	O	O
promoter	NN	O	O
activity	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
troglitazone	NN	O	O
reduces	NN	O	O
osteopontin	NN	O	O
gene	NN	O	O
expression	NN	O	O
at	NN	O	O
transcriptional	NN	O	O
level	NN	O	O
through	NN	O	O
PPARgamma	NN	O	B-protein
activation	NN	O	O
,	NN	O	O
and	NN	O	O
suggest	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
troglitazone	NN	O	O
in	NN	O	O
inhibiting	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
macrophages	NN	O	B-cell_type
to	NN	O	O
produce	NN	O	O
extracellular	NN	O	O
matrix	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
particularly	NN	O	O
relevant	NN	O	O
to	NN	O	O
atherosclerotic	NN	O	O
plaque	NN	O	O
formation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
regulates	NN	O	O
Ig	NN	O	B-protein
lambda	NN	O	I-protein
light	NN	O	O
chain	NN	O	O
gene	NN	O	O
rearrangement	NN	O	O
.	NN	O	O

The	NN	O	O
tissue-	NN	O	O
and	NN	O	O
stage-specific	NN	O	O
assembly	NN	O	O
of	NN	O	O
Ig	NN	O	B-DNA
and	NN	O	I-DNA
TCR	NN	O	I-DNA
genes	NN	O	I-DNA
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
a	NN	O	O
common	NN	O	O
V	NN	O	B-protein
(	NN	O	I-protein
D	NN	O	I-protein
)	NN	O	I-protein
J	NN	O	I-protein
recombinase	NN	O	I-protein
complex	NN	O	I-protein
in	NN	O	O
precursor	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Directed	NN	O	O
alterations	NN	O	O
in	NN	O	O
the	NN	O	O
accessibility	NN	O	O
of	NN	O	O
V	NN	O	B-DNA
,	NN	O	I-DNA
D	NN	O	I-DNA
,	NN	O	I-DNA
and	NN	O	I-DNA
J	NN	O	I-DNA
gene	NN	O	I-DNA
segments	NN	O	I-DNA
target	NN	O	O
the	NN	O	O
recombinase	NN	O	B-protein
to	NN	O	O
specific	NN	O	O
Ag	NN	O	B-DNA
receptor	NN	O	I-DNA
loci	NN	O	I-DNA
.	NN	O	O

Accessibility	NN	O	O
within	NN	O	O
a	NN	O	O
given	NN	O	O
locus	NN	O	B-DNA
is	NN	O	O
regulated	NN	O	O
by	NN	O	O
the	NN	O	O
functional	NN	O	O
interaction	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
with	NN	O	O
cognate	NN	O	B-DNA
enhancer	NN	O	I-DNA
elements	NN	O	I-DNA
and	NN	O	O
correlates	NN	O	O
with	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
unrearranged	NN	O	O
gene	NN	O	B-DNA
segments	NN	O	I-DNA
.	NN	O	O

As	NN	O	O
demonstrated	NN	O	O
in	NN	O	O
our	NN	O	O
prior	NN	O	O
studies	NN	O	O
,	NN	O	O
rearrangement	NN	O	O
of	NN	O	O
the	NN	O	O
Igkappa	NN	O	B-DNA
locus	NN	O	I-DNA
is	NN	O	O
regulated	NN	O	O
by	NN	O	O
the	NN	O	O
inducible	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
NF-kappaB	NN	O	B-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
the	NN	O	O
Igkappa	NN	O	B-DNA
locus	NN	O	I-DNA
,	NN	O	O
known	NN	O	O
transcriptional	NN	O	B-DNA
control	NN	O	I-DNA
elements	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
Iglambda	NN	O	B-DNA
locus	NN	O	I-DNA
lack	NN	O	O
functional	NN	O	O
NF-kappaB	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
.	NN	O	O

Consistent	NN	O	O
with	NN	O	O
this	NN	O	O
observation	NN	O	O
,	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
assembled	NN	O	O
Iglambda	NN	O	B-DNA
genes	NN	O	I-DNA
in	NN	O	O
mature	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
NF-kappaB	NN	O	B-protein
independent	NN	O	O
.	NN	O	O

Nonetheless	NN	O	O
,	NN	O	O
we	NN	O	O
now	NN	O	O
show	NN	O	O
that	NN	O	O
specific	NN	O	O
repression	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
inhibits	NN	O	O
germline	NN	O	O
transcription	NN	O	O
and	NN	O	O
recombination	NN	O	O
of	NN	O	O
Iglambda	NN	O	B-DNA
gene	NN	O	I-DNA
segments	NN	O	I-DNA
in	NN	O	O
precursor	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Molecular	NN	O	O
analyses	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
block	NN	O	O
in	NN	O	O
NF-kappaB	NN	O	B-protein
impairs	NN	O	O
Iglambda	NN	O	O
rearrangement	NN	O	O
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
recombinase	NN	O	B-protein
accessibility	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
the	NN	O	O
activities	NN	O	O
of	NN	O	O
known	NN	O	O
Iglambda	NN	O	B-DNA
promoter	NN	O	I-DNA
and	NN	O	O
enhancer	NN	O	B-DNA
elements	NN	O	I-DNA
are	NN	O	O
unaffected	NN	O	O
in	NN	O	O
the	NN	O	O
same	NN	O	O
cellular	NN	O	O
background	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
expand	NN	O	O
the	NN	O	O
range	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
action	NN	O	O
in	NN	O	O
precursor	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
beyond	NN	O	O
Igkappa	NN	O	B-protein
to	NN	O	O
include	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
recombinational	NN	O	O
accessibility	NN	O	O
at	NN	O	O
both	NN	O	O
L	NN	O	B-DNA
chain	NN	O	I-DNA
loci	NN	O	I-DNA
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
our	NN	O	O
results	NN	O	O
strongly	NN	O	O
suggest	NN	O	O
the	NN	O	O
existence	NN	O	O
of	NN	O	O
a	NN	O	O
novel	NN	O	O
Iglambda	NN	O	B-DNA
regulatory	NN	O	I-DNA
element	NN	O	I-DNA
that	NN	O	O
is	NN	O	O
either	NN	O	O
directly	NN	O	O
or	NN	O	O
indirectly	NN	O	O
activated	NN	O	O
by	NN	O	O
NF-kappaB	NN	O	B-protein
during	NN	O	O
the	NN	O	O
early	NN	O	O
stages	NN	O	O
of	NN	O	O
B	NN	O	O
cell	NN	O	O
development	NN	O	O
.	NN	O	O

-DOCSTART-	O

Transcription	NN	O	B-protein
factor	NN	O	I-protein
STAT5A	NN	O	B-protein
is	NN	O	O
a	NN	O	O
substrate	NN	O	O
of	NN	O	O
Bruton	NN	O	B-protein
's	NN	O	I-protein
tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

STAT5A	NN	O	B-protein
is	NN	O	O
a	NN	O	O
molecular	NN	O	O
regulator	NN	O	O
of	NN	O	O
proliferation	NN	O	O
,	NN	O	O
differentiation	NN	O	O
,	NN	O	O
and	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
lymphohematopoietic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
STAT5A	NN	O	B-protein
can	NN	O	O
serve	NN	O	O
as	NN	O	O
a	NN	O	O
functional	NN	O	O
substrate	NN	O	O
of	NN	O	O
Bruton	NN	O	B-protein
's	NN	O	I-protein
tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	O
BTK	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Purified	NN	O	B-protein
recombinant	NN	O	I-protein
BTK	NN	O	I-protein
was	NN	O	O
capable	NN	O	O
of	NN	O	O
directly	NN	O	O
binding	NN	O	O
purified	NN	O	B-protein
recombinant	NN	O	I-protein
STAT5A	NN	O	I-protein
with	NN	O	O
high	NN	O	O
affinity	NN	O	O
(	NN	O	O
K	NN	O	O
(	NN	O	O
d	NN	O	O
)	NN	O	O
=	NN	O	O
44	NN	O	O
nm	NN	O	O
)	NN	O	O
,	NN	O	O
as	NN	O	O
determined	NN	O	O
by	NN	O	O
surface	NN	O	O
plasmon	NN	O	O
resonance	NN	O	O
using	NN	O	O
a	NN	O	O
BIAcore	NN	O	O
biosensor	NN	O	O
system	NN	O	O
.	NN	O	O

BTK	NN	O	B-protein
was	NN	O	O
also	NN	O	O
capable	NN	O	O
of	NN	O	O
tyrosine-phosphorylating	NN	O	O
ectopically	NN	O	O
expressed	NN	O	O
recombinant	NN	O	B-protein
STAT5A	NN	O	I-protein
on	NN	O	O
Tyr	NN	O	O
(	NN	O	O
694	NN	O	O
)	NN	O	O
both	NN	O	O
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
in	NN	O	O
vivo	NN	O	O
in	NN	O	O
a	NN	O	O
Janus	NN	O	B-protein
kinase	NN	O	I-protein
3	NN	O	I-protein
-independent	NN	O	O
fashion	NN	O	O
.	NN	O	O

BTK	NN	O	B-protein
phosphorylated	NN	O	O
the	NN	O	O
Y665F	NN	O	B-protein
,	NN	O	I-protein
Y668F	NN	O	I-protein
,	NN	O	I-protein
and	NN	O	I-protein
Y682F	NN	O	I-protein
,	NN	O	I-protein
Y683F	NN	O	I-protein
mutants	NN	O	I-protein
but	NN	O	O
not	NN	O	O
the	NN	O	O
Y694F	NN	O	B-protein
mutant	NN	O	I-protein
of	NN	O	O
STAT5A	NN	O	B-protein
.	NN	O	O

STAT5A	NN	O	O
mutations	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
and	NN	O	I-protein
SH3	NN	O	I-protein
domains	NN	O	I-protein
did	NN	O	O
not	NN	O	O
alter	NN	O	O
the	NN	O	O
BTK	NN	O	B-protein
-mediated	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
.	NN	O	O

Recombinant	NN	O	B-protein
BTK	NN	O	I-protein
proteins	NN	O	I-protein
with	NN	O	O
mutant	NN	O	O
pleckstrin	NN	O	O
homology	NN	O	O
,	NN	O	O
SH2	NN	O	B-protein
,	NN	O	I-protein
or	NN	O	I-protein
SH3	NN	O	I-protein
domains	NN	O	I-protein
were	NN	O	O
capable	NN	O	O
of	NN	O	O
phosphorylating	NN	O	O
STAT5A	NN	O	B-protein
,	NN	O	O
whereas	NN	O	O
recombinant	NN	O	B-protein
BTK	NN	O	I-protein
proteins	NN	O	I-protein
with	NN	O	O
SH1/kinase	NN	O	O
domain	NN	O	O
mutations	NN	O	O
were	NN	O	O
not	NN	O	O
.	NN	O	O

In	NN	O	O
pull-down	NN	O	O
experiments	NN	O	O
,	NN	O	O
only	NN	O	O
full-length	NN	O	B-protein
BTK	NN	O	I-protein
and	NN	O	O
its	NN	O	O
SH1/kinase	NN	O	B-protein
domain	NN	O	I-protein
(	NN	O	O
but	NN	O	O
not	NN	O	O
the	NN	O	O
pleckstrin	NN	O	O
homology	NN	O	O
,	NN	O	O
SH2	NN	O	B-protein
,	NN	O	I-protein
or	NN	O	I-protein
SH3	NN	O	I-protein
domains	NN	O	I-protein
)	NN	O	O
were	NN	O	O
capable	NN	O	O
of	NN	O	O
binding	NN	O	O
STAT5A	NN	O	B-protein
.	NN	O	O

Ectopically	NN	O	O
expressed	NN	O	O
BTK	NN	O	B-protein
kinase	NN	O	I-protein
domain	NN	O	I-protein
was	NN	O	O
capable	NN	O	O
of	NN	O	O
tyrosine-phosphorylating	NN	O	O
STAT5A	NN	O	B-protein
both	NN	O	O
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

BTK	NN	O	B-protein
-mediated	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
ectopically	NN	O	O
expressed	NN	O	O
wild	NN	O	O
type	NN	O	O
(	NN	O	O
but	NN	O	O
not	NN	O	O
Tyr	NN	O	B-protein
(	NN	O	I-protein
694	NN	O	I-protein
)	NN	O	I-protein
mutant	NN	O	I-protein
)	NN	O	O
STAT5A	NN	O	B-protein
enhanced	NN	O	O
its	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
.	NN	O	O

In	NN	O	O
BTK	NN	O	B-protein
-competent	NN	O	O
chicken	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
anti-IgM-stimulated	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
STAT5	NN	O	B-protein
protein	NN	O	I-protein
was	NN	O	O
prevented	NN	O	O
by	NN	O	O
pretreatment	NN	O	O
with	NN	O	O
the	NN	O	O
BTK	NN	O	B-protein
inhibitor	NN	O	O
LFM-A13	NN	O	O
but	NN	O	O
not	NN	O	O
by	NN	O	O
pretreatment	NN	O	O
with	NN	O	O
the	NN	O	O
JAK3	NN	O	O
inhibitor	NN	O	O
HI-P131	NN	O	O
.	NN	O	O

B	NN	O	O
cell	NN	O	O
antigen	NN	O	O
receptor	NN	O	O
ligation	NN	O	O
resulted	NN	O	O
in	NN	O	O
enhanced	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
STAT5	NN	O	B-protein
in	NN	O	O
BTK	NN	O	B-protein
-deficient	NN	O	O
chicken	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
reconstituted	NN	O	O
with	NN	O	O
wild	NN	O	B-protein
type	NN	O	I-protein
human	NN	O	I-protein
BTK	NN	O	I-protein
but	NN	O	O
not	NN	O	O
in	NN	O	O
BTK	NN	O	B-protein
-deficient	NN	O	O
chicken	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
reconstituted	NN	O	O
with	NN	O	O
kinase-inactive	NN	O	B-protein
mutant	NN	O	I-protein
BTK	NN	O	I-protein
.	NN	O	O

Similarly	NN	O	O
,	NN	O	O
anti-IgM	NN	O	O
stimulation	NN	O	O
resulted	NN	O	O
in	NN	O	O
enhanced	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
STAT5A	NN	O	B-protein
in	NN	O	O
BTK	NN	O	B-protein
-competent	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
wild	NN	O	O
type	NN	O	O
mice	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
BTK	NN	O	B-protein
-deficient	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
XID	NN	O	O
mice	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
XID	NN	O	O
mice	NN	O	O
,	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
JAK3	NN	O	O
knockout	NN	O	O
mice	NN	O	O
showed	NN	O	O
a	NN	O	O
normal	NN	O	O
STAT5A	NN	O	O
phosphorylation	NN	O	O
response	NN	O	O
to	NN	O	O
anti-IgM	NN	O	O
stimulation	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
provide	NN	O	O
unprecedented	NN	O	O
experimental	NN	O	O
evidence	NN	O	O
that	NN	O	O
BTK	NN	O	B-protein
plays	NN	O	O
a	NN	O	O
nonredundant	NN	O	O
and	NN	O	O
pivotal	NN	O	O
role	NN	O	O
in	NN	O	O
B	NN	O	O
cell	NN	O	O
antigen	NN	O	O
receptor-mediated	NN	O	O
STAT5A	NN	O	O
activation	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type

-DOCSTART-	O

Role	NN	O	O
of	NN	O	O
T-bet	NN	O	B-protein
in	NN	O	O
commitment	NN	O	O
of	NN	O	O
TH1	NN	O	B-cell_type
cells	NN	O	I-cell_type
before	NN	O	O
IL-12	NN	O	B-protein
-dependent	NN	O	O
selection	NN	O	O
.	NN	O	O

How	NN	O	O
cytokines	NN	O	B-protein
control	NN	O	O
differentiation	NN	O	O
of	NN	O	O
helper	NN	O	B-cell_type
T	NN	O	I-cell_type
(	NN	O	I-cell_type
TH	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
controversial	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
T-bet	NN	O	B-protein
,	NN	O	O
without	NN	O	O
apparent	NN	O	O
assistance	NN	O	O
from	NN	O	O
interleukin	NN	O	B-protein
12	NN	O	I-protein
(	NN	O	I-protein
IL-12	NN	O	I-protein
)	NN	O	I-protein
/	NN	O	O
STAT4	NN	O	B-protein
,	NN	O	O
specifies	NN	O	O
TH1	NN	O	B-cell_type
effector	NN	O	O
fate	NN	O	O
by	NN	O	O
targeting	NN	O	O
chromatin	NN	O	O
remodeling	NN	O	O
to	NN	O	O
individual	NN	O	O
interferon-gamma	NN	O	B-DNA
(	NN	O	I-DNA
IFN-gamma	NN	O	I-DNA
)	NN	O	I-DNA
alleles	NN	O	I-DNA
and	NN	O	O
by	NN	O	O
inducing	NN	O	O
IL-12	NN	O	B-DNA
receptor	NN	O	I-DNA
beta2	NN	O	I-DNA
expression	NN	O	O
.	NN	O	O

Subsequently	NN	O	O
,	NN	O	O
it	NN	O	O
appears	NN	O	O
that	NN	O	O
IL-12	NN	O	B-protein
/	NN	O	O
STAT4	NN	O	B-protein
serves	NN	O	O
two	NN	O	O
essential	NN	O	O
functions	NN	O	O
in	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
TH1	NN	O	B-cell_type
cells	NN	O	I-cell_type
:	NN	O	O
as	NN	O	O
growth	NN	O	B-protein
signal	NN	O	I-protein
,	NN	O	O
inducing	NN	O	O
survival	NN	O	O
and	NN	O	O
cell	NN	O	O
division	NN	O	O
;	NN	O	O
and	NN	O	O
as	NN	O	O
trans-activator	NN	O	B-protein
,	NN	O	O
prolonging	NN	O	O
IFN-gamma	NN	O	B-protein
synthesis	NN	O	O
through	NN	O	O
a	NN	O	O
genetic	NN	O	O
interaction	NN	O	O
with	NN	O	O
the	NN	O	O
coactivator	NN	O	B-protein
,	NN	O	O
CREB-binding	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
a	NN	O	O
cytokine	NN	O	B-protein
does	NN	O	O
not	NN	O	O
simply	NN	O	O
induce	NN	O	O
TH	NN	O	O
fate	NN	O	O
choice	NN	O	O
but	NN	O	O
instead	NN	O	O
may	NN	O	O
act	NN	O	O
as	NN	O	O
an	NN	O	O
essential	NN	O	O
secondary	NN	O	O
stimulus	NN	O	O
that	NN	O	O
mediates	NN	O	O
selective	NN	O	O
survival	NN	O	O
of	NN	O	O
a	NN	O	O
lineage	NN	O	O
.	NN	O	O

-DOCSTART-	O

Stat6	NN	O	B-protein
is	NN	O	O
necessary	NN	O	O
and	NN	O	O
sufficient	NN	O	O
for	NN	O	O
IL-4	NN	O	O
's	NN	O	O
role	NN	O	O
in	NN	O	O
Th2	NN	O	O
differentiation	NN	O	O
and	NN	O	O
cell	NN	O	O
expansion	NN	O	O
.	NN	O	O

IL-4	NN	O	B-protein
plays	NN	O	O
a	NN	O	O
critical	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
differentiation	NN	O	O
of	NN	O	O
T	NN	O	O
CR-stimulated	NN	O	B-cell_type
naive	NN	O	I-cell_type
CD4	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
the	NN	O	O
Th2	NN	O	O
phenotype	NN	O	O
.	NN	O	O

In	NN	O	O
response	NN	O	O
to	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
the	NN	O	O
IL-4R	NN	O	B-protein
activates	NN	O	O
a	NN	O	O
set	NN	O	O
of	NN	O	O
phosphotyrosine	NN	O	B-protein
binding	NN	O	I-protein
domain-containing	NN	O	I-protein
proteins	NN	O	I-protein
,	NN	O	O
including	NN	O	O
insulin	NN	O	B-protein
receptor	NN	O	I-protein
substrate	NN	O	I-protein
1/2	NN	O	I-protein
,	NN	O	O
Shc	NN	O	B-protein
,	NN	O	O
and	NN	O	O
IL-4R	NN	O	B-protein
interacting	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
Stat6	NN	O	B-protein
.	NN	O	O

Stat6	NN	O	B-protein
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
Th2	NN	O	O
differentiation	NN	O	O
.	NN	O	O

To	NN	O	O
determine	NN	O	O
the	NN	O	O
roles	NN	O	O
of	NN	O	O
the	NN	O	O
phosphotyrosine	NN	O	B-protein
binding	NN	O	I-protein
adaptors	NN	O	I-protein
in	NN	O	O
Th2	NN	O	O
differentiation	NN	O	O
,	NN	O	O
we	NN	O	O
prepared	NN	O	O
a	NN	O	O
retrovirus	NN	O	O
containing	NN	O	O
a	NN	O	O
mutant	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-protein
(	NN	O	I-protein
h	NN	O	I-protein
)	NN	O	I-protein
IL-4R	NN	O	I-protein
alpha-chain	NN	O	I-protein
,	NN	O	O
Y497F	NN	O	B-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
unable	NN	O	O
to	NN	O	O
recruit	NN	O	O
these	NN	O	O
adaptors	NN	O	O
.	NN	O	O

The	NN	O	O
mutant	NN	O	B-protein
hIL-4Ralpha	NN	O	I-protein
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
wild-type	NN	O	B-protein
(	NN	O	I-protein
WT	NN	O	I-protein
)	NN	O	I-protein
hIL-4Ralpha	NN	O	I-protein
,	NN	O	O
was	NN	O	O
introduced	NN	O	O
into	NN	O	O
naive	NN	O	B-cell_type
CD4	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Upon	NN	O	O
hIL-4	NN	O	O
stimulation	NN	O	O
,	NN	O	O
Y497F	NN	O	B-protein
worked	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
WT	NN	O	B-protein
hIL-4Ralpha	NN	O	I-protein
in	NN	O	O
driving	NN	O	O
Th2	NN	O	O
differentiation	NN	O	O
,	NN	O	O
as	NN	O	O
measured	NN	O	O
by	NN	O	O
Gata3	NN	O	O
up-regulation	NN	O	O
and	NN	O	O
IL-4	NN	O	O
production	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
IL-4-driven	NN	O	O
cell	NN	O	O
expansion	NN	O	O
was	NN	O	O
also	NN	O	O
normal	NN	O	O
in	NN	O	O
the	NN	O	O
cells	NN	O	O
infected	NN	O	O
with	NN	O	O
Y497F	NN	O	B-protein
,	NN	O	O
although	NN	O	O
cells	NN	O	O
infected	NN	O	O
with	NN	O	O
Y497F	NN	O	B-protein
were	NN	O	O
not	NN	O	O
capable	NN	O	O
of	NN	O	O
phosphorylating	NN	O	O
insulin	NN	O	B-protein
receptor	NN	O	I-protein
substrate	NN	O	I-protein
2	NN	O	I-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
signal	NN	O	O
pathway	NN	O	O
mediated	NN	O	O
by	NN	O	O
Y497	NN	O	B-protein
is	NN	O	O
dispensable	NN	O	O
for	NN	O	O
both	NN	O	O
IL-4	NN	O	B-protein
-driven	NN	O	O
Th2	NN	O	O
differentiation	NN	O	O
and	NN	O	O
cell	NN	O	O
expansion	NN	O	O
.	NN	O	O

Both	NN	O	O
WT	NN	O	B-protein
and	NN	O	I-protein
Y497F	NN	O	I-protein
hIL-4Ralpha	NN	O	I-protein
lose	NN	O	O
the	NN	O	O
ability	NN	O	O
to	NN	O	O
drive	NN	O	O
Th2	NN	O	O
differentiation	NN	O	O
and	NN	O	O
cell	NN	O	O
expansion	NN	O	O
in	NN	O	O
Stat6-knockout	NN	O	B-cell_type
CD4	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

A	NN	O	O
constitutively	NN	O	O
activated	NN	O	O
form	NN	O	O
of	NN	O	O
Stat6	NN	O	B-protein
introduced	NN	O	O
into	NN	O	O
CD4	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
resulted	NN	O	O
in	NN	O	O
both	NN	O	O
Th2	NN	O	O
differentiation	NN	O	O
and	NN	O	O
enhanced	NN	O	O
cell	NN	O	O
expansion	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
activated	NN	O	B-protein
Stat6	NN	O	I-protein
is	NN	O	O
necessary	NN	O	O
and	NN	O	O
sufficient	NN	O	O
to	NN	O	O
mediate	NN	O	O
both	NN	O	O
IL-4	NN	O	B-protein
-driven	NN	O	O
Th2	NN	O	O
differentiation	NN	O	O
and	NN	O	O
cell	NN	O	O
expansion	NN	O	O
in	NN	O	O
CD4	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
effect	NN	O	O
of	NN	O	O
HIV-1	NN	O	B-protein
regulatory	NN	O	I-protein
proteins	NN	O	I-protein
on	NN	O	O
cellular	NN	O	O
genes	NN	O	O
:	NN	O	O
derepression	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
by	NN	O	O
Tat	NN	O	B-protein
.	NN	O	O

In	NN	O	O
HIV-infected	NN	O	O
individuals	NN	O	O
dysregulation	NN	O	O
of	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
is	NN	O	O
characterized	NN	O	O
by	NN	O	O
severe	NN	O	O
disorders	NN	O	O
of	NN	O	O
the	NN	O	O
cytokine	NN	O	O
network	NN	O	O
.	NN	O	O

Increase	NN	O	O
secretion	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
the	NN	O	O
major	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
growth	NN	O	O
and	NN	O	O
differentiation	NN	O	O
cytokine	NN	O	B-protein
,	NN	O	O
may	NN	O	O
play	NN	O	O
a	NN	O	O
decisive	NN	O	O
role	NN	O	O
in	NN	O	O
sensitization	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
for	NN	O	O
activation	NN	O	O
induced	NN	O	O
apoptosis	NN	O	O
and	NN	O	O
indirect	NN	O	O
death	NN	O	O
of	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
through	NN	O	O
augmented	NN	O	O
virus	NN	O	O
replication	NN	O	O
.	NN	O	O

We	NN	O	O
investigated	NN	O	O
the	NN	O	O
cause	NN	O	O
of	NN	O	O
enhanced	NN	O	O
IL-2	NN	O	O
secretion	NN	O	O
and	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
HIV	NN	O	B-protein
Tat	NN	O	I-protein
induces	NN	O	O
this	NN	O	O
effect	NN	O	O
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
increased	NN	O	O
IL-2	NN	O	O
secretion	NN	O	O
is	NN	O	O
due	NN	O	O
to	NN	O	O
Tat	NN	O	B-protein
-enhanced	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
activation	NN	O	O
.	NN	O	O

Tat	NN	O	B-protein
derepresses	NN	O	O
and	NN	O	O
activates	NN	O	O
the	NN	O	O
distal	NN	O	B-DNA
AP-1	NN	O	I-DNA
site	NN	O	I-DNA
(	NN	O	O
position	NN	O	B-DNA
-185	NN	O	I-DNA
to	NN	O	I-DNA
-177	NN	O	I-DNA
)	NN	O	O
in	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
nonstimulated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
a	NN	O	O
repressor	NN	O	O
complex	NN	O	O
containing	NN	O	O
NF-IL6	NN	O	B-protein
,	NN	O	O
JunB	NN	O	B-protein
,	NN	O	O
c-Fos	NN	O	B-protein
and	NN	O	O
Fra-1	NN	O	B-protein
is	NN	O	O
formed	NN	O	O
on	NN	O	O
the	NN	O	O
AP-1	NN	O	B-DNA
(	NN	O	I-DNA
IL-2/d	NN	O	I-DNA
)	NN	O	I-DNA
site	NN	O	I-DNA
and	NN	O	O
represses	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
.	NN	O	O

After	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
,	NN	O	O
a	NN	O	O
heterodimeric	NN	O	B-protein
activator	NN	O	I-protein
containing	NN	O	O
p65	NN	O	B-protein
and	NN	O	O
c-Jun	NN	O	B-protein
binds	NN	O	O
to	NN	O	O
the	NN	O	O
AP-1	NN	O	B-DNA
(	NN	O	I-DNA
IL-2/d	NN	O	I-DNA
)	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

HIV	NN	O	B-protein
Tat	NN	O	I-protein
enhances	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
and	NN	O	O
consequently	NN	O	O
,	NN	O	O
activates	NN	O	O
the	NN	O	O
AP-1	NN	O	B-DNA
(	NN	O	I-DNA
IL-2/d	NN	O	I-DNA
)	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

Our	NN	O	O
data	NN	O	O
provide	NN	O	O
evidence	NN	O	O
for	NN	O	O
a	NN	O	O
novel	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
HIV	NN	O	B-protein
Tat	NN	O	I-protein
dysregulates	NN	O	O
IL-2	NN	O	O
production	NN	O	O
and	NN	O	O
therefore	NN	O	O
may	NN	O	O
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
HIV-1	NN	O	O
infection	NN	O	O
in	NN	O	O
a	NN	O	O
way	NN	O	O
yet	NN	O	O
to	NN	O	O
be	NN	O	O
clarified	NN	O	O
.	NN	O	O

-DOCSTART-	O

Transforming	NN	O	B-protein
growth	NN	O	I-protein
factor-beta1	NN	O	I-protein
interferes	NN	O	O
with	NN	O	O
thrombopoietin-induced	NN	O	O
signal	NN	O	O
transduction	NN	O	O
in	NN	O	O
megakaryoblastic	NN	O	B-cell_type
and	NN	O	I-cell_type
erythroleukemic	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

OBJECTIVE	NN	O	O
:	NN	O	O
Thrombopoietin	NN	O	B-protein
(	NN	O	O
TPO	NN	O	B-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	I-protein
1	NN	O	I-protein
)	NN	O	I-protein
(	NN	O	O
TGF-beta	NN	O	B-protein
(	NN	O	I-protein
1	NN	O	I-protein
)	NN	O	I-protein
)	NN	O	O
have	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
exert	NN	O	O
opposite	NN	O	O
effects	NN	O	O
on	NN	O	O
proliferation	NN	O	O
and	NN	O	O
megakaryocytic	NN	O	O
differentiation	NN	O	O
of	NN	O	O
hematopoietic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

To	NN	O	O
determine	NN	O	O
whether	NN	O	O
TGF-beta	NN	O	B-protein
(	NN	O	I-protein
1	NN	O	I-protein
)	NN	O	I-protein
interferes	NN	O	O
directly	NN	O	O
with	NN	O	O
TPO	NN	O	B-protein
-induced	NN	O	O
signal	NN	O	O
transduction	NN	O	O
in	NN	O	O
hematopoietic	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
we	NN	O	O
compared	NN	O	O
the	NN	O	O
regulatory	NN	O	O
effects	NN	O	O
in	NN	O	O
the	NN	O	O
TPO	NN	O	B-protein
-responsive	NN	O	O
cell	NN	O	O
lines	NN	O	O
Mo-7e	NN	O	O
and	NN	O	O
HEL	NN	O	O
.	NN	O	O

MATERIALS	NN	O	O
AND	NN	O	O
METHODS	NN	O	O
:	NN	O	O
The	NN	O	O
cells	NN	O	O
were	NN	O	O
stimulated	NN	O	O
by	NN	O	O
100	NN	O	O
ng/mL	NN	O	O
TPO	NN	O	B-protein
and/or	NN	O	O
100	NN	O	O
ng/mL	NN	O	O
TGF-beta1	NN	O	B-protein
and	NN	O	O
analyzed	NN	O	O
for	NN	O	O
proliferation	NN	O	O
(	NN	O	O
3H	NN	O	O
thymidine	NN	O	O
incorporation	NN	O	O
)	NN	O	O
,	NN	O	O
viability	NN	O	O
(	NN	O	O
trypan	NN	O	O
blue	NN	O	O
exclusion	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
protein	NN	O	O
expression	NN	O	O
and	NN	O	O
phosphorylation	NN	O	O
(	NN	O	O
Western	NN	O	O
blot	NN	O	O
)	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
TPO	NN	O	B-protein
enhanced	NN	O	O
the	NN	O	O
proliferation	NN	O	O
of	NN	O	O
Mo-7e	NN	O	B-cell_line
cells	NN	O	I-cell_line
as	NN	O	O
determined	NN	O	O
by	NN	O	O
3H-thymidine	NN	O	O
incorporation	NN	O	O
,	NN	O	O
whereas	NN	O	O
TGF-beta1	NN	O	B-protein
suppressed	NN	O	O
baseline	NN	O	O
cell	NN	O	O
growth	NN	O	O
and	NN	O	O
antagonized	NN	O	O
the	NN	O	O
proliferative	NN	O	O
effect	NN	O	O
of	NN	O	O
TPO	NN	O	B-protein
.	NN	O	O

TPO	NN	O	B-protein
-induced	NN	O	O
proliferation	NN	O	O
also	NN	O	O
was	NN	O	O
reduced	NN	O	O
by	NN	O	O
a	NN	O	O
specific	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
the	NN	O	O
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	O
MAPK	NN	O	B-protein
)	NN	O	O
pathway	NN	O	O
(	NN	O	O
PD098059	NN	O	O
)	NN	O	O
,	NN	O	O
which	NN	O	O
inhibits	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
MAPK	NN	O	B-protein
extracellular	NN	O	I-protein
signal-regulated	NN	O	I-protein
kinases	NN	O	I-protein
(	NN	O	O
ERK	NN	O	B-protein
)	NN	O	O
ERK1	NN	O	B-protein
and	NN	O	O
ERK2	NN	O	B-protein
,	NN	O	O
and	NN	O	O
AG490	NN	O	O
,	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
Janus	NN	O	B-protein
kinase-2	NN	O	I-protein
,	NN	O	O
which	NN	O	O
completely	NN	O	O
blocked	NN	O	O
TPO	NN	O	B-protein
-induced	NN	O	O
proliferation	NN	O	O
.	NN	O	O

As	NN	O	O
demonstrated	NN	O	O
by	NN	O	O
Western	NN	O	O
blotting	NN	O	O
,	NN	O	O
TGF-beta1	NN	O	B-protein
reduced	NN	O	O
the	NN	O	O
TPO	NN	O	B-protein
-stimulated	NN	O	O
ERK1	NN	O	B-protein
/	NN	O	O
ERK2	NN	O	B-protein
and	NN	O	O
STAT5	NN	O	O
phosphorylation	NN	O	O
in	NN	O	O
Mo-7e	NN	O	B-cell_line
and	NN	O	I-cell_line
HEL	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

This	NN	O	O
effect	NN	O	O
was	NN	O	O
completely	NN	O	O
reversed	NN	O	O
by	NN	O	O
preincubation	NN	O	O
with	NN	O	O
a	NN	O	O
tyrosine	NN	O	O
phosphatase	NN	O	O
inhibitor	NN	O	O
(	NN	O	O
Na3VO4	NN	O	O
)	NN	O	O
,	NN	O	O
which	NN	O	O
suggests	NN	O	O
that	NN	O	O
TGF-beta1	NN	O	B-protein
activated	NN	O	O
a	NN	O	O
phosphatase	NN	O	B-protein
.	NN	O	O

Although	NN	O	O
STAT3	NN	O	B-protein
also	NN	O	O
was	NN	O	O
activated	NN	O	O
by	NN	O	O
TPO	NN	O	B-protein
,	NN	O	O
STAT3	NN	O	B-protein
activation	NN	O	O
remained	NN	O	O
unaltered	NN	O	O
by	NN	O	O
TGF-beta1	NN	O	B-protein
.	NN	O	O

CONCLUSION	NN	O	O
:	NN	O	O
Taken	NN	O	O
together	NN	O	O
,	NN	O	O
these	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
TGF-beta1	NN	O	B-protein
modulates	NN	O	O
TPO	NN	O	B-protein
-mediated	NN	O	O
effects	NN	O	O
on	NN	O	O
megakaryocytic	NN	O	O
proliferation	NN	O	O
by	NN	O	O
interfering	NN	O	O
with	NN	O	O
TPO	NN	O	B-protein
-induced	NN	O	O
signal	NN	O	O
transduction	NN	O	O
,	NN	O	O
particularly	NN	O	O
by	NN	O	O
reducing	NN	O	O
the	NN	O	O
activities	NN	O	O
of	NN	O	O
MAPK	NN	O	B-protein
ERK1/ERK2	NN	O	I-protein
and	NN	O	I-protein
STAT5	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Invariant	NN	O	B-protein
chain	NN	O	I-protein
induces	NN	O	O
B	NN	O	O
cell	NN	O	O
maturation	NN	O	O
by	NN	O	O
activating	NN	O	O
a	NN	O	O
TAF	NN	O	B-protein
(	NN	O	I-protein
II	NN	O	I-protein
)	NN	O	I-protein
105-NF-kappaB	NN	O	I-protein
-dependent	NN	O	O
transcription	NN	O	O
program	NN	O	O
.	NN	O	O

Early	NN	O	O
stages	NN	O	O
of	NN	O	O
B	NN	O	O
cell	NN	O	O
development	NN	O	O
occur	NN	O	O
in	NN	O	O
the	NN	O	O
bone	NN	O	O
marrow	NN	O	O
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
formation	NN	O	O
of	NN	O	O
immature	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

From	NN	O	O
there	NN	O	O
these	NN	O	O
immature	NN	O	B-cell_type
cells	NN	O	I-cell_type
migrate	NN	O	O
to	NN	O	O
the	NN	O	O
spleen	NN	O	O
where	NN	O	O
they	NN	O	O
differentiate	NN	O	O
to	NN	O	O
mature	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

This	NN	O	O
final	NN	O	O
maturation	NN	O	O
step	NN	O	O
is	NN	O	O
crucial	NN	O	O
for	NN	O	O
the	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
become	NN	O	O
responsive	NN	O	O
to	NN	O	O
antigens	NN	O	B-protein
and	NN	O	O
to	NN	O	O
participate	NN	O	O
in	NN	O	O
the	NN	O	O
immune	NN	O	O
response	NN	O	O
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
invariant	NN	O	B-protein
chain	NN	O	I-protein
(	NN	O	O
Ii	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
major	NN	O	B-protein
histocompatibility	NN	O	I-protein
complex	NN	O	I-protein
class	NN	O	I-protein
II	NN	O	I-protein
chaperone	NN	O	I-protein
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
c-Rel	NN	O	B-protein
and	NN	O	O
p65/RelA	NN	O	B-protein
,	NN	O	O
were	NN	O	O
found	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
final	NN	O	O
antigen-independent	NN	O	O
differentiation	NN	O	O
stage	NN	O	O
of	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
spleen	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
a	NN	O	O
possible	NN	O	O
link	NN	O	O
between	NN	O	O
Ii	NN	O	B-protein
-dependent	NN	O	O
B	NN	O	O
cell	NN	O	O
maturation	NN	O	O
and	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	O
pathway	NN	O	O
.	NN	O	O

Our	NN	O	O
studies	NN	O	O
indicate	NN	O	O
that	NN	O	O
Ii	NN	O	B-protein
-induced	NN	O	O
B	NN	O	O
cell	NN	O	O
maturation	NN	O	O
involves	NN	O	O
activation	NN	O	O
of	NN	O	O
transcription	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-protein
p65/RelA	NN	O	I-protein
homodimer	NN	O	I-protein
and	NN	O	O
requires	NN	O	O
the	NN	O	O
B	NN	O	B-protein
cell-enriched	NN	O	I-protein
coactivator	NN	O	I-protein
TBP-associated	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	I-protein
II	NN	O	I-protein
)	NN	O	I-protein
105	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Androgens	NN	O	O
indirectly	NN	O	O
accelerate	NN	O	O
thymocyte	NN	O	B-cell_type
apoptosis	NN	O	O
.	NN	O	O

Apoptotic	NN	O	O
processes	NN	O	O
,	NN	O	O
or	NN	O	O
the	NN	O	O
disturbance	NN	O	O
of	NN	O	O
the	NN	O	O
natural	NN	O	O
regulation	NN	O	O
of	NN	O	O
these	NN	O	O
processes	NN	O	O
,	NN	O	O
may	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
autoimmune	NN	O	O
diseases	NN	O	O
(	NN	O	O
AID	NN	O	O
)	NN	O	O
.	NN	O	O

Women	NN	O	O
are	NN	O	O
,	NN	O	O
in	NN	O	O
general	NN	O	O
,	NN	O	O
more	NN	O	O
susceptible	NN	O	O
than	NN	O	O
men	NN	O	O
to	NN	O	O
develop	NN	O	O
AID	NN	O	O
like	NN	O	O
rheumatoid	NN	O	O
arthritis	NN	O	O
.	NN	O	O

Androgens	NN	O	O
and	NN	O	O
glucocorticoids	NN	O	O
,	NN	O	O
in	NN	O	O
contrast	NN	O	O
to	NN	O	O
oestrogens	NN	O	O
,	NN	O	O
have	NN	O	O
favourable	NN	O	O
effects	NN	O	O
in	NN	O	O
AID	NN	O	O
models	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
in	NN	O	O
human	NN	O	O
AID	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
known	NN	O	O
that	NN	O	O
glucocorticoids	NN	O	O
(	NN	O	O
GC	NN	O	O
)	NN	O	O
,	NN	O	O
used	NN	O	O
for	NN	O	O
treatment	NN	O	O
of	NN	O	O
AID	NN	O	O
,	NN	O	O
increase	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
the	NN	O	O
thymus	NN	O	O
resulting	NN	O	O
in	NN	O	O
decreased	NN	O	O
numbers	NN	O	O
of	NN	O	O
CD4+	NN	O	B-cell_type
CD8+	NN	O	I-cell_type
thymocytes	NN	O	I-cell_type
.	NN	O	O

It	NN	O	O
was	NN	O	O
asked	NN	O	O
whether	NN	O	O
androgens	NN	O	O
,	NN	O	O
in	NN	O	O
contrast	NN	O	O
to	NN	O	O
oestrogens	NN	O	O
,	NN	O	O
exert	NN	O	O
their	NN	O	O
favourable	NN	O	O
effects	NN	O	O
in	NN	O	O
the	NN	O	O
treatment	NN	O	O
of	NN	O	O
AID	NN	O	O
by	NN	O	O
a	NN	O	O
mechanism	NN	O	O
comparable	NN	O	O
to	NN	O	O
that	NN	O	O
described	NN	O	O
for	NN	O	O
GC	NN	O	O
by	NN	O	O
eliminating	NN	O	O
the	NN	O	O
apoptosis	NN	O	O
prone	NN	O	O
CD4+	NN	O	B-cell_type
CD8+	NN	O	I-cell_type
population	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
thymus	NN	O	O
.	NN	O	O

Although	NN	O	O
both	NN	O	O
androgens	NN	O	O
and	NN	O	O
oestrogens	NN	O	O
proved	NN	O	O
thymolytic	NN	O	O
,	NN	O	O
a	NN	O	O
significantly	NN	O	O
decreased	NN	O	O
percentage	NN	O	O
of	NN	O	O
CD4+	NN	O	B-cell_type
CD8+	NN	O	I-cell_type
thymocytes	NN	O	I-cell_type
was	NN	O	O
observed	NN	O	O
by	NN	O	O
flow	NN	O	O
cytometry	NN	O	O
after	NN	O	O
treatment	NN	O	O
of	NN	O	O
mice	NN	O	O
with	NN	O	O
the	NN	O	O
androgen	NN	O	O
methyltestosterone	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
with	NN	O	O
the	NN	O	O
oestrogen	NN	O	O
ethinylestradiol	NN	O	O
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
whether	NN	O	O
the	NN	O	O
observed	NN	O	O
thymolytic	NN	O	O
effects	NN	O	O
were	NN	O	O
due	NN	O	O
to	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
hormone	NN	O	B-protein
receptors	NN	O	I-protein
on	NN	O	O
thymocytes	NN	O	B-cell_type
,	NN	O	O
cells	NN	O	O
were	NN	O	O
isolated	NN	O	O
from	NN	O	O
the	NN	O	O
thymus	NN	O	O
and	NN	O	O
incubated	NN	O	O
with	NN	O	O
androgens	NN	O	O
or	NN	O	O
oestrogens	NN	O	O
to	NN	O	O
measure	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

Several	NN	O	O
techniques	NN	O	O
were	NN	O	O
used	NN	O	O
to	NN	O	O
determine	NN	O	O
thymocyte	NN	O	B-cell_type
apoptosis	NN	O	O
in	NN	O	O
vitro	NN	O	O
,	NN	O	O
but	NN	O	O
no	NN	O	O
enhanced	NN	O	O
apoptotic	NN	O	O
signal	NN	O	O
was	NN	O	O
observed	NN	O	O
.	NN	O	O

Using	NN	O	O
the	NN	O	O
very	NN	O	O
sensitive	NN	O	O
TUNEL	NN	O	O
assay	NN	O	O
,	NN	O	O
no	NN	O	O
direct	NN	O	O
effect	NN	O	O
of	NN	O	O
androgens	NN	O	O
on	NN	O	O
thymocytes	NN	O	B-cell_type
in	NN	O	O
vitro	NN	O	O
could	NN	O	O
be	NN	O	O
observed	NN	O	O
.	NN	O	O

This	NN	O	O
is	NN	O	O
in	NN	O	O
sharp	NN	O	O
contrast	NN	O	O
to	NN	O	O
the	NN	O	O
high	NN	O	O
signal	NN	O	O
observed	NN	O	O
with	NN	O	O
GC	NN	O	O
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
upon	NN	O	O
in	NN	O	O
vivo	NN	O	O
androgen	NN	O	O
treatment	NN	O	O
,	NN	O	O
other	NN	O	O
cells	NN	O	O
containing	NN	O	O
androgen	NN	O	B-protein
receptors	NN	O	I-protein
than	NN	O	O
thymocytes	NN	O	B-cell_type
are	NN	O	O
probably	NN	O	O
involved	NN	O	O
in	NN	O	O
inducing	NN	O	O
the	NN	O	O
increase	NN	O	O
in	NN	O	O
thymic	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

To	NN	O	O
study	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
the	NN	O	O
androgen	NN	O	B-protein
receptor	NN	O	I-protein
on	NN	O	O
thymocyte	NN	O	B-cell_type
apoptosis	NN	O	O
,	NN	O	O
androgen	NN	O	B-protein
receptor	NN	O	I-protein
mutant	NN	O	O
(	NN	O	O
Tfm/Y	NN	O	O
)	NN	O	O
mice	NN	O	O
were	NN	O	O
treated	NN	O	O
with	NN	O	O
androgens	NN	O	O
.	NN	O	O

No	NN	O	O
alterations	NN	O	O
of	NN	O	O
thymocyte	NN	O	B-cell_type
subpopulations	NN	O	I-cell_type
were	NN	O	O
seen	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
changes	NN	O	O
in	NN	O	O
the	NN	O	O
percentage	NN	O	O
of	NN	O	O
CD4+	NN	O	B-cell_type
CD8+	NN	O	I-cell_type
thymocytes	NN	O	I-cell_type
after	NN	O	O
administration	NN	O	O
of	NN	O	O
androgens	NN	O	O
depend	NN	O	O
on	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
functional	NN	O	O
androgen	NN	O	B-protein
receptors	NN	O	I-protein
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
it	NN	O	O
is	NN	O	O
concluded	NN	O	O
that	NN	O	O
androgens	NN	O	O
indirectly	NN	O	O
accelerate	NN	O	O
thymocyte	NN	O	B-cell_type
apoptosis	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

-DOCSTART-	O

Inhibition	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
AP-1	NN	O	B-protein
and	NN	O	O
NF-kappaB	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
by	NN	O	O
peroxisome	NN	O	B-protein
proliferator-activated	NN	O	I-protein
receptor	NN	O	I-protein
gamma	NN	O	I-protein
ligands	NN	O	I-protein
.	NN	O	O

The	NN	O	O
peroxisome	NN	O	B-protein
proliferator-activated	NN	O	I-protein
receptor	NN	O	I-protein
gamma	NN	O	I-protein
(	NN	O	O
PPARgamma	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
hormone	NN	O	I-protein
receptor	NN	O	I-protein
superfamily	NN	O	I-protein
,	NN	O	O
is	NN	O	O
essential	NN	O	O
for	NN	O	O
adipocyte	NN	O	O
differentiation	NN	O	O
and	NN	O	O
glucose	NN	O	O
homeostasis	NN	O	O
.	NN	O	O

PPARgamma	NN	O	B-protein
has	NN	O	O
been	NN	O	O
found	NN	O	O
recently	NN	O	O
to	NN	O	O
regulate	NN	O	O
macrophage	NN	O	O
activation	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
mitogens	NN	O	B-protein
and	NN	O	O
inflammation	NN	O	O
.	NN	O	O

Our	NN	O	O
study	NN	O	O
shows	NN	O	O
PPARgamma	NN	O	B-protein
to	NN	O	O
be	NN	O	O
preferentially	NN	O	O
expressed	NN	O	O
in	NN	O	O
the	NN	O	O
nuclei	NN	O	O
of	NN	O	O
resting	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
to	NN	O	O
increase	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
by	NN	O	O
either	NN	O	O
anti-CD3	NN	O	B-protein
and	NN	O	O
anti-CD28	NN	O	B-protein
or	NN	O	O
phorbol	NN	O	O
myristyl	NN	O	O
acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
also	NN	O	O
found	NN	O	O
the	NN	O	O
PPARgamma	NN	O	B-protein
ligand	NN	O	I-protein
ciglitizone	NN	O	O
to	NN	O	O
attenuate	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
inhibiting	NN	O	O
cytokine	NN	O	O
gene	NN	O	O
expression	NN	O	O
and	NN	O	O
anti-CD3	NN	O	B-protein
and	NN	O	O
anti-CD28	NN	O	B-protein
or	NN	O	O
PMA-induced	NN	O	O
proliferative	NN	O	O
responses	NN	O	O
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
both	NN	O	O
the	NN	O	O
proliferative	NN	O	O
response	NN	O	O
and	NN	O	O
inflammatory	NN	O	O
cytokine	NN	O	O
expression	NN	O	O
in	NN	O	O
CD4	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
correlated	NN	O	O
with	NN	O	O
suppression	NN	O	O
of	NN	O	O
the	NN	O	O
activated	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
AP1	NN	O	B-protein
and	NN	O	O
NF-kappaB	NN	O	B-protein
.	NN	O	O

PPARgamma	NN	O	B-protein
ligands	NN	O	I-protein
also	NN	O	O
strongly	NN	O	O
inhibited	NN	O	O
SEA-induced	NN	O	O
Vbeta3	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
,	NN	O	O
together	NN	O	O
with	NN	O	O
previous	NN	O	O
findings	NN	O	O
of	NN	O	O
the	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
of	NN	O	O
PPARgamma	NN	O	B-protein
ligands	NN	O	I-protein
on	NN	O	O
activated	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
,	NN	O	O
provide	NN	O	O
clear	NN	O	O
evidence	NN	O	O
for	NN	O	O
PPARgamma	NN	O	B-protein
as	NN	O	O
a	NN	O	O
negative	NN	O	B-protein
regulator	NN	O	I-protein
of	NN	O	O
the	NN	O	O
inflammatory	NN	O	O
activation	NN	O	O
of	NN	O	O
both	NN	O	O
macrophage	NN	O	B-cell_type
and	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

PPARgamma	NN	O	B-protein
may	NN	O	O
thus	NN	O	O
be	NN	O	O
a	NN	O	O
potential	NN	O	O
therapeutic	NN	O	O
target	NN	O	O
for	NN	O	O
the	NN	O	O
treatment	NN	O	O
of	NN	O	O
autoimmunity	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
prominent	NN	O	O
role	NN	O	O
for	NN	O	O
activator	NN	O	B-protein
protein-1	NN	O	I-protein
in	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
2B4	NN	O	I-DNA
(	NN	O	I-DNA
CD244	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
in	NN	O	O
NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
cell	NN	O	B-protein
surface	NN	O	I-protein
glycoprotein	NN	O	I-protein
2B4	NN	O	I-protein
(	NN	O	O
CD244	NN	O	B-protein
)	NN	O	O
of	NN	O	O
the	NN	O	O
Ig	NN	O	B-protein
superfamily	NN	O	I-protein
is	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
NK	NN	O	B-cell_type
and	NN	O	O
T	NN	O	B-cell_type
lymphocyte	NN	O	I-cell_type
functions	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
recently	NN	O	O
identified	NN	O	O
CD48	NN	O	B-protein
as	NN	O	O
the	NN	O	O
high	NN	O	B-protein
affinity	NN	O	I-protein
counterreceptor	NN	O	I-protein
for	NN	O	O
2B4	NN	O	B-protein
in	NN	O	O
both	NN	O	O
mice	NN	O	O
and	NN	O	O
humans	NN	O	O
.	NN	O	O

The	NN	O	O
cytoplasmic	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
2B4	NN	O	B-protein
associates	NN	O	O
with	NN	O	O
src	NN	O	B-protein
homology	NN	O	I-protein
2	NN	O	I-protein
domain-containing	NN	O	I-protein
protein	NN	O	I-protein
or	NN	O	O
signaling	NN	O	B-protein
lymphocyte	NN	O	I-protein
activation	NN	O	I-protein
molecule-associated	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
whose	NN	O	O
mutation	NN	O	O
is	NN	O	O
the	NN	O	O
underlying	NN	O	O
genetic	NN	O	O
defect	NN	O	O
in	NN	O	O
the	NN	O	O
X-linked	NN	O	O
lymphoproliferative	NN	O	O
syndrome	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
report	NN	O	O
the	NN	O	O
molecular	NN	O	O
cloning	NN	O	O
and	NN	O	O
characterization	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
2B4	NN	O	I-DNA
(	NN	O	I-DNA
h2B4	NN	O	I-DNA
)	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Through	NN	O	O
primer	NN	O	O
extension	NN	O	O
analysis	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
h2B4	NN	O	B-DNA
gene	NN	O	I-DNA
initiates	NN	O	O
at	NN	O	O
multiple	NN	O	O
start	NN	O	O
sites	NN	O	O
.	NN	O	O

We	NN	O	O
isolated	NN	O	O
h2B4	NN	O	B-DNA
genomic	NN	O	I-DNA
clones	NN	O	I-DNA
and	NN	O	O
PCR	NN	O	O
amplified	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
containing	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
elements	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
have	NN	O	O
identified	NN	O	O
a	NN	O	O
functional	NN	O	O
AP-1	NN	O	B-DNA
site	NN	O	I-DNA
that	NN	O	O
lies	NN	O	O
between	NN	O	O
(	NN	O	O
-106	NN	O	O
to	NN	O	O
-100	NN	O	O
)	NN	O	O
through	NN	O	O
transient	NN	O	O
transfection	NN	O	O
analysis	NN	O	O
in	NN	O	O
YT	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
NK	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

EMSAs	NN	O	O
with	NN	O	O
Abs	NN	O	B-protein
specific	NN	O	O
for	NN	O	O
various	NN	O	O
protein	NN	O	B-protein
factors	NN	O	I-protein
of	NN	O	O
the	NN	O	O
AP-1	NN	O	B-protein
family	NN	O	I-protein
revealed	NN	O	O
that	NN	O	O
multiple	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
Jun	NN	O	B-protein
family	NN	O	I-protein
are	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
h2B4	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

Mutation	NN	O	O
of	NN	O	O
the	NN	O	O
AP-1	NN	O	B-DNA
site	NN	O	I-DNA
not	NN	O	O
only	NN	O	O
abolishes	NN	O	O
protein/DNA	NN	O	O
interactions	NN	O	O
but	NN	O	O
also	NN	O	O
promoter	NN	O	O
activity	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
a	NN	O	O
significant	NN	O	O
role	NN	O	O
for	NN	O	O
AP-1	NN	O	B-protein
in	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
h2B4	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Functional	NN	O	O
correction	NN	O	O
of	NN	O	O
FA-C	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
FANCC	NN	O	B-protein
suppresses	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
interferon	NN	O	B-DNA
gamma-inducible	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

Because	NN	O	O
hematopoietic	NN	O	B-cell_type
cells	NN	O	I-cell_type
derived	NN	O	O
from	NN	O	O
Fanconi	NN	O	O
anemia	NN	O	O
(	NN	O	O
FA	NN	O	O
)	NN	O	O
patients	NN	O	O
of	NN	O	O
the	NN	O	O
C-complementation	NN	O	O
group	NN	O	O
(	NN	O	O
FA-C	NN	O	O
)	NN	O	O
are	NN	O	O
hypersensitive	NN	O	O
to	NN	O	O
the	NN	O	O
inhibitory	NN	O	O
effects	NN	O	O
of	NN	O	O
interferon	NN	O	B-protein
gamma	NN	O	I-protein
(	NN	O	O
IFNgamma	NN	O	B-protein
)	NN	O	O
,	NN	O	O
the	NN	O	O
products	NN	O	O
of	NN	O	O
certain	NN	O	O
IFNgamma-inducible	NN	O	B-DNA
genes	NN	O	I-DNA
known	NN	O	O
to	NN	O	O
influence	NN	O	O
hematopoietic	NN	O	O
cell	NN	O	O
survival	NN	O	O
were	NN	O	O
quantified	NN	O	O
.	NN	O	O

High	NN	O	O
constitutive	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
IFNgamma-inducible	NN	O	B-DNA
genes	NN	O	I-DNA
,	NN	O	O
IFN-stimulated	NN	O	B-protein
gene	NN	O	I-protein
factor	NN	O	I-protein
3	NN	O	I-protein
gamma	NN	O	I-protein
subunit	NN	O	I-protein
(	NN	O	O
ISGF3gamma	NN	O	B-protein
)	NN	O	O
,	NN	O	O
IFN	NN	O	B-protein
regulatory	NN	O	I-protein
factor-1	NN	O	I-protein
(	NN	O	O
IRF-1	NN	O	B-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
cyclin-dependent	NN	O	B-protein
kinase	NN	O	I-protein
inhibitor	NN	O	I-protein
p21	NN	O	I-protein
(	NN	O	O
WAF1	NN	O	B-protein
)	NN	O	O
was	NN	O	O
found	NN	O	O
in	NN	O	O
FANCC	NN	O	B-protein
mutant	NN	O	O
B	NN	O	B-cell_type
lymphoblasts	NN	O	I-cell_type
,	NN	O	O
low-density	NN	O	B-cell_type
bone	NN	O	I-cell_type
marrow	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
murine	NN	O	B-cell_type
embryonic	NN	O	I-cell_type
fibroblasts	NN	O	I-cell_type
.	NN	O	O

Paradoxically	NN	O	O
,	NN	O	O
these	NN	O	O
cells	NN	O	O
do	NN	O	O
not	NN	O	O
activate	NN	O	B-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
STAT	NN	O	I-protein
)	NN	O	I-protein
1	NN	O	I-protein
properly	NN	O	O
.	NN	O	O

In	NN	O	O
an	NN	O	O
attempt	NN	O	O
to	NN	O	O
clarify	NN	O	O
mechanisms	NN	O	O
by	NN	O	O
which	NN	O	O
FA-C	NN	O	B-cell_type
cells	NN	O	I-cell_type
overexpress	NN	O	O
IFNgamma-inducible	NN	O	B-DNA
genes	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
face	NN	O	O
of	NN	O	O
defective	NN	O	O
STAT1	NN	O	O
phosphorylation	NN	O	O
,	NN	O	O
it	NN	O	O
was	NN	O	O
reasoned	NN	O	O
that	NN	O	O
decreased	NN	O	O
levels	NN	O	O
of	NN	O	O
activated	NN	O	B-protein
STAT1	NN	O	I-protein
might	NN	O	O
result	NN	O	O
in	NN	O	O
reduced	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
hematopoietic	NN	O	B-protein
IFNgamma-responsive	NN	O	I-protein
protein	NN	O	I-protein
that	NN	O	O
normally	NN	O	O
modulates	NN	O	O
expression	NN	O	O
of	NN	O	O
other	NN	O	O
IFNgamma-responsive	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

Levels	NN	O	O
of	NN	O	O
the	NN	O	O
IFNgamma	NN	O	B-protein
-inducible	NN	O	O
factor	NN	O	O
IFN	NN	O	B-protein
consensus	NN	O	I-protein
sequence	NN	O	I-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
ICSBP	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
negative	NN	O	B-protein
trans-acting	NN	O	I-protein
regulator	NN	O	I-protein
of	NN	O	O
some	NN	O	O
IFNgamma-inducible	NN	O	B-DNA
genes	NN	O	I-DNA
,	NN	O	O
were	NN	O	O
quantified	NN	O	O
.	NN	O	O

ICSBP	NN	O	B-protein
levels	NN	O	O
were	NN	O	O
reduced	NN	O	O
in	NN	O	O
FA-C	NN	O	B-cell_type
B	NN	O	I-cell_type
lymphoblasts	NN	O	I-cell_type
and	NN	O	O
MEFs	NN	O	B-cell_type
.	NN	O	O

However	NN	O	O
,	NN	O	O
enforced	NN	O	O
expression	NN	O	O
of	NN	O	O
ICSBP	NN	O	B-protein
failed	NN	O	O
to	NN	O	O
down-regulate	NN	O	O
IRF-1	NN	O	B-protein
,	NN	O	O
ISGF3gamma	NN	O	B-protein
,	NN	O	O
and	NN	O	O
p21	NN	O	B-protein
(	NN	O	I-protein
WAF1	NN	O	I-protein
)	NN	O	I-protein
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
FANCC	NN	O	B-protein
protein	NN	O	I-protein
functions	NN	O	O
to	NN	O	O
modulate	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
family	NN	O	O
of	NN	O	O
genes	NN	O	O
that	NN	O	O
in	NN	O	O
normal	NN	O	O
cells	NN	O	O
are	NN	O	O
inducible	NN	O	O
only	NN	O	O
by	NN	O	O
specific	NN	O	O
environmental	NN	O	O
cues	NN	O	O
for	NN	O	O
apoptosis	NN	O	O
or	NN	O	O
mitogenic	NN	O	O
inhibition	NN	O	O
,	NN	O	O
but	NN	O	O
it	NN	O	O
does	NN	O	O
so	NN	O	O
independently	NN	O	O
of	NN	O	O
the	NN	O	O
classic	NN	O	O
IFN-STAT1	NN	O	O
pathway	NN	O	O
and	NN	O	O
is	NN	O	O
not	NN	O	O
the	NN	O	O
direct	NN	O	O
result	NN	O	O
of	NN	O	O
reduced	NN	O	O
ICSBP	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
genetic	NN	O	O
investigation	NN	O	O
of	NN	O	O
E2A	NN	O	O
function	NN	O	O
in	NN	O	O
lymphocyte	NN	O	O
development	NN	O	O
.	NN	O	O

Lymphocytes	NN	O	B-cell_type
are	NN	O	O
derived	NN	O	O
from	NN	O	O
hematopoietic	NN	O	B-cell_type
stem	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
HSC	NN	O	B-cell_type
)	NN	O	O
following	NN	O	O
a	NN	O	O
series	NN	O	O
of	NN	O	O
regulated	NN	O	O
differentiation	NN	O	O
events	NN	O	O
.	NN	O	O

Multipotent	NN	O	B-cell_type
HSCs	NN	O	I-cell_type
become	NN	O	O
committed	NN	O	O
to	NN	O	O
the	NN	O	O
B	NN	O	B-cell_line
cell	NN	O	I-cell_line
lineage	NN	O	I-cell_line
in	NN	O	O
bone	NN	O	O
marrow	NN	O	O
and	NN	O	O
the	NN	O	O
T	NN	O	B-cell_line
cell	NN	O	I-cell_line
lineage	NN	O	I-cell_line
in	NN	O	O
the	NN	O	O
thymus	NN	O	O
after	NN	O	O
receiving	NN	O	O
appropriate	NN	O	O
signals	NN	O	O
from	NN	O	O
the	NN	O	O
corresponding	NN	O	O
microenvironment	NN	O	O
.	NN	O	O

These	NN	O	O
committed	NN	O	O
lymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
must	NN	O	O
then	NN	O	O
undergo	NN	O	O
V	NN	O	O
(	NN	O	O
D	NN	O	O
)	NN	O	O
J	NN	O	O
recombination	NN	O	O
at	NN	O	O
the	NN	O	O
immunoglobulin	NN	O	B-DNA
gene	NN	O	I-DNA
or	NN	O	O
T	NN	O	B-DNA
cell	NN	O	I-DNA
receptor	NN	O	I-DNA
gene	NN	O	I-DNA
locus	NN	O	I-DNA
resulting	NN	O	O
in	NN	O	O
clonal	NN	O	O
production	NN	O	O
of	NN	O	O
functional	NN	O	O
B	NN	O	B-cell_type
or	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

Lymphocyte	NN	O	O
commitment	NN	O	O
and	NN	O	O
differentiation	NN	O	O
are	NN	O	O
accompanied	NN	O	O
by	NN	O	O
programmed	NN	O	O
gene	NN	O	O
expression	NN	O	O
or	NN	O	O
repression	NN	O	O
events	NN	O	O
which	NN	O	O
are	NN	O	O
driven	NN	O	O
by	NN	O	O
lineage	NN	O	B-protein
and	NN	O	I-protein
stage	NN	O	I-protein
specific	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

The	NN	O	O
basic-helix-loop-helix	NN	O	B-protein
(	NN	O	I-protein
bHLH	NN	O	I-protein
)	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
encoded	NN	O	O
by	NN	O	O
the	NN	O	O
E2A	NN	O	B-DNA
gene	NN	O	I-DNA
are	NN	O	O
involved	NN	O	O
in	NN	O	O
several	NN	O	O
differentiation	NN	O	O
events	NN	O	O
during	NN	O	O
B	NN	O	O
and	NN	O	O
T	NN	O	O
cell	NN	O	O
development	NN	O	O
,	NN	O	O
including	NN	O	O
lineage	NN	O	O
commitment	NN	O	O
,	NN	O	O
initiation	NN	O	O
of	NN	O	O
V	NN	O	O
(	NN	O	O
D	NN	O	O
)	NN	O	O
J	NN	O	O
recombination	NN	O	O
,	NN	O	O
and	NN	O	O
antigen	NN	O	O
receptor	NN	O	O
mediated	NN	O	O
proliferation	NN	O	O
and	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Several	NN	O	O
recent	NN	O	O
reviews	NN	O	O
have	NN	O	O
provided	NN	O	O
a	NN	O	O
comprehensive	NN	O	O
discussion	NN	O	O
of	NN	O	O
biochemical	NN	O	O
,	NN	O	O
cellular	NN	O	O
,	NN	O	O
and	NN	O	O
genetic	NN	O	O
research	NN	O	O
on	NN	O	O
E2A	NN	O	O
function	NN	O	O
in	NN	O	O
lymphocyte	NN	O	O
development	NN	O	O
(	NN	O	O
1	NN	O	O
,	NN	O	O
2	NN	O	O
)	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
only	NN	O	O
discuss	NN	O	O
some	NN	O	O
of	NN	O	O
the	NN	O	O
genetic	NN	O	O
approaches	NN	O	O
our	NN	O	O
laboratory	NN	O	O
(	NN	O	O
except	NN	O	O
where	NN	O	O
it	NN	O	O
is	NN	O	O
noted	NN	O	O
)	NN	O	O
has	NN	O	O
undertaken	NN	O	O
to	NN	O	O
investigate	NN	O	O
the	NN	O	O
molecular	NN	O	O
pathways	NN	O	O
mediated	NN	O	O
by	NN	O	O
E2A	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
in	NN	O	O
lymphocyte	NN	O	O
development	NN	O	O

-DOCSTART-	O

D609	NN	O	O
inhibits	NN	O	O
ionizing	NN	O	O
radiation-induced	NN	O	O
oxidative	NN	O	O
damage	NN	O	O
by	NN	O	O
acting	NN	O	O
as	NN	O	O
a	NN	O	O
potent	NN	O	O
antioxidant	NN	O	O
.	NN	O	O

Tricyclodecan-9-yl-xanthogenate	NN	O	O
(	NN	O	O
D609	NN	O	O
)	NN	O	O
has	NN	O	O
been	NN	O	O
extensively	NN	O	O
studied	NN	O	O
in	NN	O	O
biological	NN	O	O
systems	NN	O	O
and	NN	O	O
exhibits	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
biological	NN	O	O
functions	NN	O	O
,	NN	O	O
including	NN	O	O
antiviral	NN	O	O
,	NN	O	O
antitumor	NN	O	O
,	NN	O	O
and	NN	O	O
anti-inflammatory	NN	O	O
activities	NN	O	O
.	NN	O	O

Most	NN	O	O
of	NN	O	O
these	NN	O	O
activities	NN	O	O
have	NN	O	O
been	NN	O	O
largely	NN	O	O
attributed	NN	O	O
to	NN	O	O
the	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
of	NN	O	O
D609	NN	O	O
on	NN	O	O
phosphatidylcholine-specific	NN	O	O
phospholipase	NN	O	B-protein
C	NN	O	I-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
as	NN	O	O
a	NN	O	O
xanthate	NN	O	O
derivative	NN	O	O
,	NN	O	O
D609	NN	O	O
is	NN	O	O
a	NN	O	O
strong	NN	O	O
electrolyte	NN	O	O
and	NN	O	O
readily	NN	O	O
dissociates	NN	O	O
to	NN	O	O
xanthate	NN	O	O
anions	NN	O	O
and	NN	O	O
cations	NN	O	O
of	NN	O	O
alkali	NN	O	O
metals	NN	O	O
in	NN	O	O
solution	NN	O	O
.	NN	O	O

Xanthate	NN	O	O
anions	NN	O	O
and	NN	O	O
protonated	NN	O	O
xanthic	NN	O	O
acid	NN	O	O
contain	NN	O	O
a	NN	O	O
free	NN	O	O
thiol	NN	O	O
moiety	NN	O	O
and	NN	O	O
are	NN	O	O
highly	NN	O	O
reductive	NN	O	O
.	NN	O	O

This	NN	O	O
implies	NN	O	O
that	NN	O	O
D609	NN	O	O
and	NN	O	O
other	NN	O	O
xanthate	NN	O	O
derivatives	NN	O	O
may	NN	O	O
function	NN	O	O
as	NN	O	O
potent	NN	O	O
antioxidants	NN	O	O
.	NN	O	O

Indeed	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
D609	NN	O	O
inhibited	NN	O	O
the	NN	O	O
Fenton	NN	O	O
reaction-induced	NN	O	O
oxidation	NN	O	O
of	NN	O	O
dihydrorhodamine	NN	O	O
123	NN	O	O
in	NN	O	O
a	NN	O	O
dose-dependent	NN	O	O
manner	NN	O	O
similar	NN	O	O
to	NN	O	O
that	NN	O	O
of	NN	O	O
pyrrolidinedithiocarbamate	NN	O	O
,	NN	O	O
a	NN	O	O
well	NN	O	O
known	NN	O	O
antioxidant	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
D609	NN	O	O
inhibited	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
the	NN	O	O
alpha-phenyl-tert-butylnitrone-free	NN	O	O
radical	NN	O	O
spin	NN	O	O
adducts	NN	O	O
and	NN	O	O
lipid	NN	O	O
peroxidation	NN	O	O
of	NN	O	O
synaptosomal	NN	O	O
membranes	NN	O	O
by	NN	O	O
the	NN	O	O
Fenton	NN	O	O
reagents	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
preincubation	NN	O	O
of	NN	O	O
lymphocytes	NN	O	B-cell_type
with	NN	O	O
D609	NN	O	O
resulted	NN	O	O
in	NN	O	O
a	NN	O	O
significant	NN	O	O
diminution	NN	O	O
of	NN	O	O
ionizing	NN	O	O
radiation	NN	O	O
(	NN	O	O
IR	NN	O	O
)	NN	O	O
-induced	NN	O	O
1	NN	O	O
)	NN	O	O
production	NN	O	O
of	NN	O	O
reactive	NN	O	O
oxygen	NN	O	O
species	NN	O	O
;	NN	O	O
2	NN	O	O
)	NN	O	O
decrease	NN	O	O
in	NN	O	O
intracellular	NN	O	O
reduced	NN	O	O
glutathione	NN	O	O
;	NN	O	O
3	NN	O	O
)	NN	O	O
oxidative	NN	O	O
damage	NN	O	O
to	NN	O	O
proteins	NN	O	O
and	NN	O	O
lipids	NN	O	O
;	NN	O	O
and	NN	O	O
4	NN	O	O
)	NN	O	O
activation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
when	NN	O	O
D609	NN	O	O
(	NN	O	O
50	NN	O	O
mg/kg	NN	O	O
i.v.	NN	O	O
)	NN	O	O
was	NN	O	O
administered	NN	O	O
to	NN	O	O
mice	NN	O	O
10	NN	O	O
min	NN	O	O
prior	NN	O	O
to	NN	O	O
total	NN	O	O
body	NN	O	O
IR	NN	O	O
(	NN	O	O
6.5	NN	O	O
and	NN	O	O
8.5	NN	O	O
Gy	NN	O	O
)	NN	O	O
,	NN	O	O
it	NN	O	O
protected	NN	O	O
the	NN	O	O
mice	NN	O	O
from	NN	O	O
IR-induced	NN	O	O
lethality	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
these	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
D609	NN	O	O
is	NN	O	O
a	NN	O	O
potent	NN	O	O
antioxidant	NN	O	O
and	NN	O	O
has	NN	O	O
the	NN	O	O
ability	NN	O	O
to	NN	O	O
inhibit	NN	O	O
IR-induced	NN	O	O
cellular	NN	O	O
oxidative	NN	O	O
stress	NN	O	O
.	NN	O	O

-DOCSTART-	O

Inhibition	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
by	NN	O	O
the	NN	O	O
glucocorticoid-inducible	NN	O	O
protein	NN	O	B-protein
GILZ	NN	O	I-protein
.	NN	O	O

The	NN	O	O
immunosuppressive	NN	O	O
effects	NN	O	O
of	NN	O	O
glucocorticoids	NN	O	O
arise	NN	O	O
largely	NN	O	O
by	NN	O	O
inhibition	NN	O	O
of	NN	O	O
cytokine	NN	O	O
gene	NN	O	O
expression	NN	O	O
,	NN	O	O
which	NN	O	O
has	NN	O	O
been	NN	O	O
ascribed	NN	O	O
to	NN	O	O
interference	NN	O	O
between	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
and	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
such	NN	O	O
as	NN	O	O
AP-1	NN	O	B-protein
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
as	NN	O	O
well	NN	O	O
as	NN	O	O
by	NN	O	O
competition	NN	O	O
for	NN	O	O
common	NN	O	O
coactivators	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
glucocorticoid-induced	NN	O	O
inhibition	NN	O	O
of	NN	O	O
interleukin-2	NN	O	O
mRNA	NN	O	O
expression	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
normal	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
required	NN	O	O
new	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
this	NN	O	O
phenomenon	NN	O	O
is	NN	O	O
secondary	NN	O	O
to	NN	O	O
expression	NN	O	O
of	NN	O	O
glucocorticoid-regulated	NN	O	O
genes	NN	O	B-DNA
.	NN	O	O

One	NN	O	O
of	NN	O	O
the	NN	O	O
most	NN	O	O
prominent	NN	O	O
glucocorticoid-induced	NN	O	O
genes	NN	O	B-DNA
is	NN	O	O
glucocorticoid-induced	NN	O	B-DNA
leucine	NN	O	I-DNA
zipper	NN	O	I-DNA
(	NN	O	O
GILZ	NN	O	B-DNA
)	NN	O	O
,	NN	O	O
which	NN	O	O
has	NN	O	O
been	NN	O	O
reported	NN	O	O
to	NN	O	O
inhibit	NN	O	O
activation-induced	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
Fas	NN	O	B-RNA
ligand	NN	O	I-RNA
(	NN	O	I-RNA
FasL	NN	O	I-RNA
)	NN	O	I-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

Indeed	NN	O	O
,	NN	O	O
transient	NN	O	O
expression	NN	O	O
of	NN	O	O
GILZ	NN	O	B-DNA
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
blocked	NN	O	O
induction	NN	O	O
of	NN	O	O
a	NN	O	O
reporter	NN	O	B-DNA
construct	NN	O	I-DNA
driven	NN	O	O
by	NN	O	O
the	NN	O	O
FasL	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
could	NN	O	O
be	NN	O	O
accounted	NN	O	O
for	NN	O	O
by	NN	O	O
GILZ	NN	O	B-protein
-mediated	NN	O	O
inhibition	NN	O	O
of	NN	O	O
Egr-2	NN	O	B-protein
and	NN	O	O
Egr-3	NN	O	B-protein
,	NN	O	O
NFAT/AP-1	NN	O	B-protein
-inducible	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
that	NN	O	O
bind	NN	O	O
a	NN	O	O
regulatory	NN	O	B-DNA
element	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
FasL	NN	O	B-DNA
promoter	NN	O	I-DNA
and	NN	O	O
up-regulate	NN	O	O
FasL	NN	O	O
expression	NN	O	O
.	NN	O	O

GILZ	NN	O	B-protein
also	NN	O	O
potently	NN	O	O
inhibited	NN	O	O
AP-1-driven	NN	O	B-DNA
and	NN	O	I-DNA
IL-2	NN	O	I-DNA
promoter-driven	NN	O	I-DNA
reporter	NN	O	I-DNA
constructs	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
recombinant	NN	O	O
GILZ	NN	O	B-protein
specifically	NN	O	O
interacted	NN	O	O
with	NN	O	O
c-Fos	NN	O	B-protein
and	NN	O	O
c-Jun	NN	O	B-protein
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
inhibited	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
active	NN	O	B-protein
AP-1	NN	O	I-protein
to	NN	O	O
its	NN	O	O
target	NN	O	O
DNA	NN	O	O
.	NN	O	O

Whereas	NN	O	O
homodimerization	NN	O	O
of	NN	O	O
GILZ	NN	O	B-protein
required	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
its	NN	O	O
leucine	NN	O	B-protein
zipper	NN	O	I-protein
,	NN	O	O
the	NN	O	O
interaction	NN	O	O
with	NN	O	O
c-Fos	NN	O	B-protein
and	NN	O	O
c-Jun	NN	O	B-protein
occurred	NN	O	O
through	NN	O	O
the	NN	O	O
N-terminal	NN	O	B-protein
60-amino	NN	O	B-protein
acid	NN	O	I-protein
region	NN	O	I-protein
of	NN	O	O
GILZ	NN	O	B-protein
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
GILZ	NN	O	B-protein
represents	NN	O	O
a	NN	O	O
glucocorticoid-induced	NN	O	O
gene	NN	O	O
product	NN	O	O
that	NN	O	O
can	NN	O	O
inhibit	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
activation-induced	NN	O	O
events	NN	O	O
,	NN	O	O
at	NN	O	O
least	NN	O	O
in	NN	O	O
part	NN	O	O
by	NN	O	O
direct	NN	O	O
interference	NN	O	O
with	NN	O	O
AP-1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
is	NN	O	O
therefore	NN	O	O
a	NN	O	O
candidate	NN	O	O
for	NN	O	O
a	NN	O	O
mediator	NN	O	O
of	NN	O	O
glucocorticoid-induced	NN	O	O
immunosuppression	NN	O	O
.	NN	O	O

-DOCSTART-	O

Pharmacokinetic	NN	O	O
differences	NN	O	O
between	NN	O	O
a	NN	O	O
T	NN	O	O
cell-tolerizing	NN	O	O
and	NN	O	O
a	NN	O	O
T	NN	O	O
cell-activating	NN	O	O
peptide	NN	O	O
.	NN	O	O

Vaccination	NN	O	O
with	NN	O	O
a	NN	O	O
peptide	NN	O	O
representing	NN	O	O
a	NN	O	O
CTL	NN	O	O
epitope	NN	O	O
from	NN	O	O
the	NN	O	O
human	NN	O	O
papillomavirus	NN	O	O
(	NN	O	O
HPV	NN	O	O
)	NN	O	O
16	NN	O	O
E7	NN	O	B-protein
protein	NN	O	I-protein
induces	NN	O	O
a	NN	O	O
specific	NN	O	O
CTL	NN	O	O
response	NN	O	O
that	NN	O	O
prevents	NN	O	O
the	NN	O	O
outgrowth	NN	O	O
of	NN	O	O
HPV16	NN	O	O
E7-expressing	NN	O	O
tumors	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
vaccination	NN	O	O
with	NN	O	O
a	NN	O	O
peptide	NN	O	O
encoding	NN	O	O
an	NN	O	O
adenovirus	NN	O	O
type	NN	O	O
5	NN	O	O
(	NN	O	O
Ad5	NN	O	O
)	NN	O	O
E1A	NN	O	O
CTL	NN	O	O
epitope	NN	O	O
results	NN	O	O
in	NN	O	O
CTL	NN	O	O
tolerance	NN	O	O
and	NN	O	O
enhanced	NN	O	O
growth	NN	O	O
of	NN	O	O
an	NN	O	O
Ad5	NN	O	O
E1A-expressing	NN	O	O
tumor	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
unclear	NN	O	O
why	NN	O	O
these	NN	O	O
peptides	NN	O	O
induce	NN	O	O
such	NN	O	O
opposite	NN	O	O
effects	NN	O	O
.	NN	O	O

To	NN	O	O
determine	NN	O	O
whether	NN	O	O
a	NN	O	O
difference	NN	O	O
in	NN	O	O
pharmacokinetics	NN	O	O
can	NN	O	O
explain	NN	O	O
the	NN	O	O
functional	NN	O	O
contrasts	NN	O	O
,	NN	O	O
tritiated	NN	O	O
Ad5	NN	O	O
E1A	NN	O	O
and	NN	O	O
HPV16	NN	O	O
E7	NN	O	O
peptides	NN	O	O
were	NN	O	O
injected	NN	O	O
into	NN	O	O
mice	NN	O	O
.	NN	O	O

Results	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
tolerizing	NN	O	O
peptide	NN	O	O
spread	NN	O	O
through	NN	O	O
the	NN	O	O
body	NN	O	O
16	NN	O	O
times	NN	O	O
faster	NN	O	O
than	NN	O	O
the	NN	O	O
activating	NN	O	O
peptide	NN	O	O
and	NN	O	O
was	NN	O	O
cleared	NN	O	O
at	NN	O	O
least	NN	O	O
2	NN	O	O
times	NN	O	O
faster	NN	O	O
.	NN	O	O

The	NN	O	O
HPV16	NN	O	O
E7	NN	O	O
peptide	NN	O	O
kinetics	NN	O	O
correlated	NN	O	O
with	NN	O	O
the	NN	O	O
kinetics	NN	O	O
of	NN	O	O
HPV16	NN	O	B-protein
E7	NN	O	I-protein
-specific	NN	O	O
CTL	NN	O	O
induction	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
Ad5	NN	O	O
E1A	NN	O	O
peptide	NN	O	O
injection	NN	O	O
resulted	NN	O	O
in	NN	O	O
physical	NN	O	O
deletion	NN	O	O
of	NN	O	O
preexisting	NN	O	O
Ad5	NN	O	B-protein
E1A	NN	O	I-protein
-specific	NN	O	O
CTLs	NN	O	B-protein
within	NN	O	O
24	NN	O	O
h	NN	O	O
after	NN	O	O
injection	NN	O	O
.	NN	O	O

This	NN	O	O
tolerization	NN	O	O
occurred	NN	O	O
at	NN	O	O
the	NN	O	O
time	NN	O	O
when	NN	O	O
the	NN	O	O
peptide	NN	O	O
reached	NN	O	O
its	NN	O	O
maximum	NN	O	O
peptide	NN	O	O
concentration	NN	O	O
in	NN	O	O
the	NN	O	O
organs	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
ubiquitous	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
tolerizing	NN	O	O
Ad5	NN	O	O
E1A	NN	O	O
peptide	NN	O	O
within	NN	O	O
a	NN	O	O
short	NN	O	O
period	NN	O	O
of	NN	O	O
time	NN	O	O
causes	NN	O	O
activation-induced	NN	O	O
cell	NN	O	O
death	NN	O	O
of	NN	O	O
Ad5	NN	O	B-protein
E1A	NN	O	I-protein
-specific	NN	O	O
CTLs	NN	O	B-protein
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
information	NN	O	O
on	NN	O	O
the	NN	O	O
pharmacokinetics	NN	O	O
of	NN	O	O
peptides	NN	O	O
is	NN	O	O
vital	NN	O	O
for	NN	O	O
the	NN	O	O
safety	NN	O	O
and	NN	O	O
efficacy	NN	O	O
of	NN	O	O
peptide-based	NN	O	O
vaccines	NN	O	O
.	NN	O	O

-DOCSTART-	O

Smad3	NN	O	B-protein
and	NN	O	O
Smad4	NN	O	B-protein
mediate	NN	O	O
transforming	NN	O	B-protein
growth	NN	O	I-protein
factor-beta1	NN	O	I-protein
-induced	NN	O	O
IgA	NN	O	O
expression	NN	O	O
in	NN	O	O
murine	NN	O	B-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Transforming	NN	O	B-protein
growth	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	I-protein
TGF	NN	O	I-protein
)	NN	O	I-protein
-beta1	NN	O	I-protein
is	NN	O	O
well	NN	O	O
established	NN	O	O
as	NN	O	O
a	NN	O	O
critical	NN	O	O
IgA	NN	O	B-protein
isotype	NN	O	I-protein
switching	NN	O	I-protein
factor	NN	O	I-protein
and	NN	O	O
Smad	NN	O	B-protein
molecules	NN	O	I-protein
have	NN	O	O
been	NN	O	O
reported	NN	O	O
to	NN	O	O
act	NN	O	O
as	NN	O	O
transducers	NN	O	B-protein
and	NN	O	O
transcriptional	NN	O	B-protein
factors	NN	O	I-protein
in	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
TGF-beta1	NN	O	B-protein
-targeted	NN	O	O
genes	NN	O	B-DNA
.	NN	O	O

We	NN	O	O
examined	NN	O	O
the	NN	O	O
involvement	NN	O	O
of	NN	O	O
Smad	NN	O	B-protein
proteins	NN	O	I-protein
in	NN	O	O
TGF-beta1	NN	O	B-protein
-induced	NN	O	O
IgA	NN	O	O
expression	NN	O	O
.	NN	O	O

First	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
TGF-beta1	NN	O	B-protein
significantly	NN	O	O
increases	NN	O	O
endogenous	NN	O	B-RNA
germ-line	NN	O	I-RNA
(	NN	O	I-RNA
GL	NN	O	I-RNA
)	NN	O	I-RNA
alpha	NN	O	I-RNA
transcripts	NN	O	I-RNA
by	NN	O	O
LPS-stimulated	NN	O	O
CH12.LX.4933	NN	O	B-cell_line
(	NN	O	I-cell_line
mu	NN	O	I-cell_line
(	NN	O	I-cell_line
+	NN	O	I-cell_line
)	NN	O	I-cell_line
)	NN	O	I-cell_line
B	NN	O	I-cell_line
lymphoma	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
its	NN	O	O
signaling	NN	O	O
mechanisms	NN	O	O
,	NN	O	O
the	NN	O	O
lymphoma	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
was	NN	O	O
transfected	NN	O	O
with	NN	O	O
pFL3	NN	O	B-DNA
that	NN	O	O
contains	NN	O	O
the	NN	O	O
TGF-beta-responsive	NN	O	B-DNA
element	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
GLalpha	NN	O	B-DNA
promoter	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
stimulated	NN	O	O
with	NN	O	O
TGF-beta1	NN	O	B-protein
.	NN	O	O

Similar	NN	O	O
to	NN	O	O
endogenous	NN	O	B-RNA
GLalpha	NN	O	I-RNA
transcripts	NN	O	I-RNA
,	NN	O	O
TGF-beta1	NN	O	B-protein
induces	NN	O	O
GLalpha	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
and	NN	O	O
overexpression	NN	O	O
of	NN	O	O
Smad3	NN	O	B-protein
markedly	NN	O	O
enhances	NN	O	O
the	NN	O	O
promoter	NN	O	O
activity	NN	O	O
.	NN	O	O

This	NN	O	O
activity	NN	O	O
is	NN	O	O
further	NN	O	O
augmented	NN	O	O
by	NN	O	O
cotransfected	NN	O	B-protein
Smad4	NN	O	I-protein
.	NN	O	O

On	NN	O	O
the	NN	O	O
other	NN	O	O
hand	NN	O	O
,	NN	O	O
Smad7	NN	O	B-protein
substantially	NN	O	O
abrogates	NN	O	O
the	NN	O	O
synergistic	NN	O	O
effect	NN	O	O
of	NN	O	O
Smad3/4	NN	O	B-protein
on	NN	O	O
GLalpha	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
overexpression	NN	O	O
of	NN	O	O
Smad3/4	NN	O	B-protein
enhances	NN	O	O
TGF-beta1	NN	O	B-protein
-induced	NN	O	O
endogenous	NN	O	B-RNA
GLalpha	NN	O	I-RNA
transcripts	NN	O	I-RNA
in	NN	O	O
normal	NN	O	B-cell_type
spleen	NN	O	I-cell_type
B	NN	O	I-cell_type
cell	NN	O	I-cell_type
s	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
TGF-beta1	NN	O	B-protein
,	NN	O	O
overexpression	NN	O	O
of	NN	O	O
Smad3/4	NN	O	B-protein
selectively	NN	O	O
increases	NN	O	O
both	NN	O	O
surface	NN	O	O
IgA	NN	O	O
expression	NN	O	O
and	NN	O	O
IgA	NN	O	O
production	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
from	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
indicate	NN	O	O
that	NN	O	O
Smad3	NN	O	B-protein
,	NN	O	O
Smad4	NN	O	B-protein
,	NN	O	O
and	NN	O	O
Smad7	NN	O	B-protein
,	NN	O	O
at	NN	O	O
least	NN	O	O
in	NN	O	O
part	NN	O	O
,	NN	O	O
serve	NN	O	O
as	NN	O	O
mediators	NN	O	O
linking	NN	O	O
TGF-beta1	NN	O	B-protein
to	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
IgA	NN	O	B-DNA
switching	NN	O	I-DNA
related	NN	O	I-DNA
gene	NN	O	I-DNA
and	NN	O	O
regulation	NN	O	O
of	NN	O	O
IgA	NN	O	O
class	NN	O	O
switching	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
translesion	NN	O	B-protein
DNA	NN	O	I-protein
polymerase	NN	O	I-protein
zeta	NN	O	I-protein
plays	NN	O	O
a	NN	O	O
major	NN	O	O
role	NN	O	O
in	NN	O	O
Ig	NN	O	O
and	NN	O	O
bcl-6	NN	O	O
somatic	NN	O	O
hypermutation	NN	O	O
.	NN	O	O

Ig	NN	O	O
somatic	NN	O	O
mutations	NN	O	O
would	NN	O	O
be	NN	O	O
introduced	NN	O	O
by	NN	O	O
a	NN	O	O
polymerase	NN	O	B-protein
(	NN	O	O
pol	NN	O	B-protein
)	NN	O	O
while	NN	O	O
repairing	NN	O	O
DNA	NN	O	O
outside	NN	O	O
main	NN	O	O
DNA	NN	O	O
replication	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
human	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
constitutively	NN	O	O
express	NN	O	O
the	NN	O	O
translesion	NN	O	B-protein
pol	NN	O	I-protein
zeta	NN	O	I-protein
,	NN	O	O
which	NN	O	O
effectively	NN	O	O
extends	NN	O	O
DNA	NN	O	O
past	NN	O	O
mismatched	NN	O	O
bases	NN	O	O
(	NN	O	O
mispair	NN	O	O
extender	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
pol	NN	O	B-protein
eta	NN	O	I-protein
,	NN	O	O
which	NN	O	O
bypasses	NN	O	O
DNA	NN	O	O
lesions	NN	O	O
in	NN	O	O
an	NN	O	O
error-free	NN	O	O
fashion	NN	O	O
.	NN	O	O

Upon	NN	O	O
B	NN	O	B-protein
cell	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
BCR	NN	O	B-protein
)	NN	O	O
engagement	NN	O	O
and	NN	O	O
coculture	NN	O	O
with	NN	O	O
activated	NN	O	B-cell_type
CD4+	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
these	NN	O	O
lymphocytes	NN	O	B-cell_type
upregulated	NN	O	O
pol	NN	O	B-protein
zeta	NN	O	I-protein
,	NN	O	O
downregulated	NN	O	O
pol	NN	O	B-protein
eta	NN	O	I-protein
,	NN	O	O
and	NN	O	O
mutated	NN	O	O
the	NN	O	O
Ig	NN	O	B-DNA
and	NN	O	I-DNA
bcl-6	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
the	NN	O	O
pol	NN	O	B-protein
zeta	NN	O	I-protein
REV3	NN	O	B-protein
catalytic	NN	O	I-protein
subunit	NN	O	I-protein
by	NN	O	O
specific	NN	O	O
phosphorothioate-modified	NN	O	O
oligonucleotides	NN	O	O
impaired	NN	O	O
Ig	NN	O	O
and	NN	O	O
bcl-6	NN	O	O
hypermutation	NN	O	O
and	NN	O	O
UV	NN	O	O
damage-induced	NN	O	O
DNA	NN	O	O
mutagenesis	NN	O	O
,	NN	O	O
without	NN	O	O
affecting	NN	O	O
cell	NN	O	O
cycle	NN	O	O
or	NN	O	O
viability	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
pol	NN	O	B-protein
zeta	NN	O	I-protein
plays	NN	O	O
a	NN	O	O
critical	NN	O	O
role	NN	O	O
in	NN	O	O
Ig	NN	O	O
and	NN	O	O
bcl-6	NN	O	O
hypermutation	NN	O	O
,	NN	O	O
perhaps	NN	O	O
facilitated	NN	O	O
by	NN	O	O
the	NN	O	O
downregulation	NN	O	O
of	NN	O	O
pol	NN	O	B-protein
eta	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Molecular	NN	O	O
mechanism	NN	O	O
of	NN	O	O
cell	NN	O	O
cycle	NN	O	O
progression	NN	O	O
induced	NN	O	O
by	NN	O	O
the	NN	O	O
oncogene	NN	O	B-protein
product	NN	O	I-protein
Tax	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

The	NN	O	O
trans-activator	NN	O	B-protein
protein	NN	O	I-protein
Tax	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
plays	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
adult	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
through	NN	O	O
,	NN	O	O
at	NN	O	O
least	NN	O	O
in	NN	O	O
part	NN	O	O
,	NN	O	O
its	NN	O	O
ability	NN	O	O
to	NN	O	O
stimulate	NN	O	O
cell	NN	O	O
growth	NN	O	O
.	NN	O	O

We	NN	O	O
previously	NN	O	O
reported	NN	O	O
that	NN	O	O
Tax	NN	O	B-protein
induced	NN	O	O
cell	NN	O	O
cycle	NN	O	O
progression	NN	O	O
from	NN	O	O
G0/G1	NN	O	O
phase	NN	O	O
to	NN	O	O
S	NN	O	O
and	NN	O	O
G2/M	NN	O	O
phases	NN	O	O
in	NN	O	O
human	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
line	NN	O	I-cell_line
Kit	NN	O	I-cell_line
225	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

To	NN	O	O
elucidate	NN	O	O
molecular	NN	O	O
mechanism	NN	O	O
of	NN	O	O
Tax	NN	O	B-protein
-induced	NN	O	O
cell	NN	O	O
cycle	NN	O	O
progression	NN	O	O
,	NN	O	O
we	NN	O	O
systematically	NN	O	O
examined	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
Tax	NN	O	B-protein
on	NN	O	O
biochemical	NN	O	O
events	NN	O	O
associated	NN	O	O
with	NN	O	O
cell	NN	O	O
cycle	NN	O	O
progression	NN	O	O
.	NN	O	O

Introduction	NN	O	O
of	NN	O	O
Tax	NN	O	B-protein
into	NN	O	O
resting	NN	O	B-cell_line
Kit	NN	O	I-cell_line
225	NN	O	I-cell_line
cells	NN	O	I-cell_line
induced	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
G1/S	NN	O	O
transition	NN	O	O
regulation	NN	O	O
cascade	NN	O	O
consisting	NN	O	O
of	NN	O	O
activation	NN	O	O
of	NN	O	O
cyclin	NN	O	B-protein
dependent	NN	O	I-protein
kinase	NN	O	I-protein
2	NN	O	I-protein
(	NN	O	O
CDK2	NN	O	B-protein
)	NN	O	O
and	NN	O	O
CDK4	NN	O	B-protein
,	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
the	NN	O	O
Rb	NN	O	B-protein
family	NN	O	I-protein
proteins	NN	O	I-protein
and	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
free	NN	O	O
E2F	NN	O	B-protein
.	NN	O	O

The	NN	O	O
kinase	NN	O	O
activation	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
result	NN	O	O
from	NN	O	O
Tax	NN	O	B-protein
-induced	NN	O	O
expression	NN	O	O
of	NN	O	O
genes	NN	O	O
for	NN	O	O
cell	NN	O	B-protein
cycle	NN	O	I-protein
regulatory	NN	O	I-protein
molecules	NN	O	I-protein
including	NN	O	O
cyclin	NN	O	B-protein
D2	NN	O	I-protein
,	NN	O	O
cyclin	NN	O	B-protein
E	NN	O	I-protein
,	NN	O	O
E2F1	NN	O	B-protein
,	NN	O	O
CDK2	NN	O	B-protein
,	NN	O	O
CDK4	NN	O	B-protein
and	NN	O	O
CDK6	NN	O	B-protein
,	NN	O	O
and	NN	O	O
Tax	NN	O	B-protein
-induced	NN	O	O
reduction	NN	O	O
of	NN	O	O
CDK	NN	O	B-protein
inhibitors	NN	O	I-protein
p19	NN	O	I-protein
(	NN	O	O
INK4d	NN	O	B-protein
)	NN	O	O
and	NN	O	O
p27	NN	O	B-protein
(	NN	O	I-protein
Kip1	NN	O	I-protein
)	NN	O	I-protein
.	NN	O	O

These	NN	O	O
modulations	NN	O	O
by	NN	O	O
Tax	NN	O	B-protein
always	NN	O	O
paralleled	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
Tax	NN	O	B-protein
to	NN	O	O
activate	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	O
transcription	NN	O	O
pathway	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
the	NN	O	O
important	NN	O	O
role	NN	O	O
of	NN	O	O
Tax	NN	O	B-protein
-mediated	NN	O	O
trans-activation	NN	O	O
of	NN	O	O
the	NN	O	O
genes	NN	O	O
for	NN	O	O
cell	NN	O	B-protein
cycle	NN	O	I-protein
regulatory	NN	O	I-protein
molecules	NN	O	I-protein
in	NN	O	O
Tax	NN	O	B-protein
-induced	NN	O	O
cell	NN	O	O
cycle	NN	O	O
progression	NN	O	O
.	NN	O	O

-DOCSTART-	O

Cot	NN	O	B-protein
kinase	NN	O	I-protein
induces	NN	O	O
cyclooxygenase-2	NN	O	O
expression	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
through	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
.	NN	O	O

Cyclooxygenase-2	NN	O	B-protein
(	NN	O	O
COX-2	NN	O	B-protein
)	NN	O	O
is	NN	O	O
induced	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
upon	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
receptor	NN	O	I-protein
triggering	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
Cot	NN	O	B-protein
kinase	NN	O	I-protein
,	NN	O	O
a	NN	O	O
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
kinase	NN	O	I-protein
kinase	NN	O	I-protein
involved	NN	O	O
in	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
,	NN	O	O
up-regulates	NN	O	O
COX-2	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Induction	NN	O	O
of	NN	O	O
COX-2	NN	O	O
promoter	NN	O	O
activity	NN	O	O
by	NN	O	O
Cot	NN	O	B-protein
kinase	NN	O	I-protein
occurred	NN	O	O
mainly	NN	O	O
through	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
(	NN	O	O
NFAT	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Mutation	NN	O	O
of	NN	O	O
the	NN	O	O
distal	NN	O	B-DNA
(	NN	O	I-DNA
-105/-97	NN	O	I-DNA
)	NN	O	I-DNA
and	NN	O	O
proximal	NN	O	B-DNA
(	NN	O	I-DNA
-76/-61	NN	O	I-DNA
)	NN	O	I-DNA
NFAT	NN	O	B-DNA
response	NN	O	I-DNA
elements	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
COX-2	NN	O	B-DNA
promoter	NN	O	I-DNA
abolished	NN	O	O
the	NN	O	O
activation	NN	O	O
induced	NN	O	O
by	NN	O	O
Cot	NN	O	B-protein
kinase	NN	O	I-protein
.	NN	O	O

Even	NN	O	O
more	NN	O	O
,	NN	O	O
coexpression	NN	O	O
of	NN	O	O
a	NN	O	O
dominant	NN	O	O
negative	NN	O	O
version	NN	O	O
of	NN	O	O
NFAT	NN	O	B-protein
inhibited	NN	O	O
Cot	NN	O	B-protein
kinase	NN	O	I-protein
-mediated	NN	O	O
COX-2	NN	O	B-DNA
promoter	NN	O	I-DNA
activation	NN	O	O
,	NN	O	O
whereas	NN	O	O
cotransfection	NN	O	O
of	NN	O	O
a	NN	O	O
constitutively	NN	O	O
active	NN	O	O
version	NN	O	O
of	NN	O	O
the	NN	O	O
calcium-dependent	NN	O	B-protein
phosphatase	NN	O	I-protein
calcineurin	NN	O	I-protein
synergizes	NN	O	O
with	NN	O	O
Cot	NN	O	B-protein
kinase	NN	O	I-protein
in	NN	O	O
the	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
COX-2	NN	O	B-DNA
promoter	NN	O	I-DNA
-driven	NN	O	O
transcription	NN	O	O
.	NN	O	O

Strikingly	NN	O	O
,	NN	O	O
Cot	NN	O	B-protein
kinase	NN	O	I-protein
increased	NN	O	O
transactivation	NN	O	O
mediated	NN	O	O
by	NN	O	O
a	NN	O	O
GAL4-NFAT	NN	O	B-protein
fusion	NN	O	I-protein
protein	NN	O	I-protein
containing	NN	O	O
the	NN	O	O
N-terminal	NN	O	B-protein
transactivation	NN	O	I-protein
domain	NN	O	I-protein
of	NN	O	O
NFATp	NN	O	B-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
plus	NN	O	O
calcium	NN	O	O
ionophore	NN	O	O
A23187	NN	O	O
,	NN	O	O
Cot	NN	O	B-protein
kinase	NN	O	I-protein
increases	NN	O	O
both	NN	O	O
COX-2	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
and	NN	O	O
NFAT	NN	O	B-protein
-mediated	NN	O	O
transactivation	NN	O	O
in	NN	O	O
a	NN	O	O
cyclosporin	NN	O	O
A-independent	NN	O	O
manner	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
Cot	NN	O	B-protein
kinase	NN	O	I-protein
up-regulates	NN	O	O
COX-2	NN	O	B-DNA
promoter	NN	O	I-DNA
-driven	NN	O	O
transcription	NN	O	O
through	NN	O	O
the	NN	O	O
NFAT	NN	O	B-DNA
response	NN	O	I-DNA
elements	NN	O	I-DNA
,	NN	O	O
being	NN	O	O
the	NN	O	O
Cot	NN	O	B-protein
kinase	NN	O	I-protein
-induced	NN	O	O
NFAT	NN	O	B-protein
-dependent	NN	O	O
transactivation	NN	O	O
presumably	NN	O	O
implicated	NN	O	O
in	NN	O	O
this	NN	O	O
up-regulation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Positive	NN	O	O
and	NN	O	O
negative	NN	O	O
roles	NN	O	O
of	NN	O	O
the	NN	O	O
trans-acting	NN	O	B-protein
T	NN	O	I-protein
cell	NN	O	I-protein
factor-1	NN	O	I-protein
for	NN	O	O
the	NN	O	O
acquisition	NN	O	O
of	NN	O	O
distinct	NN	O	O
Ly-49	NN	O	B-protein
MHC	NN	O	I-protein
class	NN	O	I-protein
I	NN	O	I-protein
receptors	NN	O	I-protein
by	NN	O	O
NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Members	NN	O	O
of	NN	O	O
the	NN	O	O
Ly-49	NN	O	B-DNA
gene	NN	O	I-DNA
family	NN	O	I-DNA
code	NN	O	O
for	NN	O	O
class	NN	O	B-protein
I	NN	O	I-protein
MHC-specific	NN	O	I-protein
receptors	NN	O	I-protein
that	NN	O	O
regulate	NN	O	O
NK	NN	O	O
cell	NN	O	O
function	NN	O	O
.	NN	O	O

Due	NN	O	O
to	NN	O	O
a	NN	O	O
combinatorial	NN	O	O
distribution	NN	O	O
of	NN	O	O
Ly-49	NN	O	B-protein
receptors	NN	O	I-protein
,	NN	O	O
NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
display	NN	O	O
considerable	NN	O	O
clonal	NN	O	O
heterogeneity	NN	O	O
.	NN	O	O

The	NN	O	O
acquisition	NN	O	O
of	NN	O	O
one	NN	O	O
Ly-49	NN	O	B-protein
receptor	NN	O	I-protein
,	NN	O	O
Ly-49A	NN	O	B-protein
is	NN	O	O
strictly	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
transcriptional	NN	O	B-protein
trans-acting	NN	O	I-protein
factor	NN	O	I-protein
T	NN	O	B-protein
cell-specific	NN	O	I-protein
factor-1	NN	O	I-protein
(	NN	O	O
TCF-1	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Indeed	NN	O	O
,	NN	O	O
TCF-1	NN	O	B-protein
binds	NN	O	O
to	NN	O	O
two	NN	O	O
sites	NN	O	O
in	NN	O	O
the	NN	O	O
Ly-49a	NN	O	B-DNA
promoter	NN	O	I-DNA
and	NN	O	O
regulates	NN	O	O
its	NN	O	O
activity	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
the	NN	O	O
Ly-49a	NN	O	B-DNA
gene	NN	O	I-DNA
is	NN	O	O
a	NN	O	O
direct	NN	O	O
TCF-1	NN	O	B-protein
target	NN	O	O
.	NN	O	O

TCF-1	NN	O	B-protein
deficiency	NN	O	O
resulted	NN	O	O
in	NN	O	O
the	NN	O	O
altered	NN	O	O
usage	NN	O	O
of	NN	O	O
additional	NN	O	O
Ly-49	NN	O	B-protein
receptors	NN	O	I-protein
.	NN	O	O

We	NN	O	O
show	NN	O	O
in	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
using	NN	O	O
TCF-1	NN	O	B-protein
beta	NN	O	I-protein
(	NN	O	I-protein
2	NN	O	I-protein
)	NN	O	I-protein
-microglobulin	NN	O	I-protein
double-deficient	NN	O	O
mice	NN	O	O
,	NN	O	O
that	NN	O	O
these	NN	O	O
repertoire	NN	O	O
alterations	NN	O	O
are	NN	O	O
not	NN	O	O
due	NN	O	O
to	NN	O	O
Ly-49/MHC	NN	O	O
class	NN	O	O
I	NN	O	O
interactions	NN	O	O
.	NN	O	O

Our	NN	O	O
findings	NN	O	O
rather	NN	O	O
suggest	NN	O	O
a	NN	O	O
TCF-1	NN	O	B-protein
-dependent	NN	O	O
,	NN	O	O
cell	NN	O	O
autonomous	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
acquisition	NN	O	O
of	NN	O	O
multiple	NN	O	O
Ly-49	NN	O	B-protein
receptors	NN	O	I-protein
.	NN	O	O

Besides	NN	O	O
reduced	NN	O	O
receptor	NN	O	O
usage	NN	O	O
(	NN	O	O
Ly-49A	NN	O	B-protein
and	NN	O	I-protein
D	NN	O	I-protein
)	NN	O	O
,	NN	O	O
we	NN	O	O
also	NN	O	O
observed	NN	O	O
no	NN	O	O
effect	NN	O	O
(	NN	O	O
Ly-49C	NN	O	B-protein
)	NN	O	O
and	NN	O	O
significantly	NN	O	O
expanded	NN	O	O
(	NN	O	O
Ly-49G	NN	O	B-protein
and	NN	O	I-protein
I	NN	O	I-protein
)	NN	O	O
receptor	NN	O	O
usage	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
TCF-1	NN	O	B-protein
.	NN	O	O

These	NN	O	O
effects	NN	O	O
did	NN	O	O
not	NN	O	O
in	NN	O	O
all	NN	O	O
cases	NN	O	O
correlate	NN	O	O
with	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
TCF	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
respective	NN	O	O
proximal	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
besides	NN	O	O
TCF-1	NN	O	B-protein
binding	NN	O	O
to	NN	O	O
the	NN	O	O
proximal	NN	O	B-DNA
promoter	NN	O	I-DNA
,	NN	O	O
Ly-49	NN	O	O
acquisition	NN	O	O
may	NN	O	O
also	NN	O	O
be	NN	O	O
regulated	NN	O	O
by	NN	O	O
TCF-1	NN	O	B-protein
binding	NN	O	O
to	NN	O	O
more	NN	O	O
distant	NN	O	O
cis-acting	NN	O	B-DNA
elements	NN	O	I-DNA
and/or	NN	O	O
by	NN	O	O
regulating	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
additional	NN	O	O
trans-acting	NN	O	B-DNA
factors	NN	O	I-DNA
.	NN	O	O

Consistent	NN	O	O
with	NN	O	O
the	NN	O	O
observed	NN	O	O
differential	NN	O	O
,	NN	O	O
positive	NN	O	O
or	NN	O	O
negative	NN	O	O
role	NN	O	O
of	NN	O	O
TCF-1	NN	O	B-protein
for	NN	O	O
Ly-49	NN	O	B-protein
receptor	NN	O	I-protein
acquisition	NN	O	O
,	NN	O	O
reporter	NN	O	O
gene	NN	O	O
assays	NN	O	O
revealed	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
an	NN	O	O
inducing	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
a	NN	O	O
repressing	NN	O	O
TCF	NN	O	B-DNA
site	NN	O	I-DNA
in	NN	O	O
certain	NN	O	O
proximal	NN	O	O
Ly-49	NN	O	B-DNA
promoters	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
findings	NN	O	O
reveal	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
of	NN	O	O
TCF-1	NN	O	B-protein
for	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
the	NN	O	O
NK	NN	O	B-protein
cell	NN	O	I-protein
receptor	NN	O	I-protein
repertoire	NN	O	O
.	NN	O	O

-DOCSTART-	O

Ligation	NN	O	O
of	NN	O	O
CD11b	NN	O	B-protein
and	NN	O	O
CD11c	NN	O	B-protein
beta	NN	O	I-protein
(	NN	O	I-protein
2	NN	O	I-protein
)	NN	O	I-protein
integrins	NN	O	B-protein
by	NN	O	O
antibodies	NN	O	B-protein
or	NN	O	O
soluble	NN	O	B-protein
CD23	NN	O	I-protein
induces	NN	O	O
macrophage	NN	O	B-protein
inflammatory	NN	O	I-protein
protein	NN	O	I-protein
1alpha	NN	O	I-protein
(	NN	O	O
MIP-1alpha	NN	O	B-protein
)	NN	O	O
and	NN	O	O
MIP-1beta	NN	O	B-protein
production	NN	O	O
in	NN	O	O
primary	NN	O	B-cell_type
human	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
through	NN	O	O
a	NN	O	O
pathway	NN	O	O
dependent	NN	O	O
on	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
.	NN	O	O

Chemokines	NN	O	B-protein
and	NN	O	O
adhesion	NN	O	B-protein
molecules	NN	O	I-protein
such	NN	O	O
as	NN	O	O
integrins	NN	O	B-protein
play	NN	O	O
a	NN	O	O
major	NN	O	O
part	NN	O	O
in	NN	O	O
the	NN	O	O
trafficking	NN	O	O
,	NN	O	O
extravasation	NN	O	O
,	NN	O	O
and	NN	O	O
recruitment	NN	O	O
of	NN	O	O
leukocytes	NN	O	B-cell_type
to	NN	O	O
inflammatory	NN	O	O
sites	NN	O	O
.	NN	O	O

This	NN	O	O
study	NN	O	O
investigated	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
beta	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
integrin	NN	O	O
engagement	NN	O	O
on	NN	O	O
chemokine	NN	O	O
production	NN	O	O
by	NN	O	O
freshly	NN	O	B-cell_type
isolated	NN	O	I-cell_type
human	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
ligation	NN	O	O
of	NN	O	O
CD11b	NN	O	B-protein
or	NN	O	O
CD11c	NN	O	B-protein
but	NN	O	O
not	NN	O	O
CD11a	NN	O	B-protein
alpha	NN	O	I-protein
chains	NN	O	I-protein
of	NN	O	O
beta	NN	O	B-protein
(	NN	O	I-protein
2	NN	O	I-protein
)	NN	O	I-protein
integrins	NN	O	I-protein
by	NN	O	O
antibodies	NN	O	B-protein
or	NN	O	O
soluble	NN	O	B-protein
CD23	NN	O	I-protein
(	NN	O	I-protein
sCD23	NN	O	I-protein
)	NN	O	I-protein
fusion	NN	O	I-protein
proteins	NN	O	I-protein
rapidly	NN	O	O
induced	NN	O	O
transcription	NN	O	O
and	NN	O	O
secretion	NN	O	O
of	NN	O	O
interleukin	NN	O	B-protein
8	NN	O	I-protein
,	NN	O	O
macrophage	NN	O	B-protein
inflammatory	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	I-protein
MIP	NN	O	I-protein
)	NN	O	I-protein
1alpha	NN	O	I-protein
,	NN	O	O
and	NN	O	O
MIP-1beta	NN	O	B-protein
.	NN	O	O

Because	NN	O	O
the	NN	O	O
promoters	NN	O	B-DNA
of	NN	O	O
these	NN	O	O
chemokine	NN	O	B-DNA
genes	NN	O	I-DNA
contain	NN	O	O
kappaB	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
,	NN	O	O
we	NN	O	O
assessed	NN	O	O
the	NN	O	O
possible	NN	O	O
role	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
(	NN	O	O
NF-kappaB	NN	O	B-protein
)	NN	O	O
in	NN	O	O
controlling	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
genes	NN	O	O
through	NN	O	O
beta	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
integrin	NN	O	O
engagement	NN	O	O
.	NN	O	O

Electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
showed	NN	O	O
that	NN	O	O
sCD23	NN	O	B-protein
or	NN	O	O
antibodies	NN	O	B-protein
to	NN	O	O
CD11b	NN	O	B-protein
or	NN	O	O
to	NN	O	O
CD11c	NN	O	B-protein
up-regulated	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
was	NN	O	O
accompanied	NN	O	O
by	NN	O	O
degradation	NN	O	O
of	NN	O	O
its	NN	O	O
cytosolic	NN	O	B-protein
inhibitor	NN	O	I-protein
IkappaB-alpha	NN	O	I-protein
.	NN	O	O

Blockade	NN	O	O
of	NN	O	O
depletion	NN	O	O
of	NN	O	O
IkappaB-alpha	NN	O	B-protein
by	NN	O	O
proteasome	NN	O	O
inhibitors	NN	O	O
(	NN	O	O
proteasome	NN	O	O
inhibitor	NN	O	O
I	NN	O	O
or	NN	O	O
acetyl-leucinyl-leucinyl-norleucinal	NN	O	O
)	NN	O	O
led	NN	O	O
to	NN	O	O
concomitant	NN	O	O
inhibition	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
and	NN	O	O
expression	NN	O	O
of	NN	O	O
MIP-1alpha	NN	O	B-RNA
and	NN	O	I-RNA
MIP-1beta	NN	O	I-RNA
messenger	NN	O	I-RNA
RNA	NN	O	I-RNA
induced	NN	O	O
by	NN	O	O
beta	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
integrin	NN	O	O
ligation	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
triggering	NN	O	O
of	NN	O	O
CD11b	NN	O	B-protein
or	NN	O	O
CD11c	NN	O	B-protein
beta	NN	O	I-protein
(	NN	O	I-protein
2	NN	O	I-protein
)	NN	O	I-protein
integrin	NN	O	O
on	NN	O	O
primary	NN	O	B-cell_type
human	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
provides	NN	O	O
activation	NN	O	O
signals	NN	O	O
leading	NN	O	O
to	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
and	NN	O	O
subsequent	NN	O	O
secretion	NN	O	O
of	NN	O	O
MIP-1alpha	NN	O	B-protein
and	NN	O	O
MIP-1beta	NN	O	B-protein
that	NN	O	O
may	NN	O	O
have	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
recruitment	NN	O	O
of	NN	O	O
other	NN	O	O
inflammatory	NN	O	B-cell_type
cells	NN	O	I-cell_type
during	NN	O	O
initiation	NN	O	O
of	NN	O	O
an	NN	O	O
inflammatory	NN	O	O
response	NN	O	O

-DOCSTART-	O

Synergistic	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
human	NN	O	B-DNA
Acyl-coenzyme	NN	O	I-DNA
A	NN	O	I-DNA
:	NN	O	I-DNA
cholesterol	NN	O	I-DNA
acyltransterase-1	NN	O	I-DNA
gene	NN	O	I-DNA
by	NN	O	O
interferon-gamma	NN	O	B-protein
and	NN	O	O
all-trans-retinoic	NN	O	O
acid	NN	O	O
THP-1	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Acyl-coenzyme	NN	O	B-protein
A	NN	O	I-protein
:	NN	O	I-protein
cholesterol	NN	O	I-protein
acyltransferase	NN	O	I-protein
(	NN	O	O
ACAT	NN	O	B-protein
)	NN	O	O
is	NN	O	O
an	NN	O	O
intracellular	NN	O	B-protein
enzyme	NN	O	I-protein
involved	NN	O	O
in	NN	O	O
cellular	NN	O	O
cholesterol	NN	O	O
homeostasis	NN	O	O
and	NN	O	O
in	NN	O	O
atherosclerotic	NN	O	O
foam	NN	O	O
cell	NN	O	O
formation	NN	O	O
.	NN	O	O

Human	NN	O	B-DNA
ACAT-1	NN	O	I-DNA
gene	NN	O	I-DNA
contains	NN	O	O
two	NN	O	O
promoters	NN	O	B-DNA
(	NN	O	O
P1	NN	O	B-DNA
and	NN	O	O
P7	NN	O	B-DNA
)	NN	O	O
,	NN	O	O
each	NN	O	O
located	NN	O	O
in	NN	O	O
a	NN	O	O
different	NN	O	O
chromosome	NN	O	B-DNA
(	NN	O	I-DNA
1	NN	O	I-DNA
and	NN	O	I-DNA
7	NN	O	I-DNA
)	NN	O	I-DNA
(	NN	O	O
Li	NN	O	O
,	NN	O	O
B.	NN	O	O
L.	NN	O	O
,	NN	O	O
Li	NN	O	O
,	NN	O	O
X.	NN	O	O
L.	NN	O	O
,	NN	O	O
Duan	NN	O	O
,	NN	O	O
Z.	NN	O	O
J.	NN	O	O
,	NN	O	O
Lee	NN	O	O
,	NN	O	O
O.	NN	O	O
,	NN	O	O
Lin	NN	O	O
,	NN	O	O
S.	NN	O	O
,	NN	O	O
Ma	NN	O	O
,	NN	O	O
Z.	NN	O	O
M.	NN	O	O
,	NN	O	O
Chang	NN	O	O
,	NN	O	O
C.	NN	O	O
C.	NN	O	O
,	NN	O	O
Yang	NN	O	O
,	NN	O	O
X.	NN	O	O
Y.	NN	O	O
,	NN	O	O
Park	NN	O	O
,	NN	O	O
J.	NN	O	O
P.	NN	O	O
,	NN	O	O
Mohandas	NN	O	O
,	NN	O	O
T.	NN	O	O
K.	NN	O	O
,	NN	O	O
Noll	NN	O	O
,	NN	O	O
W.	NN	O	O
,	NN	O	O
Chan	NN	O	O
,	NN	O	O
L.	NN	O	O
,	NN	O	O
and	NN	O	O
Chang	NN	O	O
,	NN	O	O
T.	NN	O	O
Y.	NN	O	O
(	NN	O	O
1999	NN	O	O
)	NN	O	O
J.	NN	O	O
Biol	NN	O	O
Chem.	NN	O	O
274	NN	O	O
,	NN	O	O
11060-11071	NN	O	O
)	NN	O	O
.	NN	O	O

Interferon-gamma	NN	O	B-protein
(	NN	O	O
IFN-gamma	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
cytokine	NN	O	B-protein
that	NN	O	O
exerts	NN	O	O
many	NN	O	O
pro-atherosclerotic	NN	O	O
effects	NN	O	O
in	NN	O	O
vivo	NN	O	O
,	NN	O	O
causes	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
ACAT-1	NN	O	B-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
human	NN	O	B-cell_type
blood	NN	O	I-cell_type
monocyte-derived	NN	O	I-cell_type
macrophages	NN	O	I-cell_type
and	NN	O	O
macrophage-like	NN	O	B-cell_type
cells	NN	O	I-cell_type
but	NN	O	O
not	NN	O	O
in	NN	O	O
other	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

To	NN	O	O
examine	NN	O	O
the	NN	O	O
molecular	NN	O	O
nature	NN	O	O
of	NN	O	O
this	NN	O	O
observation	NN	O	O
,	NN	O	O
we	NN	O	O
identified	NN	O	O
within	NN	O	O
the	NN	O	O
ACAT-1	NN	O	B-DNA
P1	NN	O	I-DNA
promoter	NN	O	I-DNA
a	NN	O	O
159-base	NN	O	B-DNA
pair	NN	O	I-DNA
core	NN	O	I-DNA
region	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
region	NN	O	O
contains	NN	O	O
4	NN	O	O
Sp1	NN	O	B-DNA
elements	NN	O	I-DNA
and	NN	O	O
an	NN	O	O
IFN-gamma	NN	O	B-DNA
activated	NN	O	I-DNA
sequence	NN	O	I-DNA
(	NN	O	O
GAS	NN	O	B-DNA
)	NN	O	O
that	NN	O	O
overlaps	NN	O	O
with	NN	O	O
the	NN	O	O
second	NN	O	O
Sp1	NN	O	B-DNA
element	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
the	NN	O	O
monocytic	NN	O	B-cell_type
cell	NN	O	I-cell_type
line	NN	O	I-cell_type
THP-1	NN	O	I-cell_type
cell	NN	O	I-cell_type
,	NN	O	O
the	NN	O	O
combination	NN	O	O
of	NN	O	O
IFN-gamma	NN	O	B-protein
and	NN	O	O
all-trans-retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
a	NN	O	O
known	NN	O	O
differentiation	NN	O	O
agent	NN	O	O
)	NN	O	O
enhances	NN	O	O
the	NN	O	O
ACAT-1	NN	O	B-DNA
P1	NN	O	I-DNA
promoter	NN	O	I-DNA
but	NN	O	O
not	NN	O	O
the	NN	O	O
P7	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Additional	NN	O	O
experiments	NN	O	O
showed	NN	O	O
that	NN	O	O
all-trans-retinoic	NN	O	O
acid	NN	O	O
causes	NN	O	O
large	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
STAT1	NN	O	I-protein
,	NN	O	O
while	NN	O	O
IFN-gamma	NN	O	B-protein
causes	NN	O	O
activation	NN	O	O
of	NN	O	O
STAT1	NN	O	B-protein
such	NN	O	O
that	NN	O	O
it	NN	O	O
binds	NN	O	O
to	NN	O	O
the	NN	O	O
GAS/Sp1	NN	O	B-DNA
site	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
ACAT-1	NN	O	B-DNA
P1	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Our	NN	O	O
work	NN	O	O
provides	NN	O	O
a	NN	O	O
molecular	NN	O	O
mechanism	NN	O	O
to	NN	O	O
account	NN	O	O
for	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
IFN-gamma	NN	O	B-protein
in	NN	O	O
causing	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
ACAT-1	NN	O	B-protein
in	NN	O	O
macrophage-like	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Inhaled	NN	O	O
nitric	NN	O	O
oxide	NN	O	O
down-regulates	NN	O	O
intrapulmonary	NN	O	O
nitric	NN	O	O
oxide	NN	O	O
production	NN	O	O
in	NN	O	O
lipopolysaccharide-induced	NN	O	O
acute	NN	O	O
lung	NN	O	O
injury	NN	O	O
.	NN	O	O

OBJECTIVE	NN	O	O
:	NN	O	O
To	NN	O	O
examine	NN	O	O
whether	NN	O	O
inhaled	NN	O	O
nitric	NN	O	O
oxide	NN	O	O
(	NN	O	O
NO	NN	O	O
)	NN	O	O
affected	NN	O	O
the	NN	O	O
intrapulmonary	NN	O	O
production	NN	O	O
of	NN	O	O
NO	NN	O	O
,	NN	O	O
reactive	NN	O	O
oxygen	NN	O	O
species	NN	O	O
,	NN	O	O
and	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
in	NN	O	O
a	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
-induced	NN	O	O
model	NN	O	O
of	NN	O	O
acute	NN	O	O
lung	NN	O	O
injury	NN	O	O
.	NN	O	O

DESIGN	NN	O	O
:	NN	O	O
Prospective	NN	O	O
,	NN	O	O
randomized	NN	O	O
,	NN	O	O
laboratory	NN	O	O
study	NN	O	O
.	NN	O	O

SETTING	NN	O	O
:	NN	O	O
Experimental	NN	O	O
laboratory	NN	O	O
at	NN	O	O
a	NN	O	O
biomedical	NN	O	O
institute	NN	O	O
.	NN	O	O

SUBJECTS	NN	O	O
:	NN	O	O
Twenty	NN	O	O
male	NN	O	O
rabbits	NN	O	O
weighing	NN	O	O
2.5-3.5	NN	O	O
kg	NN	O	O
.	NN	O	O

INTERVENTIONS	NN	O	O
:	NN	O	O
Saline	NN	O	O
or	NN	O	O
LPS	NN	O	O
(	NN	O	O
5	NN	O	O
mg/kg	NN	O	O
of	NN	O	O
body	NN	O	O
weight	NN	O	O
)	NN	O	O
was	NN	O	O
administered	NN	O	O
intravenously	NN	O	O
with	NN	O	O
or	NN	O	O
without	NN	O	O
NO	NN	O	O
inhalation	NN	O	O
(	NN	O	O
10	NN	O	O
ppm	NN	O	O
)	NN	O	O
in	NN	O	O
each	NN	O	O
group	NN	O	O
of	NN	O	O
five	NN	O	O
rabbits	NN	O	O
.	NN	O	O

MEASUREMENTS	NN	O	O
AND	NN	O	O
MAIN	NN	O	O
RESULTS	NN	O	O
:	NN	O	O
LPS	NN	O	O
increased	NN	O	O
the	NN	O	O
lung	NN	O	O
leak	NN	O	O
index	NN	O	O
,	NN	O	O
the	NN	O	O
neutrophils	NN	O	B-cell_type
and	NN	O	O
NO	NN	O	O
levels	NN	O	O
in	NN	O	O
bronchoalveolar	NN	O	O
lavage	NN	O	O
fluid	NN	O	O
,	NN	O	O
and	NN	O	O
NO	NN	O	O
levels	NN	O	O
produced	NN	O	O
by	NN	O	O
resting	NN	O	O
and	NN	O	O
stimulated	NN	O	O
alveolar	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
.	NN	O	O

Inhaled	NN	O	O
NO	NN	O	O
decreased	NN	O	O
the	NN	O	O
lung	NN	O	O
leak	NN	O	O
index	NN	O	O
,	NN	O	O
the	NN	O	O
neutrophils	NN	O	B-cell_type
and	NN	O	O
NO	NN	O	O
levels	NN	O	O
as	NN	O	O
measured	NN	O	O
by	NN	O	O
nitrite	NN	O	O
levels	NN	O	O
in	NN	O	O
the	NN	O	O
lavage	NN	O	O
fluid	NN	O	O
,	NN	O	O
and	NN	O	O
NO	NN	O	O
produced	NN	O	O
by	NN	O	O
the	NN	O	O
resting	NN	O	O
and	NN	O	O
stimulated	NN	O	O
alveolar	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
.	NN	O	O

Inhaled	NN	O	O
NO	NN	O	O
also	NN	O	O
blocked	NN	O	O
the	NN	O	O
activities	NN	O	O
of	NN	O	O
reactive	NN	O	O
oxygen	NN	O	O
species	NN	O	O
and	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
binding	NN	O	O
to	NN	O	O
DNA	NN	O	O
in	NN	O	O
lavage	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
in	NN	O	O
alveolar	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
.	NN	O	O

CONCLUSION	NN	O	O
:	NN	O	O
Inhaled	NN	O	O
NO	NN	O	O
attenuates	NN	O	O
LPS-induced	NN	O	O
acute	NN	O	O
lung	NN	O	O
injury	NN	O	O
,	NN	O	O
possibly	NN	O	O
by	NN	O	O
decreasing	NN	O	O
NO	NN	O	O
production	NN	O	O
in	NN	O	O
the	NN	O	O
lungs	NN	O	O
.	NN	O	O

The	NN	O	O
mechanism	NN	O	O
of	NN	O	O
reducing	NN	O	O
NO	NN	O	O
production	NN	O	O
resulting	NN	O	O
from	NN	O	O
inhaled	NN	O	O
NO	NN	O	O
may	NN	O	O
involve	NN	O	O
,	NN	O	O
in	NN	O	O
part	NN	O	O
,	NN	O	O
the	NN	O	O
activities	NN	O	O
of	NN	O	O
reactive	NN	O	O
oxygen	NN	O	O
species	NN	O	O
and/or	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Treatment	NN	O	O
of	NN	O	O
allergic	NN	O	O
airway	NN	O	O
inflammation	NN	O	O
and	NN	O	O
hyperresponsiveness	NN	O	O
by	NN	O	O
antisense-induced	NN	O	O
local	NN	O	O
blockade	NN	O	O
of	NN	O	O
GATA-3	NN	O	B-DNA
expression	NN	O	O
.	NN	O	O

Recent	NN	O	O
studies	NN	O	O
in	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
have	NN	O	O
revealed	NN	O	O
that	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
dominant	NN	O	O
negative	NN	O	O
form	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
GATA-3	NN	O	I-protein
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
can	NN	O	O
prevent	NN	O	O
T	NN	O	B-cell_type
helper	NN	O	I-cell_type
cell	NN	O	I-cell_type
type	NN	O	I-cell_type
2	NN	O	I-cell_type
(	NN	O	O
Th2	NN	O	B-cell_type
)	NN	O	O
-mediated	NN	O	O
allergic	NN	O	O
airway	NN	O	O
inflammation	NN	O	O
in	NN	O	O
mice	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
it	NN	O	O
remains	NN	O	O
unclear	NN	O	O
whether	NN	O	O
GATA-3	NN	O	B-protein
plays	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
effector	NN	O	O
phase	NN	O	O
of	NN	O	O
allergic	NN	O	O
airway	NN	O	O
inflammation	NN	O	O
and	NN	O	O
whether	NN	O	O
antagonizing	NN	O	O
the	NN	O	O
expression	NN	O	O
and/or	NN	O	O
function	NN	O	O
of	NN	O	O
GATA-3	NN	O	B-DNA
can	NN	O	O
be	NN	O	O
used	NN	O	O
for	NN	O	O
the	NN	O	O
therapy	NN	O	O
of	NN	O	O
allergic	NN	O	O
airway	NN	O	O
inflammation	NN	O	O
and	NN	O	O
hyperresponsiveness	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
analyzed	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
locally	NN	O	O
antagonizing	NN	O	O
GATA-3	NN	O	B-protein
function	NN	O	O
in	NN	O	O
a	NN	O	O
murine	NN	O	O
model	NN	O	O
of	NN	O	O
asthma	NN	O	O
.	NN	O	O

We	NN	O	O
could	NN	O	O
suppress	NN	O	O
GATA-3	NN	O	O
expression	NN	O	O
in	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
-producing	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
in	NN	O	O
vivo	NN	O	O
by	NN	O	O
an	NN	O	O
antisense	NN	O	O
phosphorothioate	NN	O	O
oligonucleotide	NN	O	O
overlapping	NN	O	O
the	NN	O	O
translation	NN	O	B-DNA
start	NN	O	I-DNA
site	NN	O	I-DNA
of	NN	O	O
GATA-3	NN	O	B-DNA
,	NN	O	O
whereas	NN	O	O
nonsense	NN	O	O
control	NN	O	O
oligonucleotides	NN	O	O
were	NN	O	O
virtually	NN	O	O
inactive	NN	O	O
.	NN	O	O

In	NN	O	O
a	NN	O	O
murine	NN	O	O
model	NN	O	O
of	NN	O	O
asthma	NN	O	O
associated	NN	O	O
with	NN	O	O
allergic	NN	O	O
pulmonary	NN	O	O
inflammation	NN	O	O
and	NN	O	O
hyperresponsiveness	NN	O	O
in	NN	O	O
ovalbumin	NN	O	O
(	NN	O	O
OVA	NN	O	O
)	NN	O	O
-sensitized	NN	O	O
mice	NN	O	O
,	NN	O	O
local	NN	O	O
intranasal	NN	O	O
administration	NN	O	O
of	NN	O	O
fluorescein	NN	O	O
isothiocyanate-labeled	NN	O	O
GATA-3	NN	O	O
antisense	NN	O	O
oligonucleotides	NN	O	O
led	NN	O	O
to	NN	O	O
DNA	NN	O	O
uptake	NN	O	O
in	NN	O	O
lung	NN	O	B-cell_type
cells	NN	O	I-cell_type
associated	NN	O	O
with	NN	O	O
a	NN	O	O
reduction	NN	O	O
of	NN	O	O
intracellular	NN	O	O
GATA-3	NN	O	O
expression	NN	O	O
.	NN	O	O

Such	NN	O	O
intrapulmonary	NN	O	O
blockade	NN	O	O
of	NN	O	O
GATA-3	NN	O	O
expression	NN	O	O
caused	NN	O	O
an	NN	O	O
abrogation	NN	O	O
of	NN	O	O
signs	NN	O	O
of	NN	O	O
lung	NN	O	O
inflammation	NN	O	O
including	NN	O	O
infiltration	NN	O	O
of	NN	O	O
eosinophils	NN	O	B-cell_type
and	NN	O	O
Th2	NN	O	B-cell_type
cytokine	NN	O	O
production	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
treatment	NN	O	O
with	NN	O	O
antisense	NN	O	O
but	NN	O	O
not	NN	O	O
nonsense	NN	O	O
oligonucleotides	NN	O	O
induced	NN	O	O
a	NN	O	O
significant	NN	O	O
reduction	NN	O	O
of	NN	O	O
airway	NN	O	O
hyperresponsiveness	NN	O	O
in	NN	O	O
OVA-sensitized	NN	O	O
mice	NN	O	O
to	NN	O	O
levels	NN	O	O
comparable	NN	O	O
to	NN	O	O
saline-treated	NN	O	O
control	NN	O	O
mice	NN	O	O
,	NN	O	O
as	NN	O	O
assessed	NN	O	O
by	NN	O	O
both	NN	O	O
enhanced	NN	O	O
pause	NN	O	O
(	NN	O	O
PenH	NN	O	O
)	NN	O	O
responses	NN	O	O
and	NN	O	O
pulmonary	NN	O	O
resistance	NN	O	O
determined	NN	O	O
by	NN	O	O
body	NN	O	O
plethysmography	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
indicate	NN	O	O
a	NN	O	O
critical	NN	O	O
role	NN	O	O
for	NN	O	O
GATA-3	NN	O	B-DNA
in	NN	O	O
the	NN	O	O
effector	NN	O	O
phase	NN	O	O
of	NN	O	O
a	NN	O	O
murine	NN	O	O
asthma	NN	O	O
model	NN	O	O
and	NN	O	O
suggest	NN	O	O
that	NN	O	O
local	NN	O	O
delivery	NN	O	O
of	NN	O	O
GATA-3	NN	O	O
antisense	NN	O	O
oligonucleotides	NN	O	O
may	NN	O	O
be	NN	O	O
a	NN	O	O
novel	NN	O	O
approach	NN	O	O
for	NN	O	O
the	NN	O	O
treatment	NN	O	O
of	NN	O	O
airway	NN	O	O
hyperresponsiveness	NN	O	O
such	NN	O	O
as	NN	O	O
in	NN	O	O
asthma	NN	O	O
.	NN	O	O

This	NN	O	O
approach	NN	O	O
has	NN	O	O
the	NN	O	O
potential	NN	O	O
advantage	NN	O	O
of	NN	O	O
suppressing	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
various	NN	O	O
proinflammatory	NN	O	B-protein
Th2	NN	O	I-protein
cytokines	NN	O	I-protein
simultaneously	NN	O	O
rather	NN	O	O
than	NN	O	O
suppressing	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
a	NN	O	O
single	NN	O	O
cytokine	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

T	NN	O	O
helper-cell	NN	O	O
phenotype	NN	O	O
regulates	NN	O	O
atherosclerosis	NN	O	O
in	NN	O	O
mice	NN	O	O
under	NN	O	O
conditions	NN	O	O
of	NN	O	O
mild	NN	O	O
hypercholesterolemia	NN	O	O
.	NN	O	O

BACKGROUND	NN	O	O
:	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
implicated	NN	O	O
in	NN	O	O
atherosclerosis	NN	O	O
,	NN	O	O
but	NN	O	O
little	NN	O	O
is	NN	O	O
known	NN	O	O
about	NN	O	O
the	NN	O	O
genetic	NN	O	O
control	NN	O	O
or	NN	O	O
molecular	NN	O	O
pathways	NN	O	O
,	NN	O	O
especially	NN	O	O
under	NN	O	O
conditions	NN	O	O
of	NN	O	O
mild	NN	O	O
hypercholesterolemia	NN	O	O
.	NN	O	O

METHODS	NN	O	O
AND	NN	O	O
RESULTS	NN	O	O
:	NN	O	O
BALB/c	NN	O	O
mice	NN	O	O
,	NN	O	O
making	NN	O	O
a	NN	O	O
CD4+	NN	O	O
Th2	NN	O	O
(	NN	O	O
IL-4+	NN	O	O
)	NN	O	O
cell	NN	O	O
response	NN	O	O
,	NN	O	O
express	NN	O	O
both	NN	O	O
MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
antigens	NN	O	I-protein
(	NN	O	O
IA	NN	O	B-protein
(	NN	O	I-protein
d	NN	O	I-protein
)	NN	O	I-protein
,	NN	O	O
IE	NN	O	B-protein
(	NN	O	I-protein
d	NN	O	I-protein
)	NN	O	I-protein
)	NN	O	O
and	NN	O	O
are	NN	O	O
atherosclerosis-resistant	NN	O	O
.	NN	O	O

C57Bl/6	NN	O	O
mice	NN	O	O
produce	NN	O	O
a	NN	O	O
CD4+	NN	O	O
Th1	NN	O	O
(	NN	O	O
interferon	NN	O	O
[	NN	O	O
IFN	NN	O	O
]	NN	O	O
gamma+	NN	O	O
)	NN	O	O
response	NN	O	O
,	NN	O	O
express	NN	O	O
IA	NN	O	B-protein
(	NN	O	I-protein
b	NN	O	I-protein
)	NN	O	I-protein
but	NN	O	O
no	NN	O	O
IE	NN	O	B-protein
,	NN	O	O
and	NN	O	O
are	NN	O	O
atherosclerosis-prone	NN	O	O
.	NN	O	O

To	NN	O	O
evaluate	NN	O	O
T	NN	O	O
helper-cell	NN	O	O
phenotype	NN	O	O
in	NN	O	O
fatty	NN	O	O
streak	NN	O	O
formation	NN	O	O
,	NN	O	O
wild-type	NN	O	O
C57Bl/6	NN	O	O
mice	NN	O	O
(	NN	O	O
IA	NN	O	O
(	NN	O	O
b	NN	O	O
)	NN	O	O
+IE-	NN	O	O
)	NN	O	O
and	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
,	NN	O	O
either	NN	O	O
AB	NN	O	O
(	NN	O	O
o	NN	O	O
)	NN	O	O
,	NN	O	O
IA	NN	O	O
(	NN	O	O
b	NN	O	O
)	NN	O	O
-IE-	NN	O	O
;	NN	O	O
ABEalpha	NN	O	O
,	NN	O	O
IA-IE	NN	O	O
(	NN	O	O
k	NN	O	O
)	NN	O	O
+	NN	O	O
;	NN	O	O
or	NN	O	O
BL	NN	O	O
:	NN	O	O
TG	NN	O	O
:	NN	O	O
Ealpha	NN	O	O
,	NN	O	O
IA	NN	O	O
(	NN	O	O
b	NN	O	O
)	NN	O	O
+IE	NN	O	O
(	NN	O	O
k	NN	O	O
)	NN	O	O
+	NN	O	O
,	NN	O	O
were	NN	O	O
fed	NN	O	O
a	NN	O	O
high-cholesterol	NN	O	O
diet	NN	O	O
for	NN	O	O
16	NN	O	O
weeks	NN	O	O
and	NN	O	O
evaluated	NN	O	O
histomorphometrically	NN	O	O
for	NN	O	O
aortic	NN	O	O
lesions	NN	O	O
.	NN	O	O

Lesion	NN	O	O
size	NN	O	O
in	NN	O	O
AB	NN	O	O
(	NN	O	O
o	NN	O	O
)	NN	O	O
,	NN	O	O
ABEalpha	NN	O	O
,	NN	O	O
and	NN	O	O
BL	NN	O	O
:	NN	O	O
TG	NN	O	O
:	NN	O	O
Ealpha	NN	O	O
strains	NN	O	O
was	NN	O	O
decreased	NN	O	O
by	NN	O	O
54	NN	O	O
%	NN	O	O
,	NN	O	O
79	NN	O	O
%	NN	O	O
,	NN	O	O
and	NN	O	O
82	NN	O	O
%	NN	O	O
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
compared	NN	O	O
with	NN	O	O
wild-type	NN	O	O
,	NN	O	O
correlating	NN	O	O
with	NN	O	O
decreased	NN	O	O
Th1	NN	O	B-cell_type
and	NN	O	O
increased	NN	O	O
Th2	NN	O	B-cell_type
expression	NN	O	O
and	NN	O	O
suggesting	NN	O	O
that	NN	O	O
T	NN	O	O
helper-cell	NN	O	O
phenotype	NN	O	O
is	NN	O	O
important	NN	O	O
in	NN	O	O
fatty	NN	O	O
lesion	NN	O	O
development	NN	O	O
.	NN	O	O

Decreasing	NN	O	O
Th1	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
antibodies	NN	O	B-protein
(	NN	O	O
alpha-CD4	NN	O	B-protein
)	NN	O	O
or	NN	O	O
cytokines	NN	O	B-protein
(	NN	O	O
IL-4	NN	O	B-protein
)	NN	O	O
also	NN	O	O
caused	NN	O	O
>	NN	O	O
/=80	NN	O	O
%	NN	O	O
reductions	NN	O	O
in	NN	O	O
lesion	NN	O	O
size	NN	O	O
.	NN	O	O

Immunohistology	NN	O	O
revealed	NN	O	O
IFN-gamma	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
colocalized	NN	O	O
with	NN	O	O
activated	NN	O	O
macrophages	NN	O	B-cell_type
.	NN	O	O

Confirming	NN	O	O
these	NN	O	O
findings	NN	O	O
in	NN	O	O
a	NN	O	O
different	NN	O	O
mouse	NN	O	O
strain	NN	O	O
,	NN	O	O
BALB/c	NN	O	O
Stat	NN	O	O
6	NN	O	O
knockout	NN	O	O
mice	NN	O	O
(	NN	O	O
Th2	NN	O	O
cell-deficient	NN	O	O
)	NN	O	O
developed	NN	O	O
aortic	NN	O	O
lesions	NN	O	O
comparable	NN	O	O
to	NN	O	O
C57Bl/6	NN	O	O
mice	NN	O	O
on	NN	O	O
the	NN	O	O
same	NN	O	O
diet	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
:	NN	O	O
In	NN	O	O
mildly	NN	O	O
hypercholesterolemic	NN	O	O
C57Bl/6	NN	O	O
mice	NN	O	O
,	NN	O	O
presence	NN	O	O
of	NN	O	O
IA	NN	O	B-protein
(	NN	O	I-protein
b	NN	O	I-protein
)	NN	O	I-protein
and	NN	O	O
absence	NN	O	O
of	NN	O	O
IE	NN	O	B-protein
regulated	NN	O	O
CD4+	NN	O	O
T	NN	O	O
helper-cell	NN	O	O
phenotype	NN	O	O
;	NN	O	O
fatty	NN	O	O
lesions	NN	O	O
were	NN	O	O
proportional	NN	O	O
to	NN	O	O
IFNgamma+	NN	O	B-cell_type
Th1	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
both	NN	O	O
C57Bl/6	NN	O	O
and	NN	O	O
BALB/c	NN	O	O
strains	NN	O	O
.	NN	O	O

IFN-gamma	NN	O	B-protein
may	NN	O	O
participate	NN	O	O
through	NN	O	O
macrophage	NN	O	O
activation	NN	O	O
,	NN	O	O
whereas	NN	O	O
IL-4	NN	O	B-protein
may	NN	O	O
act	NN	O	O
to	NN	O	O
limit	NN	O	O
Th1-cell	NN	O	O
response	NN	O	O
.	NN	O	O

-DOCSTART-	O

Requirement	NN	O	O
for	NN	O	O
p38	NN	O	B-protein
and	NN	O	O
p44/p42	NN	O	B-protein
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
kinases	NN	O	I-protein
in	NN	O	O
RAGE	NN	O	B-protein
-mediated	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
transcriptional	NN	O	O
activation	NN	O	O
and	NN	O	O
cytokine	NN	O	O
secretion	NN	O	O
.	NN	O	O

Advanced	NN	O	O
glycation	NN	O	O
end	NN	O	O
product	NN	O	O
(	NN	O	O
AGE	NN	O	O
)	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
signal-transducing	NN	O	B-protein
receptor	NN	O	I-protein
for	NN	O	I-protein
AGE	NN	O	I-protein
(	NN	O	O
RAGE	NN	O	B-protein
)	NN	O	O
has	NN	O	O
been	NN	O	O
linked	NN	O	O
to	NN	O	O
a	NN	O	O
proinflammatory	NN	O	O
phenotypic	NN	O	O
change	NN	O	O
within	NN	O	O
cells	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
precise	NN	O	O
intracellular	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
involved	NN	O	O
have	NN	O	O
not	NN	O	O
been	NN	O	O
elucidated	NN	O	O
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
here	NN	O	O
that	NN	O	O
human	NN	O	O
serum	NN	O	O
albumin	NN	O	O
modified	NN	O	O
with	NN	O	O
N	NN	O	O
(	NN	O	O
varepsilon	NN	O	O
)	NN	O	O
-	NN	O	O
(	NN	O	O
carboxymethyl	NN	O	O
)	NN	O	O
lysine	NN	O	O
(	NN	O	O
CML	NN	O	O
)	NN	O	O
,	NN	O	O
a	NN	O	O
major	NN	O	O
AGE	NN	O	O
adduct	NN	O	O
that	NN	O	O
progressively	NN	O	O
accumulates	NN	O	O
with	NN	O	O
aging	NN	O	O
,	NN	O	O
diabetes	NN	O	O
,	NN	O	O
and	NN	O	O
renal	NN	O	O
failure	NN	O	O
,	NN	O	O
induced	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	I-protein
NF	NN	O	I-protein
)	NN	O	I-protein
-kappaB	NN	O	I-protein
-driven	NN	O	O
reporter	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytic	NN	O	I-cell_type
THP-1	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
NF-kappaB	NN	O	O
response	NN	O	O
was	NN	O	O
blocked	NN	O	O
with	NN	O	O
a	NN	O	O
synthetic	NN	O	O
peptide	NN	O	O
corresponding	NN	O	O
to	NN	O	O
the	NN	O	O
putative	NN	O	B-protein
ligand-binding	NN	O	I-protein
domain	NN	O	I-protein
of	NN	O	O
RAGE	NN	O	B-protein
,	NN	O	O
with	NN	O	O
anti-	NN	O	O
RAGE	NN	O	B-protein
antiserum	NN	O	O
,	NN	O	O
and	NN	O	O
by	NN	O	O
coexpression	NN	O	O
of	NN	O	O
truncated	NN	O	B-protein
receptors	NN	O	I-protein
lacking	NN	O	O
the	NN	O	O
intracellular	NN	O	B-protein
domain	NN	O	I-protein
.	NN	O	O

Signal	NN	O	O
transduction	NN	O	O
from	NN	O	O
RAGE	NN	O	B-protein
to	NN	O	O
NF-kappaB	NN	O	B-protein
involved	NN	O	O
the	NN	O	O
generation	NN	O	O
of	NN	O	O
reactive	NN	O	O
oxygen	NN	O	O
species	NN	O	O
,	NN	O	O
since	NN	O	O
reporter	NN	O	O
gene	NN	O	O
expression	NN	O	O
was	NN	O	O
blocked	NN	O	O
with	NN	O	O
the	NN	O	O
antioxidant	NN	O	O
N-acetyl-L-cysteine	NN	O	O
.	NN	O	O

CML-modified	NN	O	B-protein
albumin	NN	O	I-protein
produced	NN	O	O
rapid	NN	O	O
transient	NN	O	O
activation	NN	O	O
of	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
,	NN	O	O
extracellular	NN	O	B-protein
signal-regulated	NN	O	I-protein
kinase	NN	O	I-protein
1	NN	O	I-protein
and	NN	O	I-protein
2	NN	O	I-protein
,	NN	O	O
and	NN	O	O
p38	NN	O	B-protein
mitogen-activated	NN	O	I-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	O
MAPK	NN	O	B-protein
)	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
c-Jun	NN	O	B-protein
NH	NN	O	I-protein
(	NN	O	I-protein
2	NN	O	I-protein
)	NN	O	I-protein
-terminal	NN	O	I-protein
kinase	NN	O	I-protein
.	NN	O	O

RAGE	NN	O	B-protein
-mediated	NN	O	O
NF-kappaB	NN	O	O
activation	NN	O	O
was	NN	O	O
suppressed	NN	O	O
by	NN	O	O
the	NN	O	O
selective	NN	O	O
p38	NN	O	B-protein
MAPK	NN	O	I-protein
inhibitor	NN	O	O
SB203580	NN	O	O
and	NN	O	O
by	NN	O	O
coexpression	NN	O	O
of	NN	O	O
a	NN	O	O
kinase-dead	NN	O	O
p38	NN	O	O
dominant-negative	NN	O	O
mutant	NN	O	O
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
by	NN	O	O
CML-modified	NN	O	B-protein
albumin	NN	O	I-protein
increased	NN	O	O
secretion	NN	O	O
of	NN	O	O
proinflammatory	NN	O	B-protein
cytokines	NN	O	I-protein
(	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
,	NN	O	O
interleukin-1beta	NN	O	B-protein
,	NN	O	O
and	NN	O	O
monocyte	NN	O	B-protein
chemoattractant	NN	O	I-protein
protein-1	NN	O	I-protein
)	NN	O	O
severalfold	NN	O	O
,	NN	O	O
and	NN	O	O
inhibition	NN	O	O
of	NN	O	O
p38	NN	O	B-protein
MAPK	NN	O	I-protein
blocked	NN	O	O
these	NN	O	O
increases	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
p38	NN	O	B-protein
MAPK	NN	O	I-protein
activation	NN	O	O
mediates	NN	O	O
RAGE	NN	O	B-protein
-induced	NN	O	O
NF-kappaB	NN	O	B-protein
-dependent	NN	O	O
secretion	NN	O	O
of	NN	O	O
proinflammatory	NN	O	B-protein
cytokines	NN	O	I-protein
and	NN	O	O
suggest	NN	O	O
that	NN	O	O
accelerated	NN	O	O
inflammation	NN	O	O
may	NN	O	O
be	NN	O	O
a	NN	O	O
consequence	NN	O	O
of	NN	O	O
cellular	NN	O	O
activation	NN	O	O
induced	NN	O	O
by	NN	O	O
this	NN	O	O
receptor	NN	O	O
.	NN	O	O

-DOCSTART-	O

Antigen-receptor	NN	O	O
cross-linking	NN	O	O
and	NN	O	O
lipopolysaccharide	NN	O	O
trigger	NN	O	O
distinct	NN	O	O
phosphoinositide	NN	O	B-protein
3-kinase	NN	O	I-protein
-dependent	NN	O	O
pathways	NN	O	O
to	NN	O	O
NF-kappa	NN	O	O
B	NN	O	O
activation	NN	O	O
in	NN	O	O
primary	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
NF-kappaB/Rel	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
genes	NN	O	O
involved	NN	O	O
in	NN	O	O
B	NN	O	O
cell	NN	O	O
development	NN	O	O
,	NN	O	O
differentiation	NN	O	O
and	NN	O	O
function	NN	O	O
.	NN	O	O

Nuclear	NN	O	B-protein
NF-kappaB	NN	O	I-protein
is	NN	O	O
induced	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
engagement	NN	O	O
of	NN	O	O
either	NN	O	O
the	NN	O	O
BCR	NN	O	B-protein
or	NN	O	O
CD40	NN	O	B-protein
or	NN	O	O
by	NN	O	O
stimulation	NN	O	O
with	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
.	NN	O	O

Despite	NN	O	O
the	NN	O	O
importance	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
to	NN	O	O
B	NN	O	O
cell	NN	O	O
function	NN	O	O
,	NN	O	O
little	NN	O	O
is	NN	O	O
known	NN	O	O
about	NN	O	O
the	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
leading	NN	O	O
to	NN	O	O
NF-kappaB	NN	O	O
activation	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
we	NN	O	O
address	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
phosphoinositide	NN	O	B-protein
3'-kinase	NN	O	I-protein
(	NN	O	O
PI	NN	O	B-protein
3-kinase	NN	O	I-protein
)	NN	O	O
in	NN	O	O
BCR	NN	O	B-protein
-	NN	O	O
and	NN	O	O
LPS-induced	NN	O	O
NF-kappaB	NN	O	O
activation	NN	O	O
using	NN	O	O
populations	NN	O	O
of	NN	O	O
primary	NN	O	B-cell_type
murine	NN	O	I-cell_type
resting	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Using	NN	O	O
the	NN	O	O
specific	NN	O	O
pharmacological	NN	O	O
inhibitors	NN	O	O
of	NN	O	O
PI	NN	O	B-protein
3-kinase	NN	O	I-protein
,	NN	O	O
Wortmannin	NN	O	O
and	NN	O	O
LY294002	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
PI	NN	O	B-protein
3-kinase	NN	O	I-protein
activity	NN	O	O
is	NN	O	O
vital	NN	O	O
for	NN	O	O
BCR	NN	O	B-protein
-induced	NN	O	O
NF-kappaB	NN	O	B-protein
DNA-binding	NN	O	O
activity	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
this	NN	O	O
is	NN	O	O
achieved	NN	O	O
via	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
-dependent	NN	O	O
degradation	NN	O	O
of	NN	O	O
IkappaBalpha	NN	O	B-protein
.	NN	O	O

Similar	NN	O	O
analyses	NN	O	O
reveal	NN	O	O
that	NN	O	O
PI	NN	O	B-protein
3-kinase	NN	O	I-protein
is	NN	O	O
also	NN	O	O
critical	NN	O	O
in	NN	O	O
triggering	NN	O	O
NF-kappaB	NN	O	B-protein
DNA-binding	NN	O	O
activity	NN	O	O
and	NN	O	O
IkappaBalpha	NN	O	B-protein
degradation	NN	O	O
following	NN	O	O
LPS	NN	O	O
stimulation	NN	O	O
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
a	NN	O	O
PKC	NN	O	O
inhibitor	NN	O	O
which	NN	O	O
blocked	NN	O	O
the	NN	O	O
BCR	NN	O	B-protein
-induced	NN	O	O
IkappaBalpha	NN	O	B-protein
degradation	NN	O	O
had	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
degradation	NN	O	O
of	NN	O	O
IkappaBalpha	NN	O	B-protein
after	NN	O	O
LPS	NN	O	O
stimulation	NN	O	O
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
our	NN	O	O
results	NN	O	O
indicate	NN	O	O
the	NN	O	O
involvement	NN	O	O
of	NN	O	O
PI	NN	O	B-protein
3-kinase	NN	O	I-protein
in	NN	O	O
at	NN	O	O
least	NN	O	O
two	NN	O	O
distinct	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
leading	NN	O	O
to	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Tetramer-guided	NN	O	O
epitope	NN	O	O
mapping	NN	O	O
:	NN	O	O
rapid	NN	O	O
identification	NN	O	O
and	NN	O	O
characterization	NN	O	O
of	NN	O	O
immunodominant	NN	O	B-cell_type
CD4+	NN	O	I-cell_type
T	NN	O	I-cell_type
cell	NN	O	I-cell_type
epitopes	NN	O	I-cell_type
from	NN	O	O
complex	NN	O	O
antigens	NN	O	B-protein
.	NN	O	O

T	NN	O	O
cell	NN	O	O
responses	NN	O	O
to	NN	O	O
Ags	NN	O	B-protein
involve	NN	O	O
recognition	NN	O	O
of	NN	O	O
selected	NN	O	O
peptide	NN	O	B-protein
epitopes	NN	O	I-protein
contained	NN	O	O
within	NN	O	O
the	NN	O	O
antigenic	NN	O	B-protein
protein	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
describe	NN	O	O
a	NN	O	O
new	NN	O	O
approach	NN	O	O
for	NN	O	O
direct	NN	O	O
identification	NN	O	O
of	NN	O	O
CD4+	NN	O	B-cell_type
T	NN	O	I-cell_type
cell	NN	O	I-cell_type
epitopes	NN	O	I-cell_type
of	NN	O	O
complex	NN	O	O
Ags	NN	O	B-protein
that	NN	O	O
uses	NN	O	O
human	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
tetramers	NN	O	I-protein
to	NN	O	O
identify	NN	O	O
reactive	NN	O	O
cells	NN	O	O
.	NN	O	O

With	NN	O	O
a	NN	O	O
panel	NN	O	O
of	NN	O	O
60	NN	O	O
overlapping	NN	O	O
peptides	NN	O	O
covering	NN	O	O
the	NN	O	O
entire	NN	O	O
sequence	NN	O	O
of	NN	O	O
the	NN	O	O
VP16	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
a	NN	O	O
major	NN	O	O
Ag	NN	O	B-protein
for	NN	O	O
HSV-2	NN	O	O
,	NN	O	O
we	NN	O	O
generated	NN	O	O
a	NN	O	O
panel	NN	O	O
of	NN	O	O
class	NN	O	B-protein
II	NN	O	I-protein
MHC	NN	O	I-protein
tetramers	NN	O	I-protein
loaded	NN	O	O
with	NN	O	O
peptide	NN	O	O
pools	NN	O	O
that	NN	O	O
were	NN	O	O
used	NN	O	O
to	NN	O	O
stain	NN	O	O
peripheral	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
of	NN	O	O
an	NN	O	O
HSV-2	NN	O	O
infected	NN	O	O
individual	NN	O	O
.	NN	O	O

With	NN	O	O
this	NN	O	O
approach	NN	O	O
,	NN	O	O
we	NN	O	O
identified	NN	O	O
four	NN	O	O
new	NN	O	O
DRA1*0101/DRB1*0401-	NN	O	O
and	NN	O	O
two	NN	O	O
DRA1*0101/DRB1*0404-restricted	NN	O	O
,	NN	O	O
VP16-specific	NN	O	B-protein
epitopes	NN	O	I-protein
.	NN	O	O

By	NN	O	O
using	NN	O	O
tetramers	NN	O	B-protein
to	NN	O	O
sort	NN	O	O
individual	NN	O	O
cells	NN	O	O
,	NN	O	O
we	NN	O	O
easily	NN	O	O
obtained	NN	O	O
a	NN	O	O
large	NN	O	O
number	NN	O	O
of	NN	O	O
clones	NN	O	O
specific	NN	O	O
to	NN	O	O
these	NN	O	O
epitopes	NN	O	B-protein
.	NN	O	O

Although	NN	O	O
DRA1*0101/DRB1*0401	NN	O	O
and	NN	O	O
DRA1*0101/DRB1*0404	NN	O	O
are	NN	O	O
structurally	NN	O	O
very	NN	O	O
similar	NN	O	O
,	NN	O	O
nonoverlapping	NN	O	O
VP16	NN	O	B-protein
epitopes	NN	O	I-protein
were	NN	O	O
identified	NN	O	O
,	NN	O	O
illustrating	NN	O	O
high	NN	O	O
selectivity	NN	O	O
of	NN	O	O
individual	NN	O	O
allele	NN	O	O
polymorphisms	NN	O	O
within	NN	O	O
common	NN	O	O
MHC	NN	O	O
variants	NN	O	O
.	NN	O	O

This	NN	O	O
rapid	NN	O	O
approach	NN	O	O
to	NN	O	O
detecting	NN	O	O
CD4+	NN	O	B-cell_type
T	NN	O	I-cell_type
cell	NN	O	I-cell_type
epitopes	NN	O	I-cell_type
from	NN	O	O
complex	NN	O	O
Ags	NN	O	B-protein
can	NN	O	O
be	NN	O	O
applied	NN	O	O
to	NN	O	O
any	NN	O	O
known	NN	O	O
Ag	NN	O	B-protein
that	NN	O	O
gives	NN	O	O
a	NN	O	O
T	NN	O	O
cell	NN	O	O
response	NN	O	O
.	NN	O	O

-DOCSTART-	O

Localized	NN	O	O
pancreatic	NN	O	O
NF-kappaB	NN	O	O
activation	NN	O	O
and	NN	O	O
inflammatory	NN	O	O
response	NN	O	O
in	NN	O	O
taurocholate-induced	NN	O	O
pancreatitis	NN	O	O
.	NN	O	O

Transcription	NN	O	B-protein
factor	NN	O	I-protein
nuclear	NN	O	I-protein
factor-kappaB	NN	O	I-protein
(	NN	O	O
NF-kappaB	NN	O	B-protein
)	NN	O	O
is	NN	O	O
activated	NN	O	O
in	NN	O	O
cerulein	NN	O	O
pancreatitis	NN	O	O
and	NN	O	O
mediates	NN	O	O
cytokine	NN	O	O
expression	NN	O	O
.	NN	O	O

The	NN	O	O
role	NN	O	O
of	NN	O	O
transcription	NN	O	O
factor	NN	O	O
activation	NN	O	O
in	NN	O	O
other	NN	O	O
models	NN	O	O
of	NN	O	O
pancreatitis	NN	O	O
has	NN	O	O
not	NN	O	O
been	NN	O	O
established	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
report	NN	O	O
upregulation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
and	NN	O	O
inflammatory	NN	O	B-protein
molecules	NN	O	I-protein
,	NN	O	O
and	NN	O	O
their	NN	O	O
correlation	NN	O	O
with	NN	O	O
local	NN	O	O
pancreatic	NN	O	O
injury	NN	O	O
,	NN	O	O
in	NN	O	O
a	NN	O	O
model	NN	O	O
of	NN	O	O
severe	NN	O	O
pancreatitis	NN	O	O
.	NN	O	O

Rats	NN	O	O
received	NN	O	O
intraductal	NN	O	O
infusion	NN	O	O
of	NN	O	O
taurocholate	NN	O	O
or	NN	O	O
saline	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
pancreatic	NN	O	O
head	NN	O	O
and	NN	O	O
tail	NN	O	O
were	NN	O	O
analyzed	NN	O	O
separately	NN	O	O
.	NN	O	O

NF-kappaB	NN	O	B-protein
and	NN	O	O
activator	NN	O	B-protein
protein-1	NN	O	I-protein
(	NN	O	O
AP-1	NN	O	B-protein
)	NN	O	O
activation	NN	O	O
were	NN	O	O
assessed	NN	O	O
by	NN	O	O
gel	NN	O	O
shift	NN	O	O
assay	NN	O	O
,	NN	O	O
and	NN	O	O
mRNA	NN	O	O
expression	NN	O	O
of	NN	O	O
interleukin-6	NN	O	B-RNA
,	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
,	NN	O	O
KC	NN	O	O
,	NN	O	O
monocyte	NN	O	B-protein
chemoattractant	NN	O	I-protein
protein-1	NN	O	I-protein
,	NN	O	O
and	NN	O	O
inducible	NN	O	B-protein
nitric	NN	O	I-protein
oxide	NN	O	I-protein
synthase	NN	O	I-protein
was	NN	O	O
assessed	NN	O	O
by	NN	O	O
semiquantitative	NN	O	O
RT-PCR	NN	O	O
.	NN	O	O

Morphological	NN	O	O
damage	NN	O	O
and	NN	O	O
trypsin	NN	O	O
activation	NN	O	O
were	NN	O	O
much	NN	O	O
greater	NN	O	O
in	NN	O	O
the	NN	O	O
pancreatic	NN	O	O
head	NN	O	O
than	NN	O	O
tail	NN	O	O
,	NN	O	O
in	NN	O	O
parallel	NN	O	O
with	NN	O	O
a	NN	O	O
stronger	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-RNA
and	NN	O	I-RNA
cytokine	NN	O	I-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

Saline	NN	O	O
infusion	NN	O	O
mildly	NN	O	O
affected	NN	O	O
these	NN	O	O
parameters	NN	O	O
.	NN	O	O

AP-1	NN	O	B-protein
was	NN	O	O
strongly	NN	O	O
activated	NN	O	O
in	NN	O	O
both	NN	O	O
pancreatic	NN	O	O
segments	NN	O	O
after	NN	O	O
either	NN	O	O
taurocholate	NN	O	O
or	NN	O	O
saline	NN	O	O
infusion	NN	O	O
.	NN	O	O

NF-kappaB	NN	O	B-protein
inhibition	NN	O	O
with	NN	O	O
N-acetylcysteine	NN	O	O
ameliorated	NN	O	O
the	NN	O	O
local	NN	O	O
inflammatory	NN	O	O
response	NN	O	O
.	NN	O	O

Correlation	NN	O	O
between	NN	O	O
localized	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
,	NN	O	O
cytokine	NN	O	O
upregulation	NN	O	O
,	NN	O	O
and	NN	O	O
tissue	NN	O	O
damage	NN	O	O
suggests	NN	O	O
a	NN	O	O
key	NN	O	O
role	NN	O	O
for	NN	O	O
NF-kappaB	NN	O	B-protein
in	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
the	NN	O	O
inflammatory	NN	O	O
response	NN	O	O
of	NN	O	O
acute	NN	O	O
pancreatitis	NN	O	O
.	NN	O	O

-DOCSTART-	O

CD45	NN	O	B-protein
tyrosine	NN	O	I-protein
phosphatase	NN	O	I-protein
controls	NN	O	O
common	NN	O	O
gamma-chain	NN	O	B-protein
cytokine	NN	O	I-protein
-mediated	NN	O	O
STAT	NN	O	B-protein
and	NN	O	O
extracellular	NN	O	O
signal-related	NN	O	O
kinase	NN	O	O
phosphorylation	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
human	NN	O	I-cell_type
lymphoblasts	NN	O	I-cell_type
:	NN	O	O
inhibition	NN	O	O
of	NN	O	O
proliferation	NN	O	O
without	NN	O	O
induction	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

The	NN	O	O
objective	NN	O	O
of	NN	O	O
this	NN	O	O
study	NN	O	O
was	NN	O	O
to	NN	O	O
test	NN	O	O
whether	NN	O	O
CD45	NN	O	O
signals	NN	O	O
can	NN	O	O
influence	NN	O	O
signaling	NN	O	O
processes	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
human	NN	O	I-cell_type
lymphoblasts	NN	O	I-cell_type
.	NN	O	O

To	NN	O	O
this	NN	O	O
end	NN	O	O
,	NN	O	O
we	NN	O	O
generated	NN	O	O
lymphoblasts	NN	O	B-cell_type
which	NN	O	O
proliferate	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
common	NN	O	O
gamma-chain	NN	O	B-protein
cytokines	NN	O	I-protein
,	NN	O	O
but	NN	O	O
readily	NN	O	O
undergo	NN	O	O
apoptosis	NN	O	O
after	NN	O	O
cytokine	NN	O	O
withdrawal	NN	O	O
.	NN	O	O

In	NN	O	O
experiments	NN	O	O
with	NN	O	O
the	NN	O	O
CD45R0	NN	O	B-protein
mAb	NN	O	I-protein
UCHL-1	NN	O	I-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
control	NN	O	B-protein
CD45	NN	O	I-protein
mAbs	NN	O	I-protein
,	NN	O	O
we	NN	O	O
found	NN	O	O
significant	NN	O	O
inhibition	NN	O	O
of	NN	O	O
proliferation	NN	O	O
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
the	NN	O	O
pan-CD45	NN	O	B-protein
mAb	NN	O	I-protein
GAP8.3	NN	O	I-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
most	NN	O	O
effective	NN	O	O
in	NN	O	O
inhibition	NN	O	O
of	NN	O	O
OKT-3-mediated	NN	O	O
proliferation	NN	O	O
in	NN	O	O
quiescent	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
was	NN	O	O
ineffective	NN	O	O
in	NN	O	O
lymphoblasts	NN	O	B-cell_type
.	NN	O	O

Addition	NN	O	O
of	NN	O	O
CD3	NN	O	B-protein
mAb	NN	O	I-protein
OKT-3	NN	O	I-protein
had	NN	O	O
no	NN	O	O
influence	NN	O	O
on	NN	O	O
IL-2-mediated	NN	O	O
proliferation	NN	O	O
(	NN	O	O
with	NN	O	O
or	NN	O	O
without	NN	O	O
UCHL-1	NN	O	B-protein
)	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
after	NN	O	O
addition	NN	O	O
of	NN	O	O
OKT-3	NN	O	B-protein
to	NN	O	O
IL-4	NN	O	B-protein
-	NN	O	O
and	NN	O	O
IL-7	NN	O	B-protein
-stimulated	NN	O	O
proliferation	NN	O	O
assays	NN	O	O
,	NN	O	O
UCHL-1	NN	O	B-protein
signals	NN	O	O
could	NN	O	O
not	NN	O	O
significantly	NN	O	O
alter	NN	O	O
cellular	NN	O	O
proliferation	NN	O	O
.	NN	O	O

We	NN	O	O
did	NN	O	O
not	NN	O	O
find	NN	O	O
induction	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
following	NN	O	O
CD45R0	NN	O	O
signaling	NN	O	O
.	NN	O	O

In	NN	O	O
Western	NN	O	O
blots	NN	O	O
using	NN	O	O
mAbs	NN	O	B-protein
detecting	NN	O	O
phosphorylated	NN	O	B-protein
STAT-3	NN	O	I-protein
,	NN	O	I-protein
STAT-5	NN	O	I-protein
,	NN	O	I-protein
STAT-6	NN	O	I-protein
,	NN	O	O
or	NN	O	O
extracellular	NN	O	B-protein
signal-related	NN	O	I-protein
kinase	NN	O	I-protein
1/2	NN	O	I-protein
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
CD45R0	NN	O	O
signaling	NN	O	O
could	NN	O	O
effectively	NN	O	O
diminish	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
these	NN	O	O
intracellular	NN	O	O
signaling	NN	O	O
components	NN	O	O
.	NN	O	O

Using	NN	O	O
RT-PCR	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
CD45R0	NN	O	O
signaling	NN	O	O
inhibited	NN	O	O
IL-2	NN	O	O
mRNA	NN	O	O
production	NN	O	O
without	NN	O	O
major	NN	O	O
influence	NN	O	O
on	NN	O	O
IL-13	NN	O	B-protein
,	NN	O	O
IL-5	NN	O	B-protein
,	NN	O	O
or	NN	O	O
IFN-gamma	NN	O	B-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
.	NN	O	O

Costimulation	NN	O	O
with	NN	O	O
OKT-3	NN	O	B-protein
and	NN	O	O
IL-2	NN	O	B-protein
optimally	NN	O	O
induced	NN	O	O
secretion	NN	O	O
of	NN	O	O
IFN-gamma	NN	O	B-protein
,	NN	O	O
TNF-alpha	NN	O	B-protein
,	NN	O	O
and	NN	O	O
IL-5	NN	O	B-protein
,	NN	O	O
which	NN	O	O
was	NN	O	O
not	NN	O	O
decreased	NN	O	O
by	NN	O	O
CD45	NN	O	O
signals	NN	O	O
.	NN	O	O

In	NN	O	O
conclusion	NN	O	O
,	NN	O	O
we	NN	O	O
illustrate	NN	O	O
that	NN	O	O
CD45R0	NN	O	O
signals	NN	O	O
control	NN	O	O
early	NN	O	O
cytokine	NN	O	O
receptor-associated	NN	O	O
signaling	NN	O	O
processes	NN	O	O
and	NN	O	O
mRNA	NN	O	O
and	NN	O	O
DNA	NN	O	O
synthesis	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
human	NN	O	I-cell_type
lymphoblasts	NN	O	I-cell_type
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
the	NN	O	O
existence	NN	O	O
of	NN	O	O
CD45	NN	O	B-protein
epitopes	NN	O	I-protein
(	NN	O	O
GAP8.3	NN	O	B-protein
)	NN	O	O
,	NN	O	O
which	NN	O	O
are	NN	O	O
active	NN	O	O
and	NN	O	O
critical	NN	O	O
for	NN	O	O
signaling	NN	O	O
in	NN	O	O
quiescent	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
but	NN	O	O
are	NN	O	O
nonfunctional	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
human	NN	O	I-cell_type
lymphoblasts	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Pax5	NN	O	B-DNA
determines	NN	O	O
the	NN	O	O
identity	NN	O	O
of	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
the	NN	O	O
beginning	NN	O	O
to	NN	O	O
the	NN	O	O
end	NN	O	O
of	NN	O	O
B-lymphopoiesis	NN	O	O
.	NN	O	O

Despite	NN	O	O
being	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
most	NN	O	O
intensively	NN	O	O
studied	NN	O	O
cell	NN	O	O
types	NN	O	O
,	NN	O	O
the	NN	O	O
molecular	NN	O	O
basis	NN	O	O
of	NN	O	O
B	NN	O	O
cell	NN	O	O
specification	NN	O	O
is	NN	O	O
largely	NN	O	O
unknown	NN	O	O
.	NN	O	O

The	NN	O	O
Pax5	NN	O	B-DNA
gene	NN	O	I-DNA
encoding	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
BSAP	NN	O	I-protein
is	NN	O	O
required	NN	O	O
for	NN	O	O
progression	NN	O	O
of	NN	O	O
B-lymphopoiesis	NN	O	O
beyond	NN	O	O
the	NN	O	O
pro-B	NN	O	O
cell	NN	O	O
stage	NN	O	O
.	NN	O	O

Pax5-deficient	NN	O	B-cell_type
pro-B	NN	O	I-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
not	NN	O	O
yet	NN	O	O
committed	NN	O	O
to	NN	O	O
the	NN	O	O
B-lymphoid	NN	O	B-cell_type
lineage	NN	O	I-cell_type
,	NN	O	O
but	NN	O	O
instead	NN	O	O
have	NN	O	O
a	NN	O	O
broad	NN	O	O
lymphomyeloid	NN	O	O
developmental	NN	O	O
potential	NN	O	O
.	NN	O	O

Pax5	NN	O	B-DNA
appears	NN	O	O
to	NN	O	O
mediate	NN	O	O
B-lineage	NN	O	O
commitment	NN	O	O
by	NN	O	O
repressing	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
non-B-lymphoid	NN	O	B-DNA
genes	NN	O	I-DNA
and	NN	O	O
by	NN	O	O
simultaneously	NN	O	O
activating	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
B-lineage-specific	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

Pax5	NN	O	B-DNA
thus	NN	O	O
functions	NN	O	O
both	NN	O	O
as	NN	O	O
a	NN	O	O
transcriptional	NN	O	O
repressor	NN	O	O
and	NN	O	O
activator	NN	O	O
,	NN	O	O
depending	NN	O	O
on	NN	O	O
its	NN	O	O
interactions	NN	O	O
with	NN	O	O
corepressors	NN	O	B-protein
of	NN	O	O
the	NN	O	O
Groucho	NN	O	B-protein
protein	NN	O	I-protein
family	NN	O	I-protein
or	NN	O	O
with	NN	O	O
positive	NN	O	O
regulators	NN	O	O
such	NN	O	O
as	NN	O	O
the	NN	O	O
TATA-binding	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

Once	NN	O	O
committed	NN	O	O
to	NN	O	O
the	NN	O	O
B-lineage	NN	O	B-cell_type
,	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
require	NN	O	O
Pax5	NN	O	B-DNA
function	NN	O	O
to	NN	O	O
maintain	NN	O	O
their	NN	O	O
B-lymphoid	NN	O	O
identity	NN	O	O
throughout	NN	O	O
B	NN	O	O
cell	NN	O	O
development	NN	O	O

-DOCSTART-	O

Partners	NN	O	O
in	NN	O	O
transcription	NN	O	O
:	NN	O	O
NFAT	NN	O	B-protein
and	NN	O	O
AP-1	NN	O	B-protein
.	NN	O	O

Combinatorial	NN	O	O
regulation	NN	O	O
is	NN	O	O
a	NN	O	O
powerful	NN	O	O
mechanism	NN	O	O
that	NN	O	O
enables	NN	O	O
tight	NN	O	O
control	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
,	NN	O	O
via	NN	O	O
integration	NN	O	O
of	NN	O	O
multiple	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
that	NN	O	O
induce	NN	O	O
different	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
required	NN	O	O
for	NN	O	O
enhanceosome	NN	O	O
assembly	NN	O	O
.	NN	O	O

The	NN	O	O
four	NN	O	O
calcium-regulated	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
of	NN	O	O
the	NN	O	O
NFAT	NN	O	B-protein
family	NN	O	I-protein
act	NN	O	O
synergistically	NN	O	O
with	NN	O	O
AP-1	NN	O	B-protein
(	NN	O	I-protein
Fos/Jun	NN	O	I-protein
)	NN	O	I-protein
proteins	NN	O	I-protein
on	NN	O	O
composite	NN	O	O
DNA	NN	O	O
elements	NN	O	O
which	NN	O	O
contain	NN	O	O
adjacent	NN	O	O
NFAT	NN	O	B-DNA
and	NN	O	I-DNA
AP-1	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
,	NN	O	O
where	NN	O	O
they	NN	O	O
form	NN	O	O
highly	NN	O	O
stable	NN	O	O
ternary	NN	O	O
complexes	NN	O	O
to	NN	O	O
regulate	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
diverse	NN	O	O
inducible	NN	O	O
genes	NN	O	O
.	NN	O	O

Concomitant	NN	O	O
induction	NN	O	O
of	NN	O	O
NFAT	NN	O	B-protein
and	NN	O	O
AP-1	NN	O	B-protein
requires	NN	O	O
concerted	NN	O	O
activation	NN	O	O
of	NN	O	O
two	NN	O	O
different	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
:	NN	O	O
calcium/calcineurin	NN	O	O
,	NN	O	O
which	NN	O	O
promotes	NN	O	O
NFAT	NN	O	O
dephosphorylation	NN	O	O
,	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
and	NN	O	O
activation	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
(	NN	O	I-protein
PKC	NN	O	I-protein
)	NN	O	I-protein
/Ras	NN	O	I-protein
,	NN	O	O
which	NN	O	O
promotes	NN	O	O
the	NN	O	O
synthesis	NN	O	O
,	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
Fos	NN	O	B-protein
and	NN	O	I-protein
Jun	NN	O	I-protein
families	NN	O	I-protein
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

A	NN	O	O
fifth	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
NFAT	NN	O	B-protein
family	NN	O	I-protein
,	NN	O	O
NFAT5	NN	O	B-protein
,	NN	O	O
controls	NN	O	O
the	NN	O	O
cellular	NN	O	O
response	NN	O	O
to	NN	O	O
osmotic	NN	O	O
stress	NN	O	O
,	NN	O	O
by	NN	O	O
a	NN	O	O
mechanism	NN	O	O
that	NN	O	O
requires	NN	O	O
dimer	NN	O	O
formation	NN	O	O
and	NN	O	O
is	NN	O	O
independent	NN	O	O
of	NN	O	O
calcineurin	NN	O	B-protein
or	NN	O	O
of	NN	O	O
interaction	NN	O	O
with	NN	O	O
AP-1	NN	O	B-protein
.	NN	O	O

Pharmacological	NN	O	O
interference	NN	O	O
with	NN	O	O
theNFAT	NN	O	O
:	NN	O	O
AP-1	NN	O	O
interaction	NN	O	O
may	NN	O	O
be	NN	O	O
useful	NN	O	O
in	NN	O	O
selective	NN	O	O
manipulation	NN	O	O
of	NN	O	O
the	NN	O	O
immune	NN	O	O
response	NN	O	O
.	NN	O	O

Balanced	NN	O	O
activation	NN	O	O
of	NN	O	O
NFAT	NN	O	B-protein
and	NN	O	O
AP-1	NN	O	B-protein
is	NN	O	O
known	NN	O	O
to	NN	O	O
be	NN	O	O
required	NN	O	O
for	NN	O	O
productive	NN	O	O
immune	NN	O	O
responses	NN	O	O
,	NN	O	O
but	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
NFAT	NN	O	O
:	NN	O	O
AP-1	NN	O	O
interactions	NN	O	O
in	NN	O	O
other	NN	O	O
cell	NN	O	O
types	NN	O	O
and	NN	O	O
biological	NN	O	O
processes	NN	O	O
remains	NN	O	O
to	NN	O	O
be	NN	O	O
understood	NN	O	O
.	NN	O	O

-DOCSTART-	O

Cytokine	NN	O	O
production	NN	O	O
by	NN	O	O
Vgamma	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
-T-cell	NN	O	I-cell_type
subsets	NN	O	I-cell_type
is	NN	O	O
an	NN	O	O
important	NN	O	O
factor	NN	O	O
determining	NN	O	O
CD4	NN	O	O
(	NN	O	O
+	NN	O	O
)	NN	O	O
-Th-cell	NN	O	O
phenotype	NN	O	O
and	NN	O	O
susceptibility	NN	O	O
of	NN	O	O
BALB/c	NN	O	O
mice	NN	O	O
to	NN	O	O
coxsackievirus	NN	O	O
B3-induced	NN	O	O
myocarditis	NN	O	O
.	NN	O	O

Two	NN	O	O
coxsackievirus	NN	O	O
B3	NN	O	O
(	NN	O	O
CVB3	NN	O	O
)	NN	O	O
variants	NN	O	O
(	NN	O	O
H3	NN	O	O
and	NN	O	O
H310A1	NN	O	O
)	NN	O	O
differ	NN	O	O
by	NN	O	O
a	NN	O	O
single	NN	O	O
amino	NN	O	O
acid	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
VP2	NN	O	B-protein
capsid	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

H3	NN	O	O
induces	NN	O	O
severe	NN	O	O
myocarditis	NN	O	O
in	NN	O	O
BALB/c	NN	O	O
mice	NN	O	O
,	NN	O	O
but	NN	O	O
H310A1	NN	O	O
is	NN	O	O
amyocarditic	NN	O	O
.	NN	O	O

Infection	NN	O	O
with	NN	O	O
H3	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
H310A1	NN	O	O
,	NN	O	O
preferentially	NN	O	O
activates	NN	O	O
Vgamma4	NN	O	B-cell_type
Vdelta4	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
which	NN	O	O
are	NN	O	O
strongly	NN	O	O
positive	NN	O	O
for	NN	O	O
gamma	NN	O	B-protein
interferon	NN	O	I-protein
(	NN	O	O
IFN-gamma	NN	O	B-protein
)	NN	O	O
,	NN	O	O
whereas	NN	O	O
Vgamma1	NN	O	B-cell_type
Vdelta4	NN	O	I-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
increased	NN	O	O
in	NN	O	O
both	NN	O	O
H3	NN	O	O
and	NN	O	O
H310A1	NN	O	O
virus-infected	NN	O	O
animals	NN	O	O
.	NN	O	O

Depletion	NN	O	O
of	NN	O	O
Vgamma1	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
using	NN	O	O
monoclonal	NN	O	B-protein
anti-Vgamma1	NN	O	I-protein
antibody	NN	O	I-protein
enhanced	NN	O	O
myocarditis	NN	O	O
and	NN	O	O
CD4	NN	O	O
(	NN	O	O
+	NN	O	O
)	NN	O	O
-	NN	O	O
,	NN	O	O
IFN-gamma	NN	O	B-protein
(	NN	O	O
+	NN	O	O
)	NN	O	O
-cell	NN	O	O
responses	NN	O	O
in	NN	O	O
both	NN	O	O
H3-	NN	O	O
and	NN	O	O
H310A1-infected	NN	O	O
mice	NN	O	O
yet	NN	O	O
decreased	NN	O	O
the	NN	O	O
CD4	NN	O	O
(	NN	O	O
+	NN	O	O
)	NN	O	O
-	NN	O	O
,	NN	O	O
IL-4	NN	O	O
(	NN	O	O
+	NN	O	O
)	NN	O	O
-cell	NN	O	O
response	NN	O	O
.	NN	O	O

Depleting	NN	O	O
Vgamma4	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
suppressed	NN	O	O
myocarditis	NN	O	O
and	NN	O	O
reduced	NN	O	O
CD4	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
IFN-gamma	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
but	NN	O	O
increased	NN	O	O
CD4	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
IL-4	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
role	NN	O	O
of	NN	O	O
cytokine	NN	O	O
production	NN	O	O
by	NN	O	O
Vgamma1	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
and	NN	O	I-cell_type
Vgamma4	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
investigated	NN	O	O
by	NN	O	O
adoptively	NN	O	O
transferring	NN	O	O
these	NN	O	O
cells	NN	O	O
isolated	NN	O	O
from	NN	O	O
H3-infected	NN	O	O
BALB/c	NN	O	O
Stat4	NN	O	O
knockout	NN	O	O
(	NN	O	O
Stat4ko	NN	O	O
)	NN	O	O
(	NN	O	O
defective	NN	O	O
in	NN	O	O
IFN-gamma	NN	O	B-protein
expression	NN	O	O
)	NN	O	O
or	NN	O	O
BALB/c	NN	O	O
Stat6ko	NN	O	O
(	NN	O	O
defective	NN	O	O
in	NN	O	O
IL-4	NN	O	O
expression	NN	O	O
)	NN	O	O
mice	NN	O	O
into	NN	O	O
H3	NN	O	O
virus-infected	NN	O	O
wild-type	NN	O	O
BALB/c	NN	O	O
recipients	NN	O	O
.	NN	O	O

Vgamma4	NN	O	B-cell_type
and	NN	O	I-cell_type
Vgamma1	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
Stat4ko	NN	O	O
mice	NN	O	O
expressed	NN	O	O
IL-4	NN	O	B-protein
but	NN	O	O
no	NN	O	O
or	NN	O	O
minimal	NN	O	O
IFN-gamma	NN	O	B-protein
,	NN	O	O
whereas	NN	O	O
these	NN	O	O
cell	NN	O	O
populations	NN	O	O
derived	NN	O	O
from	NN	O	O
Stat6ko	NN	O	O
mice	NN	O	O
expressed	NN	O	O
IFN-gamma	NN	O	B-protein
but	NN	O	O
no	NN	O	O
IL-4	NN	O	B-protein
.	NN	O	O

Stat4ko	NN	O	B-cell_type
Vgamma1	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	I-cell_type
IL-4	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
)	NN	O	O
suppress	NN	O	O
myocarditis	NN	O	O
.	NN	O	O

Stat6ko	NN	O	B-cell_type
Vgamma1	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	I-cell_type
IFN-gamma	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
)	NN	O	I-cell_type
were	NN	O	O
not	NN	O	O
inhibitory	NN	O	O
.	NN	O	O

Stat6ko	NN	O	B-cell_type
Vgamma4	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	I-cell_type
IFN-gamma	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
)	NN	O	I-cell_type
significantly	NN	O	O
enhanced	NN	O	O
myocarditis	NN	O	O
.	NN	O	O

Stat4ko	NN	O	B-cell_type
Vgamma4	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	I-cell_type
IL-4	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
)	NN	O	I-cell_type
neither	NN	O	O
inhibited	NN	O	O
nor	NN	O	O
enhanced	NN	O	O
disease	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
show	NN	O	O
that	NN	O	O
distinct	NN	O	O
gammadelta-T-cell	NN	O	B-cell_type
subsets	NN	O	I-cell_type
control	NN	O	O
myocarditis	NN	O	O
susceptibility	NN	O	O
and	NN	O	O
bias	NN	O	O
the	NN	O	O
CD4	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
-Th-cell	NN	O	I-cell_type
response	NN	O	O
.	NN	O	O

The	NN	O	O
cytokines	NN	O	B-protein
produced	NN	O	O
by	NN	O	O
the	NN	O	O
Vgamma	NN	O	B-cell_type
subpopulation	NN	O	I-cell_type
have	NN	O	O
a	NN	O	O
significant	NN	O	O
influence	NN	O	O
on	NN	O	O
the	NN	O	O
CD4	NN	O	B-protein
(	NN	O	O
+	NN	O	O
)	NN	O	O
-Th-cell	NN	O	O
phenotype	NN	O	O
.	NN	O	O

-DOCSTART-	O

Plasmin	NN	O	B-protein
-induced	NN	O	O
expression	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
and	NN	O	O
tissue	NN	O	B-protein
factor	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
involves	NN	O	O
AP-1	NN	O	B-protein
and	NN	O	O
IKKbeta	NN	O	B-protein
-mediated	NN	O	O
NF-kappaB	NN	O	O
activation	NN	O	O
.	NN	O	O

It	NN	O	O
was	NN	O	O
previously	NN	O	O
shown	NN	O	O
that	NN	O	O
plasmin	NN	O	B-protein
activates	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
in	NN	O	O
terms	NN	O	O
of	NN	O	O
lipid	NN	O	O
mediator	NN	O	O
release	NN	O	O
and	NN	O	O
chemotactic	NN	O	O
migration	NN	O	O
.	NN	O	O

Here	NN	O	O
it	NN	O	O
is	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
plasmin	NN	O	B-protein
induces	NN	O	O
proinflammatory	NN	O	B-protein
cytokine	NN	O	I-protein
release	NN	O	O
and	NN	O	O
tissue	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	O
TF	NN	O	B-protein
)	NN	O	O
expression	NN	O	O
by	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

Plasmin	NN	O	B-protein
0.043	NN	O	O
to	NN	O	O
1.43	NN	O	O
CTA	NN	O	O
U/mL	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
active	NN	O	B-protein
site-blocked	NN	O	I-protein
plasmin	NN	O	I-protein
,	NN	O	O
triggered	NN	O	O
concentration-dependent	NN	O	O
expression	NN	O	O
of	NN	O	O
mRNA	NN	O	O
for	NN	O	O
interleukin-1alpha	NN	O	B-protein
(	NN	O	O
IL-1alpha	NN	O	B-protein
)	NN	O	O
,	NN	O	O
IL-1beta	NN	O	B-protein
,	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
TF	NN	O	B-protein
with	NN	O	O
maximum	NN	O	O
responses	NN	O	O
after	NN	O	O
4	NN	O	O
hours	NN	O	O
.	NN	O	O

Plasmin	NN	O	B-protein
-mediated	NN	O	O
mRNA	NN	O	O
expression	NN	O	O
was	NN	O	O
inhibited	NN	O	O
in	NN	O	O
a	NN	O	O
concentration-dependent	NN	O	O
manner	NN	O	O
by	NN	O	O
the	NN	O	O
lysine	NN	O	O
analogue	NN	O	O
trans-4-	NN	O	O
(	NN	O	O
aminomethyl	NN	O	O
)	NN	O	O
cyclohexane-1-carboxylic	NN	O	O
acid	NN	O	O
(	NN	O	O
t-AMCA	NN	O	O
)	NN	O	O
.	NN	O	O

Increases	NN	O	O
in	NN	O	O
mRNA	NN	O	O
levels	NN	O	O
were	NN	O	O
followed	NN	O	O
by	NN	O	O
concentration-	NN	O	O
and	NN	O	O
time-dependent	NN	O	O
release	NN	O	O
of	NN	O	O
IL-1alpha	NN	O	B-protein
,	NN	O	O
IL-1beta	NN	O	B-protein
and	NN	O	O
TNF-alpha	NN	O	B-protein
and	NN	O	O
by	NN	O	O
TF	NN	O	B-protein
expression	NN	O	O
on	NN	O	O
monocyte	NN	O	O
surfaces	NN	O	O
.	NN	O	O

Neither	NN	O	O
cytokines	NN	O	B-protein
nor	NN	O	O
TF	NN	O	B-protein
could	NN	O	O
be	NN	O	O
detected	NN	O	O
when	NN	O	O
monocytes	NN	O	B-cell_type
were	NN	O	O
preincubated	NN	O	O
with	NN	O	O
actinomycin	NN	O	O
D	NN	O	O
or	NN	O	O
cycloheximide	NN	O	O
.	NN	O	O

Electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
indicated	NN	O	O
plasmin	NN	O	B-protein
-induced	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
;	NN	O	O
DNA-binding	NN	O	B-protein
complexes	NN	O	I-protein
were	NN	O	O
composed	NN	O	O
of	NN	O	O
p50	NN	O	B-protein
,	NN	O	O
p65	NN	O	B-protein
,	NN	O	O
and	NN	O	O
c-Rel	NN	O	B-protein
,	NN	O	O
as	NN	O	O
shown	NN	O	O
by	NN	O	O
supershift	NN	O	O
experiments	NN	O	O
.	NN	O	O

Nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
NF-kappaB/Rel	NN	O	B-protein
proteins	NN	O	I-protein
coincided	NN	O	O
with	NN	O	O
IkappaBalpha	NN	O	O
degradation	NN	O	O
.	NN	O	O

At	NN	O	O
variance	NN	O	O
with	NN	O	O
endotoxic	NN	O	O
lipopolysaccharide	NN	O	O
,	NN	O	O
plasmin	NN	O	B-protein
elicited	NN	O	O
the	NN	O	O
rapid	NN	O	O
degradation	NN	O	O
of	NN	O	O
another	NN	O	O
cytoplasmic	NN	O	B-protein
NF-kappaB	NN	O	I-protein
inhibitor	NN	O	I-protein
,	NN	O	I-protein
p105	NN	O	I-protein
.	NN	O	O

Proteolysis	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	O
inhibitors	NN	O	O
was	NN	O	O
apparently	NN	O	O
due	NN	O	O
to	NN	O	O
transient	NN	O	O
activation	NN	O	O
of	NN	O	O
IkappaB	NN	O	B-protein
kinase	NN	O	I-protein
(	NN	O	I-protein
IKK	NN	O	I-protein
)	NN	O	I-protein
beta	NN	O	I-protein
that	NN	O	O
reached	NN	O	O
maximum	NN	O	O
activity	NN	O	O
at	NN	O	O
1	NN	O	O
hour	NN	O	O
after	NN	O	O
plasmin	NN	O	B-protein
stimulation	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
AP-1	NN	O	O
binding	NN	O	O
was	NN	O	O
increased	NN	O	O
in	NN	O	O
plasmin	NN	O	B-protein
-treated	NN	O	O
monocytes	NN	O	B-cell_type
,	NN	O	O
with	NN	O	O
most	NN	O	O
complexes	NN	O	O
composed	NN	O	O
of	NN	O	O
JunD	NN	O	B-protein
,	NN	O	O
c-Fos	NN	O	B-protein
,	NN	O	O
and	NN	O	O
FosB	NN	O	B-protein
.	NN	O	O

These	NN	O	O
findings	NN	O	O
further	NN	O	O
substantiate	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
plasmin	NN	O	B-protein
as	NN	O	O
a	NN	O	O
proinflammatory	NN	O	O
activator	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
and	NN	O	O
reveal	NN	O	O
an	NN	O	O
important	NN	O	O
new	NN	O	O
link	NN	O	O
between	NN	O	O
the	NN	O	O
plasminogen-plasmin	NN	O	O
system	NN	O	O
and	NN	O	O
inflammation	NN	O	O
.	NN	O	O

(	NN	O	O
Blood.	NN	O	O
2001	NN	O	O
;	NN	O	O
97	NN	O	O
:	NN	O	O
3941-3950	NN	O	O
)	NN	O	O

-DOCSTART-	O

STAT3	NN	O	B-protein
is	NN	O	O
constitutively	NN	O	O
active	NN	O	O
in	NN	O	O
some	NN	O	O
patients	NN	O	O
with	NN	O	O
Polycythemia	NN	O	O
rubra	NN	O	O
vera	NN	O	O
.	NN	O	O

OBJECTIVE	NN	O	O
:	NN	O	O
Polycythemia	NN	O	O
vera	NN	O	O
is	NN	O	O
a	NN	O	O
clonal	NN	O	O
stem	NN	O	O
cell	NN	O	O
disorder	NN	O	O
characterized	NN	O	O
by	NN	O	O
hyperproliferation	NN	O	O
of	NN	O	O
the	NN	O	O
erythroid	NN	O	B-cell_type
,	NN	O	I-cell_type
myeloid	NN	O	I-cell_type
,	NN	O	I-cell_type
and	NN	O	I-cell_type
megakaryocytic	NN	O	I-cell_type
lineages	NN	O	I-cell_type
.	NN	O	O

While	NN	O	O
it	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
that	NN	O	O
progenitor	NN	O	B-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
P.	NN	O	O
vera	NN	O	O
patients	NN	O	O
are	NN	O	O
hypersensitive	NN	O	O
to	NN	O	O
several	NN	O	O
growth	NN	O	O
factors	NN	O	O
including	NN	O	O
erythropoietin	NN	O	B-protein
,	NN	O	O
insulin-like	NN	O	B-protein
growth	NN	O	I-protein
factor-1	NN	O	I-protein
,	NN	O	O
thrombopoietin	NN	O	B-protein
,	NN	O	O
interleukin-3	NN	O	B-protein
,	NN	O	O
and	NN	O	O
granulocyte/monocyte	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
the	NN	O	O
molecular	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
this	NN	O	O
disease	NN	O	O
remains	NN	O	O
unknown	NN	O	O
.	NN	O	O

Growth	NN	O	O
factor	NN	O	O
hypersensitivity	NN	O	O
could	NN	O	O
be	NN	O	O
mediated	NN	O	O
by	NN	O	O
changes	NN	O	O
in	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathways	NN	O	O
.	NN	O	O

We	NN	O	O
therefore	NN	O	O
investigated	NN	O	O
a	NN	O	O
common	NN	O	O
downstream	NN	O	O
effector	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
,	NN	O	O
the	NN	O	O
signal	NN	O	B-protein
transducers	NN	O	I-protein
and	NN	O	I-protein
activators	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
(	NN	O	O
STATs	NN	O	B-protein
)	NN	O	O
.	NN	O	O

A	NN	O	O
constitutive	NN	O	O
activation	NN	O	O
of	NN	O	O
STAT	NN	O	B-protein
factors	NN	O	I-protein
could	NN	O	O
explain	NN	O	O
the	NN	O	O
increased	NN	O	O
proliferation	NN	O	O
of	NN	O	O
P.	NN	O	B-cell_type
vera	NN	O	I-cell_type
cells	NN	O	I-cell_type
even	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
growth	NN	O	O
factor	NN	O	O
stimulation	NN	O	O
.	NN	O	O

METHODS	NN	O	O
:	NN	O	O
Peripheral	NN	O	B-cell_type
granulocytes	NN	O	I-cell_type
from	NN	O	O
patients	NN	O	O
with	NN	O	O
P.	NN	O	O
vera	NN	O	O
and	NN	O	O
from	NN	O	O
healthy	NN	O	O
volunteers	NN	O	O
were	NN	O	O
assayed	NN	O	O
for	NN	O	O
STAT1	NN	O	B-protein
,	NN	O	I-protein
3	NN	O	I-protein
,	NN	O	I-protein
and	NN	O	I-protein
5	NN	O	I-protein
DNA	NN	O	O
binding	NN	O	O
by	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
Four	NN	O	O
of	NN	O	O
14	NN	O	O
P.	NN	O	O
vera	NN	O	O
patients	NN	O	O
analyzed	NN	O	O
showed	NN	O	O
constitutive	NN	O	O
STAT3	NN	O	O
DNA	NN	O	O
binding	NN	O	O
in	NN	O	O
unstimulated	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
granulocytes	NN	O	I-cell_type
,	NN	O	O
while	NN	O	O
none	NN	O	O
of	NN	O	O
the	NN	O	O
17	NN	O	O
healthy	NN	O	O
volunteers	NN	O	O
tested	NN	O	O
did	NN	O	O
.	NN	O	O

None	NN	O	O
of	NN	O	O
the	NN	O	O
subjects	NN	O	O
showed	NN	O	O
constitutive	NN	O	O
STAT1	NN	O	B-protein
or	NN	O	O
STAT5	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

Western	NN	O	O
blotting	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
,	NN	O	O
in	NN	O	O
the	NN	O	O
three	NN	O	O
patients	NN	O	O
,	NN	O	O
STAT3	NN	O	B-protein
is	NN	O	O
constitutively	NN	O	O
phosphorylated	NN	O	O
on	NN	O	O
Tyr	NN	O	O
705	NN	O	O
,	NN	O	O
whereas	NN	O	O
it	NN	O	O
is	NN	O	O
unphosphorylated	NN	O	O
in	NN	O	O
the	NN	O	O
other	NN	O	O
patients	NN	O	O
and	NN	O	O
in	NN	O	O
controls	NN	O	O
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
constitutive	NN	O	O
STAT3	NN	O	B-protein
activity	NN	O	O
did	NN	O	O
not	NN	O	O
correlate	NN	O	O
with	NN	O	O
the	NN	O	O
duration	NN	O	O
of	NN	O	O
disease	NN	O	O
or	NN	O	O
the	NN	O	O
treatment	NN	O	O
regimen	NN	O	O
.	NN	O	O

It	NN	O	O
was	NN	O	O
observed	NN	O	O
in	NN	O	O
a	NN	O	O
recently	NN	O	O
diagnosed	NN	O	O
patient	NN	O	O
and	NN	O	O
in	NN	O	O
two	NN	O	O
patients	NN	O	O
treated	NN	O	O
only	NN	O	O
with	NN	O	O
phlebotomy	NN	O	O
.	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
constitutive	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
activation	NN	O	O
of	NN	O	O
STAT3	NN	O	B-protein
is	NN	O	O
not	NN	O	O
a	NN	O	O
secondary	NN	O	O
event	NN	O	O
induced	NN	O	O
by	NN	O	O
mutagenizing	NN	O	O
agents	NN	O	O
or	NN	O	O
by	NN	O	O
prolonged	NN	O	O
hyperproliferation	NN	O	O
of	NN	O	O
hematopoietic	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
but	NN	O	O
rather	NN	O	O
represents	NN	O	O
a	NN	O	O
primary	NN	O	O
molecular	NN	O	O
aberration	NN	O	O
.	NN	O	O

Constitutively	NN	O	B-protein
active	NN	O	I-protein
STAT3	NN	O	I-protein
may	NN	O	O
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
growth	NN	O	O
factor	NN	O	O
hypersensitivity	NN	O	O
of	NN	O	O
P.	NN	O	B-cell_type
vera	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Identification	NN	O	O
of	NN	O	O
phosphorylation	NN	O	B-protein
sites	NN	O	I-protein
for	NN	O	O
Bruton	NN	O	B-protein
's	NN	O	I-protein
tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
within	NN	O	O
the	NN	O	O
transcriptional	NN	O	B-protein
regulator	NN	O	I-protein
BAP/TFII-I	NN	O	I-protein
.	NN	O	O

Bruton	NN	O	B-protein
's	NN	O	I-protein
tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	O
Btk	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
Tec	NN	O	B-protein
family	NN	O	I-protein
of	NN	O	I-protein
cytosolic	NN	O	I-protein
kinases	NN	O	I-protein
,	NN	O	O
is	NN	O	O
essential	NN	O	O
for	NN	O	O
B	NN	O	B-cell_type
cell	NN	O	I-cell_type
development	NN	O	O
and	NN	O	O
function	NN	O	O
.	NN	O	O

BAP/TFII-I	NN	O	B-protein
,	NN	O	O
a	NN	O	O
protein	NN	O	O
implicated	NN	O	O
in	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
,	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
Btk	NN	O	B-protein
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
is	NN	O	O
transiently	NN	O	O
phosphorylated	NN	O	O
on	NN	O	O
tyrosine	NN	O	O
following	NN	O	O
B	NN	O	B-cell_type
cell	NN	O	I-cell_type
receptor	NN	O	O
engagement	NN	O	O
.	NN	O	O

BAP/TFII-I	NN	O	B-protein
is	NN	O	O
a	NN	O	O
substrate	NN	O	O
for	NN	O	O
Btk	NN	O	B-protein
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
is	NN	O	O
hyperphosphorylated	NN	O	O
on	NN	O	O
tyrosine	NN	O	O
upon	NN	O	O
coexpression	NN	O	O
with	NN	O	O
Btk	NN	O	B-protein
in	NN	O	O
mammalian	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
an	NN	O	O
effort	NN	O	O
to	NN	O	O
understand	NN	O	O
the	NN	O	O
physiologic	NN	O	O
consequences	NN	O	O
of	NN	O	O
BAP/TFII-I	NN	O	B-protein
tyrosine	NN	O	O
phosphorylation	NN	O	O
following	NN	O	O
B	NN	O	O
cell	NN	O	O
receptor	NN	O	O
stimulation	NN	O	O
,	NN	O	O
site-directed	NN	O	O
mutagenesis	NN	O	O
and	NN	O	O
phosphopeptide	NN	O	O
mapping	NN	O	O
were	NN	O	O
used	NN	O	O
to	NN	O	O
locate	NN	O	O
the	NN	O	O
predominant	NN	O	O
sites	NN	O	O
of	NN	O	O
BAP/TFII-I	NN	O	B-protein
phosphorylation	NN	O	O
by	NN	O	O
Btk	NN	O	B-protein
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

These	NN	O	O
residues	NN	O	O
,	NN	O	O
Tyr248	NN	O	O
,	NN	O	O
Tyr357	NN	O	O
,	NN	O	O
and	NN	O	O
Tyr462	NN	O	O
,	NN	O	O
were	NN	O	O
also	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
the	NN	O	O
major	NN	O	O
sites	NN	O	O
for	NN	O	O
Btk	NN	O	B-protein
-dependent	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
BAP/TFII-I	NN	O	B-protein
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

Residues	NN	O	O
Tyr357	NN	O	O
and	NN	O	O
Tyr462	NN	O	O
are	NN	O	O
contained	NN	O	O
within	NN	O	O
the	NN	O	O
loop	NN	O	B-protein
regions	NN	O	I-protein
of	NN	O	O
adjacent	NN	O	O
helix-loop-helix-like	NN	O	B-protein
repeats	NN	O	I-protein
within	NN	O	O
BAP/TFII-I	NN	O	B-protein
.	NN	O	O

Mutation	NN	O	O
of	NN	O	O
either	NN	O	O
Tyr248	NN	O	O
,	NN	O	O
Tyr357	NN	O	O
,	NN	O	O
or	NN	O	O
Tyr462	NN	O	O
to	NN	O	O
phenylalanine	NN	O	O
reduced	NN	O	O
transcription	NN	O	O
from	NN	O	O
a	NN	O	O
c-fos	NN	O	B-DNA
promoter	NN	O	I-DNA
relative	NN	O	O
to	NN	O	O
wild-type	NN	O	B-DNA
BAP/TFII-I	NN	O	I-DNA
in	NN	O	O
transfected	NN	O	O
COS-7	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
consistent	NN	O	O
with	NN	O	O
the	NN	O	O
interpretation	NN	O	O
that	NN	O	O
phosphorylation	NN	O	O
at	NN	O	O
these	NN	O	O
sites	NN	O	O
contributes	NN	O	O
to	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
.	NN	O	O

Phosphorylation	NN	O	O
of	NN	O	O
BAP/TFII-I	NN	O	B-protein
by	NN	O	O
Btk	NN	O	B-protein
may	NN	O	O
link	NN	O	O
engagement	NN	O	O
of	NN	O	O
receptors	NN	O	O
such	NN	O	O
as	NN	O	O
surface	NN	O	O
immunoglobulin	NN	O	B-protein
to	NN	O	O
modulation	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

Expression	NN	O	O
of	NN	O	O
interferon	NN	O	B-protein
consensus	NN	O	I-protein
sequence	NN	O	I-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
induces	NN	O	O
potent	NN	O	O
immunity	NN	O	O
against	NN	O	O
BCR/ABL-induced	NN	O	O
leukemia	NN	O	O
.	NN	O	O

Mice	NN	O	O
deficient	NN	O	O
in	NN	O	O
the	NN	O	O
interferon	NN	O	B-protein
consensus	NN	O	I-protein
sequence	NN	O	I-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
ICSBP	NN	O	B-protein
)	NN	O	O
develop	NN	O	O
a	NN	O	O
disease	NN	O	O
resembling	NN	O	O
chronic	NN	O	O
myeloid	NN	O	O
leukemia	NN	O	O
(	NN	O	O
CML	NN	O	O
)	NN	O	O
,	NN	O	O
which	NN	O	O
in	NN	O	O
humans	NN	O	O
is	NN	O	O
caused	NN	O	O
by	NN	O	O
the	NN	O	O
BCR/ABL	NN	O	B-protein
oncoprotein	NN	O	I-protein
.	NN	O	O

Interferon-alpha	NN	O	B-protein
(	NN	O	O
IFN-alpha	NN	O	B-protein
)	NN	O	O
induces	NN	O	O
ICSBP	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
is	NN	O	O
an	NN	O	O
effective	NN	O	O
therapy	NN	O	O
for	NN	O	O
CML	NN	O	O
.	NN	O	O

This	NN	O	O
study	NN	O	O
examined	NN	O	O
whether	NN	O	O
enforced	NN	O	O
expression	NN	O	O
of	NN	O	O
ICSBP	NN	O	B-protein
might	NN	O	O
antagonize	NN	O	O
BCR/ABL-induced	NN	O	O
leukemia	NN	O	O
;	NN	O	O
results	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
ICSBP-modified	NN	O	B-cell_type
cells	NN	O	I-cell_type
generated	NN	O	O
a	NN	O	O
protective	NN	O	O
CD8	NN	O	O
(	NN	O	O
+	NN	O	O
)	NN	O	O
cytotoxic	NN	O	O
T-cell	NN	O	O
response	NN	O	O
against	NN	O	O
BCR/ABL-transformed	NN	O	B-cell_type
BaF3	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
a	NN	O	O
murine	NN	O	O
leukemia	NN	O	O
model	NN	O	O
.	NN	O	O

ICSBP	NN	O	B-protein
expression	NN	O	O
represents	NN	O	O
a	NN	O	O
novel	NN	O	O
means	NN	O	O
of	NN	O	O
stimulating	NN	O	O
a	NN	O	O
host	NN	O	O
immune	NN	O	O
response	NN	O	O
to	NN	O	O
BCR/ABL	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
leukemia	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
a	NN	O	O
potential	NN	O	O
strategy	NN	O	O
for	NN	O	O
immunotherapy	NN	O	O
of	NN	O	O
CML	NN	O	O
.	NN	O	O

(	NN	O	O
Blood.	NN	O	O
2001	NN	O	O
;	NN	O	O
97	NN	O	O
:	NN	O	O
3491-3497	NN	O	O
)	NN	O	O

-DOCSTART-	O

A	NN	O	O
transcriptional	NN	O	O
block	NN	O	O
in	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
at	NN	O	O
the	NN	O	O
-150	NN	O	B-DNA
AP-1	NN	O	I-DNA
site	NN	O	I-DNA
in	NN	O	O
effector	NN	O	B-cell_type
CD8+	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Both	NN	O	O
CD4+	NN	O	B-cell_type
and	NN	O	I-cell_type
CD8+	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
that	NN	O	O
produce	NN	O	O
IL-2	NN	O	B-protein
in	NN	O	O
response	NN	O	O
to	NN	O	O
Ag	NN	O	O
recognition	NN	O	O
have	NN	O	O
been	NN	O	O
isolated	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
most	NN	O	O
effector	NN	O	B-cell_type
CD8+	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
recovered	NN	O	O
after	NN	O	O
exposure	NN	O	O
to	NN	O	O
Ag	NN	O	B-protein
do	NN	O	O
not	NN	O	O
produce	NN	O	O
sufficient	NN	O	O
IL-2	NN	O	B-protein
to	NN	O	O
sustain	NN	O	O
growth	NN	O	O
,	NN	O	O
and	NN	O	O
depend	NN	O	O
on	NN	O	O
CD4+	NN	O	B-cell_type
T	NN	O	I-cell_type
helper	NN	O	I-cell_type
cells	NN	O	I-cell_type
for	NN	O	O
this	NN	O	O
obligate	NN	O	O
growth	NN	O	O
factor	NN	O	O
.	NN	O	O

IL-2	NN	O	O
expression	NN	O	O
in	NN	O	O
CD4+	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
primarily	NN	O	O
controlled	NN	O	O
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
transcription	NN	O	O
,	NN	O	O
but	NN	O	O
mechanisms	NN	O	O
restricting	NN	O	O
IL-2	NN	O	O
production	NN	O	O
in	NN	O	O
CD8+	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
have	NN	O	O
not	NN	O	O
been	NN	O	O
elucidated	NN	O	O
.	NN	O	O

To	NN	O	O
evaluate	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
CD8+	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
we	NN	O	O
stably	NN	O	O
transfected	NN	O	O
reporter	NN	O	B-DNA
genes	NN	O	I-DNA
into	NN	O	O
Ag	NN	O	B-protein
-specific	NN	O	O
CD8+	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
clones	NN	O	I-cell_line
.	NN	O	O

CD28+	NN	O	B-cell_type
CD8	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
unable	NN	O	O
to	NN	O	O
transcribe	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
response	NN	O	O
to	NN	O	O
antigenic	NN	O	O
stimulation	NN	O	O
had	NN	O	O
a	NN	O	O
block	NN	O	O
in	NN	O	O
transactivation	NN	O	O
of	NN	O	O
the	NN	O	O
-150	NN	O	B-DNA
CD28	NN	O	I-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	I-DNA
CD28RE	NN	O	I-DNA
)	NN	O	I-DNA
/AP-1	NN	O	I-DNA
site	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
,	NN	O	O
but	NN	O	O
did	NN	O	O
transactivate	NN	O	O
the	NN	O	O
composite	NN	O	O
NFAT/AP-1	NN	O	B-DNA
and	NN	O	I-DNA
OCT/AP-1	NN	O	I-DNA
sites	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
a	NN	O	O
consensus	NN	O	B-DNA
AP-1	NN	O	I-DNA
motif	NN	O	I-DNA
.	NN	O	O

Mutation	NN	O	O
of	NN	O	O
the	NN	O	O
nonconsensus	NN	O	B-DNA
-150	NN	O	I-DNA
AP-1	NN	O	I-DNA
site	NN	O	I-DNA
to	NN	O	O
a	NN	O	O
consensus	NN	O	B-DNA
AP-1	NN	O	I-DNA
site	NN	O	I-DNA
,	NN	O	O
or	NN	O	O
insertion	NN	O	O
of	NN	O	O
a	NN	O	O
CD28RE/AP-1	NN	O	B-DNA
consensus	NN	O	I-DNA
site	NN	O	I-DNA
upstream	NN	O	O
of	NN	O	O
the	NN	O	O
native	NN	O	O
-150	NN	O	B-DNA
CD28RE/AP-1	NN	O	I-DNA
site	NN	O	I-DNA
restored	NN	O	O
transactivation	NN	O	O
of	NN	O	O
the	NN	O	O
altered	NN	O	O
promoter	NN	O	B-DNA
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
defect	NN	O	O
at	NN	O	O
the	NN	O	O
-150	NN	O	B-DNA
site	NN	O	I-DNA
may	NN	O	O
reflect	NN	O	O
the	NN	O	O
absence	NN	O	O
or	NN	O	O
inactivity	NN	O	O
of	NN	O	O
a	NN	O	O
required	NN	O	O
factor	NN	O	O
rather	NN	O	O
than	NN	O	O
repression	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Stem	NN	O	B-protein
cell	NN	O	I-protein
factor	NN	O	I-protein
and	NN	O	O
interleukin-3	NN	O	B-protein
induce	NN	O	O
stepwise	NN	O	O
generation	NN	O	O
of	NN	O	O
erythroid	NN	O	B-cell_type
precursor	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
a	NN	O	O
basic	NN	O	O
fibroblast	NN	O	B-protein
growth	NN	O	I-protein
factor	NN	O	I-protein
-dependent	NN	O	O
hematopoietic	NN	O	B-cell_line
stem	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	I-cell_line
A-6	NN	O	I-cell_line
.	NN	O	O

A	NN	O	O
m	NN	O	O
ultipotent	NN	O	B-cell_type
immature	NN	O	I-cell_type
myeloid	NN	O	I-cell_type
cell	NN	O	I-cell_type
population	NN	O	I-cell_type
was	NN	O	O
produced	NN	O	O
from	NN	O	O
a	NN	O	O
basic	NN	O	B-protein
fibroblast	NN	O	I-protein
growth	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
bFGF	NN	O	B-protein
)	NN	O	O
-dependent	NN	O	O
hematopoietic	NN	O	B-cell_line
stem	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	I-cell_line
A-6	NN	O	I-cell_line
,	NN	O	O
when	NN	O	O
cultured	NN	O	O
with	NN	O	O
stem	NN	O	B-protein
cell	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
SCF	NN	O	B-protein
)	NN	O	O
replacing	NN	O	O
bFGF	NN	O	B-protein
.	NN	O	O

Those	NN	O	O
cells	NN	O	O
were	NN	O	O
positive	NN	O	O
for	NN	O	O
stem	NN	O	B-protein
cell	NN	O	I-protein
markers	NN	O	I-protein
,	NN	O	O
c-kit	NN	O	B-protein
and	NN	O	O
CD34	NN	O	B-protein
,	NN	O	O
and	NN	O	O
a	NN	O	O
myeloid	NN	O	B-protein
cell	NN	O	I-protein
marker	NN	O	I-protein
,	NN	O	I-protein
F4/80	NN	O	I-protein
.	NN	O	O

Some	NN	O	O
cell	NN	O	O
fractions	NN	O	O
were	NN	O	O
also	NN	O	O
positive	NN	O	O
for	NN	O	O
Mac-1	NN	O	B-protein
,	NN	O	I-protein
a	NN	O	I-protein
macrophage	NN	O	I-protein
marker	NN	O	I-protein
or	NN	O	O
Gr-1	NN	O	B-protein
,	NN	O	I-protein
a	NN	O	I-protein
granulocytic	NN	O	I-protein
maker	NN	O	I-protein
,	NN	O	O
but	NN	O	O
negative	NN	O	O
for	NN	O	O
an	NN	O	O
erythroid	NN	O	B-protein
marker	NN	O	I-protein
TER119	NN	O	I-protein
.	NN	O	O

They	NN	O	O
also	NN	O	O
showed	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
mRNA	NN	O	O
for	NN	O	O
the	NN	O	O
myeloid-specific	NN	O	B-protein
PU.1	NN	O	I-protein
but	NN	O	O
did	NN	O	O
not	NN	O	O
that	NN	O	O
for	NN	O	O
the	NN	O	O
erythroid-specific	NN	O	B-protein
GATA-1	NN	O	I-protein
.	NN	O	O

Among	NN	O	O
various	NN	O	O
cytokines	NN	O	B-protein
,	NN	O	O
interleukin-3	NN	O	B-protein
(	NN	O	O
IL-3	NN	O	B-protein
)	NN	O	O
induced	NN	O	O
erythroid	NN	O	B-cell_type
precursor	NN	O	I-cell_type
cells	NN	O	I-cell_type
that	NN	O	O
expressed	NN	O	O
the	NN	O	O
erythroid-specific	NN	O	B-protein
GATA-1	NN	O	I-protein
and	NN	O	O
beta-major	NN	O	B-protein
globin	NN	O	I-protein
.	NN	O	O

The	NN	O	O
quantitative	NN	O	O
analysis	NN	O	O
showed	NN	O	O
that	NN	O	O
erythroid	NN	O	B-cell_type
precursor	NN	O	I-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
newly	NN	O	O
produced	NN	O	O
from	NN	O	O
the	NN	O	O
immature	NN	O	B-cell_type
myeloid	NN	O	I-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
cultivation	NN	O	O
with	NN	O	O
IL-3	NN	O	B-protein
.	NN	O	O

SCF	NN	O	B-protein
and	NN	O	O
IL-3	NN	O	B-protein
induced	NN	O	O
stepwise	NN	O	O
generation	NN	O	O
of	NN	O	O
erythroid	NN	O	B-cell_type
precursor	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
an	NN	O	O
A-6	NN	O	B-cell_line
hematopoietic	NN	O	I-cell_line
stem	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

Copyright	NN	O	O
2001	NN	O	O
Academic	NN	O	O
Press	NN	O	O
.	NN	O	O

-DOCSTART-	O

Distinct	NN	O	O
BMI-1	NN	O	B-protein
and	NN	O	O
EZH2	NN	O	B-protein
expression	NN	O	O
patterns	NN	O	O
in	NN	O	O
thymocytes	NN	O	B-cell_type
and	NN	O	O
mature	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
suggest	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
Polycomb	NN	O	B-DNA
genes	NN	O	I-DNA
in	NN	O	O
human	NN	O	O
T	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
.	NN	O	O

BMI-1	NN	O	B-protein
and	NN	O	O
EZH2	NN	O	B-protein
Polycomb-group	NN	O	B-protein
(	NN	O	I-protein
PcG	NN	O	I-protein
)	NN	O	I-protein
proteins	NN	O	I-protein
belong	NN	O	O
to	NN	O	O
two	NN	O	O
distinct	NN	O	O
protein	NN	O	O
complexes	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
hematopoiesis	NN	O	O
.	NN	O	O

Using	NN	O	O
unique	NN	O	O
PcG-specific	NN	O	O
antisera	NN	O	O
and	NN	O	O
triple	NN	O	O
immunofluorescence	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
mature	NN	O	B-cell_type
resting	NN	O	I-cell_type
peripheral	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
expressed	NN	O	O
BMI-1	NN	O	B-protein
,	NN	O	O
whereas	NN	O	O
dividing	NN	O	B-cell_type
blasts	NN	O	I-cell_type
were	NN	O	O
EZH2	NN	O	O
(	NN	O	O
+	NN	O	O
)	NN	O	O
.	NN	O	O

By	NN	O	O
contrast	NN	O	O
,	NN	O	O
subcapsular	NN	O	B-cell_type
immature	NN	O	I-cell_type
double-negative	NN	O	I-cell_type
(	NN	O	I-cell_type
DN	NN	O	I-cell_type
)	NN	O	I-cell_type
(	NN	O	I-cell_type
CD4	NN	O	I-cell_type
(	NN	O	I-cell_type
-	NN	O	I-cell_type
)	NN	O	I-cell_type
/CD8	NN	O	I-cell_type
(	NN	O	I-cell_type
-	NN	O	I-cell_type
)	NN	O	I-cell_type
)	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
thymus	NN	O	O
coexpressed	NN	O	O
BMI-1	NN	O	B-protein
and	NN	O	O
EZH2	NN	O	B-protein
or	NN	O	O
were	NN	O	O
BMI-1	NN	O	O
single	NN	O	O
positive	NN	O	O
.	NN	O	O

Their	NN	O	O
descendants	NN	O	O
,	NN	O	O
double-positive	NN	O	B-cell_type
(	NN	O	I-cell_type
DP	NN	O	I-cell_type
;	NN	O	I-cell_type
CD4	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
/CD8	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
)	NN	O	I-cell_type
cortical	NN	O	I-cell_type
thymocytes	NN	O	I-cell_type
,	NN	O	O
expressed	NN	O	O
EZH2	NN	O	B-protein
without	NN	O	O
BMI-1	NN	O	B-protein
.	NN	O	O

Most	NN	O	O
EZH2	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
DN	NN	O	I-cell_type
and	NN	O	I-cell_type
DP	NN	O	I-cell_type
thymocytes	NN	O	I-cell_type
were	NN	O	O
dividing	NN	O	O
,	NN	O	O
while	NN	O	O
DN	NN	O	B-cell_type
BMI-1	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
/EZH2	NN	O	I-cell_type
(	NN	O	I-cell_type
-	NN	O	I-cell_type
)	NN	O	I-cell_type
thymocytes	NN	O	I-cell_type
were	NN	O	O
resting	NN	O	O
and	NN	O	O
proliferation	NN	O	O
was	NN	O	O
occasionally	NN	O	O
noted	NN	O	O
in	NN	O	O
DN	NN	O	B-cell_type
BMI-1	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
/EZH2	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Maturation	NN	O	O
of	NN	O	O
DP	NN	O	B-cell_type
cortical	NN	O	I-cell_type
thymocytes	NN	O	I-cell_type
to	NN	O	O
single-positive	NN	O	B-cell_type
(	NN	O	I-cell_type
CD4	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
/CD8	NN	O	I-cell_type
(	NN	O	I-cell_type
-	NN	O	I-cell_type
)	NN	O	I-cell_type
or	NN	O	O
CD8	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
/CD4	NN	O	I-cell_type
(	NN	O	I-cell_type
-	NN	O	I-cell_type
)	NN	O	I-cell_type
)	NN	O	I-cell_type
medullar	NN	O	I-cell_type
thymocytes	NN	O	I-cell_type
correlated	NN	O	O
with	NN	O	O
decreased	NN	O	O
detectability	NN	O	O
of	NN	O	O
EZH2	NN	O	B-protein
and	NN	O	O
continued	NN	O	O
relative	NN	O	O
absence	NN	O	O
of	NN	O	O
BMI-1	NN	O	B-protein
.	NN	O	O

Our	NN	O	O
data	NN	O	O
show	NN	O	O
that	NN	O	O
BMI-1	NN	O	B-protein
and	NN	O	O
EZH2	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
mature	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
mutually	NN	O	O
exclusive	NN	O	O
and	NN	O	O
linked	NN	O	O
to	NN	O	O
proliferation	NN	O	O
status	NN	O	O
,	NN	O	O
and	NN	O	O
that	NN	O	O
this	NN	O	O
pattern	NN	O	O
is	NN	O	O
not	NN	O	O
yet	NN	O	O
established	NN	O	O
in	NN	O	O
thymocytes	NN	O	B-cell_type
of	NN	O	O
the	NN	O	O
cortex	NN	O	O
and	NN	O	O
medulla	NN	O	O
.	NN	O	O

T	NN	O	B-cell_type
cell	NN	O	I-cell_type
stage-specific	NN	O	O
PcG	NN	O	O
expression	NN	O	O
profiles	NN	O	O
suggest	NN	O	O
that	NN	O	O
PcG	NN	O	B-DNA
genes	NN	O	I-DNA
contribute	NN	O	O
to	NN	O	O
regulation	NN	O	O
of	NN	O	O
T	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
.	NN	O	O

They	NN	O	O
probably	NN	O	O
reflect	NN	O	O
stabilization	NN	O	O
of	NN	O	O
cell	NN	O	O
type-specific	NN	O	O
gene	NN	O	O
expression	NN	O	O
and	NN	O	O
irreversibility	NN	O	O
of	NN	O	O
lineage	NN	O	O
choice	NN	O	O
.	NN	O	O

The	NN	O	O
difference	NN	O	O
in	NN	O	O
PcG	NN	O	O
expression	NN	O	O
between	NN	O	O
medullar	NN	O	B-cell_type
thymocytes	NN	O	I-cell_type
and	NN	O	O
mature	NN	O	B-cell_type
interfollicular	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
indicates	NN	O	O
that	NN	O	O
additional	NN	O	O
maturation	NN	O	O
processes	NN	O	O
occur	NN	O	O
after	NN	O	O
thymocyte	NN	O	O
transportation	NN	O	O
from	NN	O	O
the	NN	O	O
thymus	NN	O	O
.	NN	O	O

-DOCSTART-	O

Stepwise	NN	O	O
lineage	NN	O	O
restriction	NN	O	O
of	NN	O	O
progenitors	NN	O	B-cell_type
in	NN	O	O
lympho-myelopoiesis	NN	O	O
.	NN	O	O

It	NN	O	O
has	NN	O	O
long	NN	O	O
been	NN	O	O
controversial	NN	O	O
whether	NN	O	O
hematopoiesis	NN	O	O
progresses	NN	O	O
through	NN	O	O
ordered	NN	O	O
stages	NN	O	O
of	NN	O	O
determination	NN	O	O
as	NN	O	O
in	NN	O	O
embryonic	NN	O	O
development	NN	O	O
.	NN	O	O

This	NN	O	O
is	NN	O	O
due	NN	O	O
to	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
a	NN	O	O
methodology	NN	O	O
capable	NN	O	O
of	NN	O	O
exactly	NN	O	O
determining	NN	O	O
the	NN	O	O
developmental	NN	O	O
potential	NN	O	O
of	NN	O	O
hematopoietic	NN	O	B-cell_type
stem/progenitor	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
multilineage	NN	O	O
progenitor	NN	O	O
(	NN	O	O
MLP	NN	O	O
)	NN	O	O
assay	NN	O	O
enabled	NN	O	O
us	NN	O	O
to	NN	O	O
discriminate	NN	O	O
among	NN	O	O
seven	NN	O	O
types	NN	O	O
of	NN	O	O
hematopoietic	NN	O	O
progenitors	NN	O	O
,	NN	O	O
which	NN	O	O
are	NN	O	O
multipotent	NN	O	O
progenitor	NN	O	O
p-MTB	NN	O	O
(	NN	O	O
capable	NN	O	O
of	NN	O	O
generating	NN	O	O
myeloid	NN	O	B-cell_type
,	NN	O	I-cell_type
T	NN	O	I-cell_type
and	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
)	NN	O	O
,	NN	O	O
bipotent	NN	O	O
progenitors	NN	O	O
p-MT	NN	O	O
,	NN	O	O
p-MB	NN	O	O
and	NN	O	O
p-TB	NN	O	O
,	NN	O	O
and	NN	O	O
unipotent	NN	O	O
progenitors	NN	O	O
p-M	NN	O	O
,	NN	O	O
p-T	NN	O	O
and	NN	O	O
p-B	NN	O	O
.	NN	O	O

Among	NN	O	O
these	NN	O	O
seven	NN	O	O
types	NN	O	O
,	NN	O	O
the	NN	O	O
p-TB	NN	O	O
type	NN	O	O
progenitor	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
absent	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
process	NN	O	O
of	NN	O	O
lineage	NN	O	O
commitment	NN	O	O
proceeds	NN	O	O
through	NN	O	O
an	NN	O	O
ordered	NN	O	O
but	NN	O	O
not	NN	O	O
random	NN	O	O
process	NN	O	O
.	NN	O	O

By	NN	O	O
extending	NN	O	O
the	NN	O	O
area	NN	O	O
of	NN	O	O
investigation	NN	O	O
to	NN	O	O
include	NN	O	O
the	NN	O	O
erythroid	NN	O	B-cell_type
lineage	NN	O	I-cell_type
,	NN	O	O
more	NN	O	O
convincing	NN	O	O
evidence	NN	O	O
for	NN	O	O
the	NN	O	O
ordered	NN	O	O
process	NN	O	O
was	NN	O	O
obtained	NN	O	O
.	NN	O	O

Detailed	NN	O	O
and	NN	O	O
exact	NN	O	O
illustration	NN	O	O
of	NN	O	O
the	NN	O	O
process	NN	O	O
of	NN	O	O
hematopoiesis	NN	O	O
will	NN	O	O
provide	NN	O	O
an	NN	O	O
opportunity	NN	O	O
to	NN	O	O
revive	NN	O	O
hematopoiesis	NN	O	O
as	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
most	NN	O	O
fascinating	NN	O	O
targets	NN	O	O
of	NN	O	O
research	NN	O	O
in	NN	O	O
developmental	NN	O	O
biology	NN	O	O

-DOCSTART-	O

Epstein-Barr	NN	O	O
Virus	NN	O	O
and	NN	O	O
its	NN	O	O
glycoprotein-350	NN	O	B-protein
upregulate	NN	O	O
IL-6	NN	O	B-protein
in	NN	O	O
human	NN	O	B-cell_type
B-lymphocytes	NN	O	I-cell_type
via	NN	O	O
CD21	NN	O	B-protein
,	NN	O	O
involving	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
and	NN	O	O
different	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
.	NN	O	O

Epstein-Barr	NN	O	O
virus	NN	O	O
(	NN	O	O
EBV	NN	O	O
)	NN	O	O
is	NN	O	O
a	NN	O	O
ubiquitous	NN	O	O
and	NN	O	O
highly	NN	O	O
immunotropic	NN	O	O
gamma	NN	O	O
herpesvirus	NN	O	O
that	NN	O	O
infects	NN	O	O
more	NN	O	O
than	NN	O	O
90	NN	O	O
%	NN	O	O
of	NN	O	O
humans	NN	O	O
worldwide	NN	O	O
.	NN	O	O

Its	NN	O	O
pathogenicity	NN	O	O
leads	NN	O	O
to	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
diseases	NN	O	O
including	NN	O	O
tumors	NN	O	O
that	NN	O	O
result	NN	O	O
from	NN	O	O
EBV	NN	O	O
's	NN	O	O
ability	NN	O	O
to	NN	O	O
readily	NN	O	O
transform	NN	O	O
B-lymphocytes	NN	O	B-cell_type
and	NN	O	O
,	NN	O	O
to	NN	O	O
a	NN	O	O
lesser	NN	O	O
extent	NN	O	O
,	NN	O	O
epithelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

EBV	NN	O	O
utilizes	NN	O	O
CD21/CR2	NN	O	B-protein
as	NN	O	O
its	NN	O	O
receptor	NN	O	O
on	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
initiate	NN	O	O
the	NN	O	O
infection	NN	O	O
process	NN	O	O
.	NN	O	O

EBV	NN	O	O
binds	NN	O	O
to	NN	O	O
CR2	NN	O	B-protein
through	NN	O	O
its	NN	O	O
major	NN	O	B-protein
envelope	NN	O	I-protein
glycoprotein-350	NN	O	I-protein
(	NN	O	O
gp350	NN	O	B-protein
)	NN	O	O
and	NN	O	O
is	NN	O	O
also	NN	O	O
a	NN	O	O
remarkable	NN	O	O
immunomodulating	NN	O	O
agent	NN	O	O
.	NN	O	O

We	NN	O	O
had	NN	O	O
previously	NN	O	O
shown	NN	O	O
that	NN	O	O
EBV	NN	O	O
is	NN	O	O
capable	NN	O	O
of	NN	O	O
modulating	NN	O	O
the	NN	O	O
synthesis	NN	O	O
of	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
.	NN	O	O

We	NN	O	O
now	NN	O	O
show	NN	O	O
that	NN	O	O
while	NN	O	O
both	NN	O	O
purified	NN	O	O
recombinant	NN	O	B-protein
gp350	NN	O	I-protein
(	NN	O	O
rgp350	NN	O	B-protein
)	NN	O	O
and	NN	O	O
EBV	NN	O	O
upregulate	NN	O	O
IL-6	NN	O	B-RNA
mRNA	NN	O	I-RNA
synthesis	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
EBV-induced	NN	O	O
IL-6	NN	O	O
gene	NN	O	O
activation	NN	O	O
occurs	NN	O	O
for	NN	O	O
a	NN	O	O
significantly	NN	O	O
longer	NN	O	O
period	NN	O	O
of	NN	O	O
time	NN	O	O
(	NN	O	O
i.e.	NN	O	O
12	NN	O	O
hours	NN	O	O
for	NN	O	O
EBV	NN	O	O
as	NN	O	O
compared	NN	O	O
to	NN	O	O
6	NN	O	O
hours	NN	O	O
for	NN	O	O
rgp350	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
the	NN	O	O
half-life	NN	O	O
of	NN	O	O
EBV-induced	NN	O	O
IL-6	NN	O	B-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
also	NN	O	O
significantly	NN	O	O
longer	NN	O	O
(	NN	O	O
10	NN	O	O
hours	NN	O	O
)	NN	O	O
than	NN	O	O
that	NN	O	O
of	NN	O	O
mRNA	NN	O	O
induced	NN	O	O
by	NN	O	O
rgp350	NN	O	B-protein
(	NN	O	O
about	NN	O	O
6	NN	O	O
hours	NN	O	O
)	NN	O	O
.	NN	O	O

Both	NN	O	O
EBV	NN	O	O
and	NN	O	O
gp350	NN	O	B-protein
enhance	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
as	NN	O	O
determined	NN	O	O
by	NN	O	O
band-shift	NN	O	O
and	NN	O	O
augment	NN	O	O
NF-kappaB	NN	O	B-protein
-mediated	NN	O	O
activation	NN	O	O
of	NN	O	O
a	NN	O	O
CAT	NN	O	B-DNA
reporter	NN	O	I-DNA
plasmid	NN	O	I-DNA
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
while	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
IL-6	NN	O	O
gene	NN	O	O
expression	NN	O	O
by	NN	O	O
gp350	NN	O	B-protein
is	NN	O	O
mediated	NN	O	O
primarily	NN	O	O
by	NN	O	O
the	NN	O	O
protein	NN	O	O
kinase	NN	O	O
C	NN	O	O
pathway	NN	O	O
,	NN	O	O
EBV	NN	O	O
can	NN	O	O
mediate	NN	O	O
its	NN	O	O
effects	NN	O	O
through	NN	O	O
multiple	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
.	NN	O	O

To	NN	O	O
our	NN	O	O
knowledge	NN	O	O
this	NN	O	O
is	NN	O	O
the	NN	O	O
first	NN	O	O
report	NN	O	O
showing	NN	O	O
that	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
a	NN	O	O
herpesvirus	NN	O	B-protein
envelope	NN	O	I-protein
glycoprotein	NN	O	I-protein
to	NN	O	O
CR2	NN	O	B-protein
on	NN	O	O
human	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
results	NN	O	O
in	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
leading	NN	O	O
to	NN	O	O
the	NN	O	O
upregulation	NN	O	O
of	NN	O	O
IL-6	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
these	NN	O	O
lymphocytes	NN	O	B-cell_type
.	NN	O	O

Copyright	NN	O	O
2001	NN	O	O
Academic	NN	O	O
Press	NN	O	O
.	NN	O	O

-DOCSTART-	O

Gene-	NN	O	O
and	NN	O	O
tissue-specificity	NN	O	O
of	NN	O	O
mutation	NN	O	O
in	NN	O	O
Big	NN	O	O
Blue	NN	O	O
rats	NN	O	O
treated	NN	O	O
with	NN	O	O
the	NN	O	O
hepatocarcinogen	NN	O	O
N-hydroxy-2-acetylaminofluorene	NN	O	O
.	NN	O	O

In	NN	O	O
a	NN	O	O
previous	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
treating	NN	O	O
transgenic	NN	O	O
Big	NN	O	O
Blue	NN	O	O
rats	NN	O	O
with	NN	O	O
the	NN	O	O
hepatocarcinogen	NN	O	O
N-hydroxy-2-acetylaminofluorene	NN	O	O
(	NN	O	O
N-OH-AAF	NN	O	O
)	NN	O	O
produced	NN	O	O
the	NN	O	O
same	NN	O	O
major	NN	O	O
DNA	NN	O	O
adduct	NN	O	O
in	NN	O	O
the	NN	O	O
target	NN	O	O
liver	NN	O	O
and	NN	O	O
the	NN	O	O
nontarget	NN	O	B-cell_type
spleen	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
bone	NN	O	B-cell_type
marrow	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
induced	NN	O	O
lacI	NN	O	B-cell_type
mutants	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
liver	NN	O	O
,	NN	O	O
and	NN	O	O
induced	NN	O	O
much	NN	O	O
lower	NN	O	O
frequencies	NN	O	O
of	NN	O	O
l	NN	O	O
acI	NN	O	B-cell_type
and	NN	O	I-cell_type
hprt	NN	O	I-cell_type
mutants	NN	O	I-cell_type
in	NN	O	O
spleen	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
,	NN	O	O
sequence	NN	O	O
analysis	NN	O	O
was	NN	O	O
conducted	NN	O	O
on	NN	O	O
lacI	NN	O	B-DNA
DNA	NN	O	I-DNA
and	NN	O	O
hprt	NN	O	B-DNA
cDNA	NN	O	I-DNA
from	NN	O	O
the	NN	O	O
mutants	NN	O	O
,	NN	O	O
to	NN	O	O
determine	NN	O	O
the	NN	O	O
mutational	NN	O	O
specificity	NN	O	O
of	NN	O	O
N-OH-AAF	NN	O	O
in	NN	O	O
the	NN	O	O
rat	NN	O	O
.	NN	O	O

All	NN	O	O
the	NN	O	O
mutation	NN	O	O
spectra	NN	O	O
from	NN	O	O
N-OH-AAF-treated	NN	O	O
rats	NN	O	O
differed	NN	O	O
significantly	NN	O	O
from	NN	O	O
corresponding	NN	O	O
mutation	NN	O	O
profiles	NN	O	O
from	NN	O	O
untreated	NN	O	O
animals	NN	O	O
(	NN	O	O
P	NN	O	O
=	NN	O	O
0.02	NN	O	O
to	NN	O	O
P	NN	O	O
<	NN	O	O
0.0001	NN	O	O
)	NN	O	O
.	NN	O	O

Although	NN	O	O
there	NN	O	O
were	NN	O	O
similarities	NN	O	O
among	NN	O	O
the	NN	O	O
mutational	NN	O	O
patterns	NN	O	O
derived	NN	O	O
from	NN	O	O
N-OH-AAF-treated	NN	O	O
rats	NN	O	O
(	NN	O	O
e.g.	NN	O	O
,	NN	O	O
G	NN	O	O
:	NN	O	O
C	NN	O	O
--	NN	O	O
>	NN	O	O
T	NN	O	O
:	NN	O	O
A	NN	O	O
transversion	NN	O	O
was	NN	O	O
the	NN	O	O
most	NN	O	O
common	NN	O	O
mutation	NN	O	O
in	NN	O	O
all	NN	O	O
mutation	NN	O	O
sets	NN	O	O
)	NN	O	O
,	NN	O	O
there	NN	O	O
were	NN	O	O
significant	NN	O	O
differences	NN	O	O
in	NN	O	O
the	NN	O	O
patterns	NN	O	O
of	NN	O	O
basepair	NN	O	O
substitution	NN	O	O
and	NN	O	O
frameshift	NN	O	O
mutation	NN	O	O
between	NN	O	O
the	NN	O	O
liver	NN	O	O
and	NN	O	O
spleen	NN	O	B-cell_type
lymphocyte	NN	O	I-cell_type
lacI	NN	O	I-cell_type
mutants	NN	O	I-cell_type
(	NN	O	O
P	NN	O	O
=	NN	O	O
0.02	NN	O	O
)	NN	O	O
and	NN	O	O
between	NN	O	O
the	NN	O	O
spleen	NN	O	B-cell_type
lymphocyte	NN	O	I-cell_type
lacI	NN	O	B-cell_type
and	NN	O	I-cell_type
hprt	NN	O	I-cell_type
mutants	NN	O	I-cell_type
(	NN	O	O
P	NN	O	O
=	NN	O	O
0.04	NN	O	O
)	NN	O	O
.	NN	O	O

Also	NN	O	O
,	NN	O	O
multiplex	NN	O	O
PCR	NN	O	O
analysis	NN	O	O
of	NN	O	O
genomic	NN	O	B-DNA
DNA	NN	O	I-DNA
from	NN	O	O
the	NN	O	O
hprt	NN	O	B-cell_type
mutants	NN	O	I-cell_type
indicated	NN	O	O
that	NN	O	O
12	NN	O	O
%	NN	O	O
of	NN	O	O
mutants	NN	O	O
from	NN	O	O
treated	NN	O	O
rats	NN	O	O
had	NN	O	O
major	NN	O	O
deletions	NN	O	O
in	NN	O	O
the	NN	O	O
hprt	NN	O	B-DNA
gene	NN	O	I-DNA
;	NN	O	O
no	NN	O	O
corresponding	NN	O	O
incidence	NN	O	O
of	NN	O	O
large	NN	O	O
deletions	NN	O	O
was	NN	O	O
evident	NN	O	O
among	NN	O	O
lacI	NN	O	O
mutations	NN	O	O
.	NN	O	O

All	NN	O	O
the	NN	O	O
mutation	NN	O	O
profiles	NN	O	O
reflect	NN	O	O
the	NN	O	O
general	NN	O	O
mutational	NN	O	O
specificity	NN	O	O
of	NN	O	O
the	NN	O	O
major	NN	O	O
DNA	NN	O	O
adduct	NN	O	O
formed	NN	O	O
by	NN	O	O
N-OH-AAF	NN	O	O
.	NN	O	O

The	NN	O	O
differences	NN	O	O
between	NN	O	O
N-OH-AAF	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
endogenous	NN	O	B-DNA
gene	NN	O	I-DNA
and	NN	O	O
transgene	NN	O	B-DNA
can	NN	O	O
be	NN	O	O
partially	NN	O	O
explained	NN	O	O
by	NN	O	O
the	NN	O	O
structures	NN	O	O
of	NN	O	O
the	NN	O	O
two	NN	O	O
genes	NN	O	O
.	NN	O	O

The	NN	O	O
tissue-specificity	NN	O	O
of	NN	O	O
the	NN	O	O
mutation	NN	O	O
spectra	NN	O	O
may	NN	O	O
contribute	NN	O	O
to	NN	O	O
targeting	NN	O	O
tumor	NN	O	O
formation	NN	O	O
to	NN	O	O
the	NN	O	O
liver	NN	O	O
.	NN	O	O

Environ.	NN	O	O
Mol.	NN	O	O
Mutagen.	NN	O	O
37	NN	O	O
:	NN	O	O
203-214	NN	O	O
,	NN	O	O
2001	NN	O	O
.	NN	O	O

Published	NN	O	O
2001	NN	O	O
Wiley-Liss	NN	O	O
,	NN	O	O
Inc	NN	O	O
.	NN	O	O

-DOCSTART-	O

Caspase	NN	O	B-protein
-dependent	NN	O	O
cleavage	NN	O	O
of	NN	O	O
the	NN	O	O
hematopoietic	NN	O	B-protein
specific	NN	O	I-protein
adaptor	NN	O	I-protein
protein	NN	O	I-protein
Gads	NN	O	I-protein
alters	NN	O	O
signalling	NN	O	O
from	NN	O	O
the	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
receptor	NN	O	I-protein
.	NN	O	O

Gads	NN	O	B-protein
is	NN	O	O
a	NN	O	O
SH2	NN	O	B-protein
and	NN	O	I-protein
SH3	NN	O	I-protein
domain	NN	O	I-protein
-containing	NN	O	O
,	NN	O	O
hematopoietic-specific	NN	O	B-protein
adaptor	NN	O	I-protein
protein	NN	O	I-protein
that	NN	O	O
functions	NN	O	O
in	NN	O	O
signalling	NN	O	O
from	NN	O	O
the	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
receptor	NN	O	I-protein
.	NN	O	O

Gads	NN	O	B-protein
acts	NN	O	O
by	NN	O	O
linking	NN	O	O
SLP-76	NN	O	B-protein
,	NN	O	O
bound	NN	O	O
by	NN	O	O
the	NN	O	O
carboxy-terminal	NN	O	B-protein
Gads	NN	O	I-protein
SH3	NN	O	I-protein
domain	NN	O	I-protein
,	NN	O	O
to	NN	O	O
tyrosine	NN	O	B-protein
phosphorylated	NN	O	I-protein
LAT	NN	O	I-protein
which	NN	O	O
contains	NN	O	O
binding	NN	O	O
sites	NN	O	O
for	NN	O	O
the	NN	O	O
Gads	NN	O	B-protein
SH2	NN	O	I-protein
domain	NN	O	I-protein
.	NN	O	O

Gads	NN	O	B-protein
is	NN	O	O
distinguished	NN	O	O
from	NN	O	O
Grb2	NN	O	B-protein
and	NN	O	O
the	NN	O	O
closely	NN	O	O
related	NN	O	O
Grap	NN	O	B-protein
protein	NN	O	I-protein
by	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
a	NN	O	O
120	NN	O	B-protein
amino	NN	O	I-protein
acid	NN	O	I-protein
unique	NN	O	I-protein
region	NN	O	I-protein
between	NN	O	O
the	NN	O	O
SH2	NN	O	B-protein
domain	NN	O	I-protein
and	NN	O	O
the	NN	O	O
carboxy	NN	O	B-protein
terminal	NN	O	I-protein
SH3	NN	O	I-protein
domain	NN	O	I-protein
.	NN	O	O

Here	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
unique	NN	O	O
region	NN	O	O
of	NN	O	O
Gads	NN	O	B-protein
contains	NN	O	O
a	NN	O	O
capase	NN	O	B-protein
cleavage	NN	O	I-protein
site	NN	O	I-protein
.	NN	O	O

Induction	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
results	NN	O	O
in	NN	O	O
detectable	NN	O	O
Gads	NN	O	B-protein
cleavage	NN	O	O
by	NN	O	O
60	NN	O	O
min	NN	O	O
.	NN	O	O

Gads	NN	O	B-protein
cleavage	NN	O	O
is	NN	O	O
blocked	NN	O	O
in	NN	O	O
vivo	NN	O	O
by	NN	O	O
treating	NN	O	O
cells	NN	O	O
with	NN	O	O
a	NN	O	O
caspase	NN	O	O
3	NN	O	O
inhibitor	NN	O	O
.	NN	O	O

A	NN	O	O
putative	NN	O	B-protein
caspase	NN	O	I-protein
3	NN	O	I-protein
cleavage	NN	O	I-protein
site	NN	O	I-protein
was	NN	O	O
identified	NN	O	O
within	NN	O	O
the	NN	O	O
unique	NN	O	O
region	NN	O	O
and	NN	O	O
mutation	NN	O	O
of	NN	O	O
this	NN	O	O
site	NN	O	O
prevented	NN	O	O
Gads	NN	O	B-protein
cleavage	NN	O	O
in	NN	O	O
vitro	NN	O	O
,	NN	O	O
and	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

The	NN	O	O
Gads	NN	O	B-protein
cleavage	NN	O	O
products	NN	O	O
retained	NN	O	O
the	NN	O	O
predicted	NN	O	O
binding	NN	O	O
specificity	NN	O	O
for	NN	O	O
SLP-76	NN	O	B-protein
and	NN	O	O
LAT	NN	O	B-protein
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
the	NN	O	O
Gads	NN	O	B-protein
cleavage	NN	O	O
products	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
inhibited	NN	O	O
NFAT	NN	O	O
activation	NN	O	O
following	NN	O	O
TCR	NN	O	O
cross	NN	O	O
linking	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
indicate	NN	O	O
that	NN	O	O
cleavage	NN	O	O
of	NN	O	O
Gads	NN	O	B-protein
in	NN	O	O
vivo	NN	O	O
could	NN	O	O
function	NN	O	O
to	NN	O	O
alter	NN	O	O
signalling	NN	O	O
downstream	NN	O	O
of	NN	O	O
the	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
receptor	NN	O	I-protein
by	NN	O	O
disrupting	NN	O	O
cross	NN	O	O
talk	NN	O	O
between	NN	O	O
SLP-76	NN	O	B-protein
and	NN	O	O
LAT	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Targeting	NN	O	O
of	NN	O	O
p300	NN	O	B-protein
to	NN	O	O
the	NN	O	O
interleukin-2	NN	O	B-DNA
promoter	NN	O	I-DNA
via	NN	O	O
CREB-Rel	NN	O	O
cross-talk	NN	O	O
during	NN	O	O
mitogen	NN	O	B-protein
and	NN	O	O
oncogenic	NN	O	O
molecular	NN	O	O
signaling	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
T-cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
,	NN	O	O
we	NN	O	O
explore	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
targeting	NN	O	O
of	NN	O	O
p300	NN	O	B-protein
to	NN	O	O
the	NN	O	O
interleukin-2	NN	O	B-DNA
(	NN	O	I-DNA
IL-2	NN	O	I-DNA
)	NN	O	I-DNA
promoter	NN	O	I-DNA
in	NN	O	O
response	NN	O	O
to	NN	O	O
mitogenic	NN	O	O
and	NN	O	O
oncogenic	NN	O	O
molecular	NN	O	O
signals	NN	O	O
.	NN	O	O

Recruitment	NN	O	O
of	NN	O	O
p300	NN	O	B-protein
by	NN	O	O
cAMP-responsive	NN	O	B-protein
element-binding	NN	O	I-protein
protein-Rel	NN	O	I-protein
cross-talk	NN	O	O
at	NN	O	O
the	NN	O	O
composite	NN	O	B-protein
CD28	NN	O	I-protein
response	NN	O	I-protein
element	NN	O	I-protein
(	NN	O	O
CD28RE	NN	O	B-protein
)	NN	O	O
-	NN	O	O
TRE	NN	O	B-DNA
element	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
is	NN	O	O
essential	NN	O	O
for	NN	O	O
promoter	NN	O	O
inducibility	NN	O	O
during	NN	O	O
T-cell	NN	O	O
activation	NN	O	O
,	NN	O	O
and	NN	O	O
CD28RE-TRE	NN	O	B-DNA
is	NN	O	O
the	NN	O	O
exclusive	NN	O	O
target	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-cell_type
T-cell	NN	O	I-cell_type
lymphotropic	NN	O	O
virus	NN	O	O
type	NN	O	O
I	NN	O	O
oncoprotein	NN	O	B-protein
Tax	NN	O	I-protein
.	NN	O	O

The	NN	O	O
intrinsic	NN	O	O
histone	NN	O	O
acetyltransferase	NN	O	O
activity	NN	O	O
of	NN	O	O
p300	NN	O	B-protein
is	NN	O	O
dispensable	NN	O	O
for	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
the	NN	O	O
N-terminal	NN	O	B-protein
743	NN	O	I-protein
residues	NN	O	I-protein
contain	NN	O	O
the	NN	O	O
minimal	NN	O	O
structural	NN	O	O
requirements	NN	O	O
for	NN	O	O
synergistic	NN	O	O
transactivation	NN	O	O
of	NN	O	O
the	NN	O	O
CD28RE-TRE	NN	O	B-DNA
,	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
endogenous	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
.	NN	O	O

Mutational	NN	O	O
analysis	NN	O	O
of	NN	O	O
p300	NN	O	B-protein
reveals	NN	O	O
differential	NN	O	O
structural	NN	O	O
requirements	NN	O	O
for	NN	O	O
the	NN	O	O
N-terminal	NN	O	O
p300	NN	O	O
module	NN	O	O
by	NN	O	O
individual	NN	O	O
cis-elements	NN	O	B-DNA
within	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
findings	NN	O	O
provide	NN	O	O
evidence	NN	O	O
that	NN	O	O
p300	NN	O	B-protein
assembles	NN	O	O
at	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
to	NN	O	O
form	NN	O	O
an	NN	O	O
enhanceosome-like	NN	O	O
signal	NN	O	O
transduction	NN	O	O
target	NN	O	O
that	NN	O	O
is	NN	O	O
centrally	NN	O	O
integrated	NN	O	O
at	NN	O	O
the	NN	O	O
CD28RE-TRE	NN	O	B-DNA
element	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
through	NN	O	O
specific	NN	O	O
protein	NN	O	O
module-targeted	NN	O	O
associations	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
T-cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
cytokine	NN	O	O
production	NN	O	O
in	NN	O	O
T-cell	NN	O	B-cell_type
responses	NN	O	O
to	NN	O	O
inhalant	NN	O	O
allergen	NN	O	O
:	NN	O	O
GATA-3	NN	O	B-protein
expression	NN	O	O
distinguishes	NN	O	O
between	NN	O	O
Th1-	NN	O	O
and	NN	O	O
Th2-polarized	NN	O	O
immunity	NN	O	O
.	NN	O	O

BACKGROUND	NN	O	O
:	NN	O	O
The	NN	O	O
precise	NN	O	O
nature	NN	O	O
of	NN	O	O
allergen-specific	NN	O	O
cytokine	NN	O	O
responses	NN	O	O
in	NN	O	O
atopics	NN	O	O
versus	NN	O	O
non-atopics	NN	O	O
,	NN	O	O
in	NN	O	O
particular	NN	O	O
the	NN	O	O
'Th1	NN	O	O
polarity	NN	O	O
'	NN	O	O
of	NN	O	O
responses	NN	O	O
in	NN	O	O
non-atopics	NN	O	O
,	NN	O	O
remains	NN	O	O
controversial	NN	O	O
.	NN	O	O

This	NN	O	O
is	NN	O	O
due	NN	O	O
in	NN	O	O
part	NN	O	O
to	NN	O	O
the	NN	O	O
relative	NN	O	O
insensitivity	NN	O	O
of	NN	O	O
cytokine	NN	O	O
detection	NN	O	O
systems	NN	O	O
,	NN	O	O
and	NN	O	O
associated	NN	O	O
variations	NN	O	O
in	NN	O	O
kinetics	NN	O	O
of	NN	O	O
cytokine	NN	O	O
production	NN	O	O
and	NN	O	O
catabolism	NN	O	O
in	NN	O	O
in	NN	O	O
vitro	NN	O	O
culture	NN	O	O
systems	NN	O	O
.	NN	O	O

As	NN	O	O
an	NN	O	O
alternative	NN	O	O
to	NN	O	O
cytokine	NN	O	O
measurement	NN	O	O
,	NN	O	O
this	NN	O	O
study	NN	O	O
focuses	NN	O	O
on	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
GATA-3	NN	O	I-protein
for	NN	O	O
analysis	NN	O	O
of	NN	O	O
allergen-specific	NN	O	O
Th	NN	O	O
cell	NN	O	O
responses	NN	O	O
.	NN	O	O

METHODS	NN	O	O
:	NN	O	O
Cord	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
Th1-	NN	O	O
or	NN	O	O
Th2-polarized	NN	O	O
by	NN	O	O
culture	NN	O	O
in	NN	O	O
IL-12-	NN	O	O
or	NN	O	O
IL-4-employing	NN	O	O
established	NN	O	O
methods	NN	O	O
;	NN	O	O
PBMC	NN	O	B-cell_type
from	NN	O	O
house	NN	O	O
dust	NN	O	O
mite	NN	O	O
(	NN	O	O
HDM	NN	O	O
)	NN	O	O
-sensitive	NN	O	O
atopics	NN	O	O
and	NN	O	O
controls	NN	O	O
were	NN	O	O
stimulated	NN	O	O
overnight	NN	O	O
with	NN	O	O
HDM	NN	O	O
;	NN	O	O
cytokine	NN	O	O
production	NN	O	O
was	NN	O	O
measured	NN	O	O
by	NN	O	O
ELISA	NN	O	O
and	NN	O	O
GATA-3	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
by	NN	O	O
PCR	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
Cytokine	NN	O	B-protein
-driven	NN	O	O
Th2	NN	O	O
polarization	NN	O	O
of	NN	O	O
naive	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
associated	NN	O	O
with	NN	O	O
marked	NN	O	O
upregulation	NN	O	O
of	NN	O	O
GATA-3	NN	O	B-protein
expression	NN	O	O
,	NN	O	O
whereas	NN	O	O
a	NN	O	O
reciprocal	NN	O	O
expression	NN	O	O
pattern	NN	O	O
accompanies	NN	O	O
differentiation	NN	O	O
towards	NN	O	O
the	NN	O	O
Th1	NN	O	O
cytokine	NN	O	O
phenotype	NN	O	O
.	NN	O	O

In	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
HDM	NN	O	O
skin	NN	O	O
prick	NN	O	O
test-positive	NN	O	O
(	NN	O	O
HDM-SPT+/HDM-IgE+	NN	O	O
)	NN	O	O
volunteers	NN	O	O
,	NN	O	O
overnight	NN	O	O
stimulation	NN	O	O
results	NN	O	O
in	NN	O	O
marked	NN	O	O
upregulation	NN	O	O
of	NN	O	O
GATA-3	NN	O	B-protein
expression	NN	O	O
,	NN	O	O
compared	NN	O	O
to	NN	O	O
an	NN	O	O
equally	NN	O	O
marked	NN	O	O
downregulation	NN	O	O
of	NN	O	O
expression	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
SPT-/IgE-	NN	O	O
subjects	NN	O	O
.	NN	O	O

In	NN	O	O
subjects	NN	O	O
who	NN	O	O
are	NN	O	O
HDM-SPT+	NN	O	O
but	NN	O	O
IgE-	NN	O	O
,	NN	O	O
GATA-3	NN	O	B-protein
expression	NN	O	O
levels	NN	O	O
remained	NN	O	O
relatively	NN	O	O
stable	NN	O	O
during	NN	O	O
culture	NN	O	O
with	NN	O	O
HDM	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
:	NN	O	O
Upregulation	NN	O	O
of	NN	O	O
GATA-3	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
PBMC	NN	O	B-cell_type
is	NN	O	O
a	NN	O	O
hallmark	NN	O	O
of	NN	O	O
the	NN	O	O
early	NN	O	O
phase	NN	O	O
of	NN	O	O
Th2	NN	O	O
recall	NN	O	O
responses	NN	O	O
to	NN	O	O
specific	NN	O	O
allergen	NN	O	O
in	NN	O	O
atopics	NN	O	O
.	NN	O	O

The	NN	O	O
reciprocal	NN	O	O
expression	NN	O	O
pattern	NN	O	O
observed	NN	O	O
in	NN	O	O
HDM-specific	NN	O	O
recall	NN	O	O
responses	NN	O	O
of	NN	O	O
non-atopics	NN	O	O
provides	NN	O	O
independent	NN	O	O
confirmation	NN	O	O
of	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
underlying	NN	O	O
Th1-like	NN	O	O
immunity	NN	O	O
in	NN	O	O
these	NN	O	O
subjects	NN	O	O
.	NN	O	O

The	NN	O	O
parallel	NN	O	O
findings	NN	O	O
in	NN	O	O
neonatal	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
same	NN	O	O
approach	NN	O	O
may	NN	O	O
be	NN	O	O
utilized	NN	O	O
for	NN	O	O
monitoring	NN	O	O
the	NN	O	O
progress	NN	O	O
of	NN	O	O
allergen-specific	NN	O	O
Th1/Th2	NN	O	O
memory	NN	O	O
development	NN	O	O
during	NN	O	O
early	NN	O	O
childhood	NN	O	O
,	NN	O	O
and	NN	O	O
hence	NN	O	O
in	NN	O	O
assessment	NN	O	O
of	NN	O	O
risk	NN	O	O
for	NN	O	O
future	NN	O	O
allergic	NN	O	O
disease	NN	O	O
.	NN	O	O

Copyright	NN	O	O
2001	NN	O	O
S.	NN	O	O
Karger	NN	O	O
AG	NN	O	O
,	NN	O	O
Basel	NN	O	O

-DOCSTART-	O

The	NN	O	O
heat	NN	O	O
shock	NN	O	O
response	NN	O	O
reduces	NN	O	O
myelin	NN	O	B-protein
oligodendrocyte	NN	O	I-protein
glycoprotein	NN	O	I-protein
-induced	NN	O	O
experimental	NN	O	O
autoimmune	NN	O	O
encephalomyelitis	NN	O	O
in	NN	O	O
mice	NN	O	O
.	NN	O	O

The	NN	O	O
stress	NN	O	O
response	NN	O	O
(	NN	O	O
SR	NN	O	O
)	NN	O	O
can	NN	O	O
block	NN	O	O
inflammatory	NN	O	O
gene	NN	O	O
expression	NN	O	O
by	NN	O	O
preventing	NN	O	O
activation	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
NF-kappaB	NN	O	B-protein
)	NN	O	O
.	NN	O	O

As	NN	O	O
inflammatory	NN	O	O
gene	NN	O	O
expression	NN	O	O
contributes	NN	O	O
to	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
demyelinating	NN	O	O
diseases	NN	O	O
,	NN	O	O
we	NN	O	O
tested	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
the	NN	O	O
SR	NN	O	O
on	NN	O	O
the	NN	O	O
progression	NN	O	O
of	NN	O	O
the	NN	O	O
demyelinating	NN	O	O
disease	NN	O	O
experimental	NN	O	O
autoimmune	NN	O	O
encephalomyelitis	NN	O	O
(	NN	O	O
EAE	NN	O	O
)	NN	O	O
.	NN	O	O

EAE	NN	O	O
was	NN	O	O
actively	NN	O	O
induced	NN	O	O
in	NN	O	O
C57BL/6	NN	O	O
mice	NN	O	O
using	NN	O	O
an	NN	O	O
encephalitogenic	NN	O	O
myelin	NN	O	B-protein
oligodendrocyte	NN	O	I-protein
glycoprotein	NN	O	I-protein
(	NN	O	O
MOG	NN	O	B-protein
(	NN	O	I-protein
35-55	NN	O	I-protein
)	NN	O	I-protein
)	NN	O	O
peptide	NN	O	O
.	NN	O	O

Whole	NN	O	O
body	NN	O	O
hyperthermia	NN	O	O
was	NN	O	O
used	NN	O	O
to	NN	O	O
induce	NN	O	O
a	NN	O	O
heat	NN	O	O
shock	NN	O	O
response	NN	O	O
(	NN	O	O
HSR	NN	O	O
)	NN	O	O
in	NN	O	O
immunized	NN	O	O
mice	NN	O	O
2	NN	O	O
days	NN	O	O
after	NN	O	O
the	NN	O	O
booster	NN	O	O
MOG	NN	O	O
(	NN	O	O
35-55	NN	O	O
)	NN	O	O
peptide	NN	O	O
injection	NN	O	O
.	NN	O	O

The	NN	O	O
HSR	NN	O	O
reduced	NN	O	O
the	NN	O	O
incidence	NN	O	O
of	NN	O	O
EAE	NN	O	O
by	NN	O	O
70	NN	O	O
%	NN	O	O
,	NN	O	O
delayed	NN	O	O
disease	NN	O	O
onset	NN	O	O
by	NN	O	O
6	NN	O	O
days	NN	O	O
,	NN	O	O
and	NN	O	O
attenuated	NN	O	O
disease	NN	O	O
severity	NN	O	O
.	NN	O	O

The	NN	O	O
HSR	NN	O	O
attenuated	NN	O	O
leukocyte	NN	O	B-cell_type
infiltration	NN	O	O
into	NN	O	O
CNS	NN	O	O
assessed	NN	O	O
by	NN	O	O
quantitation	NN	O	O
of	NN	O	O
perivascular	NN	O	O
infiltrates	NN	O	O
,	NN	O	O
and	NN	O	O
by	NN	O	O
reduced	NN	O	O
staining	NN	O	O
for	NN	O	O
CD4	NN	O	B-protein
and	NN	O	O
CD25	NN	O	B-protein
immunopositive	NN	O	O
T-cells	NN	O	B-cell_type
.	NN	O	O

T-cell	NN	O	O
activation	NN	O	O
,	NN	O	O
assessed	NN	O	O
by	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
interferon	NN	O	B-protein
gamma	NN	O	I-protein
(	NN	O	O
IFNgamma	NN	O	B-protein
)	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
MOG	NN	O	B-protein
(	NN	O	I-protein
35-55	NN	O	I-protein
)	NN	O	I-protein
,	NN	O	O
was	NN	O	O
also	NN	O	O
decreased	NN	O	O
by	NN	O	O
the	NN	O	O
HSR	NN	O	O
.	NN	O	O

The	NN	O	O
HSR	NN	O	O
reduced	NN	O	O
inflammatory	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
the	NN	O	O
brain	NN	O	O
that	NN	O	O
normally	NN	O	O
occurs	NN	O	O
during	NN	O	O
EAE	NN	O	O
,	NN	O	O
including	NN	O	O
the	NN	O	O
early	NN	O	O
increase	NN	O	O
in	NN	O	O
RANTES	NN	O	B-protein
(	NN	O	O
regulated	NN	O	B-protein
on	NN	O	I-protein
activation	NN	O	I-protein
of	NN	O	I-protein
normal	NN	O	I-protein
T-cell	NN	O	I-protein
expressed	NN	O	I-protein
and	NN	O	I-protein
secreted	NN	O	I-protein
)	NN	O	O
expression	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
later	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
inducible	NN	O	O
form	NN	O	O
of	NN	O	O
nitric	NN	O	B-protein
oxide	NN	O	I-protein
synthase	NN	O	I-protein
.	NN	O	O

The	NN	O	O
early	NN	O	O
activation	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappaB	NN	O	B-protein
was	NN	O	O
also	NN	O	O
blocked	NN	O	O
by	NN	O	O
the	NN	O	O
HSR	NN	O	O
.	NN	O	O

The	NN	O	O
finding	NN	O	O
that	NN	O	O
the	NN	O	O
SR	NN	O	O
reduces	NN	O	O
inflammation	NN	O	O
in	NN	O	O
the	NN	O	O
brain	NN	O	O
and	NN	O	O
the	NN	O	O
clinical	NN	O	O
severity	NN	O	O
of	NN	O	O
EAE	NN	O	O
opens	NN	O	O
a	NN	O	O
novel	NN	O	O
therapeutic	NN	O	O
approach	NN	O	O
for	NN	O	O
prevention	NN	O	O
of	NN	O	O
autoimmune	NN	O	O
diseases	NN	O	O
.	NN	O	O

-DOCSTART-	O

In	NN	O	O
vivo	NN	O	O
detection	NN	O	O
of	NN	O	O
intracellular	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
in	NN	O	O
developing	NN	O	O
thymocytes	NN	O	B-cell_type
.	NN	O	O

Information	NN	O	O
regarding	NN	O	O
the	NN	O	O
intracellular	NN	O	O
signaling	NN	O	O
processes	NN	O	O
that	NN	O	O
occur	NN	O	O
during	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
has	NN	O	O
largely	NN	O	O
been	NN	O	O
obtained	NN	O	O
with	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
transgenic	NN	O	O
mouse	NN	O	O
models	NN	O	O
,	NN	O	O
which	NN	O	O
although	NN	O	O
providing	NN	O	O
invaluable	NN	O	O
information	NN	O	O
are	NN	O	O
time	NN	O	O
consuming	NN	O	O
and	NN	O	O
costly	NN	O	O
.	NN	O	O

To	NN	O	O
this	NN	O	O
end	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
developed	NN	O	O
a	NN	O	O
novel	NN	O	O
system	NN	O	O
that	NN	O	O
facilitates	NN	O	O
the	NN	O	O
in	NN	O	O
vivo	NN	O	O
analysis	NN	O	O
of	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathways	NN	O	O
during	NN	O	O
T-lymphocyte	NN	O	B-cell_type
development	NN	O	O
.	NN	O	O

This	NN	O	O
approach	NN	O	O
uses	NN	O	O
reporter-plasmids	NN	O	B-DNA
for	NN	O	O
the	NN	O	O
detection	NN	O	O
of	NN	O	O
intracellular	NN	O	O
signals	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
or	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

Reporter-plasmids	NN	O	B-DNA
are	NN	O	O
transfected	NN	O	O
into	NN	O	O
thymocytes	NN	O	B-cell_type
in	NN	O	O
fetal	NN	O	O
thymic	NN	O	O
organ	NN	O	O
culture	NN	O	O
by	NN	O	O
accelerated	NN	O	O
DNA/particle	NN	O	O
bombardment	NN	O	O
(	NN	O	O
gene	NN	O	O
gun	NN	O	O
)	NN	O	O
,	NN	O	O
and	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
is	NN	O	O
determined	NN	O	O
in	NN	O	O
the	NN	O	O
form	NN	O	O
of	NN	O	O
a	NN	O	O
standard	NN	O	O
luciferase	NN	O	O
assay	NN	O	O
.	NN	O	O

Importantly	NN	O	O
,	NN	O	O
this	NN	O	O
powerful	NN	O	O
technique	NN	O	O
preserves	NN	O	O
the	NN	O	O
structural	NN	O	O
integrity	NN	O	O
of	NN	O	O
the	NN	O	O
thymus	NN	O	O
,	NN	O	O
and	NN	O	O
will	NN	O	O
provide	NN	O	O
an	NN	O	O
invaluable	NN	O	O
tool	NN	O	O
to	NN	O	O
study	NN	O	O
how	NN	O	O
thymocytes	NN	O	B-cell_type
respond	NN	O	O
to	NN	O	O
normal	NN	O	O
environmental	NN	O	O
stimuli	NN	O	O
encountered	NN	O	O
during	NN	O	O
differentiation	NN	O	O
within	NN	O	O
the	NN	O	O
thymic	NN	O	O
milieu	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
this	NN	O	O
method	NN	O	O
allows	NN	O	O
for	NN	O	O
the	NN	O	O
monitoring	NN	O	O
of	NN	O	O
signals	NN	O	O
that	NN	O	O
occur	NN	O	O
in	NN	O	O
a	NN	O	O
biological	NN	O	O
time	NN	O	O
frame	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
during	NN	O	O
differentiation	NN	O	O
,	NN	O	O
and	NN	O	O
within	NN	O	O
the	NN	O	O
natural	NN	O	O
environment	NN	O	O
of	NN	O	O
differentiating	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Core-binding	NN	O	B-protein
factor	NN	O	I-protein
beta	NN	O	I-protein
(	NN	O	O
CBFbeta	NN	O	B-protein
)	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
CBFbeta-smooth	NN	O	B-protein
muscle	NN	O	I-protein
myosin	NN	O	I-protein
heavy	NN	O	I-protein
chain	NN	O	I-protein
,	NN	O	O
rescues	NN	O	O
definitive	NN	O	O
hematopoiesis	NN	O	O
in	NN	O	O
CBFbeta	NN	O	B-protein
-deficient	NN	O	O
embryonic	NN	O	B-cell_type
stem	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Core-binding	NN	O	B-protein
factor	NN	O	I-protein
beta	NN	O	I-protein
(	NN	O	O
CBFbeta	NN	O	B-protein
)	NN	O	O
is	NN	O	O
the	NN	O	O
non-DNA-binding	NN	O	B-protein
subunit	NN	O	I-protein
of	NN	O	O
the	NN	O	O
heterodimeric	NN	O	B-protein
CBFs	NN	O	I-protein
.	NN	O	O

Genes	NN	O	B-DNA
encoding	NN	O	I-DNA
CBFbeta	NN	O	I-DNA
(	NN	O	O
CBFB	NN	O	B-DNA
)	NN	O	O
,	NN	O	O
and	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
DNA-binding	NN	O	B-protein
CBFalpha	NN	O	I-protein
subunits	NN	O	I-protein
,	NN	O	I-protein
Runx1	NN	O	I-protein
(	NN	O	O
also	NN	O	O
known	NN	O	O
as	NN	O	O
CBFalpha2	NN	O	B-protein
,	NN	O	O
AML1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
PEBP2alphaB	NN	O	B-protein
)	NN	O	O
,	NN	O	O
are	NN	O	O
required	NN	O	O
for	NN	O	O
normal	NN	O	O
hematopoiesis	NN	O	O
and	NN	O	O
are	NN	O	O
also	NN	O	O
frequent	NN	O	O
targets	NN	O	O
of	NN	O	O
chromosomal	NN	O	O
translocations	NN	O	O
in	NN	O	O
acute	NN	O	O
leukemias	NN	O	O
in	NN	O	O
humans	NN	O	O
.	NN	O	O

Homozygous	NN	O	O
disruption	NN	O	O
of	NN	O	O
either	NN	O	O
the	NN	O	O
Runx1	NN	O	B-DNA
or	NN	O	I-DNA
Cbfb	NN	O	I-DNA
gene	NN	O	I-DNA
in	NN	O	O
mice	NN	O	O
results	NN	O	O
in	NN	O	O
embryonic	NN	O	O
lethality	NN	O	O
at	NN	O	O
midgestation	NN	O	O
due	NN	O	O
to	NN	O	O
hemorrhaging	NN	O	O
in	NN	O	O
the	NN	O	O
central	NN	O	O
nervous	NN	O	O
system	NN	O	O
,	NN	O	O
and	NN	O	O
severely	NN	O	O
impairs	NN	O	O
fetal	NN	O	O
liver	NN	O	O
hematopoiesis	NN	O	O
.	NN	O	O

Results	NN	O	O
of	NN	O	O
this	NN	O	O
study	NN	O	O
show	NN	O	O
that	NN	O	O
Cbfb-deficient	NN	O	O
mouse	NN	O	O
embryonic	NN	O	B-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
can	NN	O	O
differentiate	NN	O	O
into	NN	O	O
primitive	NN	O	B-cell_type
erythroid	NN	O	I-cell_type
colonies	NN	O	I-cell_type
in	NN	O	O
vitro	NN	O	O
,	NN	O	O
but	NN	O	O
are	NN	O	O
impaired	NN	O	O
in	NN	O	O
their	NN	O	O
ability	NN	O	O
to	NN	O	O
produce	NN	O	O
definitive	NN	O	O
erythroid	NN	O	B-cell_type
and	NN	O	I-cell_type
myeloid	NN	O	I-cell_type
colonies	NN	O	I-cell_type
,	NN	O	O
mimicking	NN	O	O
the	NN	O	O
in	NN	O	O
vivo	NN	O	O
defect	NN	O	O
.	NN	O	O

Definitive	NN	O	O
hematopoiesis	NN	O	O
is	NN	O	O
restored	NN	O	O
by	NN	O	O
ectopic	NN	O	O
expression	NN	O	O
of	NN	O	O
full-length	NN	O	B-DNA
Cbfb	NN	O	I-DNA
transgenes	NN	O	I-DNA
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
by	NN	O	O
a	NN	O	O
transgene	NN	O	O
encoding	NN	O	O
only	NN	O	O
the	NN	O	O
heterodimerization	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
CBFbeta	NN	O	B-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
the	NN	O	O
CBFbeta	NN	O	B-protein
-	NN	O	O
smooth	NN	O	B-protein
muscle	NN	O	I-protein
myosin	NN	O	I-protein
heavy	NN	O	I-protein
chain	NN	O	I-protein
(	NN	O	I-protein
SMMHC	NN	O	I-protein
)	NN	O	I-protein
fusion	NN	O	I-protein
protein	NN	O	I-protein
generated	NN	O	O
by	NN	O	O
the	NN	O	O
inv	NN	O	B-protein
(	NN	O	I-protein
16	NN	O	I-protein
)	NN	O	I-protein
associated	NN	O	O
with	NN	O	O
acute	NN	O	B-cell_line
myeloid	NN	O	I-cell_line
leukemias	NN	O	I-cell_line
(	NN	O	O
M4Eo	NN	O	B-cell_line
)	NN	O	O
can	NN	O	O
not	NN	O	O
rescue	NN	O	O
definitive	NN	O	O
hematopoiesis	NN	O	O
by	NN	O	O
Cbfb-deficient	NN	O	B-cell_type
ES	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Sequences	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
inability	NN	O	O
of	NN	O	O
CBFbeta	NN	O	B-protein
-SMMHC	NN	O	O
to	NN	O	O
rescue	NN	O	O
definitive	NN	O	O
hematopoiesis	NN	O	O
reside	NN	O	O
in	NN	O	O
the	NN	O	O
SMMHC	NN	O	B-protein
portion	NN	O	I-protein
of	NN	O	O
the	NN	O	O
fusion	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

Results	NN	O	O
also	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
CBFbeta-SMMHC	NN	O	B-protein
fusion	NN	O	I-protein
protein	NN	O	I-protein
transdominantly	NN	O	O
inhibits	NN	O	O
definitive	NN	O	O
hematopoiesis	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
to	NN	O	O
the	NN	O	O
same	NN	O	O
extent	NN	O	O
as	NN	O	O
homozygous	NN	O	O
loss	NN	O	O
of	NN	O	O
Runx1	NN	O	B-protein
or	NN	O	O
Cbfb	NN	O	B-protein
.	NN	O	O

CBFbeta-SMMHC	NN	O	B-protein
preferentially	NN	O	O
inhibits	NN	O	O
the	NN	O	O
differentiation	NN	O	O
of	NN	O	O
myeloid	NN	O	B-cell_type
lineage	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
while	NN	O	O
increasing	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
blastlike	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
culture	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
latency	NN	O	O
pattern	NN	O	O
of	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
infection	NN	O	O
and	NN	O	O
viral	NN	O	O
IL-10	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
cutaneous	NN	O	B-cell_type
natural	NN	O	I-cell_type
killer/T-cell	NN	O	I-cell_type
lymphomas	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
nasal	NN	O	B-cell_type
type	NN	O	I-cell_type
,	NN	O	I-cell_type
extranodal	NN	O	I-cell_type
natural	NN	O	I-cell_type
killer	NN	O	I-cell_type
or	NN	O	I-cell_type
T	NN	O	I-cell_type
(	NN	O	I-cell_type
NK/T	NN	O	I-cell_type
)	NN	O	I-cell_type
-cell	NN	O	I-cell_type
lymphoma	NN	O	I-cell_type
is	NN	O	O
usually	NN	O	O
associated	NN	O	O
with	NN	O	O
latent	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
(	NN	O	O
EBV	NN	O	O
)	NN	O	O
infection	NN	O	O
.	NN	O	O

In	NN	O	O
order	NN	O	O
to	NN	O	O
elucidate	NN	O	O
the	NN	O	O
EBV	NN	O	O
gene	NN	O	O
expression	NN	O	O
patterns	NN	O	O
in	NN	O	O
vivo	NN	O	O
,	NN	O	O
we	NN	O	O
examined	NN	O	O
eight	NN	O	O
patients	NN	O	O
with	NN	O	O
cutaneous	NN	O	O
EBV-related	NN	O	O
NK/T-cell	NN	O	O
lymphomas	NN	O	O
,	NN	O	O
including	NN	O	O
six	NN	O	O
patients	NN	O	O
with	NN	O	O
a	NN	O	O
NK-cell	NN	O	O
phenotype	NN	O	O
and	NN	O	O
two	NN	O	O
patients	NN	O	O
with	NN	O	O
a	NN	O	O
T-cell	NN	O	O
phenotype	NN	O	O
.	NN	O	O

The	NN	O	O
implication	NN	O	O
of	NN	O	O
EBV	NN	O	O
in	NN	O	O
the	NN	O	O
skin	NN	O	O
lesions	NN	O	O
was	NN	O	O
determined	NN	O	O
by	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
EBV-DNA	NN	O	B-DNA
,	NN	O	O
EBV-encoded	NN	O	B-RNA
nuclear	NN	O	I-RNA
RNA	NN	O	I-RNA
(	NN	O	O
EBER	NN	O	B-RNA
)	NN	O	O
and	NN	O	O
a	NN	O	O
clonality	NN	O	O
of	NN	O	O
EBV-DNA	NN	O	B-DNA
fragments	NN	O	I-DNA
containing	NN	O	O
the	NN	O	O
terminal	NN	O	B-DNA
repeats	NN	O	I-DNA
.	NN	O	O

Transcripts	NN	O	O
of	NN	O	O
EBV-encoded	NN	O	B-DNA
genes	NN	O	I-DNA
were	NN	O	O
screened	NN	O	O
by	NN	O	O
reverse	NN	O	O
transcription-	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
(	NN	O	O
RT-PCR	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
confirmed	NN	O	O
by	NN	O	O
Southern	NN	O	O
blot	NN	O	O
hybridization	NN	O	O
.	NN	O	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
EBV-related	NN	O	B-protein
antigens	NN	O	I-protein
was	NN	O	O
examined	NN	O	O
by	NN	O	O
immunostaining	NN	O	O
using	NN	O	O
paraffin-embedded	NN	O	O
tissue	NN	O	O
sections	NN	O	O
and	NN	O	O
cell	NN	O	O
pellets	NN	O	O
of	NN	O	O
EBV-positive	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Our	NN	O	O
study	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
all	NN	O	O
samples	NN	O	O
from	NN	O	O
the	NN	O	O
patients	NN	O	O
contained	NN	O	O
EBV	NN	O	B-RNA
nuclear	NN	O	I-RNA
antigen	NN	O	I-RNA
(	NN	O	I-RNA
EBNA	NN	O	I-RNA
)	NN	O	I-RNA
-1	NN	O	I-RNA
mRNA	NN	O	I-RNA
which	NN	O	O
was	NN	O	O
transcribed	NN	O	O
using	NN	O	O
the	NN	O	O
Q	NN	O	B-DNA
promoter	NN	O	I-DNA
,	NN	O	O
whereas	NN	O	O
both	NN	O	O
the	NN	O	O
Q	NN	O	B-DNA
promoter	NN	O	I-DNA
and	NN	O	O
another	NN	O	O
upstream	NN	O	B-DNA
promoter	NN	O	I-DNA
(	NN	O	O
Cp/Wp	NN	O	B-DNA
)	NN	O	O
were	NN	O	O
used	NN	O	O
in	NN	O	O
EBV-positive	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
B95.8	NN	O	B-cell_line
,	NN	O	O
Raji	NN	O	B-cell_line
and	NN	O	O
Jiyoye	NN	O	B-cell_line
.	NN	O	O

Latent	NN	O	B-RNA
membrane	NN	O	I-RNA
protein-1	NN	O	I-RNA
(	NN	O	I-RNA
LMP-1	NN	O	I-RNA
)	NN	O	I-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
detected	NN	O	O
in	NN	O	O
seven	NN	O	O
of	NN	O	O
eight	NN	O	O
patients	NN	O	O
and	NN	O	O
all	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
whereas	NN	O	O
EBNA-2	NN	O	B-RNA
transcripts	NN	O	I-RNA
were	NN	O	O
found	NN	O	O
only	NN	O	O
in	NN	O	O
the	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Immunostaining	NN	O	O
showed	NN	O	O
no	NN	O	O
LMP-1	NN	O	B-protein
,	NN	O	I-protein
EBNA-2	NN	O	I-protein
or	NN	O	I-protein
ZEBRA	NN	O	I-protein
antigens	NN	O	I-protein
in	NN	O	O
the	NN	O	O
paraffin-embedded	NN	O	O
tissue	NN	O	O
sections	NN	O	O
,	NN	O	O
although	NN	O	O
they	NN	O	O
were	NN	O	O
positive	NN	O	O
in	NN	O	O
the	NN	O	O
cell	NN	O	B-cell_line
line	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Latent	NN	O	B-RNA
BHRF1	NN	O	I-RNA
transcripts	NN	O	I-RNA
encoding	NN	O	O
bcl-2	NN	O	B-RNA
homologue	NN	O	I-RNA
and	NN	O	O
BCRF1	NN	O	B-RNA
transcripts	NN	O	I-RNA
encoding	NN	O	O
viral	NN	O	B-protein
interleukin	NN	O	I-protein
(	NN	O	I-protein
vIL	NN	O	I-protein
)	NN	O	I-protein
-10	NN	O	I-protein
were	NN	O	O
detected	NN	O	O
in	NN	O	O
one	NN	O	O
and	NN	O	O
two	NN	O	O
of	NN	O	O
eight	NN	O	O
patients	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

A	NN	O	O
patient	NN	O	O
with	NN	O	O
NK-cell	NN	O	O
lymphoma	NN	O	O
expressing	NN	O	O
both	NN	O	O
transcripts	NN	O	O
died	NN	O	O
of	NN	O	O
rapid	NN	O	O
progression	NN	O	O
of	NN	O	O
the	NN	O	O
illness	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
restricted	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
latency-associated	NN	O	B-DNA
EBV	NN	O	I-DNA
genes	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
vIL-10	NN	O	B-RNA
and	NN	O	I-RNA
bcl-2	NN	O	I-RNA
homologue	NN	O	I-RNA
may	NN	O	O
favour	NN	O	O
tumour	NN	O	O
growth	NN	O	O
,	NN	O	O
evading	NN	O	O
the	NN	O	O
host	NN	O	O
immune	NN	O	O
surveillance	NN	O	O
.	NN	O	O

Copyright	NN	O	O
2001	NN	O	O
Cancer	NN	O	O
Research	NN	O	O
Campaign	NN	O	O
.	NN	O	O

-DOCSTART-	O

Oxidized	NN	O	O
alkyl	NN	O	O
phospholipids	NN	O	O
are	NN	O	O
specific	NN	O	O
,	NN	O	O
high	NN	O	O
affinity	NN	O	O
peroxisome	NN	O	O
proliferator-activated	NN	O	O
receptor	NN	O	O
gamma	NN	O	O
ligands	NN	O	O
and	NN	O	O
agonists	NN	O	O
.	NN	O	O

Synthetic	NN	O	O
high	NN	O	O
affinity	NN	O	O
peroxisome	NN	O	O
proliferator-activated	NN	O	O
receptor	NN	O	O
(	NN	O	O
PPAR	NN	O	O
)	NN	O	O
agonists	NN	O	O
are	NN	O	O
known	NN	O	O
,	NN	O	O
but	NN	O	O
biologic	NN	O	O
ligands	NN	O	O
are	NN	O	O
of	NN	O	O
low	NN	O	O
affinity	NN	O	O
.	NN	O	O

Oxidized	NN	O	B-protein
low	NN	O	I-protein
density	NN	O	I-protein
lipoprotein	NN	O	I-protein
(	NN	O	O
oxLDL	NN	O	B-protein
)	NN	O	O
is	NN	O	O
inflammatory	NN	O	O
and	NN	O	O
signals	NN	O	O
through	NN	O	O
PPARs	NN	O	B-protein
.	NN	O	O

We	NN	O	O
showed	NN	O	O
,	NN	O	O
by	NN	O	O
phospholipase	NN	O	O
A	NN	O	O
(	NN	O	O
1	NN	O	O
)	NN	O	O
digestion	NN	O	O
,	NN	O	O
that	NN	O	O
PPARgamma	NN	O	O
agonists	NN	O	O
in	NN	O	O
oxLDL	NN	O	B-protein
arise	NN	O	O
from	NN	O	O
the	NN	O	O
small	NN	O	O
pool	NN	O	O
of	NN	O	O
alkyl	NN	O	O
phosphatidylcholines	NN	O	O
in	NN	O	O
LDL	NN	O	B-protein
.	NN	O	O

We	NN	O	O
identified	NN	O	O
an	NN	O	O
abundant	NN	O	O
oxidatively	NN	O	O
fragmented	NN	O	O
alkyl	NN	O	O
phospholipid	NN	O	O
in	NN	O	O
oxLDL	NN	O	B-protein
,	NN	O	O
hexadecyl	NN	O	O
azelaoyl	NN	O	O
phosphatidylcholine	NN	O	O
(	NN	O	O
azPC	NN	O	O
)	NN	O	O
,	NN	O	O
as	NN	O	O
a	NN	O	O
high	NN	O	O
affinity	NN	O	O
ligand	NN	O	O
and	NN	O	O
agonist	NN	O	O
for	NN	O	O
PPARgamma	NN	O	B-protein
.	NN	O	O

[	NN	O	O
(	NN	O	O
3	NN	O	O
)	NN	O	O
H	NN	O	O
]	NN	O	O
azPC	NN	O	O
bound	NN	O	O
recombinant	NN	O	B-protein
PPARgamma	NN	O	I-protein
with	NN	O	O
an	NN	O	O
affinity	NN	O	O
(	NN	O	O
K	NN	O	O
(	NN	O	O
d	NN	O	O
)	NN	O	O
(	NN	O	O
(	NN	O	O
app	NN	O	O
)	NN	O	O
)	NN	O	O
approximately	NN	O	O
40	NN	O	O
nm	NN	O	O
)	NN	O	O
that	NN	O	O
was	NN	O	O
equivalent	NN	O	O
to	NN	O	O
rosiglitazone	NN	O	O
(	NN	O	O
BRL49653	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
competition	NN	O	O
with	NN	O	O
rosiglitazone	NN	O	O
showed	NN	O	O
that	NN	O	O
binding	NN	O	O
occurred	NN	O	O
in	NN	O	O
the	NN	O	O
ligand-binding	NN	O	B-protein
pocket	NN	O	I-protein
.	NN	O	O

azPC	NN	O	O
induced	NN	O	O
PPRE	NN	O	O
reporter	NN	O	O
gene	NN	O	O
expression	NN	O	O
,	NN	O	O
as	NN	O	O
did	NN	O	O
rosiglitazone	NN	O	O
,	NN	O	O
with	NN	O	O
a	NN	O	O
half-maximal	NN	O	O
effect	NN	O	O
at	NN	O	O
100	NN	O	O
nm	NN	O	O
.	NN	O	O

Overexpression	NN	O	O
of	NN	O	O
PPARalpha	NN	O	B-protein
or	NN	O	O
PPARgamma	NN	O	B-protein
revealed	NN	O	O
that	NN	O	O
azPC	NN	O	O
was	NN	O	O
a	NN	O	O
specific	NN	O	O
PPARgamma	NN	O	O
agonist	NN	O	O
.	NN	O	O

The	NN	O	O
scavenger	NN	O	B-protein
receptor	NN	O	I-protein
CD36	NN	O	I-protein
is	NN	O	O
encoded	NN	O	O
by	NN	O	O
a	NN	O	O
PPRE-responsive	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
azPC	NN	O	O
enhanced	NN	O	O
expression	NN	O	O
of	NN	O	O
CD36	NN	O	B-protein
in	NN	O	O
primary	NN	O	B-cell_type
human	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
anti-CD36	NN	O	B-protein
inhibited	NN	O	O
azPC	NN	O	O
uptake	NN	O	O
,	NN	O	O
and	NN	O	O
it	NN	O	O
inhibited	NN	O	O
PPRE	NN	O	O
reporter	NN	O	O
induction	NN	O	O
.	NN	O	O

Results	NN	O	O
with	NN	O	O
a	NN	O	O
small	NN	O	O
molecule	NN	O	O
phospholipid	NN	O	B-protein
flippase	NN	O	I-protein
mimetic	NN	O	I-protein
suggest	NN	O	O
azPC	NN	O	O
acts	NN	O	O
intracellularly	NN	O	O
and	NN	O	O
that	NN	O	O
cellular	NN	O	O
azPC	NN	O	O
accumulation	NN	O	O
was	NN	O	O
efficient	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
certain	NN	O	O
alkyl	NN	O	O
phospholipid	NN	O	O
oxidation	NN	O	O
products	NN	O	O
in	NN	O	O
oxLDL	NN	O	B-protein
are	NN	O	O
specific	NN	O	O
,	NN	O	O
high	NN	O	O
affinity	NN	O	O
extracellular	NN	O	O
ligands	NN	O	O
and	NN	O	O
agonists	NN	O	O
for	NN	O	O
PPARgamma	NN	O	B-protein
that	NN	O	O
induce	NN	O	O
PPAR-responsive	NN	O	B-DNA
genes	NN	O	I-DNA

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
MAT2B	NN	O	I-DNA
gene	NN	O	I-DNA
encoding	NN	O	O
the	NN	O	O
regulatory	NN	O	B-protein
beta	NN	O	I-protein
subunit	NN	O	I-protein
of	NN	O	O
methionine	NN	O	B-protein
adenosyltransferase	NN	O	I-protein
,	NN	O	O
MAT	NN	O	B-protein
II	NN	O	I-protein
.	NN	O	O

Methionine	NN	O	B-protein
adenosyltransferase	NN	O	I-protein
(	NN	O	O
MAT	NN	O	B-protein
)	NN	O	O
catalyzes	NN	O	O
the	NN	O	O
biosynthesis	NN	O	O
of	NN	O	O
S-adenosylmethionine	NN	O	B-protein
(	NN	O	O
AdoMet	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
key	NN	O	O
molecule	NN	O	O
in	NN	O	O
transmethylation	NN	O	O
reactions	NN	O	O
and	NN	O	O
polyamine	NN	O	O
biosynthesis	NN	O	O
.	NN	O	O

The	NN	O	O
MAT	NN	O	B-protein
II	NN	O	I-protein
isozyme	NN	O	I-protein
consists	NN	O	O
of	NN	O	O
a	NN	O	O
catalytic	NN	O	B-protein
alpha2	NN	O	I-protein
and	NN	O	I-protein
a	NN	O	I-protein
regulatory	NN	O	I-protein
beta	NN	O	I-protein
subunit	NN	O	I-protein
.	NN	O	O

Down-regulation	NN	O	O
of	NN	O	O
the	NN	O	O
MAT	NN	O	B-protein
II	NN	O	I-protein
beta	NN	O	I-protein
subunit	NN	O	I-protein
expression	NN	O	O
causes	NN	O	O
a	NN	O	O
6-10-fold	NN	O	O
increase	NN	O	O
in	NN	O	O
intracellular	NN	O	O
AdoMet	NN	O	B-protein
levels	NN	O	O
.	NN	O	O

To	NN	O	O
understand	NN	O	O
the	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
the	NN	O	O
beta	NN	O	B-protein
subunit	NN	O	I-protein
expression	NN	O	O
is	NN	O	O
regulated	NN	O	O
,	NN	O	O
we	NN	O	O
cloned	NN	O	O
the	NN	O	O
MAT2B	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	O
determined	NN	O	O
its	NN	O	O
organization	NN	O	O
,	NN	O	O
characterized	NN	O	O
its	NN	O	O
5'-flanking	NN	O	B-DNA
sequences	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
elucidated	NN	O	O
the	NN	O	O
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
in	NN	O	O
vivo	NN	O	O
regulation	NN	O	O
of	NN	O	O
its	NN	O	O
promoter	NN	O	B-DNA
.	NN	O	O

Transcription	NN	O	O
of	NN	O	O
the	NN	O	O
MAT2B	NN	O	B-DNA
gene	NN	O	I-DNA
initiates	NN	O	O
at	NN	O	O
position	NN	O	O
-203	NN	O	O
relative	NN	O	O
to	NN	O	O
the	NN	O	O
translation	NN	O	B-DNA
start	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

Promoter	NN	O	B-DNA
deletion	NN	O	I-DNA
analysis	NN	O	I-DNA
defined	NN	O	O
a	NN	O	O
minimal	NN	O	B-DNA
promoter	NN	O	I-DNA
between	NN	O	O
positions	NN	O	B-DNA
+52	NN	O	I-DNA
and	NN	O	I-DNA
+93	NN	O	I-DNA
base	NN	O	O
pairs	NN	O	O
,	NN	O	O
a	NN	O	O
GC-rich	NN	O	B-DNA
region	NN	O	I-DNA
.	NN	O	O

Inclusion	NN	O	O
of	NN	O	O
the	NN	O	O
sequences	NN	O	O
between	NN	O	O
-4	NN	O	O
and	NN	O	O
+52	NN	O	O
enhanced	NN	O	O
promoter	NN	O	O
activity	NN	O	O
;	NN	O	O
this	NN	O	O
was	NN	O	O
primarily	NN	O	O
because	NN	O	O
of	NN	O	O
an	NN	O	O
Sp1	NN	O	B-DNA
recognition	NN	O	I-DNA
site	NN	O	I-DNA
at	NN	O	O
+9/+15	NN	O	O
.	NN	O	O

The	NN	O	O
inclusion	NN	O	O
of	NN	O	O
sequences	NN	O	O
up	NN	O	O
to	NN	O	O
position	NN	O	O
-115	NN	O	O
provided	NN	O	O
full	NN	O	O
activity	NN	O	O
;	NN	O	O
this	NN	O	O
was	NN	O	O
attributed	NN	O	O
to	NN	O	O
a	NN	O	O
TATA	NN	O	B-DNA
at	NN	O	O
-32	NN	O	O
.	NN	O	O

The	NN	O	O
Sp1	NN	O	B-DNA
site	NN	O	I-DNA
at	NN	O	O
position	NN	O	O
+9	NN	O	O
was	NN	O	O
key	NN	O	O
for	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
protein.DNA	NN	O	O
complexes	NN	O	O
.	NN	O	O

Mutation	NN	O	O
of	NN	O	O
both	NN	O	O
the	NN	O	O
Sp1	NN	O	B-DNA
site	NN	O	I-DNA
at	NN	O	O
+9	NN	O	O
and	NN	O	O
the	NN	O	O
TATA	NN	O	B-DNA
at	NN	O	O
-32	NN	O	O
reduced	NN	O	O
promoter	NN	O	O
activity	NN	O	O
to	NN	O	O
its	NN	O	O
minimal	NN	O	O
level	NN	O	O
.	NN	O	O

Supershift	NN	O	O
assays	NN	O	O
showed	NN	O	O
no	NN	O	O
effect	NN	O	O
of	NN	O	O
the	NN	O	O
anti-Sp1	NN	O	B-protein
antibody	NN	O	I-protein
on	NN	O	O
complex	NN	O	O
formation	NN	O	O
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
anti-Sp3	NN	O	B-protein
antibody	NN	O	I-protein
had	NN	O	O
a	NN	O	O
strong	NN	O	O
effect	NN	O	O
on	NN	O	O
protein.DNA	NN	O	O
complex	NN	O	O
formation	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
Sp3	NN	O	B-protein
is	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
main	NN	O	O
factors	NN	O	O
binding	NN	O	O
to	NN	O	O
this	NN	O	O
Sp1	NN	O	B-DNA
site	NN	O	I-DNA
.	NN	O	O

Chromatin	NN	O	O
immunoprecipitation	NN	O	O
assays	NN	O	O
supported	NN	O	O
the	NN	O	O
involvement	NN	O	O
of	NN	O	O
both	NN	O	O
Sp1	NN	O	B-protein
and	NN	O	O
Sp3	NN	O	B-protein
in	NN	O	O
complexes	NN	O	O
formed	NN	O	O
on	NN	O	O
the	NN	O	O
MAT2B	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
data	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
5'-untranslated	NN	O	B-DNA
sequences	NN	O	I-DNA
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
regulating	NN	O	O
the	NN	O	O
MAT2B	NN	O	B-DNA
gene	NN	O	I-DNA
and	NN	O	O
identifies	NN	O	O
the	NN	O	O
Sp1	NN	O	B-DNA
site	NN	O	I-DNA
at	NN	O	O
+9	NN	O	O
as	NN	O	O
a	NN	O	O
potential	NN	O	O
target	NN	O	O
for	NN	O	O
modulating	NN	O	O
MAT2B	NN	O	O
expression	NN	O	O
,	NN	O	O
a	NN	O	O
process	NN	O	O
that	NN	O	O
can	NN	O	O
have	NN	O	O
a	NN	O	O
major	NN	O	O
effect	NN	O	O
on	NN	O	O
intracellular	NN	O	O
AdoMet	NN	O	B-protein
levels	NN	O	O
.	NN	O	O

-DOCSTART-	O

Molecular	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
influenza	NN	O	O
A	NN	O	O
virus	NN	O	O
infection	NN	O	O
and	NN	O	O
virus-induced	NN	O	O
regulation	NN	O	O
of	NN	O	O
cytokine	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
.	NN	O	O

Despite	NN	O	O
vaccines	NN	O	O
and	NN	O	O
antiviral	NN	O	O
substances	NN	O	O
influenza	NN	O	O
still	NN	O	O
causes	NN	O	O
significant	NN	O	O
morbidity	NN	O	O
and	NN	O	O
mortality	NN	O	O
world	NN	O	O
wide	NN	O	O
.	NN	O	O

Better	NN	O	O
understanding	NN	O	O
of	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
influenza	NN	O	O
virus	NN	O	O
replication	NN	O	O
,	NN	O	O
pathogenesis	NN	O	O
and	NN	O	O
host	NN	O	O
immune	NN	O	O
responses	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
more	NN	O	O
efficient	NN	O	O
means	NN	O	O
of	NN	O	O
prevention	NN	O	O
and	NN	O	O
treatment	NN	O	O
of	NN	O	O
influenza	NN	O	O
.	NN	O	O

Influenza	NN	O	O
A	NN	O	O
virus	NN	O	O
,	NN	O	O
which	NN	O	O
replicates	NN	O	O
in	NN	O	O
epithelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
leukocytes	NN	O	B-cell_type
,	NN	O	O
regulates	NN	O	O
host	NN	O	O
cell	NN	O	O
transcriptional	NN	O	O
and	NN	O	O
translational	NN	O	O
systems	NN	O	O
and	NN	O	O
activates	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
downregulates	NN	O	O
apoptotic	NN	O	O
pathways	NN	O	O
.	NN	O	O

Influenza	NN	O	O
A	NN	O	O
virus	NN	O	O
infection	NN	O	O
results	NN	O	O
in	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
chemotactic	NN	O	O
(	NN	O	O
RANTES	NN	O	B-protein
,	NN	O	O
MIP-1	NN	O	B-protein
alpha	NN	O	I-protein
,	NN	O	O
MCP-1	NN	O	B-protein
,	NN	O	O
MCP-3	NN	O	B-protein
,	NN	O	O
and	NN	O	O
IP-10	NN	O	B-protein
)	NN	O	O
,	NN	O	O
pro-inflammatory	NN	O	O
(	NN	O	O
IL-1	NN	O	B-protein
beta	NN	O	I-protein
,	NN	O	O
IL-6	NN	O	B-protein
,	NN	O	O
IL-18	NN	O	B-protein
,	NN	O	O
and	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
antiviral	NN	O	B-protein
(	NN	O	I-protein
IFN-alpha/beta	NN	O	I-protein
)	NN	O	I-protein
cytokines	NN	O	I-protein
.	NN	O	O

Cytokine	NN	O	O
gene	NN	O	O
expression	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
AP-1	NN	O	B-protein
,	NN	O	O
STAT	NN	O	B-protein
and	NN	O	O
IRF	NN	O	B-protein
signal	NN	O	I-protein
transducing	NN	O	I-protein
molecules	NN	O	I-protein
in	NN	O	O
influenza	NN	O	O
A	NN	O	O
virus-infected	NN	O	O
cells	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
of	NN	O	O
upregulating	NN	O	O
cytokine	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
,	NN	O	O
influenza	NN	O	O
A	NN	O	O
virus	NN	O	O
infection	NN	O	O
activates	NN	O	O
caspase-1	NN	O	B-protein
enzyme	NN	O	I-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
proteolytic	NN	O	O
processing	NN	O	O
of	NN	O	O
proIL-1	NN	O	B-protein
beta	NN	O	I-protein
and	NN	O	O
proIL-18	NN	O	B-protein
into	NN	O	O
their	NN	O	O
biologically	NN	O	O
active	NN	O	O
forms	NN	O	O
.	NN	O	O

Influenza	NN	O	O
A	NN	O	O
virus-induced	NN	O	O
IFN-alpha/beta	NN	O	B-protein
is	NN	O	O
essential	NN	O	O
in	NN	O	O
host	NN	O	O
's	NN	O	O
antiviral	NN	O	O
defence	NN	O	O
by	NN	O	O
activating	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
antiviral	NN	O	B-DNA
Mx	NN	O	I-DNA
,	NN	O	O
PKR	NN	O	B-DNA
and	NN	O	I-DNA
oligoadenylate	NN	O	I-DNA
synthetase	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

IFN-alpha/beta	NN	O	B-protein
also	NN	O	O
prolongs	NN	O	O
T	NN	O	O
cell	NN	O	O
survival	NN	O	O
,	NN	O	O
upregulates	NN	O	O
IL-12	NN	O	O
and	NN	O	O
IL-18	NN	O	O
receptor	NN	O	O
gene	NN	O	O
expression	NN	O	O
and	NN	O	O
together	NN	O	O
with	NN	O	O
IL-18	NN	O	B-protein
stimulates	NN	O	O
NK	NN	O	B-cell_type
and	NN	O	I-cell_type
T	NN	O	I-cell_type
cell	NN	O	I-cell_type
IFN-gamma	NN	O	O
production	NN	O	O
and	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
Th1-type	NN	O	O
immune	NN	O	O
response	NN	O	O
.	NN	O	O

-DOCSTART-	O

Comparison	NN	O	O
of	NN	O	O
hprt	NN	O	O
and	NN	O	O
lacI	NN	O	O
mutant	NN	O	O
frequency	NN	O	O
with	NN	O	O
DNA	NN	O	O
adduct	NN	O	O
formation	NN	O	O
in	NN	O	O
N-hydroxy-2-acetylaminofluorene-treated	NN	O	O
Big	NN	O	O
Blue	NN	O	O
rats	NN	O	O
.	NN	O	O

N-Hydroxy-2-acetylaminofluorene	NN	O	O
(	NN	O	O
N-OH-AAF	NN	O	O
)	NN	O	O
is	NN	O	O
the	NN	O	O
proximate	NN	O	O
carcinogenic	NN	O	O
metabolite	NN	O	O
of	NN	O	O
the	NN	O	O
powerful	NN	O	O
rat	NN	O	O
liver	NN	O	O
carcinogen	NN	O	O
2-acetylaminofluorene	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
transgenic	NN	O	O
Big	NN	O	O
Blue	NN	O	O
(	NN	O	O
R	NN	O	O
)	NN	O	O
rats	NN	O	O
were	NN	O	O
used	NN	O	O
to	NN	O	O
examine	NN	O	O
the	NN	O	O
relationship	NN	O	O
between	NN	O	O
in	NN	O	O
vivo	NN	O	O
mutagenicity	NN	O	O
and	NN	O	O
DNA	NN	O	O
adduct	NN	O	O
formation	NN	O	O
by	NN	O	O
N-OH-AAF	NN	O	O
in	NN	O	O
the	NN	O	O
target	NN	O	O
liver	NN	O	O
compared	NN	O	O
with	NN	O	O
that	NN	O	O
in	NN	O	O
nontarget	NN	O	O
tissues	NN	O	O
.	NN	O	O

Male	NN	O	O
rats	NN	O	O
were	NN	O	O
given	NN	O	O
one	NN	O	O
,	NN	O	O
two	NN	O	O
,	NN	O	O
or	NN	O	O
four	NN	O	O
doses	NN	O	O
of	NN	O	O
25	NN	O	O
mg	NN	O	O
N-OH-AAF/kg	NN	O	O
body	NN	O	O
weight	NN	O	O
by	NN	O	O
i.p.	NN	O	O
injection	NN	O	O
at	NN	O	O
4-day	NN	O	O
intervals	NN	O	O
,	NN	O	O
and	NN	O	O
groups	NN	O	O
of	NN	O	O
treated	NN	O	O
and	NN	O	O
control	NN	O	O
rats	NN	O	O
were	NN	O	O
euthanized	NN	O	O
up	NN	O	O
to	NN	O	O
10	NN	O	O
weeks	NN	O	O
after	NN	O	O
beginning	NN	O	O
the	NN	O	O
dosing	NN	O	O
.	NN	O	O

Mutant	NN	O	O
frequencies	NN	O	O
were	NN	O	O
measured	NN	O	O
in	NN	O	O
the	NN	O	O
spleen	NN	O	B-DNA
lymphocyte	NN	O	I-DNA
hprt	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
lacI	NN	O	O
mutant	NN	O	O
frequencies	NN	O	O
were	NN	O	O
determined	NN	O	O
in	NN	O	O
the	NN	O	O
liver	NN	O	O
and	NN	O	O
spleen	NN	O	O
lymphocytes	NN	O	B-cell_type
.	NN	O	O

At	NN	O	O
6	NN	O	O
weeks	NN	O	O
after	NN	O	O
beginning	NN	O	O
the	NN	O	O
dosing	NN	O	O
,	NN	O	O
the	NN	O	O
hprt	NN	O	O
mutant	NN	O	O
frequency	NN	O	O
in	NN	O	O
spleen	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
from	NN	O	O
the	NN	O	O
four-dose	NN	O	O
group	NN	O	O
was	NN	O	O
16.5	NN	O	O
x	NN	O	O
10	NN	O	O
(	NN	O	O
-6	NN	O	O
)	NN	O	O
compared	NN	O	O
with	NN	O	O
3.2	NN	O	O
x	NN	O	O
10	NN	O	O
(	NN	O	O
-6	NN	O	O
)	NN	O	O
in	NN	O	O
control	NN	O	O
animals	NN	O	O
.	NN	O	O

Also	NN	O	O
at	NN	O	O
6	NN	O	O
weeks	NN	O	O
,	NN	O	O
rats	NN	O	O
given	NN	O	O
one	NN	O	O
,	NN	O	O
two	NN	O	O
,	NN	O	O
or	NN	O	O
four	NN	O	O
doses	NN	O	O
of	NN	O	O
N-OH-AAF	NN	O	O
had	NN	O	O
lacI	NN	O	O
mutant	NN	O	O
frequencies	NN	O	O
in	NN	O	O
the	NN	O	O
liver	NN	O	O
of	NN	O	O
97.6	NN	O	O
,	NN	O	O
155.6	NN	O	O
,	NN	O	O
and	NN	O	O
406.8	NN	O	O
x	NN	O	O
10	NN	O	O
(	NN	O	O
-6	NN	O	O
)	NN	O	O
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
compared	NN	O	O
with	NN	O	O
a	NN	O	O
control	NN	O	O
frequency	NN	O	O
of	NN	O	O
25.7	NN	O	O
x	NN	O	O
10	NN	O	O
(	NN	O	O
-6	NN	O	O
)	NN	O	O
;	NN	O	O
rats	NN	O	O
given	NN	O	O
four	NN	O	O
doses	NN	O	O
had	NN	O	O
lacI	NN	O	O
mutant	NN	O	O
frequencies	NN	O	O
in	NN	O	O
spleen	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
of	NN	O	O
55.8	NN	O	O
x	NN	O	O
10	NN	O	O
(	NN	O	O
-6	NN	O	O
)	NN	O	O
compared	NN	O	O
with	NN	O	O
a	NN	O	O
control	NN	O	O
frequency	NN	O	O
of	NN	O	O
20.4	NN	O	O
x	NN	O	O
10	NN	O	O
(	NN	O	O
-6	NN	O	O
)	NN	O	O
.	NN	O	O

Additional	NN	O	O
rats	NN	O	O
were	NN	O	O
evaluated	NN	O	O
for	NN	O	O
DNA	NN	O	O
adduct	NN	O	O
formation	NN	O	O
in	NN	O	O
the	NN	O	O
liver	NN	O	O
,	NN	O	O
spleen	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
bone	NN	O	O
marrow	NN	O	O
by	NN	O	O
(	NN	O	O
32	NN	O	O
)	NN	O	O
P-postlabeling	NN	O	O
.	NN	O	O

Adduct	NN	O	O
analysis	NN	O	O
was	NN	O	O
conducted	NN	O	O
1	NN	O	O
day	NN	O	O
after	NN	O	O
one	NN	O	O
,	NN	O	O
two	NN	O	O
,	NN	O	O
and	NN	O	O
four	NN	O	O
treatments	NN	O	O
with	NN	O	O
N-OH-AAF	NN	O	O
,	NN	O	O
5	NN	O	O
days	NN	O	O
after	NN	O	O
one	NN	O	O
treatment	NN	O	O
,	NN	O	O
and	NN	O	O
9	NN	O	O
days	NN	O	O
after	NN	O	O
two	NN	O	O
treatments	NN	O	O
.	NN	O	O

N-	NN	O	O
(	NN	O	O
Deoxyguanosin-8-yl	NN	O	O
)	NN	O	O
-2-aminofluorene	NN	O	O
was	NN	O	O
the	NN	O	O
major	NN	O	O
DNA	NN	O	O
adduct	NN	O	O
identified	NN	O	O
in	NN	O	O
all	NN	O	O
the	NN	O	O
tissues	NN	O	O
examined	NN	O	O
.	NN	O	O

Adduct	NN	O	O
concentrations	NN	O	O
increased	NN	O	O
with	NN	O	O
total	NN	O	O
dose	NN	O	O
to	NN	O	O
maximum	NN	O	O
values	NN	O	O
in	NN	O	O
samples	NN	O	O
taken	NN	O	O
1	NN	O	O
day	NN	O	O
after	NN	O	O
two	NN	O	O
doses	NN	O	O
,	NN	O	O
and	NN	O	O
remained	NN	O	O
essentially	NN	O	O
the	NN	O	O
same	NN	O	O
after	NN	O	O
four	NN	O	O
doses	NN	O	O
.	NN	O	O

In	NN	O	O
samples	NN	O	O
taken	NN	O	O
after	NN	O	O
four	NN	O	O
doses	NN	O	O
,	NN	O	O
adduct	NN	O	O
levels	NN	O	O
were	NN	O	O
103	NN	O	O
,	NN	O	O
28	NN	O	O
,	NN	O	O
and	NN	O	O
7	NN	O	O
fmol/microg	NN	O	O
of	NN	O	O
DNA	NN	O	O
in	NN	O	O
liver	NN	O	O
,	NN	O	O
spleen	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
bone	NN	O	O
marrow	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
extent	NN	O	O
of	NN	O	O
both	NN	O	O
DNA	NN	O	O
adduct	NN	O	O
formation	NN	O	O
and	NN	O	O
mutant	NN	O	O
induction	NN	O	O
correlates	NN	O	O
with	NN	O	O
the	NN	O	O
organ	NN	O	O
specificity	NN	O	O
for	NN	O	O
N-OH-AAF	NN	O	O
carcinogenesis	NN	O	O
in	NN	O	O
the	NN	O	O
rat	NN	O	O
.	NN	O	O

Environ.	NN	O	O
Mol.	NN	O	O
Mutagen.	NN	O	O
37	NN	O	O
:	NN	O	O
195-202	NN	O	O
,	NN	O	O
2001	NN	O	O
.	NN	O	O

Published	NN	O	O
2001	NN	O	O
Wiley-Liss	NN	O	O
,	NN	O	O
Inc	NN	O	O
.	NN	O	O

-DOCSTART-	O

Expression	NN	O	O
of	NN	O	O
Mad1	NN	O	B-protein
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
leads	NN	O	O
to	NN	O	O
reduced	NN	O	O
thymic	NN	O	O
cellularity	NN	O	O
and	NN	O	O
impaired	NN	O	O
mitogen-induced	NN	O	O
proliferation	NN	O	O
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
Mad1	NN	O	B-protein
function	NN	O	O
in	NN	O	O
vivo	NN	O	O
,	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
were	NN	O	O
generated	NN	O	O
that	NN	O	O
express	NN	O	O
a	NN	O	O
Mad1	NN	O	B-DNA
transgene	NN	O	I-DNA
in	NN	O	O
T	NN	O	B-cell_type
lineage	NN	O	I-cell_type
cells	NN	O	I-cell_type
under	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
the	NN	O	O
proximal	NN	O	B-DNA
lck	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Thymus	NN	O	O
size	NN	O	O
in	NN	O	O
lck-Mad1	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
is	NN	O	O
drastically	NN	O	O
reduced	NN	O	O
although	NN	O	O
representation	NN	O	O
of	NN	O	O
the	NN	O	O
various	NN	O	O
thymocyte	NN	O	B-cell_type
sub	NN	O	I-cell_type
populations	NN	O	I-cell_type
appears	NN	O	O
normal	NN	O	O
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
more	NN	O	O
closely	NN	O	O
any	NN	O	O
effects	NN	O	O
of	NN	O	O
Mad1	NN	O	O
expression	NN	O	O
on	NN	O	O
thymocytes	NN	O	B-cell_type
,	NN	O	O
we	NN	O	O
examined	NN	O	O
thymic	NN	O	O
selection	NN	O	O
using	NN	O	O
MHC	NN	O	O
class	NN	O	O
I-restricted	NN	O	O
H-Y-TCR	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
.	NN	O	O

Mad1	NN	O	O
expression	NN	O	O
in	NN	O	O
vivo	NN	O	O
reduces	NN	O	O
the	NN	O	O
efficiency	NN	O	O
of	NN	O	O
positive	NN	O	O
selection	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
thymocytes	NN	O	B-cell_type
and	NN	O	O
splenic	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
lck-Mad1	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
display	NN	O	O
a	NN	O	O
profound	NN	O	O
proliferative	NN	O	O
defect	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
activation	NN	O	O
with	NN	O	O
either	NN	O	O
PMA/Ionomycin	NN	O	O
or	NN	O	O
immobilized	NN	O	B-protein
anti-CD3/CD28	NN	O	I-protein
antibody	NN	O	I-protein
.	NN	O	O

This	NN	O	O
proliferative	NN	O	O
defect	NN	O	O
is	NN	O	O
not	NN	O	O
reversed	NN	O	O
by	NN	O	O
addition	NN	O	O
of	NN	O	O
exogenous	NN	O	B-protein
IL-2	NN	O	I-protein
and	NN	O	O
is	NN	O	O
p53-independent	NN	O	O
.	NN	O	O

The	NN	O	O
growth	NN	O	O
inhibition	NN	O	O
caused	NN	O	O
by	NN	O	O
Mad1	NN	O	B-protein
is	NN	O	O
overcome	NN	O	O
by	NN	O	O
expression	NN	O	O
of	NN	O	O
active	NN	O	B-protein
c-Myc	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Differential	NN	O	O
requirement	NN	O	O
for	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
PU.1	NN	O	I-protein
in	NN	O	O
the	NN	O	O
generation	NN	O	O
of	NN	O	O
natural	NN	O	B-cell_type
killer	NN	O	I-cell_type
cells	NN	O	I-cell_type
versus	NN	O	O
B	NN	O	B-cell_type
and	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

PU.1	NN	O	B-protein
is	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
Ets	NN	O	B-protein
family	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
required	NN	O	O
for	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
various	NN	O	O
lymphoid	NN	O	B-cell_line
and	NN	O	I-cell_line
myeloid	NN	O	I-cell_line
cell	NN	O	I-cell_line
lineages	NN	O	I-cell_line
,	NN	O	O
but	NN	O	O
its	NN	O	O
role	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
cell	NN	O	I-cell_type
development	NN	O	O
is	NN	O	O
not	NN	O	O
known	NN	O	O
.	NN	O	O

The	NN	O	O
study	NN	O	O
shows	NN	O	O
that	NN	O	O
PU.1	NN	O	B-protein
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
that	NN	O	O
,	NN	O	O
on	NN	O	O
cell	NN	O	O
transfer	NN	O	O
into	NN	O	O
alymphoid	NN	O	O
Rag2/gammac	NN	O	O
(	NN	O	O
-/-	NN	O	O
)	NN	O	O
mice	NN	O	O
,	NN	O	O
hematopoietic	NN	O	B-cell_type
progenitors	NN	O	I-cell_type
of	NN	O	O
PU.1	NN	O	B-cell_type
(	NN	O	I-cell_type
-/-	NN	O	I-cell_type
)	NN	O	I-cell_type
fetal	NN	O	I-cell_type
liver	NN	O	I-cell_type
cells	NN	O	I-cell_type
could	NN	O	O
generate	NN	O	O
functional	NN	O	O
NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
but	NN	O	O
not	NN	O	O
B	NN	O	B-cell_type
or	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Nevertheless	NN	O	O
,	NN	O	O
the	NN	O	O
numbers	NN	O	O
of	NN	O	O
bone	NN	O	B-cell_type
marrow	NN	O	I-cell_type
NK	NN	O	I-cell_type
cell	NN	O	I-cell_type
precursors	NN	O	I-cell_type
and	NN	O	O
splenic	NN	O	B-cell_type
mature	NN	O	I-cell_type
NK	NN	O	I-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
reduced	NN	O	O
compared	NN	O	O
to	NN	O	O
controls	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
PU.1	NN	O	B-cell_type
(	NN	O	I-cell_type
-/-	NN	O	I-cell_type
)	NN	O	I-cell_type
NK	NN	O	I-cell_type
cells	NN	O	I-cell_type
displayed	NN	O	O
reduced	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
receptors	NN	O	O
for	NN	O	O
stem	NN	O	B-protein
cell	NN	O	I-protein
factor	NN	O	I-protein
and	NN	O	O
interleukin	NN	O	B-protein
(	NN	O	I-protein
IL	NN	O	I-protein
)	NN	O	I-protein
-7	NN	O	I-protein
,	NN	O	O
suggesting	NN	O	O
a	NN	O	O
nonredundant	NN	O	O
role	NN	O	O
for	NN	O	O
PU.1	NN	O	B-protein
in	NN	O	O
regulating	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
these	NN	O	O
cytokine	NN	O	B-DNA
receptor	NN	O	I-DNA
genes	NN	O	I-DNA
during	NN	O	O
NK	NN	O	O
cell	NN	O	O
development	NN	O	O
.	NN	O	O

PU.1	NN	O	B-cell_type
(	NN	O	I-cell_type
-/-	NN	O	I-cell_type
)	NN	O	I-cell_type
NK	NN	O	I-cell_type
cells	NN	O	I-cell_type
also	NN	O	O
showed	NN	O	O
defective	NN	O	O
expression	NN	O	O
of	NN	O	O
inhibitory	NN	O	O
and	NN	O	O
activating	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
Ly49	NN	O	B-protein
family	NN	O	I-protein
and	NN	O	O
failed	NN	O	O
to	NN	O	O
proliferate	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
IL-12	NN	O	B-protein
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
despite	NN	O	O
the	NN	O	O
less	NN	O	O
stringent	NN	O	O
requirement	NN	O	O
for	NN	O	O
PU.1	NN	O	B-protein
in	NN	O	O
NK	NN	O	O
cell	NN	O	O
development	NN	O	O
compared	NN	O	O
to	NN	O	O
B	NN	O	B-cell_type
and	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
PU.1	NN	O	B-protein
regulates	NN	O	O
NK	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
and	NN	O	O
homeostasis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Benzene-extracted	NN	O	O
components	NN	O	O
are	NN	O	O
important	NN	O	O
for	NN	O	O
the	NN	O	O
major	NN	O	O
activity	NN	O	O
of	NN	O	O
diesel	NN	O	O
exhaust	NN	O	O
particles	NN	O	O
:	NN	O	O
effect	NN	O	O
on	NN	O	O
interleukin-8	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
bronchial	NN	O	I-cell_type
epithelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Epidemiologic	NN	O	O
and	NN	O	O
experimental	NN	O	O
studies	NN	O	O
suggest	NN	O	O
that	NN	O	O
diesel	NN	O	O
exhaust	NN	O	O
particles	NN	O	O
(	NN	O	O
DEPs	NN	O	O
)	NN	O	O
may	NN	O	O
be	NN	O	O
related	NN	O	O
to	NN	O	O
increasing	NN	O	O
respiratory	NN	O	O
mortality	NN	O	O
and	NN	O	O
morbidity	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
shown	NN	O	O
that	NN	O	O
DEPs	NN	O	O
augmented	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
inflammatory	NN	O	B-protein
cytokines	NN	O	I-protein
by	NN	O	O
human	NN	O	B-cell_type
airway	NN	O	I-cell_type
epithelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

To	NN	O	O
better	NN	O	O
understand	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
their	NN	O	O
proinflammatory	NN	O	O
activities	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
several	NN	O	O
components	NN	O	O
extracted	NN	O	O
from	NN	O	O
DEPs	NN	O	O
on	NN	O	O
interleukin	NN	O	O
(	NN	O	O
IL	NN	O	O
)	NN	O	O
-8	NN	O	O
expression	NN	O	O
in	NN	O	O
human	NN	O	B-cell_line
bronchial	NN	O	I-cell_line
epithelial	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
BEAS-2B	NN	O	I-cell_line
and	NN	O	O
normal	NN	O	B-cell_type
human	NN	O	I-cell_type
airway	NN	O	I-cell_type
epithelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
obtained	NN	O	O
from	NN	O	O
very	NN	O	O
peripheral	NN	O	O
airways	NN	O	O
by	NN	O	O
an	NN	O	O
ultrathin	NN	O	O
bronchoscope	NN	O	O
.	NN	O	O

We	NN	O	O
used	NN	O	O
several	NN	O	O
agents	NN	O	O
active	NN	O	O
on	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathways	NN	O	O
in	NN	O	O
cytokine	NN	O	O
expression	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
the	NN	O	O
protein	NN	O	O
kinase	NN	O	O
C	NN	O	O
inhibitor	NN	O	O
staurosporin	NN	O	O
,	NN	O	O
antioxidant	NN	O	O
agents	NN	O	O
including	NN	O	O
N-acetyl	NN	O	O
cysteine	NN	O	O
(	NN	O	O
NAC	NN	O	O
)	NN	O	O
and	NN	O	O
pyrrolidine	NN	O	O
dithiocarbamate	NN	O	O
(	NN	O	O
PDTC	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
p38	NN	O	B-protein
mitogen-activated	NN	O	I-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	O
MAPK	NN	O	B-protein
)	NN	O	O
inhibitor	NN	O	O
SB203580	NN	O	O
.	NN	O	O

Benzene-extracted	NN	O	O
components	NN	O	O
showed	NN	O	O
effects	NN	O	O
mimicking	NN	O	O
DEPs	NN	O	O
on	NN	O	O
IL-8	NN	O	O
gene	NN	O	O
expression	NN	O	O
,	NN	O	O
release	NN	O	O
of	NN	O	O
several	NN	O	O
cytokines	NN	O	B-protein
(	NN	O	O
IL-8	NN	O	B-protein
;	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
regulated	NN	O	O
on	NN	O	O
activation	NN	O	O
,	NN	O	O
normal	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
expressed	NN	O	O
and	NN	O	O
secreted	NN	O	O
)	NN	O	O
and	NN	O	O
nuclear	NN	O	O
factor	NN	O	O
(	NN	O	O
NF	NN	O	O
)	NN	O	O
-kappa	NN	O	O
B	NN	O	O
activation	NN	O	O
.	NN	O	O

We	NN	O	O
also	NN	O	O
found	NN	O	O
that	NN	O	O
NAC	NN	O	O
,	NN	O	O
PDTC	NN	O	O
,	NN	O	O
and	NN	O	O
SB203580	NN	O	O
suppressed	NN	O	O
the	NN	O	O
activities	NN	O	O
of	NN	O	O
DEPs	NN	O	O
and	NN	O	O
their	NN	O	O
benzene	NN	O	O
extracts	NN	O	O
,	NN	O	O
suggesting	NN	O	O
the	NN	O	O
roles	NN	O	O
of	NN	O	O
oxidants-mediated	NN	O	O
NF-kappa	NN	O	O
B	NN	O	O
activation	NN	O	O
and	NN	O	O
p38MAPK	NN	O	B-protein
pathways	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
benzo	NN	O	O
[	NN	O	O
a	NN	O	O
]	NN	O	O
pyrene	NN	O	O
,	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
important	NN	O	O
compounds	NN	O	O
included	NN	O	O
in	NN	O	O
the	NN	O	O
benzene	NN	O	O
component	NN	O	O
,	NN	O	O
replicated	NN	O	O
the	NN	O	O
activities	NN	O	O
shown	NN	O	O
by	NN	O	O
DEPs	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
nuclear	NN	O	B-protein
receptor	NN	O	I-protein
PPAR	NN	O	I-protein
gamma	NN	O	I-protein
is	NN	O	O
expressed	NN	O	O
by	NN	O	O
mouse	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
PPAR	NN	O	O
gamma	NN	O	O
agonists	NN	O	O
induce	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

Peroxisome	NN	O	B-protein
proliferator-activated	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	I-protein
PPAR	NN	O	I-protein
)	NN	O	I-protein
-gamma	NN	O	I-protein
is	NN	O	O
a	NN	O	O
nuclear	NN	O	B-protein
hormone	NN	O	I-protein
receptor	NN	O	I-protein
that	NN	O	O
serves	NN	O	O
as	NN	O	O
a	NN	O	O
trans	NN	O	O
factor	NN	O	O
to	NN	O	O
regulate	NN	O	O
lipid	NN	O	O
metabolism	NN	O	O
.	NN	O	O

Intense	NN	O	O
interest	NN	O	O
is	NN	O	O
focused	NN	O	O
on	NN	O	O
PPAR-gamma	NN	O	B-protein
and	NN	O	O
its	NN	O	O
ligands	NN	O	O
owing	NN	O	O
to	NN	O	O
its	NN	O	O
putative	NN	O	O
role	NN	O	O
in	NN	O	O
adipocyte	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Little	NN	O	O
is	NN	O	O
known	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
about	NN	O	O
the	NN	O	O
functions	NN	O	O
of	NN	O	O
PPAR-gamma	NN	O	B-protein
in	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
,	NN	O	O
especially	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
demonstrate	NN	O	O
that	NN	O	O
both	NN	O	O
naive	NN	O	B-cell_type
and	NN	O	I-cell_type
activated	NN	O	I-cell_type
ovalbumin-specific	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
DO11.10-transgenic	NN	O	O
mice	NN	O	O
express	NN	O	O
PPAR-gamma	NN	O	B-RNA
mRNA	NN	O	O
and	NN	O	O
protein	NN	O	O
.	NN	O	O

In	NN	O	O
order	NN	O	O
to	NN	O	O
determine	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
PPAR-gamma	NN	O	B-protein
,	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
stimulated	NN	O	O
with	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
and	NN	O	O
ionomycin	NN	O	O
or	NN	O	O
antigen	NN	O	O
and	NN	O	O
antigen-presenting	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Simultaneous	NN	O	O
exposure	NN	O	O
to	NN	O	O
PPAR-gamma	NN	O	O
ligands	NN	O	O
(	NN	O	O
e.g.	NN	O	O
15-deoxy-Delta	NN	O	O
(	NN	O	O
12	NN	O	O
,	NN	O	O
14	NN	O	O
)	NN	O	O
-prostaglandin	NN	O	O
J	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
,	NN	O	O
troglitazone	NN	O	O
)	NN	O	O
showed	NN	O	O
drastic	NN	O	O
inhibition	NN	O	O
of	NN	O	O
proliferation	NN	O	O
and	NN	O	O
significant	NN	O	O
decreases	NN	O	O
in	NN	O	O
cell	NN	O	O
viability	NN	O	O
.	NN	O	O

The	NN	O	O
decrease	NN	O	O
in	NN	O	O
cell	NN	O	O
viability	NN	O	O
was	NN	O	O
due	NN	O	O
to	NN	O	O
apoptosis	NN	O	O
of	NN	O	O
the	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
occurred	NN	O	O
only	NN	O	O
when	NN	O	O
cells	NN	O	O
were	NN	O	O
treated	NN	O	O
with	NN	O	O
PPAR-gamma	NN	O	B-protein
,	NN	O	O
and	NN	O	O
not	NN	O	O
PPAR-alpha	NN	O	O
agonists	NN	O	O
,	NN	O	O
revealing	NN	O	O
specificity	NN	O	O
of	NN	O	O
this	NN	O	O
response	NN	O	O
for	NN	O	O
PPAR-gamma	NN	O	B-protein
.	NN	O	O

These	NN	O	O
observations	NN	O	O
suggest	NN	O	O
that	NN	O	O
PPAR-gamma	NN	O	O
agonists	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
regulating	NN	O	O
T	NN	O	O
cell-mediated	NN	O	O
immune	NN	O	O
responses	NN	O	O
by	NN	O	O
inducing	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

T	NN	O	O
cell	NN	O	O
death	NN	O	O
via	NN	O	O
PPAR-gamma	NN	O	O
ligation	NN	O	O
may	NN	O	O
act	NN	O	O
as	NN	O	O
a	NN	O	O
potent	NN	O	O
anti-inflammatory	NN	O	O
signal	NN	O	O
in	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
,	NN	O	O
and	NN	O	O
ligands	NN	O	O
could	NN	O	O
possibly	NN	O	O
be	NN	O	O
used	NN	O	O
to	NN	O	O
control	NN	O	O
disorders	NN	O	O
in	NN	O	O
which	NN	O	O
excessive	NN	O	O
inflammation	NN	O	O
occurs	NN	O	O
.	NN	O	O

-DOCSTART-	O

Suppression	NN	O	O
of	NN	O	O
lung	NN	O	O
inflammation	NN	O	O
in	NN	O	O
rats	NN	O	O
by	NN	O	O
prevention	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	O
activation	NN	O	O
in	NN	O	O
the	NN	O	O
liver	NN	O	O
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
and	NN	O	O
production	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
-dependent	NN	O	O
chemokines	NN	O	B-protein
are	NN	O	O
thought	NN	O	O
to	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
neutrophilic	NN	O	O
lung	NN	O	O
inflammation	NN	O	O
.	NN	O	O

Calpain-1	NN	O	O
inhibitor	NN	O	O
(	NN	O	O
CI-1	NN	O	O
)	NN	O	O
blocks	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
by	NN	O	O
preventing	NN	O	O
proteolysis	NN	O	O
of	NN	O	O
the	NN	O	O
inhibitory	NN	O	O
protein	NN	O	O
IkappaB-alpha	NN	O	B-protein
by	NN	O	O
the	NN	O	O
ubiquitin/proteasome	NN	O	O
pathway	NN	O	O
.	NN	O	O

We	NN	O	O
hypothesized	NN	O	O
that	NN	O	O
inhibition	NN	O	O
of	NN	O	O
proteasome	NN	O	O
function	NN	O	O
with	NN	O	O
CI-1	NN	O	O
would	NN	O	O
block	NN	O	O
NF-kappaB	NN	O	O
activation	NN	O	O
in	NN	O	O
vivo	NN	O	O
after	NN	O	O
intraperitoneal	NN	O	O
(	NN	O	O
i.p.	NN	O	O
)	NN	O	O
treatment	NN	O	O
with	NN	O	O
bacterial	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
that	NN	O	O
NF-kappaB	NN	O	O
inhibition	NN	O	O
would	NN	O	O
be	NN	O	O
associated	NN	O	O
with	NN	O	O
suppression	NN	O	O
of	NN	O	O
chemokine	NN	O	O
gene	NN	O	O
expression	NN	O	O
and	NN	O	O
attenuation	NN	O	O
of	NN	O	O
neutrophilic	NN	O	O
alveolitis	NN	O	O
.	NN	O	O

We	NN	O	O
treated	NN	O	O
rats	NN	O	O
with	NN	O	O
a	NN	O	O
single	NN	O	O
i.p.	NN	O	O
injection	NN	O	O
of	NN	O	O
CI-1	NN	O	O
(	NN	O	O
10	NN	O	O
mg/kg	NN	O	O
)	NN	O	O
two	NN	O	O
hours	NN	O	O
prior	NN	O	O
to	NN	O	O
i.p.	NN	O	O
LPS	NN	O	O
(	NN	O	O
7	NN	O	O
mg/kg	NN	O	O
)	NN	O	O
.	NN	O	O

Treatment	NN	O	O
with	NN	O	O
Cl-1	NN	O	O
prevented	NN	O	O
degradation	NN	O	O
of	NN	O	O
IkappaB-alpha	NN	O	B-protein
and	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
in	NN	O	O
the	NN	O	O
liver	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
LPS	NN	O	O
;	NN	O	O
however	NN	O	O
,	NN	O	O
Cl-1	NN	O	O
treatment	NN	O	O
had	NN	O	O
no	NN	O	O
detected	NN	O	O
effect	NN	O	O
on	NN	O	O
NF-kappaB	NN	O	O
activation	NN	O	O
in	NN	O	O
lung	NN	O	O
tissue	NN	O	O
.	NN	O	O

CI-1	NN	O	O
treatment	NN	O	O
prior	NN	O	O
to	NN	O	O
LPS	NN	O	O
resulted	NN	O	O
in	NN	O	O
40	NN	O	O
%	NN	O	O
lower	NN	O	O
MIP-2	NN	O	O
concentration	NN	O	O
in	NN	O	O
lung	NN	O	O
lavage	NN	O	O
fluid	NN	O	O
compared	NN	O	O
to	NN	O	O
rats	NN	O	O
treated	NN	O	O
with	NN	O	O
vehicle	NN	O	O
prior	NN	O	O
to	NN	O	O
LPS	NN	O	O
(	NN	O	O
502	NN	O	O
+/-	NN	O	O
112	NN	O	O
pg/ml	NN	O	O
vs.	NN	O	O
859	NN	O	O
+/-144	NN	O	O
pg/ml	NN	O	O
,	NN	O	O
P	NN	O	O
<	NN	O	O
0.05	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
CI-1	NN	O	O
treatment	NN	O	O
substantially	NN	O	O
inhibited	NN	O	O
LPS-induced	NN	O	O
neutrophilic	NN	O	O
alveolitis	NN	O	O
(	NN	O	O
2.7+	NN	O	O
/-	NN	O	O
1.2	NN	O	O
x	NN	O	O
10	NN	O	O
(	NN	O	O
5	NN	O	O
)	NN	O	O
vs.	NN	O	O
43.7	NN	O	O
+/-	NN	O	O
12.2	NN	O	O
x	NN	O	O
10	NN	O	O
(	NN	O	O
5	NN	O	O
)	NN	O	O
lung	NN	O	B-cell_type
lavage	NN	O	I-cell_type
neutrophils	NN	O	I-cell_type
,	NN	O	O
P	NN	O	O
<	NN	O	O
0.01	NN	O	O
)	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
NF-kappaB	NN	O	O
inhibition	NN	O	O
in	NN	O	O
the	NN	O	O
liver	NN	O	O
can	NN	O	O
alter	NN	O	O
lung	NN	O	O
inflammation	NN	O	O
induced	NN	O	O
by	NN	O	O
systemic	NN	O	O
LPS	NN	O	O
treatment	NN	O	O
and	NN	O	O
suggest	NN	O	O
that	NN	O	O
a	NN	O	O
liver-lung	NN	O	O
interaction	NN	O	O
contributes	NN	O	O
to	NN	O	O
the	NN	O	O
inflammatory	NN	O	O
response	NN	O	O
of	NN	O	O
the	NN	O	O
lung	NN	O	O
.	NN	O	O

-DOCSTART-	O

Notch1	NN	O	B-protein
regulates	NN	O	O
maturation	NN	O	O
of	NN	O	O
CD4+	NN	O	B-cell_type
and	NN	O	I-cell_type
CD8+	NN	O	I-cell_type
thymocytes	NN	O	I-cell_type
by	NN	O	O
modulating	NN	O	O
TCR	NN	O	O
signal	NN	O	O
strength	NN	O	O
.	NN	O	O

Notch	NN	O	O
signaling	NN	O	O
regulates	NN	O	O
cell	NN	O	O
fate	NN	O	O
decisions	NN	O	O
in	NN	O	O
multiple	NN	O	O
lineages	NN	O	O
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
in	NN	O	O
this	NN	O	O
report	NN	O	O
that	NN	O	O
retroviral	NN	O	O
expression	NN	O	O
of	NN	O	O
activated	NN	O	O
Notch1	NN	O	B-protein
in	NN	O	O
mouse	NN	O	B-cell_type
thymocytes	NN	O	I-cell_type
abrogates	NN	O	O
differentiation	NN	O	O
of	NN	O	O
immature	NN	O	B-cell_type
CD4+CD8+	NN	O	I-cell_type
thymocytes	NN	O	I-cell_type
into	NN	O	O
both	NN	O	O
CD4	NN	O	B-cell_type
and	NN	O	I-cell_type
CD8	NN	O	I-cell_type
mature	NN	O	I-cell_type
single-positive	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
ability	NN	O	O
of	NN	O	O
Notch1	NN	O	B-protein
to	NN	O	O
inhibit	NN	O	O
T	NN	O	O
cell	NN	O	O
development	NN	O	O
was	NN	O	O
observed	NN	O	O
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
in	NN	O	O
vivo	NN	O	O
with	NN	O	O
both	NN	O	O
normal	NN	O	B-cell_type
and	NN	O	I-cell_type
TCR	NN	O	I-cell_type
transgenic	NN	O	I-cell_type
thymocytes	NN	O	I-cell_type
.	NN	O	O

Notch1	NN	O	B-protein
-mediated	NN	O	O
developmental	NN	O	O
arrest	NN	O	O
was	NN	O	O
dose	NN	O	O
dependent	NN	O	O
and	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
impaired	NN	O	O
thymocyte	NN	O	O
responses	NN	O	O
to	NN	O	O
TCR	NN	O	O
stimulation	NN	O	O
.	NN	O	O

Notch1	NN	O	B-protein
also	NN	O	O
inhibited	NN	O	O
TCR-mediated	NN	O	O
signaling	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

These	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
constitutively	NN	O	O
active	NN	O	O
Notch1	NN	O	B-protein
abrogates	NN	O	O
CD4+	NN	O	O
and	NN	O	O
CD8+	NN	O	O
maturation	NN	O	O
by	NN	O	O
interfering	NN	O	O
with	NN	O	O
TCR	NN	O	O
signal	NN	O	O
strength	NN	O	O
and	NN	O	O
provide	NN	O	O
an	NN	O	O
explanation	NN	O	O
for	NN	O	O
the	NN	O	O
physiological	NN	O	O
regulation	NN	O	O
of	NN	O	O
Notch	NN	O	O
expression	NN	O	O
during	NN	O	O
thymocyte	NN	O	O
development	NN	O	O
.	NN	O	O

-DOCSTART-	O

Expression	NN	O	O
of	NN	O	O
SART3	NN	O	B-protein
antigen	NN	O	I-protein
and	NN	O	O
induction	NN	O	O
of	NN	O	O
CTLs	NN	O	B-cell_type
by	NN	O	O
SART3-derived	NN	O	O
peptides	NN	O	O
in	NN	O	O
breast	NN	O	O
cancer	NN	O	O
patients	NN	O	O
.	NN	O	O

We	NN	O	O
recently	NN	O	O
reported	NN	O	O
the	NN	O	O
SART3	NN	O	B-protein
tumour-rejection	NN	O	I-protein
antigen	NN	O	I-protein
as	NN	O	O
possessing	NN	O	O
tumour	NN	O	O
epitopes	NN	O	O
capable	NN	O	O
of	NN	O	O
inducing	NN	O	O
HLA-class	NN	O	B-cell_type
I-restricted	NN	O	I-cell_type
cytotoxic	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
(	NN	O	O
CTLs	NN	O	B-cell_type
)	NN	O	O
.	NN	O	O

This	NN	O	O
study	NN	O	O
investigated	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
SART3	NN	O	B-protein
antigen	NN	O	I-protein
in	NN	O	O
breast	NN	O	O
cancer	NN	O	O
to	NN	O	O
explore	NN	O	O
an	NN	O	O
appropriate	NN	O	O
molecule	NN	O	O
for	NN	O	O
use	NN	O	O
in	NN	O	O
specific	NN	O	O
immunotherapy	NN	O	O
of	NN	O	O
breast	NN	O	O
cancer	NN	O	O
patients	NN	O	O
.	NN	O	O

The	NN	O	O
SART3	NN	O	B-protein
antigen	NN	O	I-protein
was	NN	O	O
detected	NN	O	O
in	NN	O	O
all	NN	O	O
of	NN	O	O
the	NN	O	O
breast	NN	O	B-cell_line
cancer	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
tested	NN	O	O
,	NN	O	O
30	NN	O	O
of	NN	O	O
40	NN	O	O
(	NN	O	O
75	NN	O	O
%	NN	O	O
)	NN	O	O
breast	NN	O	O
cancer	NN	O	O
tissue	NN	O	O
samples	NN	O	O
,	NN	O	O
and	NN	O	O
0	NN	O	O
of	NN	O	O
3	NN	O	O
non-tumourous	NN	O	O
breast	NN	O	O
tissue	NN	O	O
samples	NN	O	O
.	NN	O	O

SART3	NN	O	O
derived	NN	O	O
peptides	NN	O	O
at	NN	O	O
positions	NN	O	B-DNA
109-118	NN	O	I-DNA
and	NN	O	I-DNA
315-323	NN	O	I-DNA
induced	NN	O	O
HLA-A24	NN	O	O
restricted	NN	O	O
CTLs	NN	O	B-cell_type
that	NN	O	O
reacted	NN	O	O
to	NN	O	O
breast	NN	O	B-cell_type
cancer	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
the	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
PBMCs	NN	O	B-cell_type
)	NN	O	O
of	NN	O	O
breast	NN	O	O
cancer	NN	O	O
patients	NN	O	O
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
the	NN	O	O
SART3	NN	O	B-protein
antigen	NN	O	I-protein
and	NN	O	O
its	NN	O	O
peptides	NN	O	O
could	NN	O	O
be	NN	O	O
an	NN	O	O
appropriate	NN	O	O
molecule	NN	O	O
for	NN	O	O
use	NN	O	O
in	NN	O	O
specific	NN	O	O
immunotherapy	NN	O	O
of	NN	O	O
the	NN	O	O
majority	NN	O	O
of	NN	O	O
HLA-A24-positive	NN	O	O
breast	NN	O	O
cancer	NN	O	O
patients	NN	O	O

-DOCSTART-	O

Autostimulation	NN	O	O
of	NN	O	O
the	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
BRLF1	NN	O	B-DNA
promoter	NN	O	I-DNA
is	NN	O	O
mediated	NN	O	O
through	NN	O	O
consensus	NN	O	O
Sp1	NN	O	B-DNA
and	NN	O	I-DNA
Sp3	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
.	NN	O	O

As	NN	O	O
an	NN	O	O
essential	NN	O	O
step	NN	O	O
in	NN	O	O
the	NN	O	O
lytic	NN	O	O
cascade	NN	O	O
,	NN	O	O
the	NN	O	O
Rta	NN	O	O
homologues	NN	O	O
of	NN	O	O
gammaherpesviruses	NN	O	O
all	NN	O	O
activate	NN	O	O
their	NN	O	O
own	NN	O	O
expression	NN	O	O
.	NN	O	O

Consistent	NN	O	O
with	NN	O	O
this	NN	O	O
biologic	NN	O	O
function	NN	O	O
,	NN	O	O
the	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
(	NN	O	O
EBV	NN	O	O
)	NN	O	O
Rta	NN	O	B-protein
protein	NN	O	I-protein
powerfully	NN	O	O
stimulates	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
of	NN	O	O
its	NN	O	O
own	NN	O	O
gene	NN	O	O
,	NN	O	O
Rp	NN	O	B-DNA
,	NN	O	O
in	NN	O	O
EBV-positive	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
transient-transfection	NN	O	O
reporter-based	NN	O	O
assays	NN	O	O
.	NN	O	O

We	NN	O	O
analyzed	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
RpCAT	NN	O	B-DNA
in	NN	O	O
response	NN	O	O
to	NN	O	O
Rta	NN	O	B-protein
by	NN	O	O
deletional	NN	O	O
and	NN	O	O
site-directed	NN	O	O
mutagenesis	NN	O	O
.	NN	O	O

Two	NN	O	O
cognate	NN	O	B-DNA
Sp1	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
located	NN	O	O
at	NN	O	O
-279	NN	O	O
and	NN	O	O
-45	NN	O	O
relative	NN	O	O
to	NN	O	O
the	NN	O	O
transcriptional	NN	O	B-DNA
start	NN	O	I-DNA
site	NN	O	I-DNA
proved	NN	O	O
crucial	NN	O	O
for	NN	O	O
Rta-mediated	NN	O	O
activation	NN	O	O
.	NN	O	O

Previously	NN	O	O
described	NN	O	O
binding	NN	O	O
sites	NN	O	O
for	NN	O	O
the	NN	O	O
cellular	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
Zif268	NN	O	I-protein
and	NN	O	O
the	NN	O	O
viral	NN	O	B-protein
transactivator	NN	O	I-protein
ZEBRA	NN	O	I-protein
were	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
dispensable	NN	O	O
for	NN	O	O
activation	NN	O	O
of	NN	O	O
RpCAT	NN	O	B-DNA
by	NN	O	O
Rta	NN	O	B-protein
.	NN	O	O

Gel	NN	O	O
shift	NN	O	O
analysis	NN	O	O
,	NN	O	O
using	NN	O	O
extracts	NN	O	O
of	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
latency	NN	O	O
or	NN	O	O
induced	NN	O	O
into	NN	O	O
the	NN	O	O
lytic	NN	O	O
cycle	NN	O	O
,	NN	O	O
identified	NN	O	O
Sp1	NN	O	B-protein
and	NN	O	O
Sp3	NN	O	B-protein
as	NN	O	O
the	NN	O	O
predominant	NN	O	O
cellular	NN	O	B-protein
proteins	NN	O	I-protein
bound	NN	O	O
to	NN	O	O
Rp	NN	O	B-DNA
near	NN	O	O
-45	NN	O	O
.	NN	O	O

During	NN	O	O
the	NN	O	O
lytic	NN	O	O
cycle	NN	O	O
,	NN	O	O
ZEBRA	NN	O	B-protein
bound	NN	O	O
Rp	NN	O	B-DNA
near	NN	O	O
the	NN	O	O
Sp1/Sp3	NN	O	B-DNA
site	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
binding	NN	O	O
of	NN	O	O
Sp1	NN	O	B-protein
and	NN	O	O
Sp3	NN	O	B-protein
to	NN	O	O
Rp	NN	O	B-DNA
correlated	NN	O	O
with	NN	O	O
the	NN	O	O
reporter	NN	O	O
activities	NN	O	O
in	NN	O	O
the	NN	O	O
mutagenesis	NN	O	O
study	NN	O	O
,	NN	O	O
establishing	NN	O	O
a	NN	O	O
direct	NN	O	O
link	NN	O	O
between	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
Rp	NN	O	B-DNA
by	NN	O	O
Rta	NN	O	B-protein
and	NN	O	O
DNA	NN	O	O
binding	NN	O	O
by	NN	O	O
Sp1	NN	O	B-protein
and/or	NN	O	O
Sp3	NN	O	B-protein
.	NN	O	O

The	NN	O	O
relative	NN	O	O
abundance	NN	O	O
or	NN	O	O
functional	NN	O	O
state	NN	O	O
of	NN	O	O
the	NN	O	O
cellular	NN	O	O
Sp1	NN	O	B-protein
and	NN	O	O
Sp3	NN	O	B-protein
transcription	NN	O	B-protein
factors	NN	O	I-protein
may	NN	O	O
be	NN	O	O
altered	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
stimuli	NN	O	O
that	NN	O	O
induce	NN	O	O
the	NN	O	O
BRLF1	NN	O	B-DNA
promoter	NN	O	I-DNA
and	NN	O	O
thereby	NN	O	O
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
viral	NN	O	O
lytic	NN	O	O
cycle	NN	O	O
.	NN	O	O

-DOCSTART-	O

Gene	NN	O	O
transfer	NN	O	O
of	NN	O	O
antisense	NN	O	O
hypoxia	NN	O	B-protein
inducible	NN	O	I-protein
factor-1	NN	O	I-protein
alpha	NN	O	I-protein
enhances	NN	O	O
the	NN	O	O
therapeutic	NN	O	O
efficacy	NN	O	O
of	NN	O	O
cancer	NN	O	O
immunotherapy	NN	O	O
.	NN	O	O

Solid	NN	O	O
tumors	NN	O	O
meet	NN	O	O
their	NN	O	O
demands	NN	O	O
for	NN	O	O
nascent	NN	O	O
blood	NN	O	O
vessels	NN	O	O
and	NN	O	O
increased	NN	O	O
glycolysis	NN	O	O
,	NN	O	O
to	NN	O	O
combat	NN	O	O
hypoxia	NN	O	O
,	NN	O	O
by	NN	O	O
activating	NN	O	O
multiple	NN	O	O
genes	NN	O	O
involved	NN	O	O
in	NN	O	O
angiogenesis	NN	O	O
and	NN	O	O
glucose	NN	O	O
metabolism	NN	O	O
.	NN	O	O

Hypoxia	NN	O	B-protein
inducible	NN	O	I-protein
factor-1	NN	O	I-protein
(	NN	O	O
HIF-1	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
constitutively	NN	O	O
expressed	NN	O	O
basic	NN	O	O
helix-loop-helix	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
formed	NN	O	O
by	NN	O	O
the	NN	O	O
assembly	NN	O	O
of	NN	O	O
HIF-1alpha	NN	O	B-protein
and	NN	O	O
HIF-1beta	NN	O	B-protein
(	NN	O	O
Arnt	NN	O	B-protein
)	NN	O	O
,	NN	O	O
that	NN	O	O
is	NN	O	O
stablized	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
hypoxia	NN	O	O
,	NN	O	O
and	NN	O	O
rapidly	NN	O	O
degraded	NN	O	O
under	NN	O	O
normoxic	NN	O	O
conditions	NN	O	O
.	NN	O	O

It	NN	O	O
activates	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
genes	NN	O	O
important	NN	O	O
for	NN	O	O
maintaining	NN	O	O
oxygen	NN	O	O
homeostasis	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
engineered	NN	O	O
down-regulation	NN	O	O
of	NN	O	O
HIF-1alpha	NN	O	B-protein
by	NN	O	O
intratumoral	NN	O	O
gene	NN	O	O
transfer	NN	O	O
of	NN	O	O
an	NN	O	O
antisense	NN	O	B-DNA
HIF-1alpha	NN	O	I-DNA
plasmid	NN	O	I-DNA
leads	NN	O	O
to	NN	O	O
the	NN	O	O
down-regulation	NN	O	O
of	NN	O	O
VEGF	NN	O	B-DNA
,	NN	O	O
and	NN	O	O
decreased	NN	O	O
tumor	NN	O	O
microvessel	NN	O	O
density	NN	O	O
.	NN	O	O

Antisense	NN	O	O
HIF-1alpha	NN	O	O
monotherapy	NN	O	O
resulted	NN	O	O
in	NN	O	O
the	NN	O	O
complete	NN	O	O
and	NN	O	O
permanent	NN	O	O
rejection	NN	O	O
of	NN	O	O
small	NN	O	O
(	NN	O	O
0.1	NN	O	O
cm	NN	O	O
in	NN	O	O
diameter	NN	O	O
)	NN	O	O
EL-4	NN	O	O
tumors	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
unusual	NN	O	O
for	NN	O	O
an	NN	O	O
anti-angiogenic	NN	O	O
agent	NN	O	O
where	NN	O	O
transient	NN	O	O
suppression	NN	O	O
of	NN	O	O
tumor	NN	O	O
growth	NN	O	O
is	NN	O	O
the	NN	O	O
norm	NN	O	O
.	NN	O	O

It	NN	O	O
induced	NN	O	O
NK	NN	O	B-cell_type
cell	NN	O	I-cell_type
-dependent	NN	O	O
rejection	NN	O	O
of	NN	O	O
tumors	NN	O	O
,	NN	O	O
but	NN	O	O
failed	NN	O	O
to	NN	O	O
stimulate	NN	O	O
systemic	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
-mediated	NN	O	O
anti-tumor	NN	O	O
immunity	NN	O	O
,	NN	O	O
and	NN	O	O
synergized	NN	O	O
with	NN	O	O
B7-1-mediated	NN	O	O
immunotherapy	NN	O	O
to	NN	O	O
cause	NN	O	O
the	NN	O	O
NK	NN	O	B-cell_type
cell	NN	O	I-cell_type
and	NN	O	O
CD8	NN	O	B-cell_type
T	NN	O	I-cell_type
cell	NN	O	I-cell_type
-dependent	NN	O	O
rejection	NN	O	O
of	NN	O	O
larger	NN	O	O
EL-4	NN	O	O
tumors	NN	O	O
(	NN	O	O
0.4	NN	O	O
cm	NN	O	O
in	NN	O	O
diameter	NN	O	O
)	NN	O	O
that	NN	O	O
were	NN	O	O
refractory	NN	O	O
to	NN	O	O
monotherapies	NN	O	O
.	NN	O	O

Mice	NN	O	O
cured	NN	O	O
of	NN	O	O
their	NN	O	O
tumors	NN	O	O
by	NN	O	O
combination	NN	O	O
therapy	NN	O	O
resisted	NN	O	O
a	NN	O	O
rechallenge	NN	O	O
with	NN	O	O
parental	NN	O	B-cell_type
tumor	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
indicating	NN	O	O
systemic	NN	O	O
antitumor	NN	O	O
immunity	NN	O	O
had	NN	O	O
been	NN	O	O
achieved	NN	O	O
.	NN	O	O

In	NN	O	O
summary	NN	O	O
,	NN	O	O
whilst	NN	O	O
intensive	NN	O	O
investigations	NN	O	O
are	NN	O	O
in	NN	O	O
progress	NN	O	O
to	NN	O	O
target	NN	O	O
the	NN	O	O
many	NN	O	O
HIF-1	NN	O	B-protein
effectors	NN	O	O
,	NN	O	O
the	NN	O	O
results	NN	O	O
herein	NN	O	O
indicate	NN	O	O
that	NN	O	O
blocking	NN	O	O
hypoxia-inducible	NN	O	O
pathways	NN	O	O
and	NN	O	O
enhancing	NN	O	O
NK-mediated	NN	O	O
antitumor	NN	O	O
immunity	NN	O	O
by	NN	O	O
targeting	NN	O	O
HIF-1	NN	O	B-protein
itself	NN	O	O
may	NN	O	O
be	NN	O	O
advantageous	NN	O	O
,	NN	O	O
especially	NN	O	O
when	NN	O	O
combined	NN	O	O
with	NN	O	O
cancer	NN	O	O
immunotherapy	NN	O	O
.	NN	O	O

-DOCSTART-	O

Runx2	NN	O	B-DNA
:	NN	O	O
a	NN	O	O
novel	NN	O	O
oncogenic	NN	O	O
effector	NN	O	O
revealed	NN	O	O
by	NN	O	O
in	NN	O	O
vivo	NN	O	O
complementation	NN	O	O
and	NN	O	O
retroviral	NN	O	O
tagging	NN	O	O
.	NN	O	O

The	NN	O	O
Runx2	NN	O	B-DNA
(	NN	O	O
Cbfa1	NN	O	B-DNA
,	NN	O	O
Pebp2alphaA	NN	O	B-DNA
,	NN	O	O
Aml3	NN	O	B-DNA
)	NN	O	O
gene	NN	O	O
was	NN	O	O
previously	NN	O	O
identified	NN	O	O
as	NN	O	O
a	NN	O	O
frequent	NN	O	O
target	NN	O	O
for	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
by	NN	O	O
proviral	NN	O	O
insertion	NN	O	O
in	NN	O	O
T-cell	NN	O	O
lymphomas	NN	O	O
of	NN	O	O
CD2-MYC	NN	O	O
transgenic	NN	O	O
mice	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
over-expression	NN	O	O
of	NN	O	O
the	NN	O	O
full-length	NN	O	O
,	NN	O	O
most	NN	O	O
highly	NN	O	O
expressed	NN	O	O
Runx2	NN	O	B-DNA
isoform	NN	O	O
in	NN	O	O
the	NN	O	O
thymus	NN	O	O
perturbs	NN	O	O
T-cell	NN	O	O
development	NN	O	O
,	NN	O	O
leads	NN	O	O
to	NN	O	O
development	NN	O	O
of	NN	O	O
spontaneous	NN	O	O
lymphomas	NN	O	O
at	NN	O	O
low	NN	O	O
frequency	NN	O	O
and	NN	O	O
is	NN	O	O
strongly	NN	O	O
synergistic	NN	O	O
with	NN	O	O
Myc	NN	O	B-protein
.	NN	O	O

To	NN	O	O
gain	NN	O	O
further	NN	O	O
insight	NN	O	O
into	NN	O	O
the	NN	O	O
relationship	NN	O	O
of	NN	O	O
Runx2	NN	O	B-DNA
to	NN	O	O
other	NN	O	O
lymphomagenic	NN	O	O
pathways	NN	O	O
,	NN	O	O
we	NN	O	O
tested	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
combining	NN	O	O
the	NN	O	O
CD2-Runx2	NN	O	B-DNA
transgene	NN	O	I-DNA
either	NN	O	O
with	NN	O	O
a	NN	O	O
Pim1	NN	O	B-DNA
transgene	NN	O	I-DNA
(	NN	O	O
E	NN	O	B-DNA
(	NN	O	I-DNA
mu	NN	O	I-DNA
)	NN	O	I-DNA
-Pim1	NN	O	I-DNA
)	NN	O	O
or	NN	O	O
with	NN	O	O
the	NN	O	O
p53	NN	O	B-DNA
null	NN	O	O
genotype	NN	O	O
,	NN	O	O
as	NN	O	O
each	NN	O	O
of	NN	O	O
these	NN	O	O
displays	NN	O	O
independent	NN	O	O
synergy	NN	O	O
with	NN	O	O
Myc	NN	O	B-protein
.	NN	O	O

In	NN	O	O
both	NN	O	O
cases	NN	O	O
we	NN	O	O
observed	NN	O	O
synergistic	NN	O	O
tumour	NN	O	O
development	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
Runx2	NN	O	B-protein
appeared	NN	O	O
to	NN	O	O
have	NN	O	O
a	NN	O	O
dominant	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
tumour	NN	O	O
phenotype	NN	O	O
in	NN	O	O
each	NN	O	O
case	NN	O	O
,	NN	O	O
with	NN	O	O
most	NN	O	O
tumours	NN	O	O
conforming	NN	O	O
to	NN	O	O
the	NN	O	O
CD3	NN	O	O
(	NN	O	O
+	NN	O	O
)	NN	O	O
,	NN	O	O
CD8	NN	O	O
(	NN	O	O
+	NN	O	O
)	NN	O	O
,	NN	O	O
CD4	NN	O	O
(	NN	O	O
+/-	NN	O	O
)	NN	O	O
phenotype	NN	O	O
seen	NN	O	O
in	NN	O	O
CD2-Runx2	NN	O	O
mice	NN	O	O
.	NN	O	O

Neonatal	NN	O	O
infection	NN	O	O
of	NN	O	O
CD2-Runx2	NN	O	O
mice	NN	O	O
with	NN	O	O
Moloney	NN	O	O
murine	NN	O	O
leukaemia	NN	O	O
virus	NN	O	O
(	NN	O	O
Moloney	NN	O	O
MLV	NN	O	O
)	NN	O	O
also	NN	O	O
led	NN	O	O
to	NN	O	O
a	NN	O	O
dramatic	NN	O	O
acceleration	NN	O	O
of	NN	O	O
tumour	NN	O	O
onset	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
known	NN	O	O
Moloney	NN	O	B-DNA
MLV	NN	O	I-DNA
target	NN	O	I-DNA
genes	NN	O	I-DNA
in	NN	O	O
these	NN	O	O
lymphomas	NN	O	O
showed	NN	O	O
a	NN	O	O
high	NN	O	O
frequency	NN	O	O
of	NN	O	O
rearrangement	NN	O	O
at	NN	O	O
c-Myc	NN	O	B-DNA
or	NN	O	O
N-Myc	NN	O	B-DNA
(	NN	O	O
82	NN	O	O
%	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
a	NN	O	O
significant	NN	O	O
number	NN	O	O
at	NN	O	O
Pim1	NN	O	B-DNA
or	NN	O	O
Pim2	NN	O	B-DNA
(	NN	O	O
23	NN	O	O
%	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
at	NN	O	O
Pal1/Gfi1	NN	O	B-DNA
(	NN	O	O
18	NN	O	O
%	NN	O	O
)	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
Runx2	NN	O	B-protein
makes	NN	O	O
a	NN	O	O
distinct	NN	O	O
contribution	NN	O	O
to	NN	O	O
T-cell	NN	O	O
lymphoma	NN	O	O
development	NN	O	O
which	NN	O	O
does	NN	O	O
not	NN	O	O
coincide	NN	O	O
with	NN	O	O
any	NN	O	O
of	NN	O	O
the	NN	O	O
oncogene	NN	O	O
complementation	NN	O	O
groups	NN	O	O
previously	NN	O	O
identified	NN	O	O
by	NN	O	O
retroviral	NN	O	O
tagging	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
involvement	NN	O	O
of	NN	O	O
TNF-alpha	NN	O	B-protein
-related	NN	O	O
apoptosis-inducing	NN	O	O
ligand	NN	O	O
in	NN	O	O
the	NN	O	O
enhanced	NN	O	O
cytotoxicity	NN	O	O
of	NN	O	O
IFN-beta	NN	O	B-protein
-stimulated	NN	O	O
human	NN	O	B-cell_type
dendritic	NN	O	I-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
tumor	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

TNF-alpha	NN	O	B-protein
-related	NN	O	O
apoptosis-inducing	NN	O	O
ligand	NN	O	O
(	NN	O	O
TRAIL	NN	O	O
)	NN	O	O
is	NN	O	O
characterized	NN	O	O
by	NN	O	O
its	NN	O	O
preferential	NN	O	O
induction	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
of	NN	O	O
tumor	NN	O	B-cell_type
cells	NN	O	I-cell_type
but	NN	O	O
not	NN	O	O
normal	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	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
besides	NN	O	O
their	NN	O	O
role	NN	O	O
as	NN	O	O
APCs	NN	O	O
,	NN	O	O
now	NN	O	O
have	NN	O	O
been	NN	O	O
demonstrated	NN	O	O
to	NN	O	O
exert	NN	O	O
cytotoxicity	NN	O	O
or	NN	O	O
cytostasis	NN	O	O
on	NN	O	O
some	NN	O	O
tumor	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
both	NN	O	O
human	NN	O	B-cell_type
CD34	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
stem	NN	O	I-cell_type
cell	NN	O	I-cell_type
-derived	NN	O	O
DCs	NN	O	B-cell_type
(	NN	O	O
CD34DCs	NN	O	B-cell_type
)	NN	O	O
and	NN	O	O
human	NN	O	B-cell_type
CD14	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
monocyte	NN	O	I-cell_type
-derived	NN	O	O
DCs	NN	O	B-cell_type
(	NN	O	O
MoDCs	NN	O	B-cell_type
)	NN	O	O
express	NN	O	O
TRAIL	NN	O	O
and	NN	O	O
exhibit	NN	O	O
cytotoxicity	NN	O	O
to	NN	O	O
some	NN	O	O
types	NN	O	O
of	NN	O	O
tumor	NN	O	B-cell_type
cells	NN	O	I-cell_type
partially	NN	O	O
through	NN	O	O
TRAIL	NN	O	O
.	NN	O	O

Moderate	NN	O	O
expression	NN	O	O
of	NN	O	O
TRAIL	NN	O	O
appeared	NN	O	O
on	NN	O	O
CD34DCs	NN	O	B-cell_type
from	NN	O	O
the	NN	O	O
8th	NN	O	O
day	NN	O	O
of	NN	O	O
culture	NN	O	O
and	NN	O	O
was	NN	O	O
also	NN	O	O
seen	NN	O	O
on	NN	O	O
freshly	NN	O	B-cell_type
isolated	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
level	NN	O	O
of	NN	O	O
TRAIL	NN	O	O
expression	NN	O	O
remained	NN	O	O
constant	NN	O	O
until	NN	O	O
DC	NN	O	O
maturation	NN	O	O
.	NN	O	O

TRAIL	NN	O	O
expression	NN	O	O
on	NN	O	O
immature	NN	O	B-cell_type
CD34DCs	NN	O	I-cell_type
or	NN	O	O
MoDCs	NN	O	B-cell_type
was	NN	O	O
greatly	NN	O	O
up-regulated	NN	O	O
after	NN	O	O
IFN-beta	NN	O	B-protein
stimulation	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
IFN-beta	NN	O	B-protein
could	NN	O	O
strikingly	NN	O	O
enhance	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
CD34DCs	NN	O	B-cell_type
or	NN	O	O
MoDCs	NN	O	B-cell_type
to	NN	O	O
kill	NN	O	O
TRAIL-sensitive	NN	O	O
tumor	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
but	NN	O	O
LPS	NN	O	O
did	NN	O	O
not	NN	O	O
have	NN	O	O
such	NN	O	O
an	NN	O	O
effect	NN	O	O
.	NN	O	O

The	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
TRAIL	NN	O	O
on	NN	O	O
IFN-beta	NN	O	B-protein
-stimulated	NN	O	O
DCs	NN	O	B-cell_type
partially	NN	O	O
contributed	NN	O	O
to	NN	O	O
the	NN	O	O
increased	NN	O	O
cytotoxicity	NN	O	O
of	NN	O	O
DCS	NN	O	B-cell_type
:	NN	O	O
Pretreatment	NN	O	O
of	NN	O	O
TRAIL-sensitive	NN	O	O
tumor	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
caspase-3	NN	O	O
inhibitor	NN	O	O
could	NN	O	O
significantly	NN	O	O
increase	NN	O	O
their	NN	O	O
resistance	NN	O	O
to	NN	O	O
the	NN	O	O
cytotoxicity	NN	O	O
of	NN	O	O
IFN-beta	NN	O	B-protein
-stimulated	NN	O	O
DCS	NN	O	B-cell_type
:	NN	O	O
In	NN	O	O
contrast	NN	O	O
,	NN	O	O
NF-kappaB	NN	O	O
inhibitor	NN	O	O
could	NN	O	O
significantly	NN	O	O
increase	NN	O	O
the	NN	O	O
sensitivity	NN	O	O
of	NN	O	O
tumor	NN	O	B-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
the	NN	O	O
killing	NN	O	O
by	NN	O	O
nonstimulated	NN	O	O
or	NN	O	O
LPS-stimulated	NN	O	O
DCS	NN	O	B-cell_type
:	NN	O	O
Our	NN	O	O
studies	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
IFN-beta	NN	O	B-protein
-stimulated	NN	O	O
DCs	NN	O	B-cell_type
are	NN	O	O
functionally	NN	O	O
cytotoxic	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
an	NN	O	O
innate	NN	O	O
mechanism	NN	O	O
of	NN	O	O
DC-mediated	NN	O	O
antitumor	NN	O	O
immunity	NN	O	O
might	NN	O	O
exist	NN	O	O
in	NN	O	O
vivo	NN	O	O
in	NN	O	O
which	NN	O	O
DCs	NN	O	B-cell_type
act	NN	O	O
as	NN	O	O
effectors	NN	O	O
to	NN	O	O
directly	NN	O	O
kill	NN	O	O
tumor	NN	O	B-cell_type
cells	NN	O	I-cell_type
partially	NN	O	O
via	NN	O	O
TRAIL	NN	O	O
.	NN	O	O

Subsequently	NN	O	O
,	NN	O	O
DCs	NN	O	B-cell_type
act	NN	O	O
as	NN	O	O
APCs	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
uptake	NN	O	O
,	NN	O	O
processing	NN	O	O
,	NN	O	O
and	NN	O	O
presentation	NN	O	O
of	NN	O	O
apoptotic	NN	O	B-protein
tumor	NN	O	I-protein
Ags	NN	O	I-protein
to	NN	O	O
cross-prime	NN	O	O
CD8	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
CTL	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Tumor	NN	O	B-cell_type
cells	NN	O	I-cell_type
regulate	NN	O	O
the	NN	O	O
lytic	NN	O	O
activity	NN	O	O
of	NN	O	O
tumor-specific	NN	O	B-cell_type
cytotoxic	NN	O	I-cell_type
t	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
by	NN	O	O
modulating	NN	O	O
the	NN	O	O
inhibitory	NN	O	O
natural	NN	O	O
killer	NN	O	O
receptor	NN	O	O
function	NN	O	O
.	NN	O	O

Tumor-infiltrating	NN	O	B-cell_type
p58+	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
a	NN	O	O
renal	NN	O	O
tumor	NN	O	O
were	NN	O	O
specifically	NN	O	O
expanded	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
tumor	NN	O	O
cell	NN	O	O
stimulation	NN	O	O
and	NN	O	O
cloned	NN	O	O
.	NN	O	O

These	NN	O	O
p58+	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
found	NN	O	O
to	NN	O	O
express	NN	O	O
a	NN	O	O
memory	NN	O	B-cell_type
phenotype	NN	O	I-cell_type
and	NN	O	O
corresponded	NN	O	O
to	NN	O	O
clonal	NN	O	O
TCRBV3	NN	O	O
T-cell	NN	O	O
expansion	NN	O	O
.	NN	O	O

Functionally	NN	O	O
,	NN	O	O
p58	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
CTLs	NN	O	I-cell_type
displayed	NN	O	O
a	NN	O	O
low	NN	O	O
lytic	NN	O	O
activity	NN	O	O
for	NN	O	O
HLA-A2	NN	O	B-cell_type
tumor	NN	O	I-cell_type
and	NN	O	I-cell_type
normal	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

However	NN	O	O
,	NN	O	O
this	NN	O	O
lytic	NN	O	O
activity	NN	O	O
was	NN	O	O
significantly	NN	O	O
increased	NN	O	O
after	NN	O	O
blockade	NN	O	O
of	NN	O	O
p58	NN	O	B-protein
with	NN	O	O
specific	NN	O	O
monoclonal	NN	O	B-protein
antibodies	NN	O	I-protein
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
stimulation	NN	O	O
by	NN	O	O
tumor	NN	O	B-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
required	NN	O	O
to	NN	O	O
trigger	NN	O	O
the	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
of	NN	O	O
p58	NN	O	B-protein
on	NN	O	O
the	NN	O	O
lytic	NN	O	O
activity	NN	O	O
of	NN	O	O
antigen-specific	NN	O	B-cell_type
CTLs	NN	O	I-cell_type
and	NN	O	O
that	NN	O	O
stimulation	NN	O	O
of	NN	O	O
the	NN	O	O
inhibitory	NN	O	O
function	NN	O	O
of	NN	O	O
p58	NN	O	B-protein
by	NN	O	O
tumor	NN	O	B-cell_type
cells	NN	O	I-cell_type
correlated	NN	O	O
with	NN	O	O
an	NN	O	O
inhibition	NN	O	O
of	NN	O	O
nuclear	NN	O	O
factor-kappaB	NN	O	O
activation	NN	O	O
in	NN	O	O
p58+	NN	O	B-cell_type
tumor-specific	NN	O	I-cell_type
CTLS	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

T-cell	NN	O	B-protein
factor-1	NN	O	I-protein
expression	NN	O	O
during	NN	O	O
human	NN	O	B-cell_type
natural	NN	O	I-cell_type
killer	NN	O	I-cell_type
cell	NN	O	I-cell_type
development	NN	O	O
and	NN	O	O
in	NN	O	O
circulating	NN	O	O
CD56	NN	O	B-protein
(	NN	O	I-protein
+	NN	O	I-protein
)	NN	O	I-protein
bright	NN	O	O
natural	NN	O	B-cell_type
killer	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Transcription	NN	O	B-protein
factors	NN	O	I-protein
are	NN	O	O
essential	NN	O	O
to	NN	O	O
govern	NN	O	O
differentiation	NN	O	O
along	NN	O	O
the	NN	O	O
lymphoid	NN	O	B-cell_type
lineage	NN	O	I-cell_type
from	NN	O	O
uncommitted	NN	O	B-cell_type
hematopoietic	NN	O	I-cell_type
stem	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Although	NN	O	O
many	NN	O	O
of	NN	O	O
these	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
have	NN	O	O
putative	NN	O	O
roles	NN	O	O
based	NN	O	O
on	NN	O	O
murine	NN	O	O
knockout	NN	O	O
experiments	NN	O	O
,	NN	O	O
their	NN	O	O
function	NN	O	O
in	NN	O	O
human	NN	O	O
lymphoid	NN	O	O
development	NN	O	O
is	NN	O	O
less	NN	O	O
known	NN	O	O
and	NN	O	O
was	NN	O	O
studied	NN	O	O
further	NN	O	O
.	NN	O	O

Transcription	NN	O	O
factor	NN	O	O
expression	NN	O	O
in	NN	O	O
fresh	NN	O	O
and	NN	O	O
cultured	NN	O	O
adult	NN	O	B-cell_type
human	NN	O	I-cell_type
bone	NN	O	I-cell_type
marrow	NN	O	I-cell_type
and	NN	O	I-cell_type
umbilical	NN	O	I-cell_type
cord	NN	O	I-cell_type
blood	NN	O	I-cell_type
progenitors	NN	O	I-cell_type
was	NN	O	O
evaluated	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
fresh	NN	O	B-cell_type
CD34	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
Lin	NN	O	I-cell_type
(	NN	O	I-cell_type
-	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
that	NN	O	O
are	NN	O	O
human	NN	O	O
leukocyte	NN	O	O
antigen	NN	O	O
(	NN	O	O
HLA	NN	O	O
)	NN	O	O
-DR	NN	O	O
(	NN	O	O
-	NN	O	O
)	NN	O	O
or	NN	O	O
CD38	NN	O	O
(	NN	O	O
-	NN	O	O
)	NN	O	O
constitutively	NN	O	O
express	NN	O	O
GATA-3	NN	O	B-protein
but	NN	O	O
not	NN	O	O
T-cell	NN	O	B-protein
factor-1	NN	O	I-protein
(	NN	O	O
TCF-1	NN	O	B-protein
)	NN	O	O
or	NN	O	O
Id-3	NN	O	B-protein
.	NN	O	O

Culture	NN	O	O
with	NN	O	O
the	NN	O	O
murine	NN	O	B-cell_line
fetal	NN	O	I-cell_line
liver	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
AFT024	NN	O	I-cell_line
and	NN	O	O
defined	NN	O	O
cytokines	NN	O	B-protein
was	NN	O	O
capable	NN	O	O
of	NN	O	O
inducing	NN	O	O
TCF-1	NN	O	B-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

However	NN	O	O
,	NN	O	O
no	NN	O	O
T-cell	NN	O	B-DNA
receptor	NN	O	I-DNA
gene	NN	O	I-DNA
rearrangement	NN	O	O
was	NN	O	O
identified	NN	O	O
in	NN	O	O
cultured	NN	O	O
progeny	NN	O	O
.	NN	O	O

Id-3	NN	O	B-protein
,	NN	O	O
a	NN	O	O
basic	NN	O	B-protein
helix	NN	O	I-protein
loop	NN	O	I-protein
helix	NN	O	I-protein
factor	NN	O	I-protein
with	NN	O	O
dominant	NN	O	O
negative	NN	O	O
function	NN	O	O
for	NN	O	O
T-cell	NN	O	O
differentiation	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
also	NN	O	O
was	NN	O	O
upregulated	NN	O	O
and	NN	O	O
may	NN	O	O
explain	NN	O	O
unsuccessful	NN	O	O
T-cell	NN	O	O
maturation	NN	O	O
.	NN	O	O

To	NN	O	O
better	NN	O	O
understand	NN	O	O
the	NN	O	O
developmental	NN	O	O
link	NN	O	O
between	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
derived	NN	O	O
from	NN	O	O
progenitors	NN	O	B-cell_type
,	NN	O	O
we	NN	O	O
studied	NN	O	O
NK	NN	O	B-cell_type
cell	NN	O	I-cell_type
subsets	NN	O	I-cell_type
circulating	NN	O	O
in	NN	O	O
blood	NN	O	O
.	NN	O	O

CD56	NN	O	B-cell_type
(	NN	O	I-cell_type
+bright	NN	O	I-cell_type
)	NN	O	I-cell_type
,	NN	O	O
but	NN	O	O
not	NN	O	O
CD56	NN	O	B-cell_type
(	NN	O	I-cell_type
+dim	NN	O	I-cell_type
)	NN	O	I-cell_type
,	NN	O	O
NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
constitutively	NN	O	O
express	NN	O	O
TCF-1	NN	O	B-protein
by	NN	O	O
reverse	NN	O	O
transcriptase	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
and	NN	O	O
Western	NN	O	O
blot	NN	O	O
analysis	NN	O	O
.	NN	O	O

The	NN	O	O
TCF-1	NN	O	B-protein
isoform	NN	O	I-protein
found	NN	O	O
in	NN	O	O
CD56	NN	O	B-cell_type
(	NN	O	I-cell_type
+bright	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
which	NN	O	O
express	NN	O	O
lectin	NN	O	B-protein
but	NN	O	O
not	NN	O	O
immunoglobulin	NN	O	B-protein
class	NN	O	I-protein
I	NN	O	I-protein
recognizing	NN	O	O
inhibitory	NN	O	B-protein
receptors	NN	O	I-protein
,	NN	O	O
was	NN	O	O
identical	NN	O	O
to	NN	O	O
that	NN	O	O
induced	NN	O	O
in	NN	O	O
NK	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
culture	NN	O	O
and	NN	O	O
was	NN	O	O
distinctly	NN	O	O
different	NN	O	O
from	NN	O	O
isoforms	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
TCF-1	NN	O	B-protein
does	NN	O	O
not	NN	O	O
target	NN	O	O
human	NN	O	B-DNA
killer	NN	O	I-DNA
immunoglobulin	NN	O	I-DNA
receptor	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
TCF-1	NN	O	B-protein
is	NN	O	O
uniquely	NN	O	O
expressed	NN	O	O
in	NN	O	O
circulating	NN	O	O
CD56	NN	O	B-cell_type
(	NN	O	I-cell_type
+bright	NN	O	I-cell_type
)	NN	O	I-cell_type
NK	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
specific	NN	O	O
TCF-1	NN	O	B-protein
isoforms	NN	O	I-protein
may	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
regulating	NN	O	O
NK	NN	O	O
differentiation	NN	O	O
from	NN	O	O
a	NN	O	O
common	NN	O	O
NK/T-cell	NN	O	B-cell_type
progenitor	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Transcriptional	NN	O	O
activation	NN	O	O
by	NN	O	O
a	NN	O	O
matrix	NN	O	B-protein
associating	NN	O	I-protein
region-binding	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

contextual	NN	O	O
requirements	NN	O	O
for	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
bright	NN	O	B-protein
.	NN	O	O

Bright	NN	O	B-protein
(	NN	O	O
B	NN	O	B-protein
cell	NN	O	I-protein
regulator	NN	O	I-protein
of	NN	O	I-protein
IgH	NN	O	I-protein
transcription	NN	O	I-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
B	NN	O	B-cell_type
cell	NN	O	I-cell_type
-specific	NN	O	O
,	NN	O	O
matrix	NN	O	B-protein
associating	NN	O	I-protein
region-binding	NN	O	I-protein
protein	NN	O	I-protein
that	NN	O	O
transactivates	NN	O	O
gene	NN	O	O
expression	NN	O	O
from	NN	O	O
the	NN	O	O
IgH	NN	O	B-DNA
intronic	NN	O	I-DNA
enhancer	NN	O	I-DNA
(	NN	O	O
E	NN	O	B-DNA
mu	NN	O	I-DNA
)	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
here	NN	O	O
that	NN	O	O
Bright	NN	O	B-protein
has	NN	O	O
multiple	NN	O	O
contextual	NN	O	O
requirements	NN	O	O
to	NN	O	O
function	NN	O	O
as	NN	O	O
a	NN	O	O
transcriptional	NN	O	B-protein
activator	NN	O	I-protein
.	NN	O	O

Bright	NN	O	B-protein
can	NN	O	O
not	NN	O	O
transactivate	NN	O	O
via	NN	O	O
out	NN	O	O
of	NN	O	O
context	NN	O	O
,	NN	O	O
concatenated	NN	O	B-protein
binding	NN	O	I-protein
sites	NN	O	I-protein
.	NN	O	O

Transactivation	NN	O	O
is	NN	O	O
maximal	NN	O	O
on	NN	O	O
integrated	NN	O	O
substrates	NN	O	O
.	NN	O	O

Two	NN	O	O
of	NN	O	O
the	NN	O	O
three	NN	O	O
previously	NN	O	O
identified	NN	O	O
binding	NN	O	B-DNA
sites	NN	O	I-DNA
in	NN	O	O
E	NN	O	B-DNA
mu	NN	O	I-DNA
are	NN	O	O
required	NN	O	O
for	NN	O	O
full	NN	O	O
Bright	NN	O	B-protein
transactivation	NN	O	O
.	NN	O	O

The	NN	O	O
Bright	NN	O	B-protein
DNA	NN	O	I-protein
binding	NN	O	I-protein
domain	NN	O	I-protein
defined	NN	O	O
a	NN	O	O
new	NN	O	O
family	NN	O	O
,	NN	O	O
which	NN	O	O
includes	NN	O	O
SWI1	NN	O	B-protein
,	NN	O	O
a	NN	O	O
component	NN	O	O
of	NN	O	O
the	NN	O	O
SWI.SNF	NN	O	B-protein
complex	NN	O	I-protein
shown	NN	O	O
to	NN	O	O
have	NN	O	O
high	NN	O	O
mobility	NN	O	O
group-like	NN	O	O
DNA	NN	O	O
binding	NN	O	O
characteristics	NN	O	O
.	NN	O	O

Similar	NN	O	O
to	NN	O	O
one	NN	O	O
group	NN	O	O
of	NN	O	O
high	NN	O	B-protein
mobility	NN	O	I-protein
group	NN	O	I-protein
box	NN	O	I-protein
proteins	NN	O	I-protein
,	NN	O	O
Bright	NN	O	B-protein
distorts	NN	O	O
E	NN	O	B-DNA
mu	NN	O	I-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
-containing	NN	O	O
DNA	NN	O	O
on	NN	O	O
binding	NN	O	O
,	NN	O	O
supporting	NN	O	O
the	NN	O	O
concept	NN	O	O
that	NN	O	O
it	NN	O	O
mediates	NN	O	O
E	NN	O	B-DNA
mu	NN	O	I-DNA
remodeling	NN	O	O
.	NN	O	O

Transfection	NN	O	O
studies	NN	O	O
further	NN	O	O
implicate	NN	O	O
Bright	NN	O	B-protein
in	NN	O	O
facilitating	NN	O	O
spatially	NN	O	O
separated	NN	O	O
promoter-enhancer	NN	O	O
interactions	NN	O	O
in	NN	O	O
both	NN	O	O
transient	NN	O	O
and	NN	O	O
stable	NN	O	O
assays	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
overexpression	NN	O	O
of	NN	O	O
Bright	NN	O	B-protein
leads	NN	O	O
to	NN	O	O
enhanced	NN	O	O
DNase	NN	O	O
I	NN	O	O
sensitivity	NN	O	O
of	NN	O	O
the	NN	O	O
endogenous	NN	O	O
E	NN	O	B-DNA
mu	NN	O	I-DNA
matrix	NN	O	O
associating	NN	O	O
regions	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
further	NN	O	O
suggest	NN	O	O
that	NN	O	O
Bright	NN	O	B-protein
may	NN	O	O
contribute	NN	O	O
to	NN	O	O
increased	NN	O	O
gene	NN	O	O
expression	NN	O	O
by	NN	O	O
remodeling	NN	O	O
the	NN	O	O
immunoglobulin	NN	O	B-DNA
locus	NN	O	I-DNA
during	NN	O	O
B	NN	O	B-cell_type
cell	NN	O	I-cell_type
development	NN	O	O
.	NN	O	O

-DOCSTART-	O

Analysis	NN	O	O
of	NN	O	O
BCL-6	NN	O	O
mutations	NN	O	O
in	NN	O	O
classic	NN	O	O
Hodgkin	NN	O	O
disease	NN	O	O
of	NN	O	O
the	NN	O	O
B-	NN	O	B-cell_type
and	NN	O	I-cell_type
T-cell	NN	O	I-cell_type
type	NN	O	I-cell_type
.	NN	O	O

BCL-6	NN	O	B-DNA
is	NN	O	O
essential	NN	O	O
for	NN	O	O
germinal	NN	O	O
center	NN	O	O
formation	NN	O	O
and	NN	O	O
thus	NN	O	O
for	NN	O	O
affinity	NN	O	O
maturation	NN	O	O
of	NN	O	O
immunoglobulin	NN	O	B-DNA
(	NN	O	I-DNA
Ig	NN	O	I-DNA
)	NN	O	I-DNA
genes	NN	O	I-DNA
by	NN	O	O
somatic	NN	O	O
mutations	NN	O	O
.	NN	O	O

The	NN	O	O
5'-noncoding	NN	O	B-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
BCL-6	NN	O	B-DNA
gene	NN	O	I-DNA
is	NN	O	O
even	NN	O	O
a	NN	O	O
target	NN	O	O
for	NN	O	O
the	NN	O	O
mutation	NN	O	O
machinery	NN	O	O
.	NN	O	O

Translocations	NN	O	O
of	NN	O	O
the	NN	O	O
BCL-6	NN	O	B-DNA
gene	NN	O	I-DNA
to	NN	O	O
heterologous	NN	O	B-DNA
promoters	NN	O	I-DNA
and	NN	O	O
mutations	NN	O	O
of	NN	O	O
its	NN	O	O
5'-noncoding	NN	O	B-DNA
regulatory	NN	O	I-DNA
region	NN	O	I-DNA
were	NN	O	O
reported	NN	O	O
to	NN	O	O
be	NN	O	O
potential	NN	O	O
mechanisms	NN	O	O
for	NN	O	O
deregulating	NN	O	O
BCL-6	NN	O	B-DNA
expression	NN	O	O
and	NN	O	O
for	NN	O	O
playing	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
genesis	NN	O	O
of	NN	O	O
non-Hodgkin	NN	O	O
lymphoma	NN	O	O
.	NN	O	O

In	NN	O	O
line	NN	O	O
with	NN	O	O
this	NN	O	O
hypothesis	NN	O	O
is	NN	O	O
the	NN	O	O
observation	NN	O	O
that	NN	O	O
B-cell	NN	O	O
lymphoma	NN	O	O
with	NN	O	O
somatic	NN	O	O
mutations	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
diffuse	NN	O	O
large	NN	O	O
B-cell	NN	O	O
lymphoma	NN	O	O
and	NN	O	O
follicular	NN	O	O
lymphoma	NN	O	O
,	NN	O	O
also	NN	O	O
carry	NN	O	O
BCL-6	NN	O	O
mutations	NN	O	O
,	NN	O	O
some	NN	O	O
of	NN	O	O
which	NN	O	O
are	NN	O	O
recurrently	NN	O	O
detectable	NN	O	O
.	NN	O	O

Classic	NN	O	O
Hodgkin	NN	O	O
disease	NN	O	O
(	NN	O	O
cHD	NN	O	O
)	NN	O	O
is	NN	O	O
also	NN	O	O
derived	NN	O	O
from	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
high	NN	O	O
loads	NN	O	O
of	NN	O	O
somatic	NN	O	O
mutations	NN	O	O
and	NN	O	O
thus	NN	O	O
a	NN	O	O
further	NN	O	O
candidate	NN	O	O
for	NN	O	O
BCL-6	NN	O	O
mutations	NN	O	O
.	NN	O	O

To	NN	O	O
determine	NN	O	O
the	NN	O	O
presence	NN	O	O
and	NN	O	O
potential	NN	O	O
role	NN	O	O
of	NN	O	O
BCL-6	NN	O	O
mutations	NN	O	O
in	NN	O	O
cHD	NN	O	O
,	NN	O	O
the	NN	O	O
5'-noncoding	NN	O	B-DNA
BCL-6	NN	O	I-DNA
proportion	NN	O	O
of	NN	O	O
single	NN	O	B-cell_type
Hodgkin	NN	O	I-cell_type
and	NN	O	I-cell_type
Reed-Sternberg	NN	O	I-cell_type
(	NN	O	I-cell_type
HRS	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
6	NN	O	O
cases	NN	O	O
of	NN	O	O
cHD	NN	O	O
and	NN	O	O
6	NN	O	O
cases	NN	O	O
of	NN	O	O
HD-derived	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
was	NN	O	O
analyzed	NN	O	O
.	NN	O	O

All	NN	O	O
B-cell-derived	NN	O	O
HD	NN	O	O
cases	NN	O	O
and	NN	O	O
cell	NN	O	O
lines	NN	O	O
harbored	NN	O	O
BCL-6	NN	O	O
mutations	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
both	NN	O	O
T-cell-derived	NN	O	O
HD	NN	O	O
cases	NN	O	O
and	NN	O	O
cell	NN	O	O
lines	NN	O	O
were	NN	O	O
devoid	NN	O	O
of	NN	O	O
BCL-6	NN	O	O
mutations	NN	O	O
.	NN	O	O

With	NN	O	O
only	NN	O	O
one	NN	O	O
exception	NN	O	O
,	NN	O	O
there	NN	O	O
were	NN	O	O
no	NN	O	O
lymphoma-specific	NN	O	O
recurrent	NN	O	O
BCL-6	NN	O	O
mutations	NN	O	O
detected	NN	O	O
,	NN	O	O
and	NN	O	O
BCL-6	NN	O	B-protein
protein	NN	O	I-protein
was	NN	O	O
absent	NN	O	O
from	NN	O	O
the	NN	O	O
HRS	NN	O	B-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
most	NN	O	O
cases	NN	O	O
.	NN	O	O

In	NN	O	O
conclusion	NN	O	O
,	NN	O	O
(	NN	O	O
1	NN	O	O
)	NN	O	O
somatic	NN	O	O
BCL-6	NN	O	O
mutations	NN	O	O
are	NN	O	O
restricted	NN	O	O
to	NN	O	O
cHD	NN	O	O
cases	NN	O	O
of	NN	O	O
B-cell	NN	O	B-cell_type
origin	NN	O	O
,	NN	O	O
and	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
the	NN	O	O
BCL-6	NN	O	O
mutations	NN	O	O
represent	NN	O	O
mostly	NN	O	O
irrelevant	NN	O	O
somatic	NN	O	O
base	NN	O	O
substitutions	NN	O	O
without	NN	O	O
consequences	NN	O	O
for	NN	O	O
BCL-6	NN	O	B-protein
protein	NN	O	I-protein
expression	NN	O	O
and	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
cHD	NN	O	O
.	NN	O	O

-DOCSTART-	O

Gadd45gamma	NN	O	B-DNA
is	NN	O	O
dispensable	NN	O	O
for	NN	O	O
normal	NN	O	O
mouse	NN	O	O
development	NN	O	O
and	NN	O	O
T-cell	NN	O	O
proliferation	NN	O	O
.	NN	O	O

Gadd45gamma	NN	O	B-DNA
,	NN	O	O
a	NN	O	O
family	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
growth	NN	O	B-DNA
arrest	NN	O	I-DNA
and	NN	O	I-DNA
DNA	NN	O	I-DNA
damage-inducible	NN	O	I-DNA
gene	NN	O	I-DNA
family	NN	O	I-DNA
45	NN	O	I-DNA
(	NN	O	O
Gadd45	NN	O	B-DNA
)	NN	O	O
,	NN	O	O
is	NN	O	O
strongly	NN	O	O
induced	NN	O	O
by	NN	O	O
interleukin-2	NN	O	B-protein
(	NN	O	O
IL-2	NN	O	B-protein
)	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

While	NN	O	O
in	NN	O	O
most	NN	O	O
tissues	NN	O	O
all	NN	O	O
Gadd45	NN	O	B-DNA
family	NN	O	I-DNA
members	NN	O	I-DNA
are	NN	O	O
expressed	NN	O	O
,	NN	O	O
Gadd45gamma	NN	O	B-DNA
is	NN	O	O
the	NN	O	O
only	NN	O	O
member	NN	O	O
that	NN	O	O
is	NN	O	O
induced	NN	O	O
by	NN	O	O
IL-2	NN	O	B-protein
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
IL-2	NN	O	B-protein
-induced	NN	O	O
expression	NN	O	O
of	NN	O	O
Gadd45gamma	NN	O	B-DNA
is	NN	O	O
dependent	NN	O	O
on	NN	O	O
a	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
tyrosine	NN	O	B-protein
kinase	NN	O	I-protein
Jak3	NN	O	I-protein
and	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
Stat5a	NN	O	B-protein
and	NN	O	O
Stat5b	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
.	NN	O	O

Previous	NN	O	O
studies	NN	O	O
with	NN	O	O
ectopically	NN	O	O
overexpressed	NN	O	O
Gadd45gamma	NN	O	B-DNA
in	NN	O	O
various	NN	O	O
cell	NN	O	O
lines	NN	O	O
implicated	NN	O	O
its	NN	O	O
function	NN	O	O
in	NN	O	O
negative	NN	O	O
growth	NN	O	O
control	NN	O	O
.	NN	O	O

To	NN	O	O
analyze	NN	O	O
the	NN	O	O
physiological	NN	O	O
role	NN	O	O
of	NN	O	O
Gadd45gamma	NN	O	B-DNA
we	NN	O	O
used	NN	O	O
homologous	NN	O	O
recombination	NN	O	O
to	NN	O	O
generate	NN	O	O
mice	NN	O	O
lacking	NN	O	O
Gadd45gamma	NN	O	B-DNA
.	NN	O	O

Gadd45gamma	NN	O	B-DNA
-deficient	NN	O	O
mice	NN	O	O
develop	NN	O	O
normally	NN	O	O
,	NN	O	O
are	NN	O	O
indistinguishable	NN	O	O
from	NN	O	O
their	NN	O	O
littermates	NN	O	O
,	NN	O	O
and	NN	O	O
are	NN	O	O
fertile	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
hematopoiesis	NN	O	O
in	NN	O	O
mice	NN	O	O
lacking	NN	O	O
Gadd45gamma	NN	O	B-DNA
is	NN	O	O
not	NN	O	O
impaired	NN	O	O
and	NN	O	O
Gadd45gamma	NN	O	B-DNA
-deficient	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
show	NN	O	O
normal	NN	O	O
responses	NN	O	O
to	NN	O	O
IL-2	NN	O	B-protein
.	NN	O	O

These	NN	O	O
data	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
Gadd45gamma	NN	O	B-DNA
is	NN	O	O
not	NN	O	O
essential	NN	O	O
for	NN	O	O
normal	NN	O	O
mouse	NN	O	O
development	NN	O	O
and	NN	O	O
hematopoiesis	NN	O	O
,	NN	O	O
possibly	NN	O	O
due	NN	O	O
to	NN	O	O
functional	NN	O	O
redundancy	NN	O	O
among	NN	O	O
the	NN	O	O
Gadd45	NN	O	B-DNA
family	NN	O	I-DNA
members	NN	O	I-DNA
.	NN	O	O

Gadd45gamma	NN	O	B-DNA
is	NN	O	O
also	NN	O	O
dispensable	NN	O	O
for	NN	O	O
IL-2	NN	O	B-protein
-induced	NN	O	O
T-cell	NN	O	O
proliferation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Decreased	NN	O	O
expression	NN	O	O
of	NN	O	O
c-myc	NN	O	B-DNA
family	NN	O	I-DNA
genes	NN	O	I-DNA
in	NN	O	O
thymuses	NN	O	O
from	NN	O	O
myasthenia	NN	O	O
gravis	NN	O	O
patients	NN	O	O
.	NN	O	O

The	NN	O	O
thymus	NN	O	O
is	NN	O	O
a	NN	O	O
critical	NN	O	O
organ	NN	O	O
for	NN	O	O
the	NN	O	O
elimination	NN	O	O
of	NN	O	O
autoreactive	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

We	NN	O	O
studied	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
apoptosis-associated	NN	O	O
genes	NN	O	O
,	NN	O	O
bcl-xL	NN	O	B-DNA
,	NN	O	O
bad	NN	O	B-DNA
,	NN	O	O
caspase-3	NN	O	B-DNA
,	NN	O	O
and	NN	O	O
c-myc	NN	O	B-DNA
family	NN	O	I-DNA
genes	NN	O	I-DNA
in	NN	O	O
myasthenia	NN	O	O
gravis	NN	O	O
(	NN	O	O
MG	NN	O	O
)	NN	O	O
thymuses	NN	O	O
.	NN	O	O

We	NN	O	O
observed	NN	O	O
that	NN	O	O
the	NN	O	O
mRNA	NN	O	O
levels	NN	O	O
of	NN	O	O
myc	NN	O	B-DNA
family	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
c-myc	NN	O	B-DNA
and	NN	O	O
max	NN	O	B-DNA
,	NN	O	O
were	NN	O	O
markedly	NN	O	O
reduced	NN	O	O
in	NN	O	O
MG	NN	O	O
thymuses	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
c-myc	NN	O	B-DNA
-mediated	NN	O	O
signaling	NN	O	O
is	NN	O	O
abnormal	NN	O	O
in	NN	O	O
MG	NN	O	O
thymuses	NN	O	O
.	NN	O	O

The	NN	O	O
levels	NN	O	O
of	NN	O	O
molecules	NN	O	O
whose	NN	O	O
expressions	NN	O	O
are	NN	O	O
associated	NN	O	O
with	NN	O	O
myc	NN	O	B-DNA
,	NN	O	O
such	NN	O	O
as	NN	O	O
STAM	NN	O	B-protein
,	NN	O	O
prothymosin-alpha	NN	O	B-protein
,	NN	O	O
and	NN	O	O
NFkappaB	NN	O	B-protein

-DOCSTART-	O

Immunohistochemical	NN	O	O
detection	NN	O	O
of	NN	O	O
interferon-gamma-producing	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
dermatophytosis	NN	O	O
.	NN	O	O

Skin	NN	O	O
lesions	NN	O	O
of	NN	O	O
dermatophytosis	NN	O	O
are	NN	O	O
thought	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
result	NN	O	O
of	NN	O	O
a	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
-dependent	NN	O	O
inflammatory	NN	O	O
response	NN	O	O
that	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
various	NN	O	O
cytokines	NN	O	B-protein
.	NN	O	O

We	NN	O	O
examined	NN	O	O
whether	NN	O	O
IFN-gamma-positive	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
as	NN	O	O
expression	NN	O	O
of	NN	O	O
Th1	NN	O	B-cell_type
response	NN	O	O
)	NN	O	O
were	NN	O	O
present	NN	O	O
in	NN	O	O
the	NN	O	O
skin	NN	O	O
lesions	NN	O	O
of	NN	O	O
dermatophytosis	NN	O	O
in	NN	O	O
situ	NN	O	O
by	NN	O	O
immunohistochemical	NN	O	O
techniques	NN	O	O
.	NN	O	O

Mixtures	NN	O	O
of	NN	O	O
CD4-positive	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
CD8-positive	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
present	NN	O	O
in	NN	O	O
the	NN	O	O
dermal	NN	O	O
infiltrates	NN	O	O
of	NN	O	O
the	NN	O	O
lesions	NN	O	O
.	NN	O	O

Considerable	NN	O	O
numbers	NN	O	O
of	NN	O	O
CD1a-positive	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
detected	NN	O	O
in	NN	O	O
the	NN	O	O
upper	NN	O	O
dermis	NN	O	O
and	NN	O	O
epidermis	NN	O	O
.	NN	O	O

A	NN	O	O
marked	NN	O	O
accumulation	NN	O	O
of	NN	O	O
CD68-positive	NN	O	B-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
upper	NN	O	O
dermis	NN	O	O
.	NN	O	O

IFN-gamma-positive	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
present	NN	O	O
in	NN	O	O
the	NN	O	O
upper	NN	O	O
dermis	NN	O	O
of	NN	O	O
the	NN	O	O
lesions	NN	O	O
.	NN	O	O

The	NN	O	O
pattern	NN	O	O
of	NN	O	O
IFN-gamma	NN	O	B-protein
staining	NN	O	O
appeared	NN	O	O
to	NN	O	O
be	NN	O	O
intracellular	NN	O	O
in	NN	O	O
mononuclear	NN	O	B-cell_type
lymphoid	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
staining	NN	O	O
was	NN	O	O
considered	NN	O	O
to	NN	O	O
be	NN	O	O
highly	NN	O	O
specific	NN	O	O
because	NN	O	O
it	NN	O	O
could	NN	O	O
be	NN	O	O
completely	NN	O	O
blocked	NN	O	O
by	NN	O	O
preabsorption	NN	O	O
with	NN	O	O
recombinant	NN	O	B-protein
IFN-gamma	NN	O	I-protein
.	NN	O	O

Our	NN	O	O
data	NN	O	O
support	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
the	NN	O	O
skin	NN	O	O
lesions	NN	O	O
of	NN	O	O
dermatophytosis	NN	O	O
may	NN	O	O
be	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
Th1	NN	O	B-cell_type
response	NN	O	O
.	NN	O	O

Th1	NN	O	B-cell_type
response	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
characterized	NN	O	O
by	NN	O	O
IFN-gamma	NN	O	B-protein
release	NN	O	O
,	NN	O	O
is	NN	O	O
thought	NN	O	O
to	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
host	NN	O	O
defense	NN	O	O
against	NN	O	O
dermatophytes	NN	O	O
and	NN	O	O
to	NN	O	O
reflect	NN	O	O
cutaneous	NN	O	O
reaction	NN	O	O
in	NN	O	O
dermatophytosis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
the	NN	O	O
helix-loop-helix	NN	O	B-protein
proteins	NN	O	I-protein
,	NN	O	O
E2A	NN	O	B-protein
and	NN	O	O
Id3	NN	O	B-protein
,	NN	O	O
by	NN	O	O
the	NN	O	O
Ras	NN	O	B-protein
-	NN	O	O
ERK	NN	O	B-protein
MAPK	NN	O	B-protein
cascade	NN	O	O
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	O
MAPK	NN	O	B-protein
)	NN	O	O
pathways	NN	O	O
leads	NN	O	O
to	NN	O	O
cellular	NN	O	O
differentiation	NN	O	O
and/or	NN	O	O
proliferation	NN	O	O
in	NN	O	O
a	NN	O	O
wide	NN	O	O
variety	NN	O	O
of	NN	O	O
cell	NN	O	O
types	NN	O	O
,	NN	O	O
including	NN	O	O
developing	NN	O	B-cell_type
thymocytes	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
basic	NN	O	B-protein
helix-loop-helix	NN	O	I-protein
(	NN	O	I-protein
bHLH	NN	O	I-protein
)	NN	O	I-protein
proteins	NN	O	I-protein
E12	NN	O	B-protein
and	NN	O	O
E47	NN	O	B-protein
and	NN	O	O
an	NN	O	O
inhibitor	NN	O	B-protein
HLH	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
Id3	NN	O	B-protein
,	NN	O	O
play	NN	O	O
key	NN	O	O
roles	NN	O	O
in	NN	O	O
thymocyte	NN	O	O
differentiation	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
here	NN	O	O
that	NN	O	O
E2A	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
is	NN	O	O
lowered	NN	O	O
in	NN	O	O
primary	NN	O	B-cell_type
immature	NN	O	I-cell_type
thymocytes	NN	O	I-cell_type
consequent	NN	O	O
to	NN	O	O
T	NN	O	O
cell	NN	O	O
receptor	NN	O	O
(	NN	O	O
TCR	NN	O	O
)	NN	O	O
-mediated	NN	O	O
ligation	NN	O	O
.	NN	O	O

Whereas	NN	O	O
expression	NN	O	O
of	NN	O	O
E2A	NN	O	O
mRNA	NN	O	O
and	NN	O	O
protein	NN	O	O
are	NN	O	O
unaltered	NN	O	O
,	NN	O	O
Id3	NN	O	B-RNA
transcripts	NN	O	I-RNA
are	NN	O	O
rapidly	NN	O	O
induced	NN	O	O
upon	NN	O	O
signaling	NN	O	O
from	NN	O	O
the	NN	O	O
TCR	NN	O	B-protein
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
Id3	NN	O	O
transcription	NN	O	O
is	NN	O	O
regulated	NN	O	O
in	NN	O	O
a	NN	O	O
dose-dependent	NN	O	O
manner	NN	O	O
by	NN	O	O
the	NN	O	O
extracellular	NN	O	O
signal-regulated	NN	O	O
kinase	NN	O	O
(	NN	O	O
ERK	NN	O	B-protein
)	NN	O	O
MAPK	NN	O	B-protein
module	NN	O	O
.	NN	O	O

These	NN	O	O
observations	NN	O	O
directly	NN	O	O
connect	NN	O	O
the	NN	O	O
ERK	NN	O	B-protein
MAPK	NN	O	B-protein
cascade	NN	O	O
and	NN	O	O
HLH	NN	O	B-protein
proteins	NN	O	I-protein
in	NN	O	O
a	NN	O	O
linear	NN	O	O
pathway	NN	O	O
.	NN	O	O

-DOCSTART-	O

Retinoic	NN	O	O
acid	NN	O	O
up-regulates	NN	O	O
myeloid	NN	O	O
ICAM-3	NN	O	O
expression	NN	O	O
and	NN	O	O
function	NN	O	O
in	NN	O	O
a	NN	O	O
cell-specific	NN	O	O
fashion	NN	O	O
--	NN	O	O
evidence	NN	O	O
for	NN	O	O
retinoid	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
in	NN	O	O
the	NN	O	O
mast	NN	O	B-cell_type
cell	NN	O	I-cell_type
lineage	NN	O	I-cell_type
.	NN	O	O

Investigation	NN	O	O
of	NN	O	O
mast	NN	O	O
cell	NN	O	O
responsiveness	NN	O	O
toward	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
RA	NN	O	O
)	NN	O	O
revealed	NN	O	O
selective	NN	O	O
promotion	NN	O	O
of	NN	O	O
ICAM-3	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
mast	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
HMC-1	NN	O	I-cell_line
.	NN	O	O

This	NN	O	O
process	NN	O	O
was	NN	O	O
dose-	NN	O	O
and	NN	O	O
time-dependent	NN	O	O
and	NN	O	O
detectable	NN	O	O
by	NN	O	O
flow	NN	O	O
cytometry	NN	O	O
,	NN	O	O
Western	NN	O	O
blot	NN	O	O
analysis	NN	O	O
,	NN	O	O
ELISA	NN	O	O
,	NN	O	O
and	NN	O	O
Northern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
.	NN	O	O

ICAM-3	NN	O	B-protein
modulation	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
cell-type	NN	O	O
dependent	NN	O	O
,	NN	O	O
detectable	NN	O	O
also	NN	O	O
for	NN	O	O
HL-60	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
monocytes	NN	O	B-cell_type
but	NN	O	O
not	NN	O	O
U-937	NN	O	B-cell_line
and	NN	O	O
only	NN	O	O
weakly	NN	O	O
for	NN	O	O
KU812	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Terminally	NN	O	B-cell_type
differentiated	NN	O	I-cell_type
skin	NN	O	I-cell_type
mast	NN	O	I-cell_type
cells	NN	O	I-cell_type
also	NN	O	O
failed	NN	O	O
to	NN	O	O
up-modulate	NN	O	O
their	NN	O	O
ICAM-3	NN	O	B-protein
,	NN	O	O
suggesting	NN	O	O
the	NN	O	O
requirement	NN	O	O
for	NN	O	O
some	NN	O	O
degree	NN	O	O
of	NN	O	O
immaturity	NN	O	O
for	NN	O	O
the	NN	O	O
process	NN	O	O
.	NN	O	O

RA-mediated	NN	O	O
effects	NN	O	O
on	NN	O	O
ICAM-1	NN	O	B-protein
expression	NN	O	O
,	NN	O	O
studied	NN	O	O
in	NN	O	O
parallel	NN	O	O
,	NN	O	O
were	NN	O	O
clearly	NN	O	O
distinct	NN	O	O
from	NN	O	O
those	NN	O	O
on	NN	O	O
ICAM-3	NN	O	B-protein
.	NN	O	O

Investigation	NN	O	O
of	NN	O	O
retinoid	NN	O	B-protein
receptor	NN	O	I-protein
expression	NN	O	O
,	NN	O	O
known	NN	O	O
to	NN	O	O
mediate	NN	O	O
intracellular	NN	O	O
RA	NN	O	O
signaling	NN	O	O
,	NN	O	O
revealed	NN	O	O
presence	NN	O	O
of	NN	O	O
RAR	NN	O	B-RNA
alpha	NN	O	I-RNA
,	NN	O	I-RNA
RAR	NN	O	I-RNA
gamma	NN	O	I-RNA
,	NN	O	I-RNA
RXR	NN	O	I-RNA
beta	NN	O	I-RNA
,	NN	O	I-RNA
and	NN	O	I-RNA
RXR	NN	O	I-RNA
gamma	NN	O	I-RNA
transcripts	NN	O	I-RNA
in	NN	O	O
all	NN	O	O
cell	NN	O	O
lines	NN	O	O
studied	NN	O	O
,	NN	O	O
and	NN	O	O
HMC-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
the	NN	O	O
only	NN	O	O
line	NN	O	O
lacking	NN	O	O
RXR	NN	O	B-protein
alpha	NN	O	I-protein
.	NN	O	O

RAR	NN	O	B-protein
beta	NN	O	I-protein
,	NN	O	O
not	NN	O	O
expressed	NN	O	O
at	NN	O	O
baseline	NN	O	O
,	NN	O	O
was	NN	O	O
induced	NN	O	O
by	NN	O	O
RA	NN	O	O
in	NN	O	O
a	NN	O	O
fashion	NN	O	O
obviously	NN	O	O
correlating	NN	O	O
with	NN	O	O
ICAM-3	NN	O	B-protein
up-regulation	NN	O	O
.	NN	O	O

Increased	NN	O	O
ICAM-3	NN	O	B-protein
expression	NN	O	O
was	NN	O	O
of	NN	O	O
functional	NN	O	O
significance	NN	O	O
,	NN	O	O
such	NN	O	O
that	NN	O	O
processes	NN	O	O
stimulated	NN	O	O
or	NN	O	O
co-stimulated	NN	O	O
via	NN	O	O
ICAM-3	NN	O	B-protein
(	NN	O	O
homotypic	NN	O	O
aggregation	NN	O	O
,	NN	O	O
IL-8	NN	O	O
secretion	NN	O	O
)	NN	O	O
were	NN	O	O
clearly	NN	O	O
enhanced	NN	O	O
upon	NN	O	O
RA	NN	O	O
pretreatment	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
RA	NN	O	O
may	NN	O	O
contribute	NN	O	O
via	NN	O	O
hitherto	NN	O	O
unrecognized	NN	O	O
pathways	NN	O	O
to	NN	O	O
immune	NN	O	O
function	NN	O	O
and	NN	O	O
host	NN	O	O
defense	NN	O	O
.	NN	O	O

-DOCSTART-	O

CD28	NN	O	B-protein
and	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
co-stimulation	NN	O	O
.	NN	O	O

Over	NN	O	O
the	NN	O	O
last	NN	O	O
decade	NN	O	O
the	NN	O	O
concept	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
co-stimulation	NN	O	O
has	NN	O	O
emerged	NN	O	O
to	NN	O	O
take	NN	O	O
a	NN	O	O
central	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
process	NN	O	O
of	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
exact	NN	O	O
definition	NN	O	O
of	NN	O	O
co-stimulation	NN	O	O
is	NN	O	O
still	NN	O	O
unclear	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
review	NN	O	O
,	NN	O	O
we	NN	O	O
re-examine	NN	O	O
the	NN	O	O
concept	NN	O	O
of	NN	O	O
co-stimulation	NN	O	O
.	NN	O	O

We	NN	O	O
suggest	NN	O	O
that	NN	O	O
while	NN	O	O
co-stimulation	NN	O	O
is	NN	O	O
important	NN	O	O
,	NN	O	O
there	NN	O	O
is	NN	O	O
little	NN	O	O
evidence	NN	O	O
to	NN	O	O
link	NN	O	O
co-stimulation	NN	O	O
with	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
anergy	NN	O	O
.	NN	O	O

We	NN	O	O
then	NN	O	O
suggest	NN	O	O
a	NN	O	O
framework	NN	O	O
for	NN	O	O
studying	NN	O	O
co-stimulation	NN	O	O
.	NN	O	O

Focusing	NN	O	O
on	NN	O	O
recent	NN	O	O
advances	NN	O	O
in	NN	O	O
our	NN	O	O
understanding	NN	O	O
of	NN	O	O
CD28	NN	O	B-protein
,	NN	O	O
we	NN	O	O
discuss	NN	O	O
four	NN	O	O
areas	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
activation	NN	O	O
where	NN	O	O
co-stimulation	NN	O	O
may	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
activator	NN	O	B-protein
protein-1	NN	O	I-protein
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
CD8+	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
exposed	NN	O	O
to	NN	O	O
peripheral	NN	O	B-protein
self-antigens	NN	O	I-protein
.	NN	O	O

The	NN	O	O
transcriptional	NN	O	O
events	NN	O	O
that	NN	O	O
control	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
tolerance	NN	O	O
to	NN	O	O
peripheral	NN	O	B-protein
self	NN	O	I-protein
Ags	NN	O	I-protein
are	NN	O	O
still	NN	O	O
unknown	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
analyzed	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
-	NN	O	O
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
-mediated	NN	O	O
transcription	NN	O	O
during	NN	O	O
in	NN	O	O
vivo	NN	O	O
induction	NN	O	O
of	NN	O	O
tolerance	NN	O	O
to	NN	O	O
a	NN	O	O
self	NN	O	B-protein
Ag	NN	O	I-protein
expressed	NN	O	O
exclusively	NN	O	O
on	NN	O	O
hepatocytes	NN	O	B-cell_type
.	NN	O	O

Naive	NN	O	B-cell_type
CD8	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
Desire	NN	O	I-cell_type
(	NN	O	I-cell_type
Des	NN	O	I-cell_type
)	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
isolated	NN	O	O
from	NN	O	O
the	NN	O	O
Des	NN	O	O
TCR-transgenic	NN	O	O
mice	NN	O	O
that	NN	O	O
are	NN	O	O
specific	NN	O	O
for	NN	O	O
the	NN	O	O
H-2K	NN	O	B-protein
(	NN	O	I-protein
b	NN	O	I-protein
)	NN	O	I-protein
class	NN	O	I-protein
I	NN	O	I-protein
Ag	NN	O	I-protein
were	NN	O	O
transferred	NN	O	O
into	NN	O	O
Alb-K	NN	O	O
(	NN	O	O
b	NN	O	O
)	NN	O	O
-transgenic	NN	O	O
mice	NN	O	O
that	NN	O	O
express	NN	O	O
the	NN	O	O
H-2K	NN	O	B-protein
(	NN	O	I-protein
b	NN	O	I-protein
)	NN	O	I-protein
Ag	NN	O	I-protein
on	NN	O	O
hepatocytes	NN	O	B-cell_type
only	NN	O	O
.	NN	O	O

Tolerance	NN	O	O
develops	NN	O	O
in	NN	O	O
these	NN	O	O
mice	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
self-reactive	NN	O	B-cell_type
CD8	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
Des	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
transiently	NN	O	O
activated	NN	O	O
,	NN	O	O
then	NN	O	O
became	NN	O	O
unresponsive	NN	O	O
and	NN	O	O
were	NN	O	O
further	NN	O	O
deleted	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
CD8	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
Des	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
activated	NN	O	O
in	NN	O	O
vivo	NN	O	O
with	NN	O	O
APCs	NN	O	O
,	NN	O	O
which	NN	O	O
express	NN	O	O
high	NN	O	O
AP-1	NN	O	B-protein
and	NN	O	O
high	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
transcriptional	NN	O	O
activity	NN	O	O
,	NN	O	O
the	NN	O	O
unresponsive	NN	O	O
CD8	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
Des	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
expressed	NN	O	O
no	NN	O	O
AP-1	NN	O	B-protein
and	NN	O	O
only	NN	O	O
weak	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
transcriptional	NN	O	O
activity	NN	O	O
.	NN	O	O

The	NN	O	O
differences	NN	O	O
in	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
transcriptional	NN	O	O
activity	NN	O	O
correlated	NN	O	O
with	NN	O	O
the	NN	O	O
generation	NN	O	O
of	NN	O	O
distinct	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
complexes	NN	O	I-protein
.	NN	O	O

Indeed	NN	O	O
,	NN	O	O
in	NN	O	O
vivo	NN	O	O
primed	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
predominantly	NN	O	O
express	NN	O	O
p50/p50	NN	O	B-protein
and	NN	O	I-protein
p65/p50	NN	O	I-protein
dimers	NN	O	I-protein
,	NN	O	O
whereas	NN	O	O
these	NN	O	O
p50-containing	NN	O	B-protein
complexes	NN	O	I-protein
are	NN	O	O
barely	NN	O	O
detectable	NN	O	O
in	NN	O	O
tolerant	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
that	NN	O	O
express	NN	O	O
p65-	NN	O	B-protein
and	NN	O	I-protein
c-Rel-containing	NN	O	I-protein
complexes	NN	O	I-protein
.	NN	O	O

These	NN	O	O
observations	NN	O	O
suggest	NN	O	O
that	NN	O	O
fine	NN	O	O
regulation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
complex	NN	O	I-protein
formation	NN	O	O
may	NN	O	O
determine	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
fate	NN	O	O
.	NN	O	O

-DOCSTART-	O

Transcriptional	NN	O	O
regulation	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
.	NN	O	O

Lymphocytes	NN	O	B-cell_type
have	NN	O	O
been	NN	O	O
used	NN	O	O
to	NN	O	O
investigate	NN	O	O
many	NN	O	O
cellular	NN	O	O
processes	NN	O	O
,	NN	O	O
including	NN	O	O
lineage	NN	O	O
commitment	NN	O	O
,	NN	O	O
differentiation	NN	O	O
,	NN	O	O
proliferation	NN	O	O
and	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

The	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
that	NN	O	O
mediate	NN	O	O
these	NN	O	O
processes	NN	O	O
are	NN	O	O
often	NN	O	O
expressed	NN	O	O
broadly	NN	O	O
in	NN	O	O
many	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

The	NN	O	O
emerging	NN	O	O
theme	NN	O	O
is	NN	O	O
one	NN	O	O
of	NN	O	O
cell-type-specific	NN	O	O
regulation	NN	O	O
,	NN	O	O
affecting	NN	O	O
not	NN	O	O
only	NN	O	O
the	NN	O	O
functional	NN	O	O
activation	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
but	NN	O	O
also	NN	O	O
their	NN	O	O
access	NN	O	O
to	NN	O	O
appropriate	NN	O	O
regions	NN	O	O
of	NN	O	O
DNA	NN	O	O
.	NN	O	O

-DOCSTART-	O

Existence	NN	O	O
of	NN	O	O
retinoic	NN	O	B-protein
acid-receptor	NN	O	I-protein
-independent	NN	O	O
retinoid	NN	O	B-protein
X-receptor	NN	O	I-protein
-dependent	NN	O	O
pathway	NN	O	O
in	NN	O	O
myeloid	NN	O	B-cell_type
cell	NN	O	I-cell_type
function	NN	O	O
.	NN	O	O

We	NN	O	O
previously	NN	O	O
reported	NN	O	O
that	NN	O	O
ER-27191	NN	O	O
(	NN	O	O
4-	NN	O	O
[	NN	O	O
4	NN	O	O
,	NN	O	O
5	NN	O	O
,	NN	O	O
7	NN	O	O
,	NN	O	O
8	NN	O	O
,	NN	O	O
9	NN	O	O
,	NN	O	O
10-hexahydro-7	NN	O	O
,	NN	O	O
7	NN	O	O
,	NN	O	O
10	NN	O	O
,	NN	O	O
10-tetramethyl-1-	NN	O	O
(	NN	O	O
3-pyridylmethyl	NN	O	O
)	NN	O	O
anthra	NN	O	O
[	NN	O	O
1	NN	O	O
,	NN	O	O
2-b	NN	O	O
]	NN	O	O
pyrrol-3-yl	NN	O	O
]	NN	O	O
benzoic	NN	O	O
acid	NN	O	O
)	NN	O	O
is	NN	O	O
a	NN	O	O
potent	NN	O	O
antagonist	NN	O	O
of	NN	O	O
retinoic	NN	O	B-protein
acid	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
RAR	NN	O	B-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
ER-35795	NN	O	O
(	NN	O	O
(	NN	O	O
2E	NN	O	O
,	NN	O	O
4E	NN	O	O
,	NN	O	O
6E	NN	O	O
)	NN	O	O
-7-	NN	O	O
[	NN	O	O
1-	NN	O	O
(	NN	O	O
1-methylethyl	NN	O	O
)	NN	O	O
-8-chloro-1	NN	O	O
,	NN	O	O
2	NN	O	O
,	NN	O	O
3	NN	O	O
,	NN	O	O
4-tetrahydroquinolin-6-yl	NN	O	O
]	NN	O	O
-6-fluoro-3-methyl-2	NN	O	O
,	NN	O	O
4	NN	O	O
,	NN	O	O
6-nonatrienoic	NN	O	O
acid	NN	O	O
)	NN	O	O
is	NN	O	O
a	NN	O	O
novel	NN	O	O
retinoid	NN	O	B-protein
X	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
RXR	NN	O	B-protein
)	NN	O	O
-specific	NN	O	O
agonist	NN	O	O
.	NN	O	O

By	NN	O	O
using	NN	O	O
these	NN	O	O
compounds	NN	O	O
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
whether	NN	O	O
distinct	NN	O	O
RAR	NN	O	B-protein
-dependent	NN	O	O
and	NN	O	O
RXR	NN	O	B-protein
-dependent	NN	O	O
pathways	NN	O	O
operate	NN	O	O
to	NN	O	O
mediate	NN	O	O
the	NN	O	O
diverse	NN	O	O
activities	NN	O	O
of	NN	O	O
retinoids	NN	O	O
,	NN	O	O
particularly	NN	O	O
,	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
the	NN	O	O
RXR	NN	O	B-protein
pathway	NN	O	O
on	NN	O	O
cellular	NN	O	O
function	NN	O	O
.	NN	O	O

ER-27191	NN	O	O
completely	NN	O	O
antagonized	NN	O	O
HL60	NN	O	B-cell_line
cell	NN	O	I-cell_line
differentiation	NN	O	O
induced	NN	O	O
by	NN	O	O
all-trans-retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
atRA	NN	O	O
)	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
differentiation	NN	O	O
induced	NN	O	O
by	NN	O	O
the	NN	O	O
ER-35795	NN	O	O
was	NN	O	O
not	NN	O	O
antagonized	NN	O	O
at	NN	O	O
all	NN	O	O
by	NN	O	O
the	NN	O	O
RAR	NN	O	B-protein
antagonist	NN	O	O
,	NN	O	O
but	NN	O	O
was	NN	O	O
inhibited	NN	O	O
by	NN	O	O
an	NN	O	O
RXR	NN	O	B-protein
homodimer	NN	O	I-protein
antagonist	NN	O	O
(	NN	O	O
LGD100754	NN	O	O
,	NN	O	O
(	NN	O	O
2E	NN	O	O
,	NN	O	O
4E	NN	O	O
,	NN	O	O
6Z	NN	O	O
)	NN	O	O
-7-	NN	O	O
(	NN	O	O
3-n-propoxy-5	NN	O	O
,	NN	O	O
6	NN	O	O
,	NN	O	O
7	NN	O	O
,	NN	O	O
8-tetrahydro-5	NN	O	O
,	NN	O	O
5	NN	O	O
,	NN	O	O
8	NN	O	O
,	NN	O	O
8-tetramethylnaphthalen-2-yl	NN	O	O
)	NN	O	O
-3-methylocta-2	NN	O	O
,	NN	O	O
4	NN	O	O
,	NN	O	O
6-trienoic	NN	O	O
acid	NN	O	O
)	NN	O	O
.	NN	O	O

Its	NN	O	O
agonistic	NN	O	O
action	NN	O	O
on	NN	O	O
RXR/RAR	NN	O	B-protein
heterodimer	NN	O	I-protein
,	NN	O	O
on	NN	O	O
the	NN	O	O
other	NN	O	O
hand	NN	O	O
,	NN	O	O
was	NN	O	O
neutralized	NN	O	O
by	NN	O	O
the	NN	O	O
RAR	NN	O	B-protein
antagonist	NN	O	O
.	NN	O	O

During	NN	O	O
HL60	NN	O	B-cell_line
cell	NN	O	I-cell_line
differentiation	NN	O	O
,	NN	O	O
atRA	NN	O	O
induced	NN	O	O
RARbeta	NN	O	B-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
while	NN	O	O
the	NN	O	O
RXR	NN	O	B-protein
had	NN	O	O
no	NN	O	O
effect	NN	O	O
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
a	NN	O	O
functional	NN	O	O
RXR	NN	O	B-protein
-pathway	NN	O	O
was	NN	O	O
also	NN	O	O
seen	NN	O	O
in	NN	O	O
lipopolysaccharide-induced	NN	O	O
inhibition	NN	O	O
of	NN	O	O
mouse	NN	O	B-cell_type
splenocyte	NN	O	I-cell_type
proliferation	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
strongly	NN	O	O
suggest	NN	O	O
the	NN	O	O
existence	NN	O	O
of	NN	O	O
a	NN	O	O
pharmacological	NN	O	O
RXR	NN	O	B-protein
-dependent	NN	O	O
pathway	NN	O	O
that	NN	O	O
is	NN	O	O
activated	NN	O	O
by	NN	O	O
a	NN	O	O
ligand	NN	O	O
that	NN	O	O
can	NN	O	O
bind	NN	O	O
to	NN	O	O
RXR	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Targeting	NN	O	O
Src	NN	O	B-protein
homology	NN	O	I-protein
2	NN	O	I-protein
domain-containing	NN	O	I-protein
tyrosine	NN	O	I-protein
phosphatase	NN	O	I-protein
(	NN	O	O
SHP-1	NN	O	B-protein
)	NN	O	O
into	NN	O	O
lipid	NN	O	O
rafts	NN	O	O
inhibits	NN	O	O
CD3	NN	O	B-protein
-induced	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
activation	NN	O	O
.	NN	O	O

To	NN	O	O
study	NN	O	O
the	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
protein	NN	O	B-protein
tyrosine	NN	O	I-protein
phosphatases	NN	O	I-protein
(	NN	O	O
PTPs	NN	O	B-protein
)	NN	O	O
regulate	NN	O	O
CD3	NN	O	B-protein
-induced	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
the	NN	O	O
distribution	NN	O	O
of	NN	O	O
PTPs	NN	O	B-protein
in	NN	O	O
subdomains	NN	O	O
of	NN	O	O
plasma	NN	O	O
membrane	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
bulk	NN	O	O
PTP	NN	O	B-protein
activity	NN	O	O
associated	NN	O	O
with	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
membrane	NN	O	O
is	NN	O	O
present	NN	O	O
outside	NN	O	O
the	NN	O	O
lipid	NN	O	O
rafts	NN	O	O
,	NN	O	O
as	NN	O	O
determined	NN	O	O
by	NN	O	O
sucrose	NN	O	O
density	NN	O	O
gradient	NN	O	O
sedimentation	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
approximately	NN	O	O
5	NN	O	O
--	NN	O	O
10	NN	O	O
%	NN	O	O
of	NN	O	O
Src	NN	O	B-protein
homology	NN	O	I-protein
2	NN	O	I-protein
domain-containing	NN	O	I-protein
tyrosine	NN	O	I-protein
phosphatase	NN	O	I-protein
(	NN	O	O
SHP-1	NN	O	B-protein
)	NN	O	O
is	NN	O	O
constitutively	NN	O	O
associated	NN	O	O
with	NN	O	O
plasma	NN	O	O
membrane	NN	O	O
,	NN	O	O
and	NN	O	O
nearly	NN	O	O
50	NN	O	O
%	NN	O	O
of	NN	O	O
SHP-2	NN	O	B-protein
is	NN	O	O
translocated	NN	O	O
to	NN	O	O
plasma	NN	O	O
membrane	NN	O	O
after	NN	O	O
vanadate	NN	O	O
treatment	NN	O	O
.	NN	O	O

Similar	NN	O	O
to	NN	O	O
transmembrane	NN	O	B-protein
PTP	NN	O	I-protein
,	NN	O	O
CD45	NN	O	B-protein
,	NN	O	O
the	NN	O	O
membrane-associated	NN	O	O
populations	NN	O	O
of	NN	O	O
SHP-1	NN	O	B-protein
and	NN	O	O
SHP-2	NN	O	B-protein
are	NN	O	O
essentially	NN	O	O
excluded	NN	O	O
from	NN	O	O
lipid	NN	O	O
rafts	NN	O	O
,	NN	O	O
where	NN	O	O
other	NN	O	O
signaling	NN	O	B-protein
molecules	NN	O	I-protein
such	NN	O	O
as	NN	O	O
Lck	NN	O	B-protein
,	NN	O	I-protein
linker	NN	O	I-protein
for	NN	O	I-protein
activation	NN	O	I-protein
of	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
,	NN	O	O
and	NN	O	O
CD3	NN	O	B-protein
zeta	NN	O	I-protein
are	NN	O	O
enriched	NN	O	O
.	NN	O	O

We	NN	O	O
further	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
CD3	NN	O	B-protein
-induced	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
these	NN	O	O
substrates	NN	O	O
is	NN	O	O
largely	NN	O	O
restricted	NN	O	O
to	NN	O	O
lipid	NN	O	O
rafts	NN	O	O
,	NN	O	O
unless	NN	O	O
PTPs	NN	O	B-protein
are	NN	O	O
inhibited	NN	O	O
.	NN	O	O

It	NN	O	O
suggests	NN	O	O
that	NN	O	O
a	NN	O	O
restricted	NN	O	O
partition	NN	O	O
of	NN	O	O
PTPs	NN	O	B-protein
among	NN	O	O
membrane	NN	O	O
subdomains	NN	O	O
may	NN	O	O
regulate	NN	O	O
protein	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
membrane	NN	O	O
.	NN	O	O

To	NN	O	O
test	NN	O	O
this	NN	O	O
hypothesis	NN	O	O
,	NN	O	O
we	NN	O	O
targeted	NN	O	O
SHP-1	NN	O	B-protein
into	NN	O	O
lipid	NN	O	O
rafts	NN	O	O
by	NN	O	O
using	NN	O	O
the	NN	O	O
N-terminal	NN	O	B-protein
region	NN	O	I-protein
of	NN	O	I-protein
Lck	NN	O	I-protein
(	NN	O	O
residues	NN	O	B-protein
1	NN	O	I-protein
--	NN	O	I-protein
14	NN	O	I-protein
)	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
Lck/SHP-1	NN	O	B-protein
chimera	NN	O	I-protein
inside	NN	O	O
lipid	NN	O	O
rafts	NN	O	O
profoundly	NN	O	O
inhibits	NN	O	O
CD3	NN	O	B-protein
-induced	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
CD3	NN	O	B-protein
zeta/epsilon	NN	O	I-protein
,	NN	O	O
IL-2	NN	O	O
generation	NN	O	O
,	NN	O	O
and	NN	O	O
nuclear	NN	O	O
mobilization	NN	O	O
of	NN	O	O
NF-AT	NN	O	B-protein
.	NN	O	O

Collectively	NN	O	O
,	NN	O	O
these	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
exclusion	NN	O	O
of	NN	O	O
PTPs	NN	O	B-protein
from	NN	O	O
lipid	NN	O	O
rafts	NN	O	O
may	NN	O	O
be	NN	O	O
a	NN	O	O
mechanism	NN	O	O
that	NN	O	O
potentiates	NN	O	O
TCR	NN	O	B-protein
/	NN	O	O
CD3	NN	O	B-protein
activation	NN	O	O

-DOCSTART-	O

An	NN	O	O
instructive	NN	O	O
component	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
type	NN	O	I-cell_type
2	NN	O	I-cell_type
(	NN	O	O
Th2	NN	O	B-cell_type
)	NN	O	O
development	NN	O	O
mediated	NN	O	O
by	NN	O	O
GATA-3	NN	O	B-protein
.	NN	O	O

Although	NN	O	O
interleukin	NN	O	B-protein
(	NN	O	I-protein
IL	NN	O	I-protein
)	NN	O	I-protein
-12	NN	O	I-protein
and	NN	O	O
IL-4	NN	O	B-protein
polarize	NN	O	O
naive	NN	O	B-cell_type
CD4	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
toward	NN	O	O
T	NN	O	O
helper	NN	O	O
cell	NN	O	O
type	NN	O	O
1	NN	O	O
(	NN	O	O
Th1	NN	O	O
)	NN	O	O
or	NN	O	O
Th2	NN	O	O
phenotypes	NN	O	O
,	NN	O	O
it	NN	O	O
is	NN	O	O
not	NN	O	O
known	NN	O	O
whether	NN	O	O
cytokines	NN	O	B-protein
instruct	NN	O	O
the	NN	O	O
developmental	NN	O	O
fate	NN	O	O
in	NN	O	O
uncommitted	NN	O	B-cell_type
progenitors	NN	O	I-cell_type
or	NN	O	O
select	NN	O	O
for	NN	O	O
outgrowth	NN	O	O
of	NN	O	O
cells	NN	O	O
that	NN	O	O
have	NN	O	O
stochastically	NN	O	O
committed	NN	O	O
to	NN	O	O
a	NN	O	O
particular	NN	O	O
fate	NN	O	O
.	NN	O	O

To	NN	O	O
distinguish	NN	O	O
these	NN	O	O
instructive	NN	O	O
and	NN	O	O
selective	NN	O	O
models	NN	O	O
,	NN	O	O
we	NN	O	O
used	NN	O	O
surface	NN	O	O
affinity	NN	O	O
matrix	NN	O	O
technology	NN	O	O
to	NN	O	O
isolate	NN	O	O
committed	NN	O	B-cell_type
progenitors	NN	O	I-cell_type
based	NN	O	O
on	NN	O	O
cytokine	NN	O	O
secretion	NN	O	O
phenotype	NN	O	O
and	NN	O	O
developed	NN	O	O
retroviral-based	NN	O	O
tagging	NN	O	O
approaches	NN	O	O
to	NN	O	O
directly	NN	O	O
monitor	NN	O	O
individual	NN	O	O
progenitor	NN	O	O
fate	NN	O	O
decisions	NN	O	O
at	NN	O	O
the	NN	O	O
clonal	NN	O	O
and	NN	O	O
population	NN	O	O
levels	NN	O	O
.	NN	O	O

We	NN	O	O
observe	NN	O	O
IL-4	NN	O	B-protein
-dependent	NN	O	O
redirection	NN	O	O
of	NN	O	O
phenotype	NN	O	O
in	NN	O	O
cells	NN	O	O
that	NN	O	O
have	NN	O	O
already	NN	O	O
committed	NN	O	O
to	NN	O	O
a	NN	O	O
non-IL-4-producing	NN	O	O
fate	NN	O	O
,	NN	O	O
inconsistent	NN	O	O
with	NN	O	O
predictions	NN	O	O
of	NN	O	O
the	NN	O	O
selective	NN	O	O
model	NN	O	O
.	NN	O	O

Further	NN	O	O
,	NN	O	O
retroviral	NN	O	O
tagging	NN	O	O
of	NN	O	O
naive	NN	O	B-cell_type
progenitors	NN	O	I-cell_type
with	NN	O	O
the	NN	O	O
Th2-specific	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
GATA-3	NN	O	I-protein
provided	NN	O	O
direct	NN	O	O
evidence	NN	O	O
for	NN	O	O
instructive	NN	O	O
differentiation	NN	O	O
,	NN	O	O
and	NN	O	O
no	NN	O	O
evidence	NN	O	O
for	NN	O	O
the	NN	O	O
selective	NN	O	O
outgrowth	NN	O	O
of	NN	O	O
cells	NN	O	O
committed	NN	O	O
to	NN	O	O
either	NN	O	O
the	NN	O	O
Th1	NN	O	O
or	NN	O	O
Th2	NN	O	O
fate	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
would	NN	O	O
seem	NN	O	O
to	NN	O	O
exclude	NN	O	O
selection	NN	O	O
as	NN	O	O
an	NN	O	O
exclusive	NN	O	O
mechanism	NN	O	O
in	NN	O	O
Th1	NN	O	B-cell_type
/	NN	O	O
Th2	NN	O	B-cell_type
differentiation	NN	O	O
,	NN	O	O
and	NN	O	O
support	NN	O	O
an	NN	O	O
instructive	NN	O	O
model	NN	O	O
of	NN	O	O
cytokine-driven	NN	O	O
transcriptional	NN	O	O
programming	NN	O	O
of	NN	O	O
cell	NN	O	O
fate	NN	O	O
decisions	NN	O	O
.	NN	O	O

-DOCSTART-	O

In	NN	O	O
vitro-activated	NN	O	O
human	NN	O	B-cell_type
lupus	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
express	NN	O	O
normal	NN	O	O
estrogen	NN	O	B-protein
receptor	NN	O	I-protein
proteins	NN	O	I-protein
which	NN	O	O
bind	NN	O	O
to	NN	O	O
the	NN	O	O
estrogen	NN	O	B-protein
response	NN	O	I-protein
element	NN	O	I-protein
.	NN	O	O

We	NN	O	O
have	NN	O	O
shown	NN	O	O
that	NN	O	O
estrogen	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
ERalpha	NN	O	B-protein
,	NN	O	O
ERbeta	NN	O	B-protein
)	NN	O	O
transcripts	NN	O	O
are	NN	O	O
expressed	NN	O	O
in	NN	O	O
SLE	NN	O	B-cell_type
and	NN	O	I-cell_type
normal	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
T	NN	O	O
cell	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
from	NN	O	O
female	NN	O	O
lupus	NN	O	O
patients	NN	O	O
and	NN	O	O
normal	NN	O	O
donors	NN	O	O
were	NN	O	O
tested	NN	O	O
for	NN	O	O
biologically	NN	O	O
active	NN	O	O
ER	NN	O	B-protein
proteins	NN	O	I-protein
capable	NN	O	O
of	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
estrogen	NN	O	I-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
hERE	NN	O	B-DNA
)	NN	O	O
by	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
.	NN	O	O

When	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
stimulated	NN	O	O
with	NN	O	O
17beta-estradiol	NN	O	O
(	NN	O	O
E2	NN	O	O
)	NN	O	O
,	NN	O	O
PMA	NN	O	O
and	NN	O	O
ionomycin	NN	O	O
,	NN	O	O
two	NN	O	O
major	NN	O	O
retarded	NN	O	O
bands	NN	O	O
in	NN	O	O
T	NN	O	O
cell	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
exhibited	NN	O	O
a	NN	O	O
migration	NN	O	O
pattern	NN	O	O
similar	NN	O	O
to	NN	O	O
slow	NN	O	O
migrating	NN	O	O
protein-ERE	NN	O	B-protein
complexes	NN	O	I-protein
in	NN	O	O
human	NN	O	O
breast	NN	O	O
cancer	NN	O	O
cell	NN	O	O
extracts	NN	O	O
.	NN	O	O

T	NN	O	B-cell_type
cells	NN	O	I-cell_type
cultured	NN	O	O
only	NN	O	O
with	NN	O	O
E2	NN	O	O
did	NN	O	O
not	NN	O	O
have	NN	O	O
these	NN	O	O
complexes	NN	O	O
.	NN	O	O

The	NN	O	O
formation	NN	O	O
of	NN	O	O
the	NN	O	O
complexes	NN	O	O
was	NN	O	O
inhibited	NN	O	O
by	NN	O	O
competition	NN	O	O
with	NN	O	O
the	NN	O	O
hERE	NN	O	O
cold	NN	O	O
oligonucleotide	NN	O	O
and	NN	O	O
partially	NN	O	O
with	NN	O	O
anti-ERalpha	NN	O	B-protein
antibodies	NN	O	I-protein
.	NN	O	O

There	NN	O	O
was	NN	O	O
no	NN	O	O
notable	NN	O	O
difference	NN	O	O
in	NN	O	O
the	NN	O	O
migration	NN	O	O
pattern	NN	O	O
of	NN	O	O
ERE-binding	NN	O	B-protein
proteins	NN	O	I-protein
between	NN	O	O
the	NN	O	O
SLE	NN	O	O
and	NN	O	O
normal	NN	O	O
T	NN	O	O
cell	NN	O	O
extracts	NN	O	O
.	NN	O	O

Together	NN	O	O
,	NN	O	O
these	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
activated	NN	O	B-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
whether	NN	O	O
lupus-derived	NN	O	O
or	NN	O	O
normal-derived	NN	O	O
,	NN	O	O
contain	NN	O	O
biologically	NN	O	O
active	NN	O	O
ERalpha	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

Other	NN	O	O
factors	NN	O	O
may	NN	O	O
be	NN	O	O
responsible	NN	O	O
for	NN	O	O
differential	NN	O	O
sensitivity	NN	O	O
of	NN	O	O
lupus	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
estrogen	NN	O	O
.	NN	O	O

-DOCSTART-	O

Mechanism	NN	O	O
of	NN	O	O
the	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
of	NN	O	O
protease	NN	O	O
inhibitor	NN	O	O
on	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
production	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

If	NN	O	O
the	NN	O	O
inflammatory	NN	O	O
response	NN	O	O
becomes	NN	O	O
excessive	NN	O	O
or	NN	O	O
uncontrolled	NN	O	O
by	NN	O	O
some	NN	O	O
stimuli	NN	O	O
,	NN	O	O
inappropriate	NN	O	O
inflammatory	NN	O	O
responses	NN	O	O
occur	NN	O	O
.	NN	O	O

Monocytes	NN	O	B-cell_type
are	NN	O	O
extremely	NN	O	O
important	NN	O	O
cells	NN	O	O
for	NN	O	O
regulating	NN	O	O
the	NN	O	O
cytokine	NN	O	O
network	NN	O	O
and	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
TNFalpha	NN	O	B-protein
)	NN	O	O
and	NN	O	O
interleukin-	NN	O	B-protein
(	NN	O	I-protein
IL	NN	O	I-protein
)	NN	O	I-protein
10	NN	O	I-protein
,	NN	O	O
which	NN	O	O
are	NN	O	O
mainly	NN	O	O
synthesized	NN	O	O
by	NN	O	O
monocytes	NN	O	B-cell_type
,	NN	O	O
are	NN	O	O
representative	NN	O	O
cytokines	NN	O	B-protein
that	NN	O	O
play	NN	O	O
a	NN	O	O
central	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
cytokine	NN	O	O
network	NN	O	O
.	NN	O	O

Protease	NN	O	O
inhibitors	NN	O	O
such	NN	O	O
as	NN	O	O
gabexate	NN	O	O
mesilate	NN	O	O
(	NN	O	O
GM	NN	O	O
)	NN	O	O
and	NN	O	O
ulinastatin	NN	O	O
(	NN	O	O
UTI	NN	O	O
)	NN	O	O
have	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
have	NN	O	O
various	NN	O	O
beneficial	NN	O	O
effects	NN	O	O
by	NN	O	O
inhibiting	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
leukocytes	NN	O	B-cell_type
,	NN	O	O
but	NN	O	O
the	NN	O	O
mechanism	NN	O	O
for	NN	O	O
this	NN	O	O
has	NN	O	O
yet	NN	O	O
to	NN	O	O
be	NN	O	O
fully	NN	O	O
elucidated	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
we	NN	O	O
investigated	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
the	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
of	NN	O	O
protease	NN	O	O
inhibitors	NN	O	O
on	NN	O	O
the	NN	O	O
proinflammatory	NN	O	O
cytokine	NN	O	O
production	NN	O	O
of	NN	O	O
lipopolysaccharide-	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
stimulated	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

LPS-stimulated	NN	O	O
monocytes	NN	O	B-cell_type
were	NN	O	O
treated	NN	O	O
with	NN	O	O
GM	NN	O	O
or	NN	O	O
UTI	NN	O	O
.	NN	O	O

The	NN	O	O
value	NN	O	O
of	NN	O	O
TNFalpha	NN	O	B-protein
and	NN	O	O
IL-10	NN	O	B-protein
in	NN	O	O
the	NN	O	O
culture	NN	O	O
medium	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
was	NN	O	O
measured	NN	O	O
and	NN	O	O
each	NN	O	O
mRNA	NN	O	O
expression	NN	O	O
was	NN	O	O
assayed	NN	O	O
.	NN	O	O

The	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
of	NN	O	O
protease	NN	O	O
inhibitors	NN	O	O
on	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
intracellular	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathways	NN	O	O
such	NN	O	O
as	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
(	NN	O	O
PKC	NN	O	B-protein
)	NN	O	O
and	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
NFkappaB	NN	O	B-protein
)	NN	O	O
were	NN	O	O
also	NN	O	O
evaluated	NN	O	O
.	NN	O	O

GM	NN	O	O
decreased	NN	O	O
the	NN	O	O
TNFalpha	NN	O	B-protein
production	NN	O	O
of	NN	O	O
LPS-stimulated	NN	O	O
monocytes	NN	O	B-cell_type
as	NN	O	O
shown	NN	O	O
by	NN	O	O
the	NN	O	O
inhibition	NN	O	O
of	NN	O	O
mRNA	NN	O	O
expression	NN	O	O
and	NN	O	O
increased	NN	O	O
the	NN	O	O
IL-10	NN	O	B-protein
production	NN	O	O
of	NN	O	O
LPS-stimulated	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

GM	NN	O	O
also	NN	O	O
suppressed	NN	O	O
the	NN	O	O
NFkappaB	NN	O	B-protein
activity	NN	O	O
of	NN	O	O
LPS-stimulated	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

UTI	NN	O	O
decreased	NN	O	O
the	NN	O	O
TNFalpha	NN	O	B-protein
production	NN	O	O
of	NN	O	O
LPS-stimulated	NN	O	O
monocytes	NN	O	B-cell_type
,	NN	O	O
but	NN	O	O
did	NN	O	O
not	NN	O	O
inhibit	NN	O	O
the	NN	O	O
TNFalpha	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
.	NN	O	O

The	NN	O	O
present	NN	O	O
study	NN	O	O
shows	NN	O	O
that	NN	O	O
the	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
of	NN	O	O
GM	NN	O	O
on	NN	O	O
the	NN	O	O
TNFalpha	NN	O	B-protein
production	NN	O	O
of	NN	O	O
activated	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
suppression	NN	O	O
of	NN	O	O
NFkappaB	NN	O	B-protein
activation	NN	O	O
,	NN	O	O
while	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
UTI	NN	O	O
inhibiting	NN	O	O
TNFalpha	NN	O	B-protein
production	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
may	NN	O	O
be	NN	O	O
due	NN	O	O
to	NN	O	O
the	NN	O	O
inhibition	NN	O	O
of	NN	O	O
either	NN	O	O
the	NN	O	O
translation	NN	O	O
or	NN	O	O
secretion	NN	O	O
of	NN	O	O
TNFalpha	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Homocysteine	NN	O	O
stimulates	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
monocyte	NN	O	B-protein
chemoattractant	NN	O	I-protein
protein-1	NN	O	I-protein
in	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
leading	NN	O	O
to	NN	O	O
enhanced	NN	O	O
monocyte	NN	O	O
chemotaxis	NN	O	O
.	NN	O	O

Hyperhomocysteinemia	NN	O	O
has	NN	O	O
been	NN	O	O
identified	NN	O	O
as	NN	O	O
an	NN	O	O
independent	NN	O	O
risk	NN	O	O
factor	NN	O	O
for	NN	O	O
atherosclerosis	NN	O	O
.	NN	O	O

The	NN	O	O
infiltration	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
into	NN	O	O
the	NN	O	O
arterial	NN	O	O
wall	NN	O	O
is	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
key	NN	O	O
events	NN	O	O
during	NN	O	O
atherogenesis	NN	O	O
.	NN	O	O

Monocyte	NN	O	B-protein
chemoattractant	NN	O	I-protein
protein-1	NN	O	I-protein
(	NN	O	O
MCP-1	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
potent	NN	O	O
chemokine	NN	O	B-protein
that	NN	O	O
stimulates	NN	O	O
the	NN	O	O
migration	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
into	NN	O	O
the	NN	O	O
intima	NN	O	O
of	NN	O	O
the	NN	O	O
arterial	NN	O	O
wall	NN	O	O
.	NN	O	O

The	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
increased	NN	O	O
monocyte	NN	O	O
infiltration	NN	O	O
occurs	NN	O	O
in	NN	O	O
atherosclerotic	NN	O	O
lesions	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
hyperhomocysteinemia	NN	O	O
has	NN	O	O
not	NN	O	O
been	NN	O	O
delineated	NN	O	O
.	NN	O	O

The	NN	O	O
objective	NN	O	O
of	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
was	NN	O	O
to	NN	O	O
investigate	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
homocysteine	NN	O	O
on	NN	O	O
MCP-1	NN	O	B-protein
production	NN	O	O
in	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Cells	NN	O	O
were	NN	O	O
incubated	NN	O	O
with	NN	O	O
homocysteine	NN	O	O
.	NN	O	O

The	NN	O	O
secretion	NN	O	O
of	NN	O	O
MCP-1	NN	O	B-protein
protein	NN	O	I-protein
was	NN	O	O
significantly	NN	O	O
increased	NN	O	O
(	NN	O	O
195	NN	O	O
%	NN	O	O
as	NN	O	O
compared	NN	O	O
to	NN	O	O
the	NN	O	O
control	NN	O	O
)	NN	O	O
in	NN	O	O
cells	NN	O	O
treated	NN	O	O
with	NN	O	O
pathological	NN	O	O
concentrations	NN	O	O
of	NN	O	O
homocysteine	NN	O	O
.	NN	O	O

Such	NN	O	O
effect	NN	O	O
was	NN	O	O
accompanied	NN	O	O
by	NN	O	O
an	NN	O	O
increased	NN	O	O
expression	NN	O	O
of	NN	O	O
MCP-1	NN	O	B-RNA
mRNA	NN	O	I-RNA
(	NN	O	O
176	NN	O	O
%	NN	O	O
as	NN	O	O
compared	NN	O	O
to	NN	O	O
the	NN	O	O
control	NN	O	O
)	NN	O	O
in	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
which	NN	O	O
resulted	NN	O	O
in	NN	O	O
enhanced	NN	O	O
monocyte	NN	O	O
chemotaxis	NN	O	O
.	NN	O	O

The	NN	O	O
p38	NN	O	B-protein
MAP	NN	O	I-protein
kinase	NN	O	I-protein
as	NN	O	O
well	NN	O	O
as	NN	O	O
other	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
p38	NN	O	O
MAP	NN	O	O
kinase	NN	O	O
pathway	NN	O	O
,	NN	O	O
including	NN	O	O
MKK3	NN	O	B-protein
,	NN	O	O
MKK6	NN	O	B-protein
,	NN	O	O
ATF-2	NN	O	B-protein
and	NN	O	O
Elk-1	NN	O	B-protein
,	NN	O	O
were	NN	O	O
activated	NN	O	O
in	NN	O	O
homocysteine-treated	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Homocysteine-induced	NN	O	O
MCP-1	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
subsequent	NN	O	O
monocyte	NN	O	O
chemotaxis	NN	O	O
were	NN	O	O
blocked	NN	O	O
by	NN	O	O
a	NN	O	O
p38	NN	O	B-protein
MAP	NN	O	I-protein
kinase	NN	O	I-protein
inhibitor	NN	O	O
(	NN	O	O
SB203580	NN	O	O
)	NN	O	O
suggesting	NN	O	O
that	NN	O	O
the	NN	O	O
p38	NN	O	B-protein
MAP	NN	O	I-protein
kinase	NN	O	I-protein
pathway	NN	O	O
might	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
homocysteine-induced	NN	O	O
MCP-1	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
endothelial	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
staurosporine	NN	O	O
,	NN	O	O
a	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
inhibitor	NN	O	O
,	NN	O	O
had	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
homocysteine-induced	NN	O	O
MCP-1	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

In	NN	O	O
conclusion	NN	O	O
,	NN	O	O
our	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
homocysteine	NN	O	O
stimulates	NN	O	O
MCP-1	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
leading	NN	O	O
to	NN	O	O
enhanced	NN	O	O
monocyte	NN	O	O
chemotaxis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Inducible	NN	O	O
resistance	NN	O	O
to	NN	O	O
Fas-mediated	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Apoptosis	NN	O	O
produced	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
through	NN	O	O
Fas	NN	O	B-protein
(	NN	O	O
APO-1	NN	O	B-protein
,	NN	O	O
CD95	NN	O	B-protein
)	NN	O	O
triggering	NN	O	O
is	NN	O	O
regulated	NN	O	O
by	NN	O	O
signals	NN	O	O
derived	NN	O	O
from	NN	O	O
other	NN	O	O
surface	NN	O	O
receptors	NN	O	O
:	NN	O	O
CD40	NN	O	O
engagement	NN	O	O
produces	NN	O	O
upregulation	NN	O	O
of	NN	O	O
Fas	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
marked	NN	O	O
susceptibility	NN	O	O
to	NN	O	O
Fas	NN	O	B-protein
-induced	NN	O	O
cell	NN	O	O
death	NN	O	O
,	NN	O	O
whereas	NN	O	O
antigen	NN	O	O
receptor	NN	O	O
engagement	NN	O	O
,	NN	O	O
or	NN	O	O
IL-4R	NN	O	O
engagement	NN	O	O
,	NN	O	O
inhibits	NN	O	O
Fas	NN	O	B-protein
killing	NN	O	O
and	NN	O	O
in	NN	O	O
so	NN	O	O
doing	NN	O	O
induces	NN	O	O
a	NN	O	O
state	NN	O	O
of	NN	O	O
Fas	NN	O	B-protein
-resistance	NN	O	O
,	NN	O	O
even	NN	O	O
in	NN	O	O
otherwise	NN	O	O
sensitive	NN	O	O
,	NN	O	O
CD40	NN	O	B-protein
-stimulated	NN	O	O
targets	NN	O	O
.	NN	O	O

Surface	NN	O	B-protein
immunoglobulin	NN	O	I-protein
and	NN	O	O
IL-4R	NN	O	B-protein
utilize	NN	O	O
at	NN	O	O
least	NN	O	O
partially	NN	O	O
distinct	NN	O	O
pathways	NN	O	O
to	NN	O	O
produce	NN	O	O
Fas	NN	O	B-protein
-resistance	NN	O	O
that	NN	O	O
differentially	NN	O	O
depend	NN	O	O
on	NN	O	O
PKC	NN	O	B-protein
and	NN	O	O
STAT6	NN	O	B-protein
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

Further	NN	O	O
,	NN	O	O
surface	NN	O	O
immunoglobulin	NN	O	O
signaling	NN	O	O
for	NN	O	O
inducible	NN	O	O
Fas	NN	O	B-protein
-resistance	NN	O	O
bypasses	NN	O	O
Btk	NN	O	B-protein
,	NN	O	O
requires	NN	O	O
NF-kappaB	NN	O	B-protein
,	NN	O	O
and	NN	O	O
entails	NN	O	O
new	NN	O	O
macromolecular	NN	O	O
synthesis	NN	O	O
.	NN	O	O

Terminal	NN	O	O
effectors	NN	O	O
of	NN	O	O
B	NN	O	B-cell_type
cell	NN	O	I-cell_type
Fas	NN	O	B-protein
-resistance	NN	O	O
include	NN	O	O
the	NN	O	O
known	NN	O	O
anti-apoptotic	NN	O	B-protein
gene	NN	O	I-protein
products	NN	O	I-protein
,	NN	O	O
Bcl-xL	NN	O	B-protein
and	NN	O	O
FLIP	NN	O	B-protein
,	NN	O	O
and	NN	O	O
a	NN	O	O
novel	NN	O	O
anti-apoptotic	NN	O	B-DNA
gene	NN	O	I-DNA
that	NN	O	O
encodes	NN	O	O
FAIM	NN	O	B-protein
(	NN	O	O
Fas	NN	O	B-protein
Apoptosis	NN	O	I-protein
Inhibitory	NN	O	I-protein
Molecule	NN	O	I-protein
)	NN	O	O
.	NN	O	O

faim	NN	O	B-protein
was	NN	O	O
identified	NN	O	O
by	NN	O	O
differential	NN	O	O
display	NN	O	O
and	NN	O	O
exists	NN	O	O
in	NN	O	O
two	NN	O	O
alternatively	NN	O	O
spliced	NN	O	O
forms	NN	O	O
;	NN	O	O
faim-S	NN	O	B-protein
is	NN	O	O
broadly	NN	O	O
expressed	NN	O	O
,	NN	O	O
but	NN	O	O
faim-L	NN	O	O
expression	NN	O	O
is	NN	O	O
tissue-specific	NN	O	O
.	NN	O	O

The	NN	O	O
FAIM	NN	O	B-DNA
sequence	NN	O	I-DNA
is	NN	O	O
highly	NN	O	O
evolu-	NN	O	O
tionarily	NN	O	O
conserved	NN	O	O
,	NN	O	O
suggesting	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
for	NN	O	O
this	NN	O	O
molecule	NN	O	O
throughout	NN	O	O
phylogeny	NN	O	O
.	NN	O	O

Inducible	NN	O	O
resistance	NN	O	O
to	NN	O	O
Fas	NN	O	B-protein
killing	NN	O	O
is	NN	O	O
hypothesized	NN	O	O
to	NN	O	O
protect	NN	O	O
foreign	NN	O	O
antigen-specific	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
during	NN	O	O
potentially	NN	O	O
hazardous	NN	O	O
interactions	NN	O	O
with	NN	O	O
FasL-bearing	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
whereas	NN	O	O
autoreactive	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
fail	NN	O	O
to	NN	O	O
become	NN	O	O
Fas	NN	O	B-protein
-resistant	NN	O	O
and	NN	O	O
are	NN	O	O
deleted	NN	O	O
via	NN	O	O
Fas	NN	O	B-protein
-dependent	NN	O	O
cytotoxicity	NN	O	O
.	NN	O	O

Inadvertent	NN	O	O
or	NN	O	O
aberrant	NN	O	O
acquisition	NN	O	O
of	NN	O	O
Fas	NN	O	B-protein
-resistance	NN	O	O
may	NN	O	O
permit	NN	O	O
autoreactive	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
escape	NN	O	O
Fas	NN	O	B-protein
deletion	NN	O	O
,	NN	O	O
and	NN	O	O
malignant	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
to	NN	O	O
impede	NN	O	O
anti-tumor	NN	O	O
immunity	NN	O	O
.	NN	O	O

-DOCSTART-	O

Stromal	NN	O	B-protein
cell-derived	NN	O	I-protein
factor	NN	O	I-protein
1	NN	O	I-protein
alpha	NN	O	I-protein
-induced	NN	O	O
chemotaxis	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
nitric	NN	O	O
oxide	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
.	NN	O	O

Stromal	NN	O	B-protein
cell-derived	NN	O	I-protein
factor	NN	O	I-protein
1	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
SDF1	NN	O	B-protein
alpha	NN	O	I-protein
)	NN	O	O
and	NN	O	O
its	NN	O	O
cognate	NN	O	O
chemokine	NN	O	B-protein
receptor	NN	O	I-protein
CXCR4	NN	O	I-protein
act	NN	O	O
as	NN	O	O
potent	NN	O	O
chemoattractants	NN	O	O
and	NN	O	O
regulate	NN	O	O
trafficking	NN	O	O
and	NN	O	O
homing	NN	O	O
of	NN	O	O
hematopoietic	NN	O	B-cell_type
progenitor	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
lymphocytes	NN	O	B-cell_type
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
regulating	NN	O	O
SDF1	NN	O	B-protein
alpha	NN	O	I-protein
-driven	NN	O	O
cell	NN	O	O
migration	NN	O	O
are	NN	O	O
not	NN	O	O
well	NN	O	O
defined	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
explored	NN	O	O
the	NN	O	O
roles	NN	O	O
of	NN	O	O
the	NN	O	O
second	NN	O	O
messenger	NN	O	O
NO	NN	O	O
and	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
SDF1	NN	O	B-protein
alpha	NN	O	I-protein
-induced	NN	O	O
T	NN	O	O
cell	NN	O	O
migration	NN	O	O
.	NN	O	O

SDF1	NN	O	B-protein
alpha	NN	O	I-protein
treatment	NN	O	O
of	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
increased	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
NO	NN	O	B-protein
synthase	NN	O	I-protein
,	NN	O	O
which	NN	O	O
catalyzes	NN	O	O
the	NN	O	O
generation	NN	O	O
of	NN	O	O
NO	NN	O	O
.	NN	O	O

We	NN	O	O
observed	NN	O	O
that	NN	O	O
pretreatment	NN	O	O
of	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
or	NN	O	O
activated	NN	O	O
PBLs	NN	O	B-cell_type
with	NN	O	O
several	NN	O	O
NO	NN	O	O
donors	NN	O	O
significantly	NN	O	O
enhanced	NN	O	O
the	NN	O	O
SDF1	NN	O	B-protein
alpha	NN	O	I-protein
-induced	NN	O	O
migration	NN	O	O
,	NN	O	O
whereas	NN	O	O
various	NN	O	O
inhibitors	NN	O	O
of	NN	O	O
NO	NN	O	B-protein
synthase	NN	O	I-protein
markedly	NN	O	O
abrogated	NN	O	O
the	NN	O	O
chemotactic	NN	O	O
response	NN	O	O
in	NN	O	O
a	NN	O	O
concentration-dependent	NN	O	O
manner	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
we	NN	O	O
observed	NN	O	O
that	NN	O	O
inhibitors	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
linked	NN	O	O
to	NN	O	O
NO	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
,	NN	O	O
also	NN	O	O
significantly	NN	O	O
blocked	NN	O	O
the	NN	O	O
SDF1	NN	O	B-protein
alpha	NN	O	I-protein
-induced	NN	O	O
chemotactic	NN	O	O
response	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
these	NN	O	O
compounds	NN	O	O
did	NN	O	O
not	NN	O	O
have	NN	O	O
a	NN	O	O
significant	NN	O	O
effect	NN	O	O
on	NN	O	O
SDF1	NN	O	B-protein
alpha	NN	O	I-protein
-induced	NN	O	O
mitogen-activated	NN	O	O
protein	NN	O	O
kinase	NN	O	O
activity	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
the	NN	O	O
MAP/Erk	NN	O	O
kinase	NN	O	O
kinase	NN	O	O
inhibitor	NN	O	O
PD98059	NN	O	O
did	NN	O	O
not	NN	O	O
abrogate	NN	O	O
SDF1	NN	O	B-protein
alpha	NN	O	I-protein
-induced	NN	O	O
chemotaxis	NN	O	O
.	NN	O	O

AKT	NN	O	B-protein
,	NN	O	O
which	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
mediate	NN	O	O
NO	NN	O	O
production	NN	O	O
,	NN	O	O
was	NN	O	O
also	NN	O	O
phosphorylated	NN	O	O
upon	NN	O	O
SDF1	NN	O	B-protein
alpha	NN	O	I-protein
stimulation	NN	O	O
.	NN	O	O

These	NN	O	O
studies	NN	O	O
suggest	NN	O	O
that	NN	O	O
NO-related	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
may	NN	O	O
mediate	NN	O	O
SDF1	NN	O	B-protein
alpha	NN	O	I-protein
-induced	NN	O	O
chemotaxis	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
lack	NN	O	O
of	NN	O	O
NF-kappa	NN	O	O
B	NN	O	O
transactivation	NN	O	O
and	NN	O	O
PKC	NN	O	O
epsilon	NN	O	O
expression	NN	O	O
in	NN	O	O
CD4	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
CD8	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
thymocytes	NN	O	I-cell_type
correlates	NN	O	O
with	NN	O	O
negative	NN	O	O
selection	NN	O	O
.	NN	O	O

Deletion	NN	O	O
of	NN	O	O
autoreactive	NN	O	B-cell_type
thymocytes	NN	O	I-cell_type
at	NN	O	O
the	NN	O	O
DP	NN	O	O
stage	NN	O	O
is	NN	O	O
the	NN	O	O
basis	NN	O	O
for	NN	O	O
tolerance	NN	O	O
to	NN	O	O
thymus-expressed	NN	O	B-protein
self	NN	O	I-protein
antigens	NN	O	I-protein
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
we	NN	O	O
investigated	NN	O	O
whether	NN	O	O
distinct	NN	O	O
signalling	NN	O	O
pathways	NN	O	O
are	NN	O	O
induced	NN	O	O
in	NN	O	O
DP	NN	O	B-cell_type
thymocytes	NN	O	I-cell_type
as	NN	O	O
compared	NN	O	O
to	NN	O	O
mature	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
upon	NN	O	O
stimulation	NN	O	O
with	NN	O	O
antigen	NN	O	O
.	NN	O	O

Using	NN	O	O
triple	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
expressing	NN	O	O
a	NN	O	O
TCR	NN	O	B-DNA
transgene	NN	O	I-DNA
,	NN	O	O
dominant	NN	O	B-protein
negative	NN	O	I-protein
ras/Mek	NN	O	I-protein
proteins	NN	O	I-protein
and	NN	O	O
a	NN	O	O
reporter	NN	O	B-DNA
gene	NN	O	I-DNA
construct	NN	O	I-DNA
with	NN	O	O
AP-1	NN	O	B-DNA
or	NN	O	I-DNA
NF-kappa	NN	O	I-DNA
B	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
,	NN	O	O
we	NN	O	O
showed	NN	O	O
a	NN	O	O
complete	NN	O	O
lack	NN	O	O
of	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
but	NN	O	O
not	NN	O	O
AP-1	NN	O	B-protein
in	NN	O	O
DP	NN	O	B-cell_type
thymocytes	NN	O	I-cell_type
,	NN	O	O
whereas	NN	O	O
both	NN	O	O
were	NN	O	O
transcriptionally	NN	O	O
active	NN	O	O
in	NN	O	O
mature	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
after	NN	O	O
antigenic	NN	O	O
stimulation	NN	O	O
.	NN	O	O

Lack	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
induction	NN	O	O
correlated	NN	O	O
with	NN	O	O
increased	NN	O	O
death	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
antigen	NN	O	B-protein
.	NN	O	O

AP-1	NN	O	B-protein
induction	NN	O	O
was	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
integrity	NN	O	O
of	NN	O	O
the	NN	O	O
ras/Mek	NN	O	O
pathway	NN	O	O
indicating	NN	O	O
that	NN	O	O
this	NN	O	O
pathway	NN	O	O
was	NN	O	O
activated	NN	O	O
in	NN	O	O
the	NN	O	O
DP	NN	O	B-cell_type
thymocytes	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
a	NN	O	O
complete	NN	O	O
lack	NN	O	O
of	NN	O	O
constitutive	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
epsilon	NN	O	B-protein
isoform	NN	O	I-protein
of	NN	O	I-protein
Protein	NN	O	I-protein
Kinase	NN	O	I-protein
C	NN	O	I-protein
(	NN	O	O
PKC	NN	O	B-protein
)	NN	O	O
in	NN	O	O
DP	NN	O	B-cell_type
thymocytes	NN	O	I-cell_type
,	NN	O	O
although	NN	O	O
it	NN	O	O
was	NN	O	O
present	NN	O	O
in	NN	O	O
mature	NN	O	B-cell_type
thymocytes	NN	O	I-cell_type
and	NN	O	O
peripheral	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
the	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
lack	NN	O	O
of	NN	O	O
PKC	NN	O	B-protein
epsilon	NN	O	I-protein
in	NN	O	O
DP	NN	O	B-cell_type
thymocytes	NN	O	I-cell_type
could	NN	O	O
lead	NN	O	O
to	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activity	NN	O	O
after	NN	O	O
antigenic	NN	O	O
stimulation	NN	O	O
contributing	NN	O	O
to	NN	O	O
negative	NN	O	O
selection	NN	O	O
.	NN	O	O

Cell	NN	O	O
Death	NN	O	O
and	NN	O	O
Differentiation	NN	O	O
(	NN	O	O
2000	NN	O	O
)	NN	O	O
7	NN	O	O
,	NN	O	O
1253	NN	O	O
-	NN	O	O
1262	NN	O	O
.	NN	O	O

-DOCSTART-	O

CD2	NN	O	O
stimulation	NN	O	O
leads	NN	O	O
to	NN	O	O
the	NN	O	O
delayed	NN	O	O
and	NN	O	O
prolonged	NN	O	O
activation	NN	O	O
of	NN	O	O
STAT1	NN	O	B-protein
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
but	NN	O	O
not	NN	O	O
NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

OBJECTIVE	NN	O	O
:	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
can	NN	O	O
be	NN	O	O
activated	NN	O	O
by	NN	O	O
soluble	NN	O	O
factors	NN	O	O
such	NN	O	O
as	NN	O	O
cytokines	NN	O	B-protein
or	NN	O	O
through	NN	O	O
direct	NN	O	O
cell-cell	NN	O	O
interactions	NN	O	O
.	NN	O	O

Although	NN	O	O
cytokine	NN	O	B-protein
receptors	NN	O	I-protein
are	NN	O	O
known	NN	O	O
to	NN	O	O
signal	NN	O	O
through	NN	O	O
STAT	NN	O	B-protein
family	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
,	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
by	NN	O	O
which	NN	O	O
other	NN	O	O
cell-surface	NN	O	B-protein
molecules	NN	O	I-protein
,	NN	O	O
such	NN	O	O
as	NN	O	O
CD2	NN	O	B-protein
,	NN	O	O
transduce	NN	O	O
signals	NN	O	O
is	NN	O	O
unclear	NN	O	O
.	NN	O	O

The	NN	O	O
goal	NN	O	O
of	NN	O	O
this	NN	O	O
study	NN	O	O
was	NN	O	O
to	NN	O	O
determine	NN	O	O
whether	NN	O	O
stimulation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
through	NN	O	O
CD2	NN	O	B-protein
recapitulates	NN	O	O
aspects	NN	O	O
of	NN	O	O
cytokine	NN	O	B-protein
-induced	NN	O	O
T-cell	NN	O	O
activation	NN	O	O
by	NN	O	O
use	NN	O	O
of	NN	O	O
STAT	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

MATERIALS	NN	O	O
AND	NN	O	O
METHODS	NN	O	O
:	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
treated	NN	O	O
with	NN	O	O
anti-CD2	NN	O	B-protein
antibodies	NN	O	I-protein
or	NN	O	O
cells	NN	O	O
bearing	NN	O	O
the	NN	O	O
natural	NN	O	O
CD2	NN	O	B-protein
ligand	NN	O	I-protein
CD58	NN	O	I-protein
,	NN	O	O
after	NN	O	O
which	NN	O	O
signaling	NN	O	O
through	NN	O	O
STAT	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
was	NN	O	O
assessed	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
Stimulation	NN	O	O
of	NN	O	O
CD2	NN	O	B-protein
on	NN	O	O
primary	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
leads	NN	O	O
to	NN	O	O
the	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
,	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
,	NN	O	O
and	NN	O	O
DNA	NN	O	O
binding	NN	O	O
of	NN	O	O
STAT1	NN	O	B-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
stimulation	NN	O	O
by	NN	O	O
cytokines	NN	O	B-protein
,	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
STAT1	NN	O	B-protein
in	NN	O	O
response	NN	O	O
to	NN	O	O
CD2	NN	O	O
ligation	NN	O	O
is	NN	O	O
delayed	NN	O	O
and	NN	O	O
does	NN	O	O
not	NN	O	O
involve	NN	O	O
Jak	NN	O	B-protein
kinases	NN	O	I-protein
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
while	NN	O	O
STAT	NN	O	O
phosphorylation	NN	O	O
induced	NN	O	O
by	NN	O	O
cytokines	NN	O	B-protein
is	NN	O	O
generally	NN	O	O
transient	NN	O	O
,	NN	O	O
STAT1	NN	O	O
phosphorylation	NN	O	O
following	NN	O	O
CD2	NN	O	O
stimulation	NN	O	O
persists	NN	O	O
for	NN	O	O
a	NN	O	O
period	NN	O	O
of	NN	O	O
days	NN	O	O
.	NN	O	O

Transcription	NN	O	O
of	NN	O	O
key	NN	O	O
target	NN	O	O
genes	NN	O	O
such	NN	O	O
as	NN	O	O
IRF1	NN	O	B-DNA
and	NN	O	O
c-fos	NN	O	B-DNA
proceeds	NN	O	O
with	NN	O	O
delayed	NN	O	O
kinetics	NN	O	O
following	NN	O	O
CD2	NN	O	O
stimulation	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
this	NN	O	O
unique	NN	O	O
pattern	NN	O	O
of	NN	O	O
STAT	NN	O	O
activation	NN	O	O
may	NN	O	O
lead	NN	O	O
to	NN	O	O
a	NN	O	O
distinct	NN	O	O
cellular	NN	O	O
response	NN	O	O
following	NN	O	O
CD2	NN	O	O
ligation	NN	O	O
.	NN	O	O

This	NN	O	O
pathway	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
restricted	NN	O	O
to	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
as	NN	O	O
stimulation	NN	O	O
of	NN	O	O
CD2	NN	O	B-protein
on	NN	O	O
NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
does	NN	O	O
not	NN	O	O
lead	NN	O	O
to	NN	O	O
STAT1	NN	O	O
activation	NN	O	O
.	NN	O	O

CONCLUSION	NN	O	O
:	NN	O	O
Stimulation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
through	NN	O	O
cell-surface	NN	O	B-protein
molecules	NN	O	I-protein
such	NN	O	O
as	NN	O	O
CD2	NN	O	B-protein
involves	NN	O	O
activation	NN	O	O
of	NN	O	O
STAT	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
,	NN	O	O
thus	NN	O	O
recapitulating	NN	O	O
elements	NN	O	O
of	NN	O	O
cytokine	NN	O	O
signaling	NN	O	O
.	NN	O	O

-DOCSTART-	O

NFATc1	NN	O	B-protein
and	NN	O	O
NFATc2	NN	O	B-protein
together	NN	O	O
control	NN	O	O
both	NN	O	O
T	NN	O	O
and	NN	O	O
B	NN	O	O
cell	NN	O	O
activation	NN	O	O
and	NN	O	O
differentiation	NN	O	O
.	NN	O	O

NFAT	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
play	NN	O	O
critical	NN	O	O
roles	NN	O	O
in	NN	O	O
gene	NN	O	O
transcription	NN	O	O
during	NN	O	O
immune	NN	O	O
responses	NN	O	O
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
further	NN	O	O
the	NN	O	O
two	NN	O	O
most	NN	O	O
prominent	NN	O	O
NFAT	NN	O	B-protein
family	NN	O	I-protein
members	NN	O	I-protein
,	NN	O	O
NFATc1	NN	O	B-protein
and	NN	O	O
NFATc2	NN	O	B-protein
,	NN	O	O
we	NN	O	O
generated	NN	O	O
mice	NN	O	O
bearing	NN	O	O
lymphoid	NN	O	O
systems	NN	O	O
devoid	NN	O	O
of	NN	O	O
both	NN	O	O
.	NN	O	O

Doubly	NN	O	O
deficient	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
displayed	NN	O	O
cell	NN	O	O
surface	NN	O	O
markers	NN	O	O
of	NN	O	O
activation	NN	O	O
yet	NN	O	O
were	NN	O	O
significantly	NN	O	O
deficient	NN	O	O
in	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
multiple	NN	O	O
effector	NN	O	O
functions	NN	O	O
,	NN	O	O
including	NN	O	O
Th	NN	O	B-protein
cytokine	NN	O	I-protein
production	NN	O	O
,	NN	O	O
surface	NN	O	B-protein
effector	NN	O	I-protein
molecule	NN	O	I-protein
expression	NN	O	O
,	NN	O	O
and	NN	O	O
cytolytic	NN	O	O
activity	NN	O	O
.	NN	O	O

Nevertheless	NN	O	O
,	NN	O	O
doubly	NN	O	B-cell_type
deficient	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
hyperactivated	NN	O	O
,	NN	O	O
as	NN	O	O
evidenced	NN	O	O
by	NN	O	O
extremely	NN	O	O
elevated	NN	O	O
serum	NN	O	O
IgG1	NN	O	B-protein
and	NN	O	O
IgE	NN	O	B-protein
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
plasma	NN	O	B-cell_type
cell	NN	O	I-cell_type
expansion	NN	O	O
and	NN	O	O
infiltration	NN	O	O
of	NN	O	O
end	NN	O	O
organs	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
NFATc1	NN	O	B-protein
and	NN	O	O
NFATc2	NN	O	B-protein
are	NN	O	O
dispensable	NN	O	O
for	NN	O	O
inflammatory	NN	O	O
reactivity	NN	O	O
but	NN	O	O
are	NN	O	O
required	NN	O	O
for	NN	O	O
effector	NN	O	O
differentiation	NN	O	O
,	NN	O	O
while	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
NFATs	NN	O	B-protein
regulate	NN	O	O
both	NN	O	O
normal	NN	O	O
homeostasis	NN	O	O
and	NN	O	O
differentiation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Epstein-barr	NN	O	O
virus	NN	O	O
immediate-early	NN	O	B-protein
protein	NN	O	I-protein
BZLF1	NN	O	I-protein
is	NN	O	O
SUMO-1	NN	O	O
modified	NN	O	O
and	NN	O	O
disrupts	NN	O	O
promyelocytic	NN	O	O
leukemia	NN	O	O
bodies	NN	O	O
.	NN	O	O

Although	NN	O	O
the	NN	O	O
immediate-early	NN	O	B-protein
proteins	NN	O	I-protein
of	NN	O	O
both	NN	O	O
herpes	NN	O	O
simplex	NN	O	O
virus	NN	O	O
(	NN	O	O
HSV	NN	O	O
)	NN	O	O
and	NN	O	O
cytomegalovirus	NN	O	O
(	NN	O	O
CMV	NN	O	O
)	NN	O	O
are	NN	O	O
known	NN	O	O
to	NN	O	O
modify	NN	O	O
promyelocytic	NN	O	O
leukemia	NN	O	O
(	NN	O	O
PML	NN	O	O
)	NN	O	O
(	NN	O	O
ND10	NN	O	O
)	NN	O	O
bodies	NN	O	O
in	NN	O	O
the	NN	O	O
nucleus	NN	O	O
of	NN	O	O
the	NN	O	O
host	NN	O	O
cell	NN	O	O
,	NN	O	O
it	NN	O	O
has	NN	O	O
been	NN	O	O
unclear	NN	O	O
whether	NN	O	O
lytic	NN	O	O
infection	NN	O	O
with	NN	O	O
gamma	NN	O	O
herpesviruses	NN	O	O
induces	NN	O	O
a	NN	O	O
similar	NN	O	O
effect	NN	O	O
.	NN	O	O

The	NN	O	O
PML	NN	O	B-protein
protein	NN	O	I-protein
is	NN	O	O
induced	NN	O	O
by	NN	O	O
interferon	NN	O	B-protein
,	NN	O	O
involved	NN	O	O
in	NN	O	O
major	NN	O	B-protein
histocompatibility	NN	O	I-protein
complex	NN	O	I-protein
class	NN	O	I-protein
I	NN	O	I-protein
presentation	NN	O	O
,	NN	O	O
and	NN	O	O
necessary	NN	O	O
for	NN	O	O
certain	NN	O	O
types	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
it	NN	O	O
is	NN	O	O
likely	NN	O	O
that	NN	O	O
PML	NN	O	O
bodies	NN	O	O
function	NN	O	O
in	NN	O	O
an	NN	O	O
antiviral	NN	O	O
capacity	NN	O	O
.	NN	O	O

SUMO-1	NN	O	O
modification	NN	O	O
of	NN	O	O
PML	NN	O	B-protein
is	NN	O	O
known	NN	O	O
to	NN	O	O
be	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
PML	NN	O	O
bodies	NN	O	O
.	NN	O	O

To	NN	O	O
examine	NN	O	O
whether	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
(	NN	O	O
EBV	NN	O	O
)	NN	O	O
lytic	NN	O	O
replication	NN	O	O
interferes	NN	O	O
with	NN	O	O
PML	NN	O	O
bodies	NN	O	O
,	NN	O	O
we	NN	O	O
expressed	NN	O	O
the	NN	O	O
EBV	NN	O	B-DNA
immediate-early	NN	O	I-DNA
genes	NN	O	I-DNA
BZLF1	NN	O	B-DNA
(	NN	O	I-DNA
Z	NN	O	I-DNA
)	NN	O	I-DNA
and	NN	O	O
BRLF1	NN	O	B-DNA
(	NN	O	I-DNA
R	NN	O	I-DNA
)	NN	O	I-DNA
in	NN	O	O
EBV-positive	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
and	NN	O	O
examined	NN	O	O
PML	NN	O	B-protein
localization	NN	O	O
.	NN	O	O

Both	NN	O	O
Z	NN	O	O
and	NN	O	O
R	NN	O	O
expression	NN	O	O
resulted	NN	O	O
in	NN	O	O
PML	NN	O	B-protein
dispersion	NN	O	O
in	NN	O	O
EBV-positive	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Z	NN	O	O
but	NN	O	O
not	NN	O	O
R	NN	O	O
expression	NN	O	O
is	NN	O	O
sufficient	NN	O	O
to	NN	O	O
disrupt	NN	O	O
PML	NN	O	O
bodies	NN	O	O
in	NN	O	O
EBV-negative	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
dispersion	NN	O	O
of	NN	O	O
PML	NN	O	O
bodies	NN	O	O
by	NN	O	O
Z	NN	O	B-DNA
requires	NN	O	O
a	NN	O	O
portion	NN	O	O
of	NN	O	O
the	NN	O	O
transcriptional	NN	O	B-DNA
activation	NN	O	I-DNA
domain	NN	O	I-DNA
of	NN	O	O
Z	NN	O	B-DNA
but	NN	O	O
not	NN	O	O
the	NN	O	O
DNA-binding	NN	O	O
function	NN	O	O
.	NN	O	O

As	NN	O	O
was	NN	O	O
previously	NN	O	O
reported	NN	O	O
for	NN	O	O
the	NN	O	O
HSV-1	NN	O	B-protein
ICP0	NN	O	I-protein
and	NN	O	I-protein
CMV	NN	O	I-protein
IE1	NN	O	I-protein
proteins	NN	O	I-protein
,	NN	O	O
Z	NN	O	B-DNA
reduces	NN	O	O
the	NN	O	O
amount	NN	O	O
of	NN	O	O
SUMO-1-modified	NN	O	B-protein
PML	NN	O	I-protein
.	NN	O	O

We	NN	O	O
also	NN	O	O
found	NN	O	O
that	NN	O	O
Z	NN	O	B-DNA
itself	NN	O	O
is	NN	O	O
SUMO-1	NN	O	O
modified	NN	O	O
(	NN	O	O
through	NN	O	O
amino	NN	O	O
acid	NN	O	O
12	NN	O	O
)	NN	O	O
and	NN	O	O
that	NN	O	O
Z	NN	O	B-DNA
competes	NN	O	O
with	NN	O	O
PML	NN	O	B-protein
for	NN	O	O
limiting	NN	O	O
amounts	NN	O	O
of	NN	O	O
SUMO-1	NN	O	B-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
disruption	NN	O	O
of	NN	O	O
PML	NN	O	O
bodies	NN	O	O
is	NN	O	O
important	NN	O	O
for	NN	O	O
efficient	NN	O	O
lytic	NN	O	O
replication	NN	O	O
of	NN	O	O
EBV	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
Z	NN	O	B-DNA
may	NN	O	O
potentially	NN	O	O
alter	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
cellular	NN	O	O
proteins	NN	O	O
by	NN	O	O
inhibiting	NN	O	O
SUMO-1	NN	O	O
modification	NN	O	O

-DOCSTART-	O

Suppression	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
and	NN	O	O
stimulation	NN	O	O
of	NN	O	O
inhibitor	NN	O	B-protein
kappaB	NN	O	I-protein
by	NN	O	O
troglitazone	NN	O	O
:	NN	O	O
evidence	NN	O	O
for	NN	O	O
an	NN	O	O
anti-inflammatory	NN	O	O
effect	NN	O	O
and	NN	O	O
a	NN	O	O
potential	NN	O	O
antiatherosclerotic	NN	O	O
effect	NN	O	O
in	NN	O	O
the	NN	O	O
obese	NN	O	O
.	NN	O	O

To	NN	O	O
elucidate	NN	O	O
whether	NN	O	O
troglitazone	NN	O	O
exerts	NN	O	O
an	NN	O	O
antiinflammatory	NN	O	O
effect	NN	O	O
in	NN	O	O
humans	NN	O	O
,	NN	O	O
in	NN	O	O
vivo	NN	O	O
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
the	NN	O	O
suppression	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappaB	NN	O	I-protein
(	NN	O	O
NFkappaB	NN	O	B-protein
)	NN	O	O
in	NN	O	O
mononuclear	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	I-cell_type
MNC	NN	O	I-cell_type
)	NN	O	I-cell_type
by	NN	O	O
this	NN	O	O
drug	NN	O	O
.	NN	O	O

We	NN	O	O
measured	NN	O	O
intranuclear	NN	O	B-protein
NFkappaB	NN	O	I-protein
,	NN	O	O
total	NN	O	O
cellular	NN	O	B-protein
NFkappaB	NN	O	I-protein
,	NN	O	O
inhibitor	NN	O	B-protein
kappaB	NN	O	I-protein
(	NN	O	I-protein
IkappaB	NN	O	I-protein
)	NN	O	I-protein
alpha	NN	O	I-protein
,	NN	O	O
reactive	NN	O	O
oxygen	NN	O	O
species	NN	O	O
(	NN	O	O
ROS	NN	O	O
)	NN	O	O
generation	NN	O	O
,	NN	O	O
and	NN	O	O
p47	NN	O	B-protein
(	NN	O	I-protein
phox	NN	O	I-protein
)	NN	O	I-protein
subunit	NN	O	I-protein
(	NN	O	O
a	NN	O	O
key	NN	O	O
component	NN	O	O
protein	NN	O	O
of	NN	O	O
nicotinamide	NN	O	B-protein
adenine	NN	O	I-protein
dinucleotide	NN	O	I-protein
phosphate	NN	O	I-protein
oxidase	NN	O	I-protein
)	NN	O	O
in	NN	O	O
MNC	NN	O	B-cell_type
.	NN	O	O

Plasma	NN	O	B-protein
tumor	NN	O	I-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	I-protein
TNF	NN	O	I-protein
)	NN	O	I-protein
-alpha	NN	O	I-protein
,	NN	O	O
soluble	NN	O	B-protein
intercellular	NN	O	I-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
(	NN	O	O
sICAM-1	NN	O	B-protein
)	NN	O	O
,	NN	O	O
monocyte	NN	O	B-protein
chemoattractant	NN	O	I-protein
protein-1	NN	O	I-protein
(	NN	O	I-protein
MCP-1	NN	O	I-protein
)	NN	O	I-protein
,	NN	O	O
plasminogen	NN	O	B-protein
activator	NN	O	I-protein
inhibitor	NN	O	I-protein
type	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	O
PAI-1	NN	O	B-protein
)	NN	O	O
,	NN	O	O
C-reactive	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	I-protein
CRP	NN	O	I-protein
)	NN	O	I-protein
,	NN	O	O
and	NN	O	O
interleukin	NN	O	B-protein
(	NN	O	I-protein
IL	NN	O	I-protein
)	NN	O	I-protein
-10	NN	O	I-protein
(	NN	O	O
antiinflammatory	NN	O	B-protein
cytokine	NN	O	I-protein
)	NN	O	O
concentrations	NN	O	O
were	NN	O	O
also	NN	O	O
measured	NN	O	O
as	NN	O	O
mediators	NN	O	O
of	NN	O	O
inflammatory	NN	O	O
activity	NN	O	O
that	NN	O	O
are	NN	O	O
regulated	NN	O	O
by	NN	O	O
the	NN	O	O
proinflammatory	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
NFkappaB	NN	O	I-protein
.	NN	O	O

Seven	NN	O	O
nondiabetic	NN	O	O
obese	NN	O	O
patients	NN	O	O
were	NN	O	O
given	NN	O	O
400	NN	O	O
mg	NN	O	O
troglitazone	NN	O	O
daily	NN	O	O
for	NN	O	O
4	NN	O	O
weeks	NN	O	O
.	NN	O	O

Blood	NN	O	O
samples	NN	O	O
were	NN	O	O
collected	NN	O	O
before	NN	O	O
and	NN	O	O
at	NN	O	O
weekly	NN	O	O
intervals	NN	O	O
thereafter	NN	O	O
.	NN	O	O

MNC	NN	O	B-cell_type
were	NN	O	O
separated	NN	O	O
;	NN	O	O
and	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
intranuclear	NN	O	B-protein
NFkappaB	NN	O	I-protein
,	NN	O	O
total	NN	O	O
cellular	NN	O	B-protein
NFkappaB	NN	O	I-protein
,	NN	O	O
IkappaBalpha	NN	O	B-protein
,	NN	O	O
and	NN	O	O
p47	NN	O	B-protein
(	NN	O	I-protein
phox	NN	O	I-protein
)	NN	O	I-protein
subunit	NN	O	I-protein
and	NN	O	O
ROS	NN	O	O
generation	NN	O	O
were	NN	O	O
determined	NN	O	O
.	NN	O	O

Plasma	NN	O	O
was	NN	O	O
used	NN	O	O
to	NN	O	O
measure	NN	O	O
insulin	NN	O	O
glucose	NN	O	O
,	NN	O	O
TNFalpha	NN	O	B-protein
,	NN	O	O
sICAM	NN	O	B-protein
,	NN	O	O
MCP-1	NN	O	B-protein
,	NN	O	O
PAI-1	NN	O	B-protein
,	NN	O	O
CRP	NN	O	B-protein
,	NN	O	O
and	NN	O	O
IL-10	NN	O	B-protein
.	NN	O	O

Plasma	NN	O	O
insulin	NN	O	O
concentrations	NN	O	O
fell	NN	O	O
significantly	NN	O	O
at	NN	O	O
week	NN	O	O
1	NN	O	O
,	NN	O	O
from	NN	O	O
31.2	NN	O	O
+/-	NN	O	O
29.1	NN	O	O
to	NN	O	O
14.2	NN	O	O
+/-	NN	O	O
11.4	NN	O	O
mU/L	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
remained	NN	O	O
low	NN	O	O
throughout	NN	O	O
4	NN	O	O
weeks	NN	O	O
.	NN	O	O

Plasma	NN	O	O
glucose	NN	O	O
concentrations	NN	O	O
did	NN	O	O
not	NN	O	O
alter	NN	O	O
significantly	NN	O	O
.	NN	O	O

There	NN	O	O
was	NN	O	O
a	NN	O	O
fall	NN	O	O
in	NN	O	O
intranuclear	NN	O	B-protein
NFkappaB	NN	O	I-protein
,	NN	O	O
total	NN	O	O
cellular	NN	O	B-protein
NFkappaB	NN	O	I-protein
,	NN	O	O
and	NN	O	O
p47	NN	O	B-protein
(	NN	O	I-protein
phox	NN	O	I-protein
)	NN	O	I-protein
subunit	NN	O	I-protein
,	NN	O	O
with	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
cellular	NN	O	O
IkappaBalpha	NN	O	B-protein
at	NN	O	O
week	NN	O	O
2	NN	O	O
,	NN	O	O
which	NN	O	O
persisted	NN	O	O
until	NN	O	O
week	NN	O	O
4	NN	O	O
.	NN	O	O

There	NN	O	O
was	NN	O	O
a	NN	O	O
parallel	NN	O	O
fall	NN	O	O
in	NN	O	O
ROS	NN	O	O
generation	NN	O	O
by	NN	O	O
MNC	NN	O	B-cell_type
at	NN	O	O
week	NN	O	O
1	NN	O	O
;	NN	O	O
this	NN	O	O
progressed	NN	O	O
and	NN	O	O
persisted	NN	O	O
until	NN	O	O
week	NN	O	O
4	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.001	NN	O	O
)	NN	O	O
.	NN	O	O

Plasma	NN	O	O
TNF-alpha	NN	O	B-protein
,	NN	O	O
sICAM-1	NN	O	B-protein
,	NN	O	O
MCP-1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
PAI-1	NN	O	B-protein
concentrations	NN	O	O
fell	NN	O	O
significantly	NN	O	O
at	NN	O	O
week	NN	O	O
4	NN	O	O
.	NN	O	O

Plasma	NN	O	O
IL-10	NN	O	B-protein
concentration	NN	O	O
increased	NN	O	O
significantly	NN	O	O
,	NN	O	O
whereas	NN	O	O
plasma	NN	O	B-protein
CRP	NN	O	I-protein
concentrations	NN	O	O
decreased	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
troglitazone	NN	O	O
has	NN	O	O
an	NN	O	O
antiinflammatory	NN	O	O
action	NN	O	O
that	NN	O	O
may	NN	O	O
contribute	NN	O	O
to	NN	O	O
its	NN	O	O
putative	NN	O	O
antiatherosclerotic	NN	O	O
effects	NN	O	O
.	NN	O	O

-DOCSTART-	O

Tyrosine	NN	O	O
phosphorylation-dependent	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

Requirement	NN	O	O
for	NN	O	O
p56	NN	O	B-protein
LCK	NN	O	I-protein
and	NN	O	I-protein
ZAP-70	NN	O	I-protein
protein	NN	O	I-protein
tyrosine	NN	O	I-protein
kinases	NN	O	I-protein
.	NN	O	O

Phosphorylation	NN	O	O
of	NN	O	O
the	NN	O	O
N-terminal	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
inhibitory	NN	O	I-protein
subunits	NN	O	I-protein
induces	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
.	NN	O	O

Although	NN	O	O
serine	NN	O	O
phosphorylation	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
induce	NN	O	O
ubiquitination	NN	O	O
and	NN	O	O
subsequent	NN	O	O
proteasome-mediated	NN	O	O
degradation	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B-alpha	NN	O	I-protein
,	NN	O	O
little	NN	O	O
is	NN	O	O
known	NN	O	O
about	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
that	NN	O	O
lead	NN	O	O
to	NN	O	O
release	NN	O	O
of	NN	O	O
active	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
as	NN	O	O
a	NN	O	O
consequence	NN	O	O
of	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B-alpha	NN	O	I-protein
[	NN	O	O
Imbert	NN	O	O
,	NN	O	O
V.	NN	O	O
,	NN	O	O
Rupec	NN	O	O
,	NN	O	O
R.A.	NN	O	O
,	NN	O	O
Livolsi	NN	O	O
,	NN	O	O
A.	NN	O	O
,	NN	O	O
Pahl	NN	O	O
,	NN	O	O
H.L.	NN	O	O
,	NN	O	O
Traenckner	NN	O	O
,	NN	O	O
B.M.	NN	O	O
,	NN	O	O
Mueller-Dieckmann	NN	O	O
,	NN	O	O
C.	NN	O	O
,	NN	O	O
Farahifar	NN	O	O
,	NN	O	O
D.	NN	O	O
,	NN	O	O
Rossi	NN	O	O
,	NN	O	O
B.	NN	O	O
,	NN	O	O
Auberger	NN	O	O
,	NN	O	O
P.	NN	O	O
,	NN	O	O
Baeuerle	NN	O	O
,	NN	O	O
P.	NN	O	O
&	NN	O	O
Peyron	NN	O	O
,	NN	O	O
J.F.	NN	O	O
(	NN	O	O
1996	NN	O	O
)	NN	O	O
Cell	NN	O	O
86	NN	O	O
,	NN	O	O
787	NN	O	O
--	NN	O	O
798	NN	O	O
]	NN	O	O
.	NN	O	O

The	NN	O	O
involvement	NN	O	O
of	NN	O	O
the	NN	O	O
tyrosine	NN	O	B-protein
kinases	NN	O	I-protein
p56	NN	O	B-protein
(	NN	O	I-protein
lck	NN	O	I-protein
)	NN	O	I-protein
and	NN	O	O
ZAP-70	NN	O	B-protein
in	NN	O	O
this	NN	O	O
reaction	NN	O	O
is	NN	O	O
demonstrated	NN	O	O
here	NN	O	O
using	NN	O	O
specific	NN	O	O
pharmacological	NN	O	O
inhibitors	NN	O	O
and	NN	O	O
Jurkat	NN	O	B-cell_line
mutants	NN	O	I-cell_line
unable	NN	O	O
to	NN	O	O
express	NN	O	O
these	NN	O	O
kinases	NN	O	B-protein
.	NN	O	O

Although	NN	O	O
the	NN	O	O
inhibitors	NN	O	O
prevented	NN	O	O
both	NN	O	O
pervanadate-induced	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B-alpha	NN	O	I-protein
on	NN	O	O
Tyr42	NN	O	O
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
,	NN	O	O
we	NN	O	O
observed	NN	O	O
that	NN	O	O
,	NN	O	O
in	NN	O	O
p56	NN	O	B-cell_line
(	NN	O	I-cell_line
lck	NN	O	I-cell_line
)	NN	O	I-cell_line
-deficient	NN	O	I-cell_line
Jurkat	NN	O	I-cell_line
mutants	NN	O	I-cell_line
,	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
could	NN	O	O
still	NN	O	O
associate	NN	O	O
with	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B-alpha	NN	O	I-protein
despite	NN	O	O
phosphorylation	NN	O	O
on	NN	O	O
Tyr42	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
the	NN	O	O
SH2	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
p56	NN	O	B-protein
(	NN	O	I-protein
lck	NN	O	I-protein
)	NN	O	I-protein
appeared	NN	O	O
to	NN	O	O
be	NN	O	O
required	NN	O	O
for	NN	O	O
pervanadate-induced	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
but	NN	O	O
not	NN	O	O
for	NN	O	O
Tyr42	NN	O	O
phosphorylation	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
show	NN	O	O
that	NN	O	O
p56	NN	O	B-protein
(	NN	O	I-protein
lck	NN	O	I-protein
)	NN	O	I-protein
and	NN	O	O
ZAP-70	NN	O	B-protein
are	NN	O	O
key	NN	O	O
components	NN	O	O
of	NN	O	O
the	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
that	NN	O	O
leads	NN	O	O
to	NN	O	O
phosphotyrosine-dependent	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
confirm	NN	O	O
that	NN	O	O
tyrosine	NN	O	B-protein
kinases	NN	O	I-protein
must	NN	O	O
control	NN	O	O
at	NN	O	O
least	NN	O	O
two	NN	O	O
different	NN	O	O
steps	NN	O	O
to	NN	O	O
induce	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

Finally	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
H	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
O	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
,	NN	O	O
which	NN	O	O
stimulates	NN	O	O
p56	NN	O	B-protein
(	NN	O	I-protein
lck	NN	O	I-protein
)	NN	O	I-protein
and	NN	O	O
ZAP-70	NN	O	B-protein
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
is	NN	O	O
an	NN	O	O
activator	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
through	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B-alpha	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Specific	NN	O	O
missense	NN	O	O
mutations	NN	O	O
in	NN	O	O
NEMO	NN	O	B-DNA
result	NN	O	O
in	NN	O	O
hyper-IgM	NN	O	O
syndrome	NN	O	O
with	NN	O	O
hypohydrotic	NN	O	O
ectodermal	NN	O	O
dysplasia	NN	O	O
.	NN	O	O

The	NN	O	O
gene	NN	O	O
that	NN	O	O
encodes	NN	O	O
nuclear	NN	O	B-DNA
factor	NN	O	I-DNA
kappaB	NN	O	I-DNA
(	NN	O	I-DNA
NF-kappaB	NN	O	I-DNA
)	NN	O	I-DNA
essential	NN	O	I-DNA
modulator	NN	O	I-DNA
(	NN	O	O
or	NN	O	O
NEMO	NN	O	B-DNA
,	NN	O	O
also	NN	O	O
known	NN	O	O
as	NN	O	O
IKKgamma	NN	O	B-DNA
)	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappaB	NN	O	I-protein
.	NN	O	O

We	NN	O	O
describe	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
putative	NN	O	B-protein
zinc-finger	NN	O	I-protein
domain	NN	O	I-protein
of	NN	O	O
NEMO	NN	O	B-protein
that	NN	O	O
result	NN	O	O
in	NN	O	O
an	NN	O	O
X-linked	NN	O	O
primary	NN	O	O
immunodeficiency	NN	O	O
characterized	NN	O	O
by	NN	O	O
hyper-IgM	NN	O	O
syndrome	NN	O	O
and	NN	O	O
hypohydrotic	NN	O	O
ectodermal	NN	O	O
dysplasia	NN	O	O
(	NN	O	O
XHM-ED	NN	O	O
)	NN	O	O
.	NN	O	O

These	NN	O	O
mutations	NN	O	O
prevent	NN	O	O
CD40	NN	O	B-protein
ligand	NN	O	I-protein
(	NN	O	O
CD40L	NN	O	B-protein
)	NN	O	O
-mediated	NN	O	O
degradation	NN	O	O
of	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
alpha	NN	O	I-protein
(	NN	O	O
IkappaB-alpha	NN	O	B-protein
)	NN	O	O
and	NN	O	O
account	NN	O	O
for	NN	O	O
the	NN	O	O
following	NN	O	O
observations	NN	O	O
:	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
XHM-ED	NN	O	O
patients	NN	O	O
are	NN	O	O
unable	NN	O	O
to	NN	O	O
undergo	NN	O	O
immunoglobulin	NN	O	O
class-switch	NN	O	O
recombination	NN	O	O
and	NN	O	O
antigen-presenting	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
APCs	NN	O	B-cell_type
)	NN	O	O
are	NN	O	O
unable	NN	O	O
to	NN	O	O
synthesize	NN	O	O
the	NN	O	O
NF-kappaB-regulated	NN	O	B-protein
cytokines	NN	O	I-protein
interleukin	NN	O	B-protein
12	NN	O	I-protein
(	NN	O	O
IL-12	NN	O	B-protein
)	NN	O	O
or	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
when	NN	O	O
stimulated	NN	O	O
with	NN	O	O
CD40L	NN	O	B-protein
.	NN	O	O

Nevertheless	NN	O	O
,	NN	O	O
innate	NN	O	O
immunity	NN	O	O
is	NN	O	O
preserved	NN	O	O
in	NN	O	O
XHM-ED	NN	O	O
patients	NN	O	O
because	NN	O	O
APCs	NN	O	B-cell_type
retain	NN	O	O
the	NN	O	O
capacity	NN	O	O
to	NN	O	O
respond	NN	O	O
to	NN	O	O
stimulation	NN	O	O
by	NN	O	O
lipopolysaccharide	NN	O	O
or	NN	O	O
Staphylococcus	NN	O	B-protein
aureus	NN	O	I-protein
Cowan	NN	O	I-protein
's	NN	O	I-protein
antigen	NN	O	I-protein
(	NN	O	O
SAC	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Overall	NN	O	O
,	NN	O	O
the	NN	O	O
phenotype	NN	O	O
observed	NN	O	O
in	NN	O	O
XHM-ED	NN	O	O
patients	NN	O	O
shows	NN	O	O
that	NN	O	O
the	NN	O	O
putative	NN	O	B-protein
zinc-finger	NN	O	I-protein
domain	NN	O	I-protein
of	NN	O	O
NEMO	NN	O	B-protein
has	NN	O	O
a	NN	O	O
regulatory	NN	O	O
function	NN	O	O
and	NN	O	O
demonstrates	NN	O	O
the	NN	O	O
definite	NN	O	O
requirement	NN	O	O
of	NN	O	O
CD40	NN	O	B-protein
-mediated	NN	O	O
NF-kappaB	NN	O	O
activation	NN	O	O
for	NN	O	O
B	NN	O	B-cell_type
cell	NN	O	I-cell_type
immunoglobulin	NN	O	O
class-switching	NN	O	O
.	NN	O	O

-DOCSTART-	O

Granulocytic	NN	O	O
differentiation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
NB4	NN	O	I-cell_type
promyelocytic	NN	O	I-cell_type
leukemia	NN	O	I-cell_type
cells	NN	O	I-cell_type
induced	NN	O	O
by	NN	O	O
all-trans	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
metabolites	NN	O	O
.	NN	O	O

The	NN	O	O
metabolism	NN	O	O
of	NN	O	O
all-trans	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
ATRA	NN	O	O
)	NN	O	O
has	NN	O	O
been	NN	O	O
reported	NN	O	O
to	NN	O	O
be	NN	O	O
partly	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
in	NN	O	O
vivo	NN	O	O
resistance	NN	O	O
to	NN	O	O
ATRA	NN	O	O
seen	NN	O	O
in	NN	O	O
the	NN	O	O
treatment	NN	O	O
of	NN	O	O
human	NN	O	O
acute	NN	O	O
promyelocytic	NN	O	O
leukemia	NN	O	O
(	NN	O	O
APL	NN	O	O
)	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
ATRA	NN	O	O
metabolism	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
growth	NN	O	O
inhibition	NN	O	O
of	NN	O	O
several	NN	O	O
cancer	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

The	NN	O	O
purpose	NN	O	O
of	NN	O	O
this	NN	O	O
study	NN	O	O
was	NN	O	O
to	NN	O	O
evaluate	NN	O	O
the	NN	O	O
in	NN	O	O
vitro	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
principal	NN	O	O
metabolites	NN	O	O
of	NN	O	O
ATRA	NN	O	O
[	NN	O	O
4-hydroxy-retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
4-OH-RA	NN	O	O
)	NN	O	O
,	NN	O	O
18-hydroxy-retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
18-OH-RA	NN	O	O
)	NN	O	O
,	NN	O	O
4-oxo-retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
4-oxo-RA	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
5	NN	O	O
,	NN	O	O
6-epoxy-retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
5	NN	O	O
,	NN	O	O
6-epoxy-RA	NN	O	O
)	NN	O	O
]	NN	O	O
in	NN	O	O
NB4	NN	O	B-cell_line
,	NN	O	I-cell_line
a	NN	O	I-cell_line
human	NN	O	I-cell_line
promyelocytic	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
that	NN	O	O
exhibits	NN	O	O
the	NN	O	O
APL	NN	O	O
diagnostic	NN	O	O
t	NN	O	O
(	NN	O	O
15	NN	O	O
;	NN	O	O
17	NN	O	O
)	NN	O	O
chromosomal	NN	O	O
translocation	NN	O	O
and	NN	O	O
expresses	NN	O	O
the	NN	O	O
PML-RAR	NN	O	B-protein
alpha	NN	O	I-protein
fusion	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

We	NN	O	O
established	NN	O	O
that	NN	O	O
the	NN	O	O
four	NN	O	O
ATRA	NN	O	O
metabolites	NN	O	O
were	NN	O	O
indeed	NN	O	O
formed	NN	O	O
by	NN	O	O
the	NN	O	O
NB4	NN	O	B-cell_line
cells	NN	O	I-cell_line
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

NB4	NN	O	O
cell	NN	O	O
growth	NN	O	O
was	NN	O	O
inhibited	NN	O	O
(	NN	O	O
69-78	NN	O	O
%	NN	O	O
at	NN	O	O
120	NN	O	O
h	NN	O	O
)	NN	O	O
and	NN	O	O
cell	NN	O	O
cycle	NN	O	O
progression	NN	O	O
in	NN	O	O
the	NN	O	O
G1	NN	O	O
phase	NN	O	O
(	NN	O	O
82-85	NN	O	O
%	NN	O	O
at	NN	O	O
120	NN	O	O
h	NN	O	O
)	NN	O	O
was	NN	O	O
blocked	NN	O	O
by	NN	O	O
ATRA	NN	O	O
and	NN	O	O
all	NN	O	O
of	NN	O	O
the	NN	O	O
metabolites	NN	O	O
at	NN	O	O
1	NN	O	O
microM	NN	O	O
concentration	NN	O	O
.	NN	O	O

ATRA	NN	O	O
and	NN	O	O
its	NN	O	O
metabolites	NN	O	O
could	NN	O	O
induce	NN	O	O
NB4	NN	O	B-cell_line
cells	NN	O	I-cell_line
differentiation	NN	O	O
with	NN	O	O
similar	NN	O	O
activity	NN	O	O
,	NN	O	O
as	NN	O	O
evaluated	NN	O	O
by	NN	O	O
cell	NN	O	O
morphology	NN	O	O
,	NN	O	O
by	NN	O	O
the	NN	O	O
nitroblue	NN	O	O
tetrazolium	NN	O	O
reduction	NN	O	O
test	NN	O	O
(	NN	O	O
82-88	NN	O	O
%	NN	O	O
at	NN	O	O
120	NN	O	O
h	NN	O	O
)	NN	O	O
or	NN	O	O
by	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
maturation	NN	O	B-protein
specific	NN	O	I-protein
cell	NN	O	I-protein
surface	NN	O	I-protein
marker	NN	O	I-protein
CD11c	NN	O	I-protein
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
nuclear	NN	O	O
body	NN	O	O
reorganization	NN	O	O
to	NN	O	O
macropunctated	NN	O	O
structures	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
degradation	NN	O	O
of	NN	O	O
PML-RAR	NN	O	B-protein
alpha	NN	O	I-protein
,	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
similar	NN	O	O
for	NN	O	O
ATRA	NN	O	O
and	NN	O	O
all	NN	O	O
of	NN	O	O
its	NN	O	O
metabolites	NN	O	O
.	NN	O	O

Comparison	NN	O	O
of	NN	O	O
the	NN	O	O
relative	NN	O	O
potency	NN	O	O
of	NN	O	O
the	NN	O	O
retinoids	NN	O	O
using	NN	O	O
the	NN	O	O
nitroblue	NN	O	O
tetrazolium	NN	O	O
reduction	NN	O	O
test	NN	O	O
showed	NN	O	O
effective	NN	O	O
concentrations	NN	O	O
required	NN	O	O
to	NN	O	O
differentiate	NN	O	O
50	NN	O	O
%	NN	O	O
of	NN	O	O
cells	NN	O	O
in	NN	O	O
72	NN	O	O
h	NN	O	O
as	NN	O	O
follows	NN	O	O
:	NN	O	O
ATRA	NN	O	O
,	NN	O	O
15.8	NN	O	O
+/-	NN	O	O
1.7	NN	O	O
nM	NN	O	O
;	NN	O	O
4-oxo-RA	NN	O	O
,	NN	O	O
38.3	NN	O	O
+/-	NN	O	O
1.3	NN	O	O
nM	NN	O	O
;	NN	O	O
18-OH-RA	NN	O	O
,	NN	O	O
55.5	NN	O	O
+/-	NN	O	O
1.8	NN	O	O
nM	NN	O	O
;	NN	O	O
4-OH-RA	NN	O	O
,	NN	O	O
79.8	NN	O	O
+/-	NN	O	O
1.8	NN	O	O
nM	NN	O	O
;	NN	O	O
and	NN	O	O
5	NN	O	O
,	NN	O	O
6-epoxy-RA	NN	O	O
,	NN	O	O
99.5	NN	O	O
+/-	NN	O	O
1.5	NN	O	O
nM	NN	O	O
.	NN	O	O

The	NN	O	O
ATRA	NN	O	O
metabolites	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
exert	NN	O	O
their	NN	O	O
differentiation	NN	O	O
effects	NN	O	O
via	NN	O	O
the	NN	O	O
RAR	NN	O	B-protein
alpha	NN	O	I-protein
nuclear	NN	O	I-protein
receptors	NN	O	I-protein
,	NN	O	O
because	NN	O	O
the	NN	O	O
RAR	NN	O	O
alpha-specific	NN	O	O
antagonist	NN	O	O
BMS614	NN	O	O
blocked	NN	O	O
metabolite-induced	NN	O	O
CD11c	NN	O	O
expression	NN	O	O
in	NN	O	O
NB4	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

These	NN	O	O
data	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
principal	NN	O	O
ATRA	NN	O	O
Phase	NN	O	O
1	NN	O	O
metabolites	NN	O	O
can	NN	O	O
elicit	NN	O	O
leukemia	NN	O	B-cell_type
cell	NN	O	I-cell_type
growth	NN	O	O
inhibition	NN	O	O
and	NN	O	O
differentiation	NN	O	O
in	NN	O	O
vitro	NN	O	O
through	NN	O	O
the	NN	O	O
RAR	NN	O	O
alpha	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
,	NN	O	O
and	NN	O	O
they	NN	O	O
suggest	NN	O	O
that	NN	O	O
these	NN	O	O
metabolites	NN	O	O
may	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
ATRA	NN	O	O
antileukemic	NN	O	O
activity	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

-DOCSTART-	O

Expression	NN	O	O
of	NN	O	O
oestrogen	NN	O	B-protein
and	NN	O	I-protein
progesterone	NN	O	I-protein
receptors	NN	O	I-protein
by	NN	O	O
mast	NN	O	B-cell_type
cells	NN	O	I-cell_type
alone	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
lymphocytes	NN	O	B-cell_type
,	NN	O	O
macrophages	NN	O	B-cell_type
or	NN	O	O
other	NN	O	O
immune	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
human	NN	O	O
upper	NN	O	O
airways	NN	O	O
.	NN	O	O

BACKGROUND	NN	O	O
:	NN	O	O
Nasal	NN	O	O
polyposis	NN	O	O
often	NN	O	O
coexists	NN	O	O
with	NN	O	O
asthma	NN	O	O
in	NN	O	O
airway	NN	O	O
inflammatory	NN	O	O
conditions	NN	O	O
characterised	NN	O	O
by	NN	O	O
the	NN	O	O
infiltration	NN	O	O
of	NN	O	O
a	NN	O	O
range	NN	O	O
of	NN	O	O
immune	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

A	NN	O	O
potentially	NN	O	O
important	NN	O	O
role	NN	O	O
for	NN	O	O
ovarian	NN	O	O
hormones	NN	O	O
has	NN	O	O
been	NN	O	O
implicated	NN	O	O
in	NN	O	O
airway	NN	O	O
inflammation	NN	O	O
but	NN	O	O
the	NN	O	O
cellular	NN	O	O
target	NN	O	O
for	NN	O	O
such	NN	O	O
action	NN	O	O
is	NN	O	O
not	NN	O	O
known	NN	O	O
.	NN	O	O

METHODS	NN	O	O
:	NN	O	O
Expression	NN	O	O
of	NN	O	O
oestrogen	NN	O	B-protein
receptors	NN	O	I-protein
(	NN	O	O
ER	NN	O	B-protein
)	NN	O	O
and	NN	O	O
progesterone	NN	O	B-protein
receptors	NN	O	I-protein
(	NN	O	O
PR	NN	O	B-protein
)	NN	O	O
was	NN	O	O
examined	NN	O	O
using	NN	O	O
immunohistochemistry	NN	O	O
in	NN	O	O
formalin	NN	O	O
fixed	NN	O	O
nasal	NN	O	O
polyp	NN	O	O
tissues	NN	O	O
from	NN	O	O
47	NN	O	O
subjects	NN	O	O
.	NN	O	O

The	NN	O	O
cells	NN	O	O
positive	NN	O	O
for	NN	O	O
ER	NN	O	B-protein
or	NN	O	O
PR	NN	O	B-protein
were	NN	O	O
confirmed	NN	O	O
by	NN	O	O
spatial	NN	O	O
location	NN	O	O
,	NN	O	O
dual	NN	O	O
immunolabelling	NN	O	O
,	NN	O	O
and	NN	O	O
histochemical	NN	O	O
staining	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
Consistent	NN	O	O
with	NN	O	O
the	NN	O	O
known	NN	O	O
features	NN	O	O
of	NN	O	O
nasal	NN	O	O
polyps	NN	O	O
,	NN	O	O
CD4+	NN	O	B-cell_type
(	NN	O	I-cell_type
T	NN	O	I-cell_type
helper/inducer	NN	O	I-cell_type
)	NN	O	I-cell_type
,	NN	O	O
CD8+	NN	O	B-cell_type
(	NN	O	I-cell_type
cytotoxic/suppressor	NN	O	I-cell_type
)	NN	O	I-cell_type
,	NN	O	O
CD68+	NN	O	B-cell_type
(	NN	O	O
macrophages	NN	O	B-cell_type
)	NN	O	O
,	NN	O	O
mast	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
eosinophils	NN	O	B-cell_type
and	NN	O	O
neutrophils	NN	O	B-cell_type
were	NN	O	O
all	NN	O	O
clearly	NN	O	O
detected	NN	O	O
by	NN	O	O
their	NN	O	O
relevant	NN	O	O
monoclonal	NN	O	B-protein
antibodies	NN	O	I-protein
or	NN	O	O
appropriate	NN	O	O
histochemical	NN	O	O
staining	NN	O	O
,	NN	O	O
but	NN	O	O
only	NN	O	O
mast	NN	O	B-cell_type
cells	NN	O	I-cell_type
tested	NN	O	O
positive	NN	O	O
for	NN	O	O
ER	NN	O	B-protein
/PR	NN	O	O
labelling	NN	O	O
with	NN	O	O
their	NN	O	O
polyclonal	NN	O	B-protein
and	NN	O	I-protein
monoclonal	NN	O	I-protein
antibodies	NN	O	I-protein
.	NN	O	O

The	NN	O	O
frequencies	NN	O	O
for	NN	O	O
expression	NN	O	O
were	NN	O	O
61.7	NN	O	O
%	NN	O	O
for	NN	O	O
ER	NN	O	B-cell_type
positive	NN	O	I-cell_type
and	NN	O	O
59.6	NN	O	O
%	NN	O	O
for	NN	O	O
PR	NN	O	B-cell_type
positive	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
ER	NN	O	B-protein
/	NN	O	O
PR	NN	O	B-protein
was	NN	O	O
independent	NN	O	O
of	NN	O	O
patient	NN	O	O
sex	NN	O	O
and	NN	O	O
age	NN	O	O
but	NN	O	O
was	NN	O	O
highly	NN	O	O
correlated	NN	O	O
with	NN	O	O
the	NN	O	O
numbers	NN	O	O
of	NN	O	O
mast	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
r	NN	O	O
=	NN	O	O
0.973	NN	O	O
,	NN	O	O
p	NN	O	O
<	NN	O	O
0.001	NN	O	O
for	NN	O	O
ER	NN	O	B-protein
;	NN	O	O
r	NN	O	O
=	NN	O	O
0.955	NN	O	O
,	NN	O	O
p	NN	O	O
<	NN	O	O
0.001	NN	O	O
for	NN	O	O
PR	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Fewer	NN	O	O
than	NN	O	O
5	NN	O	O
%	NN	O	O
of	NN	O	O
mast	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
negative	NN	O	O
for	NN	O	O
ER	NN	O	B-protein
/	NN	O	O
PR	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
:	NN	O	O
Mast	NN	O	B-cell_type
cells	NN	O	I-cell_type
alone	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
lymphocytes	NN	O	B-cell_type
,	NN	O	O
macrophages	NN	O	B-cell_type
,	NN	O	O
or	NN	O	O
other	NN	O	O
immune	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
express	NN	O	O
ER	NN	O	B-protein
/	NN	O	O
PR	NN	O	B-protein
in	NN	O	O
human	NN	O	O
upper	NN	O	O
airways	NN	O	O
.	NN	O	O

Numerous	NN	O	O
ER/PR	NN	O	B-cell_type
positive	NN	O	I-cell_type
mast	NN	O	I-cell_type
cells	NN	O	I-cell_type
exist	NN	O	O
in	NN	O	O
nasal	NN	O	O
polyps	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
this	NN	O	O
may	NN	O	O
be	NN	O	O
a	NN	O	O
major	NN	O	O
route	NN	O	O
for	NN	O	O
the	NN	O	O
involvement	NN	O	O
of	NN	O	O
sex	NN	O	O
hormones	NN	O	O
in	NN	O	O
airway	NN	O	O
inflammation	NN	O	O
when	NN	O	O
exposed	NN	O	O
to	NN	O	O
the	NN	O	O
higher	NN	O	O
and	NN	O	O
varying	NN	O	O
concentration	NN	O	O
of	NN	O	O
oestrogen	NN	O	O
and	NN	O	O
progesterone	NN	O	O
characteristic	NN	O	O
of	NN	O	O
females	NN	O	O
.	NN	O	O

-DOCSTART-	O

NF	NN	O	O
kappa	NN	O	O
b	NN	O	O
signaling	NN	O	O
in	NN	O	O
posthypoxic	NN	O	B-cell_type
endothelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
:	NN	O	O
relevance	NN	O	O
to	NN	O	O
E-selectin	NN	O	O
expression	NN	O	O
and	NN	O	O
neutrophil	NN	O	O
adhesion	NN	O	O
.	NN	O	O

Our	NN	O	O
previous	NN	O	O
studies	NN	O	O
have	NN	O	O
implicated	NN	O	O
the	NN	O	O
nuclear	NN	O	O
transcription	NN	O	O
factor	NN	O	O
kappa	NN	O	O
B	NN	O	O
(	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
)	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
adhesion	NN	O	B-protein
molecule	NN	O	I-protein
expression	NN	O	O
in	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
exposed	NN	O	O
to	NN	O	O
anoxia-reoxygenation	NN	O	O
(	NN	O	O
A/R	NN	O	O
)	NN	O	O
or	NN	O	O
a	NN	O	O
redox	NN	O	O
imbalance	NN	O	O
.	NN	O	O

The	NN	O	O
objectives	NN	O	O
of	NN	O	O
this	NN	O	O
study	NN	O	O
were	NN	O	O
(	NN	O	O
1	NN	O	O
)	NN	O	O
to	NN	O	O
define	NN	O	O
the	NN	O	O
kinetics	NN	O	O
of	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
activation	NN	O	O
by	NN	O	O
examining	NN	O	O
I	NN	O	O
kappa	NN	O	O
B	NN	O	O
alpha	NN	O	O
degradation	NN	O	O
and	NN	O	O
the	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
p65	NN	O	B-protein
in	NN	O	O
response	NN	O	O
to	NN	O	O
A/R	NN	O	O
or	NN	O	O
redox	NN	O	O
imbalance	NN	O	O
(	NN	O	O
induced	NN	O	O
by	NN	O	O
treatment	NN	O	O
of	NN	O	O
cells	NN	O	O
with	NN	O	O
diamide	NN	O	O
and	NN	O	O
buthionine	NN	O	O
sulfoximine	NN	O	O
)	NN	O	O
and	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
to	NN	O	O
determine	NN	O	O
whether	NN	O	O
the	NN	O	O
signal	NN	O	O
for	NN	O	O
I	NN	O	O
kappa	NN	O	O
B	NN	O	O
alpha	NN	O	O
degradation	NN	O	O
,	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
p65	NN	O	B-protein
,	NN	O	O
and	NN	O	O
E-selectin	NN	O	B-protein
-mediated	NN	O	O
neutrophil	NN	O	O
adhesion	NN	O	O
is	NN	O	O
related	NN	O	O
to	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	O
PTK	NN	O	B-protein
)	NN	O	O
,	NN	O	O
protein	NN	O	B-protein
tyrosine	NN	O	I-protein
phosphatase	NN	O	I-protein
(	NN	O	O
PTPase	NN	O	B-protein
)	NN	O	O
and/or	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
(	NN	O	O
PKC	NN	O	B-protein
)	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
both	NN	O	O
A/R	NN	O	O
and	NN	O	O
redox	NN	O	O
imbalance	NN	O	O
led	NN	O	O
to	NN	O	O
I	NN	O	O
kappa	NN	O	O
B	NN	O	O
alpha	NN	O	O
degradation	NN	O	O
within	NN	O	O
30	NN	O	O
min	NN	O	O
and	NN	O	O
the	NN	O	O
concomitant	NN	O	O
appearance	NN	O	O
of	NN	O	O
p65	NN	O	B-protein
in	NN	O	O
the	NN	O	O
nucleus	NN	O	O
,	NN	O	O
consistent	NN	O	O
with	NN	O	O
rapid	NN	O	O
cytosolic	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
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
the	NN	O	O
activated	NN	O	B-protein
p65	NN	O	I-protein
subunit	NN	O	I-protein
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
PKC	NN	O	B-protein
blocked	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
degradation	NN	O	O
and	NN	O	O
p65	NN	O	B-protein
translocation	NN	O	O
in	NN	O	O
A/R-challenged	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
redox-altered	NN	O	O
,	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

However	NN	O	O
,	NN	O	O
both	NN	O	O
A/R-	NN	O	O
and	NN	O	O
redox-induced	NN	O	O
NF	NN	O	O
kappa	NN	O	O
B	NN	O	O
activation	NN	O	O
was	NN	O	O
blocked	NN	O	O
by	NN	O	O
inhibition	NN	O	O
of	NN	O	O
PTK	NN	O	B-protein
.	NN	O	O

Similarly	NN	O	O
,	NN	O	O
A/R-induced	NN	O	O
E-selectin	NN	O	O
expression	NN	O	O
and	NN	O	O
neutrophil-endothelial	NN	O	O
cell	NN	O	O
adhesion	NN	O	O
were	NN	O	O
blocked	NN	O	O
by	NN	O	O
inhibition	NN	O	O
of	NN	O	O
PKC	NN	O	B-protein
or	NN	O	O
PTK	NN	O	B-protein
,	NN	O	O
while	NN	O	O
only	NN	O	O
PTK	NN	O	B-protein
inhibited	NN	O	O
the	NN	O	O
redox-induced	NN	O	O
adhesion	NN	O	O
response	NN	O	O
.	NN	O	O

Pretreatment	NN	O	O
of	NN	O	O
cells	NN	O	O
with	NN	O	O
N-acetyl	NN	O	O
cysteine	NN	O	O
effectively	NN	O	O
blocked	NN	O	O
A/R-	NN	O	O
or	NN	O	O
redox-induced	NN	O	O
I	NN	O	O
kappa	NN	O	O
B	NN	O	O
degradation	NN	O	O
and	NN	O	O
significantly	NN	O	O
attenuated	NN	O	O
the	NN	O	O
respective	NN	O	O
neutrophil	NN	O	O
adhesion	NN	O	O
responses	NN	O	O
.	NN	O	O

Collectively	NN	O	O
,	NN	O	O
these	NN	O	O
findings	NN	O	O
indicate	NN	O	O
that	NN	O	O
A/R-induced	NN	O	O
E-selectin	NN	O	O
expression	NN	O	O
and	NN	O	O
neutrophil-endothelial	NN	O	O
cell	NN	O	O
adhesion	NN	O	O
are	NN	O	O
mediated	NN	O	O
by	NN	O	O
both	NN	O	O
PKC	NN	O	B-protein
and	NN	O	O
PTK	NN	O	B-protein
,	NN	O	O
which	NN	O	O
signal	NN	O	O
rapid	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

This	NN	O	O
A/R-induced	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
signaling	NN	O	O
response	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
mediated	NN	O	O
,	NN	O	O
at	NN	O	O
least	NN	O	O
in	NN	O	O
part	NN	O	O
,	NN	O	O
by	NN	O	O
intracellular	NN	O	O
redox	NN	O	O
imbalance	NN	O	O
.	NN	O	O

Copyright	NN	O	O
2001	NN	O	O
S.	NN	O	O
Karger	NN	O	O
AG	NN	O	O
,	NN	O	O
Basel	NN	O	O

-DOCSTART-	O

Induction	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
by	NN	O	O
the	NN	O	O
herbicide	NN	O	O
2	NN	O	O
,	NN	O	O
4-dichlorophenoxyacetic	NN	O	O
acid	NN	O	O
.	NN	O	O

Dimethylammonium	NN	O	O
salt	NN	O	O
of	NN	O	O
2	NN	O	O
,	NN	O	O
4-dichlorophenoxyacetic	NN	O	O
acid	NN	O	O
(	NN	O	O
DMA-2	NN	O	O
,	NN	O	O
4-D	NN	O	O
)	NN	O	O
is	NN	O	O
a	NN	O	O
widely	NN	O	O
used	NN	O	O
herbicide	NN	O	O
that	NN	O	O
is	NN	O	O
considered	NN	O	O
moderately	NN	O	O
toxic	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
DMA-2	NN	O	O
,	NN	O	O
4-D	NN	O	O
is	NN	O	O
able	NN	O	O
to	NN	O	O
cause	NN	O	O
apoptosis	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
healthy	NN	O	O
individuals	NN	O	O
and	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Apoptosis	NN	O	O
induced	NN	O	O
by	NN	O	O
DMA-2	NN	O	O
,	NN	O	O
4-D	NN	O	O
was	NN	O	O
dose	NN	O	O
and	NN	O	O
time	NN	O	O
dependent	NN	O	O
,	NN	O	O
independent	NN	O	O
of	NN	O	O
Fas	NN	O	B-protein
,	NN	O	O
TNF	NN	O	B-protein
receptor	NN	O	I-protein
1	NN	O	I-protein
or	NN	O	O
the	NN	O	O
aromatic	NN	O	B-protein
hydrocarbon	NN	O	I-protein
receptor	NN	O	I-protein
,	NN	O	O
and	NN	O	O
involved	NN	O	O
disruption	NN	O	O
of	NN	O	O
the	NN	O	O
mitochondrial	NN	O	O
transmembrane	NN	O	O
potential	NN	O	O
and	NN	O	O
activation	NN	O	O
of	NN	O	O
caspase-9	NN	O	B-protein
.	NN	O	O

ZVAD-FMK	NN	O	O
,	NN	O	O
a	NN	O	O
broad-spectrum	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
caspases	NN	O	B-protein
,	NN	O	O
blocked	NN	O	O
DMA-2	NN	O	O
,	NN	O	O
4-D-induced	NN	O	O
apoptosis	NN	O	O
completely	NN	O	O
.	NN	O	O

While	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
caspase-9	NN	O	B-protein
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
caspase-9	NN	O	B-protein
and	NN	O	O
caspase-3	NN	O	O
inhibitors	NN	O	O
in	NN	O	O
combination	NN	O	O
,	NN	O	O
strongly	NN	O	O
blocked	NN	O	O
DMA-2	NN	O	O
,	NN	O	O
4-D-induced	NN	O	O
apoptosis	NN	O	O
,	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
caspase-3	NN	O	B-protein
had	NN	O	O
a	NN	O	O
moderate	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
.	NN	O	O

Unlike	NN	O	O
Fas	NN	O	B-protein
-mediated	NN	O	O
apoptosis	NN	O	O
,	NN	O	O
the	NN	O	O
initiator	NN	O	B-protein
caspase	NN	O	I-protein
,	NN	O	O
caspase-8	NN	O	B-protein
,	NN	O	O
was	NN	O	O
not	NN	O	O
involved	NN	O	O
in	NN	O	O
DMA-2	NN	O	O
,	NN	O	O
4-D-induced	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

Transfection	NN	O	O
of	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
Bcl-2	NN	O	B-protein
prevented	NN	O	O
DMA-2	NN	O	O
,	NN	O	O
4-D-induced	NN	O	O
disruption	NN	O	O
of	NN	O	O
the	NN	O	O
mitochondrial	NN	O	O
transmembrane	NN	O	O
potential	NN	O	O
and	NN	O	O
led	NN	O	O
to	NN	O	O
a	NN	O	O
complete	NN	O	O
blockage	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

Our	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
DMA-2	NN	O	O
,	NN	O	O
4-D	NN	O	O
kills	NN	O	O
human	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
by	NN	O	O
initiating	NN	O	O
apoptosis	NN	O	O
via	NN	O	O
a	NN	O	O
direct	NN	O	O
effect	NN	O	O
on	NN	O	O
mitochondria	NN	O	O
.	NN	O	O

The	NN	O	O
activation	NN	O	O
of	NN	O	O
caspases	NN	O	B-protein
occurs	NN	O	O
downstream	NN	O	O
of	NN	O	O
mitochondrial	NN	O	O
damage	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
dysfunction	NN	O	O
of	NN	O	O
mitochondria	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
sufficient	NN	O	O
for	NN	O	O
triggering	NN	O	O
all	NN	O	O
downstream	NN	O	O
events	NN	O	O
leading	NN	O	O
to	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Inhibition	NN	O	O
of	NN	O	O
Th1	NN	O	O
differentiation	NN	O	O
by	NN	O	O
IL-6	NN	O	B-protein
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
SOCS1	NN	O	B-protein
.	NN	O	O

Interleukin	NN	O	B-protein
6	NN	O	I-protein
(	NN	O	O
IL-6	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
cytokine	NN	O	B-protein
produced	NN	O	O
by	NN	O	O
immune	NN	O	B-cell_type
and	NN	O	I-cell_type
nonimmune	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
exhibits	NN	O	O
functional	NN	O	O
pleiotropy	NN	O	O
and	NN	O	O
redundancy	NN	O	O
.	NN	O	O

IL-6	NN	O	B-protein
plays	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
differentiation	NN	O	O
of	NN	O	O
several	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
describe	NN	O	O
a	NN	O	O
novel	NN	O	O
function	NN	O	O
of	NN	O	O
IL-6	NN	O	B-protein
:	NN	O	O
the	NN	O	O
negative	NN	O	O
regulation	NN	O	O
of	NN	O	O
CD4	NN	O	B-protein
+	NN	O	O
Th1	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
.	NN	O	O

While	NN	O	O
IL-6	NN	O	B-protein
-directed	NN	O	O
CD4	NN	O	B-protein
+	NN	O	O
Th2	NN	O	O
differentiation	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
inhibition	NN	O	O
of	NN	O	O
Th1	NN	O	O
differentiation	NN	O	O
by	NN	O	O
IL-6	NN	O	B-protein
is	NN	O	O
independent	NN	O	O
of	NN	O	O
IL-4	NN	O	B-protein
.	NN	O	O

IL-6	NN	O	B-protein
upregulates	NN	O	O
suppressor	NN	O	B-protein
of	NN	O	I-protein
cytokine	NN	O	I-protein
signaling	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	O
SOCS1	NN	O	B-protein
)	NN	O	O
expression	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
CD4+	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
thereby	NN	O	O
interfering	NN	O	O
with	NN	O	O
signal	NN	O	B-protein
transducer	NN	O	I-protein
and	NN	O	I-protein
activator	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	O
STAT1	NN	O	B-protein
)	NN	O	O
phosphorylation	NN	O	O
induced	NN	O	O
by	NN	O	O
interferon	NN	O	B-protein
gamma	NN	O	I-protein
(	NN	O	O
IFNgamma	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
IFNgamma	NN	O	B-protein
receptor-mediated	NN	O	O
signals	NN	O	O
by	NN	O	O
IL-6	NN	O	B-protein
prevents	NN	O	O
autoregulation	NN	O	O
of	NN	O	O
IFNgamma	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
by	NN	O	O
IFNgamma	NN	O	B-protein
during	NN	O	O
CD4+	NN	O	B-cell_type
T	NN	O	I-cell_type
cell	NN	O	I-cell_type
activation	NN	O	O
,	NN	O	O
thereby	NN	O	O
preventing	NN	O	O
Th1	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
IL-6	NN	O	B-protein
promotes	NN	O	O
CD4	NN	O	B-protein
+	NN	O	O
Th2	NN	O	O
differentiation	NN	O	O
and	NN	O	O
inhibits	NN	O	O
Th1	NN	O	O
differentiation	NN	O	O
by	NN	O	O
two	NN	O	O
independent	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
.	NN	O	O

-DOCSTART-	O

Interaction	NN	O	O
between	NN	O	O
CCAAT/enhancer	NN	O	B-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
and	NN	O	O
cyclic	NN	O	B-protein
AMP	NN	O	I-protein
response	NN	O	I-protein
element	NN	O	I-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
1	NN	O	I-protein
regulates	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
transcription	NN	O	O
in	NN	O	O
cells	NN	O	O
of	NN	O	O
the	NN	O	O
monocyte/macrophage	NN	O	B-cell_type
lineage	NN	O	I-cell_type
.	NN	O	O

Recent	NN	O	O
observations	NN	O	O
have	NN	O	O
shown	NN	O	O
two	NN	O	O
CCAAT/enhancer	NN	O	B-DNA
binding	NN	O	I-DNA
protein	NN	O	I-DNA
(	NN	O	I-DNA
C/EBP	NN	O	I-DNA
)	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
to	NN	O	O
be	NN	O	O
critically	NN	O	O
important	NN	O	O
for	NN	O	O
efficient	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
within	NN	O	O
cells	NN	O	O
of	NN	O	O
the	NN	O	O
monocyte/macrophage	NN	O	B-cell_type
lineage	NN	O	I-cell_type
,	NN	O	O
a	NN	O	O
cell	NN	O	O
type	NN	O	O
likely	NN	O	O
involved	NN	O	O
in	NN	O	O
transport	NN	O	O
of	NN	O	O
the	NN	O	O
virus	NN	O	O
to	NN	O	O
the	NN	O	O
brain	NN	O	O
.	NN	O	O

Additionally	NN	O	O
,	NN	O	O
sequence	NN	O	O
variation	NN	O	O
at	NN	O	O
C/EBP	NN	O	B-DNA
site	NN	O	I-DNA
I	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
lies	NN	O	O
immediately	NN	O	O
upstream	NN	O	O
of	NN	O	O
the	NN	O	O
distal	NN	O	B-DNA
nuclear	NN	O	I-DNA
factor	NN	O	I-DNA
kappa	NN	O	I-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
and	NN	O	O
immediately	NN	O	O
downstream	NN	O	O
of	NN	O	O
a	NN	O	O
binding	NN	O	O
site	NN	O	O
for	NN	O	O
activating	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
ATF	NN	O	B-protein
)	NN	O	O
/	NN	O	O
cyclic	NN	O	B-protein
AMP	NN	O	I-protein
response	NN	O	I-protein
element	NN	O	I-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
CREB	NN	O	B-protein
)	NN	O	O
,	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
affect	NN	O	O
HIV-1	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
(	NN	O	O
LTR	NN	O	B-DNA
)	NN	O	O
activity	NN	O	O
.	NN	O	O

Given	NN	O	O
that	NN	O	O
C/EBP	NN	O	B-protein
proteins	NN	O	I-protein
have	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
interact	NN	O	O
with	NN	O	O
many	NN	O	O
other	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
including	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
ATF/CREB	NN	O	B-protein
family	NN	O	I-protein
,	NN	O	O
we	NN	O	O
proceeded	NN	O	O
to	NN	O	O
determine	NN	O	O
whether	NN	O	O
an	NN	O	O
adjacent	NN	O	O
ATF/CREB	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
could	NN	O	O
affect	NN	O	O
C/EBP	NN	O	B-protein
protein	NN	O	I-protein
binding	NN	O	O
to	NN	O	O
C/EBP	NN	O	B-DNA
site	NN	O	I-DNA
I	NN	O	I-DNA
.	NN	O	O

Electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
analyses	NN	O	O
indicated	NN	O	O
that	NN	O	O
selected	NN	O	O
ATF/CREB	NN	O	B-DNA
site	NN	O	I-DNA
variants	NN	O	I-DNA
assisted	NN	O	O
in	NN	O	O
the	NN	O	O
recruitment	NN	O	O
of	NN	O	O
C/EBP	NN	O	B-protein
proteins	NN	O	I-protein
to	NN	O	O
an	NN	O	O
adjacent	NN	O	O
,	NN	O	O
naturally	NN	O	O
occurring	NN	O	O
,	NN	O	O
low-affinity	NN	O	O
C/EBP	NN	O	B-DNA
site	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
biophysical	NN	O	O
interaction	NN	O	O
appears	NN	O	O
to	NN	O	O
occur	NN	O	O
via	NN	O	O
at	NN	O	O
least	NN	O	O
two	NN	O	O
mechanisms	NN	O	O
.	NN	O	O

First	NN	O	O
,	NN	O	O
low	NN	O	O
amounts	NN	O	O
of	NN	O	O
CREB-1	NN	O	B-protein
and	NN	O	O
C/EBP	NN	O	B-protein
appear	NN	O	O
to	NN	O	O
heterodimerize	NN	O	O
and	NN	O	O
bind	NN	O	O
to	NN	O	O
a	NN	O	O
site	NN	O	O
consisting	NN	O	O
of	NN	O	O
a	NN	O	O
half	NN	O	O
site	NN	O	O
from	NN	O	O
both	NN	O	O
the	NN	O	O
ATF/CREB	NN	O	B-DNA
and	NN	O	I-DNA
C/EBP	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
CREB-1	NN	O	B-protein
homodimers	NN	O	I-protein
bind	NN	O	O
to	NN	O	O
the	NN	O	O
ATF/CREB	NN	O	B-DNA
site	NN	O	I-DNA
and	NN	O	O
recruit	NN	O	O
C/EBP	NN	O	B-protein
dimers	NN	O	I-protein
to	NN	O	O
their	NN	O	O
cognate	NN	O	O
weak	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
interaction	NN	O	O
is	NN	O	O
reciprocal	NN	O	O
,	NN	O	O
since	NN	O	O
C/EBP	NN	O	B-protein
dimer	NN	O	I-protein
binding	NN	O	O
to	NN	O	O
a	NN	O	O
strong	NN	O	O
C/EBP	NN	O	B-DNA
site	NN	O	I-DNA
leads	NN	O	O
to	NN	O	O
enhanced	NN	O	O
CREB-1	NN	O	B-protein
recruitment	NN	O	O
to	NN	O	O
ATF/CREB	NN	O	B-DNA
sites	NN	O	I-DNA
that	NN	O	O
are	NN	O	O
weakly	NN	O	O
bound	NN	O	O
by	NN	O	O
CREB	NN	O	B-protein
.	NN	O	O

Sequence	NN	O	O
variation	NN	O	O
at	NN	O	O
both	NN	O	O
C/EBP	NN	O	B-DNA
and	NN	O	I-DNA
ATF/CREB	NN	O	I-DNA
sites	NN	O	I-DNA
affects	NN	O	O
the	NN	O	O
molecular	NN	O	O
interactions	NN	O	O
involved	NN	O	O
in	NN	O	O
mediating	NN	O	O
both	NN	O	O
of	NN	O	O
these	NN	O	O
mechanisms	NN	O	O
.	NN	O	O

Most	NN	O	O
importantly	NN	O	O
,	NN	O	O
sequence	NN	O	O
variation	NN	O	O
at	NN	O	O
the	NN	O	O
ATF/CREB	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
affected	NN	O	O
basal	NN	O	O
LTR	NN	O	B-DNA
activity	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
LTR	NN	O	B-DNA
function	NN	O	O
following	NN	O	O
interleukin-6	NN	O	B-protein
stimulation	NN	O	O
,	NN	O	O
a	NN	O	O
treatment	NN	O	O
that	NN	O	O
leads	NN	O	O
to	NN	O	O
increases	NN	O	O
in	NN	O	O
C/EBP	NN	O	O
activation	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
HIV-1	NN	O	B-DNA
LTR	NN	O	I-DNA
ATF/CREB	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
sequence	NN	O	O
variation	NN	O	O
may	NN	O	O
modulate	NN	O	O
cellular	NN	O	O
signaling	NN	O	O
at	NN	O	O
the	NN	O	O
viral	NN	O	B-DNA
promoter	NN	O	I-DNA
through	NN	O	O
the	NN	O	O
C/EBP	NN	O	O
pathway	NN	O	O

-DOCSTART-	O

Selective	NN	O	O
inhibition	NN	O	O
of	NN	O	O
interleukin-4	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
aspirin	NN	O	O
.	NN	O	O

Previous	NN	O	O
studies	NN	O	O
indicated	NN	O	O
that	NN	O	O
aspirin	NN	O	O
(	NN	O	O
acetylsalicylic	NN	O	O
acid	NN	O	O
[	NN	O	O
ASA	NN	O	O
]	NN	O	O
)	NN	O	O
can	NN	O	O
have	NN	O	O
profound	NN	O	O
immunomodulatory	NN	O	O
effects	NN	O	O
by	NN	O	O
regulating	NN	O	O
cytokine	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
several	NN	O	O
types	NN	O	O
of	NN	O	O
cells	NN	O	O
.	NN	O	O

This	NN	O	O
study	NN	O	O
is	NN	O	O
the	NN	O	O
first	NN	O	O
in	NN	O	O
which	NN	O	O
concentrations	NN	O	O
of	NN	O	O
ASA	NN	O	O
in	NN	O	O
the	NN	O	O
therapeutic	NN	O	O
range	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
significantly	NN	O	O
reduce	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
secretion	NN	O	O
and	NN	O	O
RNA	NN	O	O
expression	NN	O	O
in	NN	O	O
freshly	NN	O	B-cell_type
isolated	NN	O	I-cell_type
and	NN	O	I-cell_type
mitogen-primed	NN	O	I-cell_type
human	NN	O	I-cell_type
CD4+	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
ASA	NN	O	O
did	NN	O	O
not	NN	O	O
affect	NN	O	O
IL-13	NN	O	B-protein
,	NN	O	O
interferon-gamma	NN	O	B-protein
,	NN	O	O
and	NN	O	O
IL-2	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

ASA	NN	O	O
inhibited	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
promoter-driven	NN	O	O
chloramphenicol	NN	O	O
acetyltransferase	NN	O	O
expression	NN	O	O
in	NN	O	O
transiently	NN	O	O
transfected	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
structurally	NN	O	O
unrelated	NN	O	O
nonsteroidal	NN	O	O
anti-inflammatory	NN	O	O
drugs	NN	O	O
indomethacin	NN	O	O
and	NN	O	O
flurbiprofen	NN	O	O
did	NN	O	O
not	NN	O	O
affect	NN	O	O
cytokine	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
weak	NN	O	O
cyclo-oxygenase	NN	O	O
inhibitor	NN	O	O
salicylic	NN	O	O
acid	NN	O	O
was	NN	O	O
at	NN	O	O
least	NN	O	O
as	NN	O	O
effective	NN	O	O
as	NN	O	O
ASA	NN	O	O
in	NN	O	O
inhibiting	NN	O	O
IL-4	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
promoter	NN	O	O
activity	NN	O	O
.	NN	O	O

The	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
of	NN	O	O
ASA	NN	O	O
on	NN	O	O
IL-4	NN	O	O
transcription	NN	O	O
was	NN	O	O
not	NN	O	O
mediated	NN	O	O
by	NN	O	O
decreased	NN	O	O
nuclear	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
known	NN	O	O
salicylate	NN	O	B-protein
target	NN	O	I-protein
nuclear	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	I-protein
NF	NN	O	I-protein
)	NN	O	I-protein
-kappaB	NN	O	I-protein
and	NN	O	O
was	NN	O	O
accompanied	NN	O	O
by	NN	O	O
reduced	NN	O	O
binding	NN	O	O
of	NN	O	O
an	NN	O	O
inducible	NN	O	O
factor	NN	O	O
to	NN	O	O
an	NN	O	O
IL-4	NN	O	B-DNA
promoter	NN	O	I-DNA
region	NN	O	I-DNA
upstream	NN	O	O
of	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
overlapping	NN	O	O
,	NN	O	O
the	NN	O	O
NF	NN	O	B-DNA
of	NN	O	I-DNA
activated	NN	O	I-DNA
T	NN	O	I-DNA
cells-	NN	O	I-DNA
and	NN	O	I-DNA
NF-kappaB-binding	NN	O	I-DNA
P1	NN	O	I-DNA
element	NN	O	I-DNA
.	NN	O	O

It	NN	O	O
is	NN	O	O
concluded	NN	O	O
that	NN	O	O
anti-inflammatory	NN	O	O
salicylates	NN	O	O
,	NN	O	O
by	NN	O	O
means	NN	O	O
of	NN	O	O
a	NN	O	O
previously	NN	O	O
unrecognized	NN	O	O
mechanism	NN	O	O
of	NN	O	O
action	NN	O	O
,	NN	O	O
can	NN	O	O
influence	NN	O	O
the	NN	O	O
nature	NN	O	O
of	NN	O	O
adaptive	NN	O	O
immune	NN	O	O
responses	NN	O	O
by	NN	O	O
selectively	NN	O	O
inhibiting	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
a	NN	O	O
critical	NN	O	O
effector	NN	O	O
of	NN	O	O
these	NN	O	O
responses	NN	O	O
,	NN	O	O
in	NN	O	O
CD4+	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Molecular	NN	O	O
and	NN	O	O
cellular	NN	O	O
mediators	NN	O	O
of	NN	O	O
interleukin-1	NN	O	B-protein
-dependent	NN	O	O
acute	NN	O	O
inflammatory	NN	O	O
arthritis	NN	O	O
.	NN	O	O

OBJECTIVE	NN	O	O
:	NN	O	O
To	NN	O	O
examine	NN	O	O
the	NN	O	O
molecular	NN	O	O
and	NN	O	O
cellular	NN	O	O
mechanisms	NN	O	O
in	NN	O	O
a	NN	O	O
model	NN	O	O
of	NN	O	O
acute	NN	O	O
inflammatory	NN	O	O
monarticular	NN	O	O
arthritis	NN	O	O
induced	NN	O	O
by	NN	O	O
methylated	NN	O	B-protein
bovine	NN	O	I-protein
serum	NN	O	I-protein
albumin	NN	O	I-protein
(	NN	O	O
mBSA	NN	O	B-protein
)	NN	O	O
and	NN	O	O
interleukin-1	NN	O	B-protein
(	NN	O	O
IL-1	NN	O	B-protein
)	NN	O	O
.	NN	O	O

METHODS	NN	O	O
:	NN	O	O
Mice	NN	O	O
were	NN	O	O
injected	NN	O	O
intraarticularly	NN	O	O
with	NN	O	O
mBSA	NN	O	B-protein
on	NN	O	O
day	NN	O	O
0	NN	O	O
and	NN	O	O
subcutaneously	NN	O	O
with	NN	O	O
recombinant	NN	O	B-protein
human	NN	O	I-protein
IL-1beta	NN	O	I-protein
on	NN	O	O
days	NN	O	O
0-2	NN	O	O
.	NN	O	O

At	NN	O	O
day	NN	O	O
7	NN	O	O
,	NN	O	O
knee	NN	O	O
joints	NN	O	O
were	NN	O	O
removed	NN	O	O
and	NN	O	O
assessed	NN	O	O
histologically	NN	O	O
.	NN	O	O

Flow	NN	O	O
cytometry	NN	O	O
and	NN	O	O
RNase	NN	O	O
protection	NN	O	O
were	NN	O	O
used	NN	O	O
to	NN	O	O
analyze	NN	O	O
IL-1	NN	O	B-protein
-dependent	NN	O	O
events	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
C57BL/6	NN	O	O
(	NN	O	O
B6	NN	O	O
)	NN	O	O
,	NN	O	O
129/Sv	NN	O	O
,	NN	O	O
and	NN	O	O
(	NN	O	O
B6	NN	O	O
x	NN	O	O
129/	NN	O	O
Sv	NN	O	O
)	NN	O	O
F1	NN	O	O
hybrid	NN	O	O
mice	NN	O	O
,	NN	O	O
all	NN	O	O
H-2b	NN	O	O
strains	NN	O	O
,	NN	O	O
were	NN	O	O
susceptible	NN	O	O
to	NN	O	O
mBSA	NN	O	B-protein
/IL-1-induced	NN	O	O
arthritis	NN	O	O
,	NN	O	O
whereas	NN	O	O
C3H/HeJ	NN	O	O
(	NN	O	O
H-2k	NN	O	O
)	NN	O	O
mice	NN	O	O
were	NN	O	O
not	NN	O	O
.	NN	O	O

B6	NN	O	O
mice	NN	O	O
lacking	NN	O	O
T	NN	O	B-cell_type
and	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
RAG1-/-	NN	O	O
)	NN	O	O
or	NN	O	O
major	NN	O	B-protein
histocompatibility	NN	O	I-protein
complex	NN	O	I-protein
(	NN	O	I-protein
MHC	NN	O	I-protein
)	NN	O	I-protein
class	NN	O	I-protein
II	NN	O	I-protein
antigens	NN	O	I-protein
(	NN	O	O
MHCII-/-	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
B6	NN	O	O
mice	NN	O	O
treated	NN	O	O
with	NN	O	O
a	NN	O	O
CD4+	NN	O	B-protein
T	NN	O	I-protein
cell-depleting	NN	O	I-protein
monoclonal	NN	O	I-protein
antibody	NN	O	I-protein
,	NN	O	O
were	NN	O	O
resistant	NN	O	O
to	NN	O	O
disease	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
B	NN	O	O
cell-deficient	NN	O	O
(	NN	O	O
muMT/	NN	O	O
muMT	NN	O	O
)	NN	O	O
mice	NN	O	O
developed	NN	O	O
arthritis	NN	O	O
at	NN	O	O
an	NN	O	O
incidence	NN	O	O
and	NN	O	O
severity	NN	O	O
similar	NN	O	O
to	NN	O	O
that	NN	O	O
of	NN	O	O
controls	NN	O	O
.	NN	O	O

RelB-deficient	NN	O	O
(	NN	O	O
RelB-/-	NN	O	O
)	NN	O	O
bone	NN	O	O
marrow	NN	O	O
chimeric	NN	O	O
mice	NN	O	O
had	NN	O	O
arthritis	NN	O	O
that	NN	O	O
was	NN	O	O
significantly	NN	O	O
reduced	NN	O	O
in	NN	O	O
incidence	NN	O	O
and	NN	O	O
severity	NN	O	O
.	NN	O	O

In	NN	O	O
B6	NN	O	O
mice	NN	O	O
,	NN	O	O
flow	NN	O	O
cytometry	NN	O	O
demonstrated	NN	O	O
an	NN	O	O
IL-1-dependent	NN	O	B-cell_type
leukocyte	NN	O	I-cell_type
infiltration	NN	O	O
into	NN	O	O
the	NN	O	O
synovial	NN	O	O
compartment	NN	O	O
and	NN	O	O
RNase	NN	O	O
protection	NN	O	O
assays	NN	O	O
revealed	NN	O	O
induction	NN	O	O
of	NN	O	O
messenger	NN	O	B-RNA
RNA	NN	O	I-RNA
(	NN	O	O
mRNA	NN	O	B-RNA
)	NN	O	O
for	NN	O	O
the	NN	O	O
chemokines	NN	O	B-protein
monocyte	NN	O	B-protein
chemoattractant	NN	O	I-protein
protein	NN	O	I-protein
1	NN	O	I-protein
,	NN	O	O
macrophage	NN	O	B-protein
inhibitory	NN	O	I-protein
protein	NN	O	I-protein
2	NN	O	I-protein
(	NN	O	O
MIP-2	NN	O	B-protein
)	NN	O	O
,	NN	O	O
RANTES	NN	O	B-protein
,	NN	O	O
MIP-1alpha	NN	O	B-protein
,	NN	O	O
and	NN	O	O
MIP-1beta	NN	O	B-protein
,	NN	O	O
in	NN	O	O
vivo	NN	O	O
and	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

CONCLUSION	NN	O	O
:	NN	O	O
Arthritis	NN	O	O
induced	NN	O	O
by	NN	O	O
mBSA	NN	O	B-protein
/	NN	O	O
IL-1	NN	O	B-protein
is	NN	O	O
strain	NN	O	O
specific	NN	O	O
and	NN	O	O
dependent	NN	O	O
on	NN	O	O
CD4+	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
at	NN	O	O
least	NN	O	O
partially	NN	O	O
on	NN	O	O
RelB	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
on	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
or	NN	O	O
antibody	NN	O	B-protein
.	NN	O	O

IL-1	NN	O	B-protein
contributes	NN	O	O
to	NN	O	O
leukocyte	NN	O	O
recruitment	NN	O	O
to	NN	O	O
the	NN	O	O
synovium	NN	O	O
and	NN	O	O
directly	NN	O	O
induces	NN	O	O
chemokine	NN	O	O
mRNA	NN	O	O
production	NN	O	O
by	NN	O	O
synovial	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

This	NN	O	O
model	NN	O	O
of	NN	O	O
acute	NN	O	O
monarticular	NN	O	O
arthritis	NN	O	O
is	NN	O	O
particularly	NN	O	O
suitable	NN	O	O
for	NN	O	O
further	NN	O	O
investigations	NN	O	O
into	NN	O	O
cell-mediated	NN	O	O
immunity	NN	O	O
in	NN	O	O
arthritis	NN	O	O
and	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
IL-1	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Positive	NN	O	O
and	NN	O	O
negative	NN	O	O
regulation	NN	O	O
of	NN	O	O
granulopoiesis	NN	O	O
by	NN	O	O
endogenous	NN	O	B-protein
RARalpha	NN	O	I-protein
.	NN	O	O

Acute	NN	O	O
promyelocytic	NN	O	O
leukemia	NN	O	O
(	NN	O	O
APL	NN	O	O
)	NN	O	O
is	NN	O	O
always	NN	O	O
associated	NN	O	O
with	NN	O	O
chromosomal	NN	O	O
translocations	NN	O	O
that	NN	O	O
disrupt	NN	O	O
the	NN	O	O
retinoic	NN	O	B-DNA
acid	NN	O	I-DNA
receptor	NN	O	I-DNA
alpha	NN	O	I-DNA
(	NN	O	I-DNA
RARalpha	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

Whether	NN	O	O
these	NN	O	O
translocations	NN	O	O
relate	NN	O	O
to	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
endogenous	NN	O	B-protein
RARalpha	NN	O	I-protein
in	NN	O	O
normal	NN	O	O
granulopoiesis	NN	O	O
remains	NN	O	O
uncertain	NN	O	O
because	NN	O	O
most	NN	O	O
studies	NN	O	O
addressing	NN	O	O
this	NN	O	O
question	NN	O	O
have	NN	O	O
used	NN	O	O
non-physiological	NN	O	O
overexpression	NN	O	O
systems	NN	O	O
.	NN	O	O

Granulocyte	NN	O	O
differentiation	NN	O	O
in	NN	O	O
cells	NN	O	O
derived	NN	O	O
from	NN	O	O
RARalpha-deficient	NN	O	O
(	NN	O	O
RARalpha	NN	O	O
(	NN	O	O
-/-	NN	O	O
)	NN	O	O
)	NN	O	O
mice	NN	O	O
was	NN	O	O
studied	NN	O	O
and	NN	O	O
evaluated	NN	O	O
in	NN	O	O
the	NN	O	O
context	NN	O	O
of	NN	O	O
agonist-bound	NN	O	B-protein
and	NN	O	I-protein
ligand-free	NN	O	I-protein
RARalpha	NN	O	I-protein
.	NN	O	O

Our	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
RARalpha	NN	O	B-protein
is	NN	O	O
dispensable	NN	O	O
for	NN	O	O
granulopoiesis	NN	O	O
,	NN	O	O
as	NN	O	O
RARalpha	NN	O	O
(	NN	O	O
-/-	NN	O	O
)	NN	O	O
mice	NN	O	O
have	NN	O	O
a	NN	O	O
normal	NN	O	B-cell_type
granulocyte	NN	O	I-cell_type
population	NN	O	I-cell_type
despite	NN	O	O
an	NN	O	O
impaired	NN	O	O
ability	NN	O	O
to	NN	O	O
respond	NN	O	O
to	NN	O	O
retinoids	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
although	NN	O	O
it	NN	O	O
is	NN	O	O
not	NN	O	O
absolutely	NN	O	O
required	NN	O	O
,	NN	O	O
RARalpha	NN	O	B-protein
can	NN	O	O
bidirectionally	NN	O	O
modulate	NN	O	O
granulopoiesis	NN	O	O
.	NN	O	O

RARalpha	NN	O	B-protein
stimulates	NN	O	O
differentiation	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
exogenous	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
endogenous	NN	O	O
retinoids	NN	O	O
control	NN	O	O
granulopoiesis	NN	O	O
in	NN	O	O
vivo	NN	O	O
,	NN	O	O
as	NN	O	O
either	NN	O	O
vitamin	NN	O	O
A-deficient	NN	O	O
mice	NN	O	O
or	NN	O	O
animals	NN	O	O
treated	NN	O	O
with	NN	O	O
an	NN	O	O
RAR	NN	O	O
antagonist	NN	O	O
accumulate	NN	O	O
more	NN	O	O
immature	NN	O	B-cell_type
granulocytes	NN	O	I-cell_type
in	NN	O	O
their	NN	O	O
bone	NN	O	O
marrow	NN	O	O
.	NN	O	O

Conversely	NN	O	O
,	NN	O	O
RARalpha	NN	O	B-protein
acts	NN	O	O
to	NN	O	O
limit	NN	O	O
differentiation	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
ligand	NN	O	B-protein
because	NN	O	O
granulocyte	NN	O	B-cell_type
precursors	NN	O	I-cell_type
from	NN	O	O
RARalpha	NN	O	O
(	NN	O	O
-/-	NN	O	O
)	NN	O	O
mice	NN	O	O
differentiate	NN	O	O
earlier	NN	O	O
in	NN	O	O
culture	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
block	NN	O	O
in	NN	O	O
granulopoiesis	NN	O	O
exerted	NN	O	O
by	NN	O	O
RARalpha	NN	O	B-protein
fusion	NN	O	B-protein
proteins	NN	O	I-protein
expressed	NN	O	O
in	NN	O	O
APL	NN	O	B-cell_type
cells	NN	O	I-cell_type
may	NN	O	O
correspond	NN	O	O
to	NN	O	O
an	NN	O	O
amplification	NN	O	O
of	NN	O	O
a	NN	O	O
normal	NN	O	O
function	NN	O	O
of	NN	O	O
unliganded	NN	O	B-protein
RARalpha	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Expression	NN	O	O
and	NN	O	O
function	NN	O	O
of	NN	O	O
a	NN	O	O
stem	NN	O	B-DNA
cell	NN	O	I-DNA
promoter	NN	O	I-DNA
for	NN	O	O
the	NN	O	O
murine	NN	O	B-DNA
CBFalpha2	NN	O	I-DNA
gene	NN	O	I-DNA
:	NN	O	O
distinct	NN	O	O
roles	NN	O	O
and	NN	O	O
regulation	NN	O	O
in	NN	O	O
natural	NN	O	B-cell_type
killer	NN	O	I-cell_type
and	NN	O	I-cell_type
T	NN	O	I-cell_type
cell	NN	O	I-cell_type
development	NN	O	O
.	NN	O	O

The	NN	O	O
Runt	NN	O	B-protein
family	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
CBFalpha2	NN	O	B-protein
(	NN	O	O
AML1	NN	O	B-protein
,	NN	O	O
PEBP2alphaB	NN	O	B-protein
,	NN	O	O
or	NN	O	O
Runx1	NN	O	B-protein
)	NN	O	O
is	NN	O	O
required	NN	O	O
by	NN	O	O
hematopoietic	NN	O	B-cell_type
stem	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
expressed	NN	O	O
at	NN	O	O
high	NN	O	O
levels	NN	O	O
in	NN	O	O
T-lineage	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	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
CBFalpha2	NN	O	B-protein
is	NN	O	O
usually	NN	O	O
transcribed	NN	O	O
from	NN	O	O
a	NN	O	O
different	NN	O	B-DNA
promoter	NN	O	I-DNA
(	NN	O	O
distal	NN	O	B-DNA
promoter	NN	O	I-DNA
)	NN	O	O
than	NN	O	O
in	NN	O	O
myeloid	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
proximal	NN	O	B-DNA
promoter	NN	O	I-DNA
)	NN	O	O
,	NN	O	O
but	NN	O	O
the	NN	O	O
developmental	NN	O	O
and	NN	O	O
functional	NN	O	O
significance	NN	O	O
of	NN	O	O
this	NN	O	O
promoter	NN	O	O
switch	NN	O	O
has	NN	O	O
not	NN	O	O
been	NN	O	O
known	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
both	NN	O	O
coding	NN	O	B-DNA
and	NN	O	I-DNA
noncoding	NN	O	I-DNA
sequences	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
distal	NN	O	B-DNA
5	NN	O	I-DNA
'	NN	O	I-DNA
end	NN	O	I-DNA
are	NN	O	O
highly	NN	O	O
conserved	NN	O	O
between	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
and	NN	O	I-DNA
the	NN	O	I-DNA
murine	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
the	NN	O	O
distal	NN	O	B-DNA
promoter	NN	O	I-DNA
is	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
overwhelming	NN	O	O
majority	NN	O	O
of	NN	O	O
CBFalpha2	NN	O	O
expression	NN	O	O
in	NN	O	O
murine	NN	O	O
hematopoietic	NN	O	B-cell_type
stem	NN	O	I-cell_type
cells	NN	O	I-cell_type
as	NN	O	O
well	NN	O	O
as	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Distal	NN	O	O
promoter	NN	O	O
activity	NN	O	O
is	NN	O	O
maintained	NN	O	O
throughout	NN	O	O
T	NN	O	O
cell	NN	O	O
development	NN	O	O
and	NN	O	O
at	NN	O	O
lower	NN	O	O
levels	NN	O	O
in	NN	O	O
B	NN	O	O
cell	NN	O	O
development	NN	O	O
,	NN	O	O
but	NN	O	O
downregulated	NN	O	O
in	NN	O	O
natural	NN	O	O
killer	NN	O	O
cell	NN	O	O
development	NN	O	O
.	NN	O	O

The	NN	O	O
distal	NN	O	B-DNA
N-terminal	NN	O	I-DNA
isoform	NN	O	I-DNA
binds	NN	O	O
to	NN	O	O
functionally	NN	O	O
important	NN	O	O
regulatory	NN	O	B-DNA
sites	NN	O	I-DNA
from	NN	O	O
known	NN	O	O
target	NN	O	B-DNA
genes	NN	O	I-DNA
with	NN	O	O
two-	NN	O	O
to	NN	O	O
threefold	NN	O	O
higher	NN	O	O
affinity	NN	O	O
than	NN	O	O
the	NN	O	O
proximal	NN	O	B-DNA
N-terminal	NN	O	I-DNA
isoform	NN	O	I-DNA
.	NN	O	O

Neither	NN	O	O
full-length	NN	O	B-DNA
isoform	NN	O	I-DNA
alters	NN	O	O
growth	NN	O	O
of	NN	O	O
a	NN	O	O
myeloid	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
under	NN	O	O
nondifferentiating	NN	O	O
conditions	NN	O	O
,	NN	O	O
but	NN	O	O
the	NN	O	O
proximal	NN	O	B-DNA
isoform	NN	O	I-DNA
selectively	NN	O	O
delays	NN	O	O
mitotic	NN	O	O
arrest	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	O
line	NN	O	O
under	NN	O	O
differentiating	NN	O	O
conditions	NN	O	O
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
the	NN	O	O
generation	NN	O	O
of	NN	O	O
greater	NN	O	O
numbers	NN	O	O
of	NN	O	O
neutrophils	NN	O	B-cell_type
.	NN	O	O

-DOCSTART-	O

Constitutive	NN	O	O
PI3-K	NN	O	O
activity	NN	O	O
is	NN	O	O
essential	NN	O	O
for	NN	O	O
proliferation	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
survival	NN	O	O
,	NN	O	O
of	NN	O	O
Theileria	NN	O	B-cell_type
parva-transformed	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Theileria	NN	O	O
is	NN	O	O
an	NN	O	O
intracellular	NN	O	O
parasite	NN	O	O
that	NN	O	O
causes	NN	O	O
lymphoproliferative	NN	O	O
disorders	NN	O	O
in	NN	O	O
cattle	NN	O	O
,	NN	O	O
and	NN	O	O
infection	NN	O	O
of	NN	O	O
leucocytes	NN	O	B-cell_type
induces	NN	O	O
a	NN	O	O
transformed	NN	O	O
phenotype	NN	O	O
similar	NN	O	O
to	NN	O	O
tumour	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
but	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
by	NN	O	O
which	NN	O	O
the	NN	O	O
parasite	NN	O	O
induces	NN	O	O
this	NN	O	O
phenotype	NN	O	O
are	NN	O	O
not	NN	O	O
understood	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
infected	NN	O	B-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
display	NN	O	O
constitutive	NN	O	O
phosphoinositide	NN	O	O
3-kinase	NN	O	O
(	NN	O	O
PI3-K	NN	O	O
)	NN	O	O
activity	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
proliferation	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
survival	NN	O	O
.	NN	O	O

Importantly	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
one	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
PI3-K	NN	O	B-protein
mediates	NN	O	O
the	NN	O	O
proliferation	NN	O	O
of	NN	O	O
infected	NN	O	B-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
is	NN	O	O
through	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
a	NN	O	O
granulocyte-monocyte	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
-dependent	NN	O	O
autocrine	NN	O	O
loop	NN	O	O
.	NN	O	O

PI3-K	NN	O	B-protein
induction	NN	O	O
of	NN	O	O
GM-CSF	NN	O	B-protein
appears	NN	O	O
to	NN	O	O
be	NN	O	O
at	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
level	NN	O	O
and	NN	O	O
,	NN	O	O
consistently	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
PI3-K	NN	O	B-protein
is	NN	O	O
also	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
constitutive	NN	O	O
induction	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
and	NN	O	O
NF-kappaB	NN	O	B-protein
,	NN	O	O
which	NN	O	O
characterizes	NN	O	O
Theileria-infected	NN	O	O
leucocytes	NN	O	B-cell_type
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
our	NN	O	O
results	NN	O	O
highlight	NN	O	O
a	NN	O	O
novel	NN	O	O
strategy	NN	O	O
exploited	NN	O	O
by	NN	O	O
the	NN	O	O
intracellular	NN	O	O
parasite	NN	O	O
Theileria	NN	O	O
to	NN	O	O
induce	NN	O	O
continued	NN	O	O
proliferation	NN	O	O
of	NN	O	O
its	NN	O	O
host	NN	O	B-cell_type
leucocyte	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Lymphokine	NN	O	B-protein
dependence	NN	O	O
of	NN	O	O
STAT3	NN	O	O
activation	NN	O	O
produced	NN	O	O
by	NN	O	O
surface	NN	O	O
immunoglobulin	NN	O	O
cross-linking	NN	O	O
and	NN	O	O
by	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
plus	NN	O	O
calcium	NN	O	O
ionophore	NN	O	O
treatment	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Stimulation	NN	O	O
of	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
surface	NN	O	B-protein
immunoglobulin	NN	O	I-protein
(	NN	O	O
sIg	NN	O	B-protein
)	NN	O	O
triggering	NN	O	O
,	NN	O	O
or	NN	O	O
through	NN	O	O
the	NN	O	O
mitogenic	NN	O	O
combination	NN	O	O
of	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
and	NN	O	O
calcium	NN	O	O
ionophore	NN	O	O
,	NN	O	O
is	NN	O	O
accompanied	NN	O	O
by	NN	O	O
activation	NN	O	O
of	NN	O	O
STAT	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

The	NN	O	O
mechanism	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
delayed	NN	O	O
nuclear	NN	O	O
accumulation	NN	O	O
of	NN	O	O
phosphorylated	NN	O	B-protein
STAT3	NN	O	I-protein
was	NN	O	O
examined	NN	O	O
in	NN	O	O
detail	NN	O	O
,	NN	O	O
focusing	NN	O	O
on	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
B	NN	O	B-protein
cell-derived	NN	O	I-protein
lymphokines	NN	O	I-protein
.	NN	O	O

sIg	NN	O	B-protein
-induced	NN	O	O
activation	NN	O	O
of	NN	O	O
STAT3	NN	O	B-protein
was	NN	O	O
partially	NN	O	O
inhibited	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
obtained	NN	O	O
from	NN	O	O
IL-6-	NN	O	O
or	NN	O	O
IL-10-deficient	NN	O	O
mice	NN	O	O
,	NN	O	O
and	NN	O	O
was	NN	O	O
partially	NN	O	O
blocked	NN	O	O
by	NN	O	O
neutralizing	NN	O	B-protein
antibodies	NN	O	I-protein
directed	NN	O	O
against	NN	O	O
either	NN	O	O
of	NN	O	O
these	NN	O	O
lymphokines	NN	O	B-protein
.	NN	O	O

sIg	NN	O	B-protein
-induced	NN	O	O
STAT3	NN	O	O
activation	NN	O	O
was	NN	O	O
completely	NN	O	O
inhibited	NN	O	O
by	NN	O	O
combining	NN	O	O
IL-6-	NN	O	B-protein
and	NN	O	I-protein
IL-10-specific	NN	O	I-protein
neutralizing	NN	O	I-protein
antibodies	NN	O	I-protein
,	NN	O	O
or	NN	O	O
by	NN	O	O
adding	NN	O	O
individual	NN	O	O
neutralizing	NN	O	B-protein
antibodies	NN	O	I-protein
to	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
obtained	NN	O	O
from	NN	O	O
lymphokine-deficient	NN	O	O
animals	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
IL-10	NN	O	B-protein
alone	NN	O	O
appeared	NN	O	O
to	NN	O	O
account	NN	O	O
for	NN	O	O
STAT3	NN	O	O
activation	NN	O	O
resulting	NN	O	O
from	NN	O	O
B	NN	O	O
cell	NN	O	O
stimulation	NN	O	O
with	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
and	NN	O	O
calcium	NN	O	O
ionophore	NN	O	O
.	NN	O	O

In	NN	O	O
keeping	NN	O	O
with	NN	O	O
these	NN	O	O
results	NN	O	O
,	NN	O	O
soluble	NN	O	O
IL-6	NN	O	B-protein
and	NN	O	O
IL-10	NN	O	B-protein
were	NN	O	O
found	NN	O	O
in	NN	O	O
supernatant	NN	O	O
fluid	NN	O	O
obtained	NN	O	O
from	NN	O	O
stimulated	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

This	NN	O	O
work	NN	O	O
indicates	NN	O	O
that	NN	O	O
a	NN	O	O
lymphokine	NN	O	O
pathway	NN	O	O
is	NN	O	O
responsible	NN	O	O
for	NN	O	O
STAT3	NN	O	O
activation	NN	O	O
that	NN	O	O
occurs	NN	O	O
late	NN	O	O
after	NN	O	O
B	NN	O	O
cell	NN	O	O
stimulation	NN	O	O
,	NN	O	O
and	NN	O	O
points	NN	O	O
out	NN	O	O
differences	NN	O	O
in	NN	O	O
B	NN	O	O
cell	NN	O	O
activation	NN	O	O
that	NN	O	O
result	NN	O	O
from	NN	O	O
stimulation	NN	O	O
through	NN	O	O
the	NN	O	O
antigen	NN	O	B-protein
receptor	NN	O	I-protein
and	NN	O	O
through	NN	O	O
pharmacological	NN	O	O
mimicry	NN	O	O
of	NN	O	O
signaling	NN	O	B-protein
mediators	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Transcription	NN	O	B-protein
factor	NN	O	I-protein
AP-4	NN	O	I-protein
is	NN	O	O
a	NN	O	O
ligand	NN	O	B-protein
for	NN	O	O
immunoglobulin-kappa	NN	O	B-DNA
promoter	NN	O	I-DNA
E-box	NN	O	I-DNA
elements	NN	O	I-DNA
.	NN	O	O

Immunoglobulin	NN	O	B-DNA
(	NN	O	I-DNA
Ig	NN	O	I-DNA
)	NN	O	I-DNA
-kappa	NN	O	I-DNA
promoters	NN	O	I-DNA
from	NN	O	O
humans	NN	O	O
and	NN	O	O
mice	NN	O	O
share	NN	O	O
conserved	NN	O	B-DNA
sequences	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
octamer	NN	O	B-DNA
element	NN	O	I-DNA
is	NN	O	O
common	NN	O	O
to	NN	O	O
all	NN	O	O
Ig	NN	O	B-DNA
promoters	NN	O	I-DNA
and	NN	O	O
pivotal	NN	O	O
for	NN	O	O
their	NN	O	O
function	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
other	NN	O	O
conserved	NN	O	B-DNA
sequence	NN	O	I-DNA
motifs	NN	O	I-DNA
,	NN	O	O
that	NN	O	O
differ	NN	O	O
between	NN	O	O
Ig	NN	O	B-DNA
variable	NN	O	I-DNA
gene	NN	O	I-DNA
families	NN	O	I-DNA
,	NN	O	O
are	NN	O	O
required	NN	O	O
for	NN	O	O
normal	NN	O	O
promoter	NN	O	O
function	NN	O	O
.	NN	O	O

These	NN	O	O
conserved	NN	O	O
motifs	NN	O	O
do	NN	O	O
not	NN	O	O
stimulate	NN	O	O
transcription	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
an	NN	O	O
octamer	NN	O	B-DNA
.	NN	O	O

One	NN	O	O
example	NN	O	O
is	NN	O	O
an	NN	O	O
E-box	NN	O	B-DNA
of	NN	O	O
the	NN	O	O
E47/E12	NN	O	B-DNA
type	NN	O	I-DNA
(	NN	O	I-DNA
5'-CAGCTG-3	NN	O	I-DNA
'	NN	O	I-DNA
)	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
is	NN	O	O
found	NN	O	O
in	NN	O	O
all	NN	O	O
promoters	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
and	NN	O	I-DNA
murine	NN	O	I-DNA
Ig-kappa	NN	O	I-DNA
gene	NN	O	I-DNA
subgroups/families	NN	O	I-DNA
,	NN	O	O
with	NN	O	O
the	NN	O	O
exception	NN	O	O
of	NN	O	O
subgroups	NN	O	O
II	NN	O	O
and	NN	O	O
VI	NN	O	O
and	NN	O	O
their	NN	O	O
related	NN	O	O
murine	NN	O	B-DNA
families	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
ubiquitously	NN	O	O
expressed	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
AP-4	NN	O	I-protein
,	NN	O	O
and	NN	O	O
not	NN	O	O
E47	NN	O	B-protein
,	NN	O	O
interacts	NN	O	O
specifically	NN	O	O
with	NN	O	O
the	NN	O	O
kappa	NN	O	B-DNA
promoter	NN	O	I-DNA
E-boxes	NN	O	I-DNA
when	NN	O	O
tested	NN	O	O
in	NN	O	O
electrophoretic	NN	O	O
mobility-shift	NN	O	O
assays	NN	O	O
using	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
derived	NN	O	O
from	NN	O	O
human	NN	O	B-cell_line
and	NN	O	I-cell_line
murine	NN	O	I-cell_line
B-cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
AP-4	NN	O	B-protein
,	NN	O	O
unlike	NN	O	O
E47	NN	O	B-protein
,	NN	O	O
did	NN	O	O
not	NN	O	O
act	NN	O	O
as	NN	O	O
a	NN	O	O
transactivator	NN	O	B-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
in	NN	O	O
agreement	NN	O	O
with	NN	O	O
previous	NN	O	O
studies	NN	O	O
on	NN	O	O
intact	NN	O	O
kappa	NN	O	B-DNA
promoters	NN	O	I-DNA
,	NN	O	O
showing	NN	O	O
that	NN	O	O
transcription	NN	O	O
is	NN	O	O
absent	NN	O	O
when	NN	O	O
the	NN	O	O
octamer	NN	O	B-DNA
element	NN	O	I-DNA
has	NN	O	O
been	NN	O	O
mutated	NN	O	O
.	NN	O	O

Based	NN	O	O
on	NN	O	O
these	NN	O	O
data	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
conservation	NN	O	O
of	NN	O	O
the	NN	O	O
5'-CAGCTG-3	NN	O	B-DNA
'	NN	O	I-DNA
motif	NN	O	I-DNA
among	NN	O	O
human	NN	O	B-DNA
and	NN	O	I-DNA
murine	NN	O	I-DNA
kappa	NN	O	I-DNA
promoters	NN	O	I-DNA
,	NN	O	O
we	NN	O	O
propose	NN	O	O
that	NN	O	O
AP-4	NN	O	B-protein
is	NN	O	O
the	NN	O	O
major	NN	O	B-protein
ligand	NN	O	I-protein
for	NN	O	O
Ig-kappa	NN	O	B-DNA
promoter	NN	O	I-DNA
E-boxes	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
16-mer	NN	O	O
peptide	NN	O	O
(	NN	O	O
RQIKIWFQNRRMKWKK	NN	O	O
)	NN	O	O
from	NN	O	O
antennapedia	NN	O	O
preferentially	NN	O	O
targets	NN	O	O
the	NN	O	O
Class	NN	O	O
I	NN	O	O
pathway	NN	O	O
.	NN	O	O

Translocation	NN	O	O
of	NN	O	O
antigenic	NN	O	O
peptides	NN	O	O
into	NN	O	O
the	NN	O	O
cytosol	NN	O	O
of	NN	O	O
antigen	NN	O	B-cell_type
presenting	NN	O	I-cell_type
cells	NN	O	I-cell_type
facilitates	NN	O	O
proteosomal	NN	O	O
processing	NN	O	O
and	NN	O	O
loading	NN	O	O
into	NN	O	O
Class	NN	O	B-protein
I	NN	O	I-protein
molecules	NN	O	I-protein
for	NN	O	O
MHC	NN	O	O
presentation	NN	O	O
on	NN	O	O
the	NN	O	O
cell	NN	O	O
surface	NN	O	O
.	NN	O	O

The	NN	O	O
DNA	NN	O	B-protein
binding	NN	O	I-protein
domain	NN	O	I-protein
of	NN	O	O
the	NN	O	O
Drosophila	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
Antennapedia	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
60	NN	O	B-protein
amino	NN	O	I-protein
acid	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
is	NN	O	O
rapidly	NN	O	O
taken	NN	O	O
up	NN	O	O
by	NN	O	O
cells	NN	O	O
and	NN	O	O
has	NN	O	O
been	NN	O	O
fused	NN	O	O
to	NN	O	O
selected	NN	O	O
antigens	NN	O	B-protein
to	NN	O	O
enhance	NN	O	O
their	NN	O	O
immunogenicity	NN	O	O
.	NN	O	O

We	NN	O	O
now	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
a	NN	O	O
16	NN	O	O
amino	NN	O	O
acid	NN	O	O
peptide	NN	O	O
from	NN	O	O
antennapedia	NN	O	O
can	NN	O	O
facilitate	NN	O	O
the	NN	O	O
cytoplasmic	NN	O	O
uptake	NN	O	O
of	NN	O	O
CTL	NN	O	O
epitope	NN	O	O
9-mer	NN	O	O
peptides	NN	O	O
.	NN	O	O

Synthetic	NN	O	O
peptides	NN	O	O
were	NN	O	O
made	NN	O	O
containing	NN	O	O
the	NN	O	O
16-mer	NN	O	O
antennapedia	NN	O	O
peptide	NN	O	O
linked	NN	O	O
in	NN	O	O
tandem	NN	O	O
to	NN	O	O
the	NN	O	O
ovalbumin	NN	O	O
SIINFEKL	NN	O	O
CTL	NN	O	O
peptide	NN	O	O
.	NN	O	O

The	NN	O	O
peptide	NN	O	O
complex	NN	O	O
was	NN	O	O
shown	NN	O	O
to	NN	O	O
rapidly	NN	O	O
internalise	NN	O	O
into	NN	O	O
APCs	NN	O	B-cell_type
by	NN	O	O
confocal	NN	O	O
microscopy	NN	O	O
.	NN	O	O

This	NN	O	O
peptide	NN	O	O
induced	NN	O	O
CTL	NN	O	B-cell_type
in	NN	O	O
C57BL/6	NN	O	O
mice	NN	O	O
and	NN	O	O
protected	NN	O	O
them	NN	O	O
against	NN	O	O
growth	NN	O	O
of	NN	O	O
an	NN	O	O
ovalbumin	NN	O	B-cell_line
expressing	NN	O	I-cell_line
tumour	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
(	NN	O	O
E.G7-OVA	NN	O	B-cell_line
)	NN	O	O
.	NN	O	O

The	NN	O	O
ability	NN	O	O
of	NN	O	O
the	NN	O	O
hybrid	NN	O	O
peptide	NN	O	O
to	NN	O	O
be	NN	O	O
processed	NN	O	O
and	NN	O	O
presented	NN	O	O
by	NN	O	O
APCs	NN	O	B-cell_type
was	NN	O	O
similar	NN	O	O
,	NN	O	O
whether	NN	O	O
the	NN	O	O
SIINFEKL	NN	O	O
sequence	NN	O	O
was	NN	O	O
appended	NN	O	O
at	NN	O	O
the	NN	O	O
C-terminus	NN	O	B-protein
or	NN	O	O
N-terminus	NN	O	B-protein
of	NN	O	O
the	NN	O	O
Antennapedia	NN	O	O
peptide	NN	O	O
.	NN	O	O

The	NN	O	O
production	NN	O	O
of	NN	O	O
synthetic	NN	O	O
peptides	NN	O	O
containing	NN	O	O
other	NN	O	O
CTL	NN	O	O
peptide	NN	O	O
epitopes	NN	O	O
may	NN	O	O
be	NN	O	O
useful	NN	O	O
for	NN	O	O
priming	NN	O	O
CTLs	NN	O	B-cell_type
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
in	NN	O	O
vivo	NN	O	O

-DOCSTART-	O

Bone	NN	O	B-cell_type
marrow	NN	O	I-cell_type
cells	NN	O	I-cell_type
promote	NN	O	O
TH2	NN	O	O
polarization	NN	O	O
and	NN	O	O
inhibit	NN	O	O
virus-specific	NN	O	O
CTL	NN	O	O
generation	NN	O	O
.	NN	O	O

This	NN	O	O
laboratory	NN	O	O
recently	NN	O	O
reported	NN	O	O
that	NN	O	O
human	NN	O	B-cell_type
bone	NN	O	I-cell_type
marrow	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
BMC	NN	O	B-cell_type
)	NN	O	O
inhibit	NN	O	O
the	NN	O	O
generation	NN	O	O
of	NN	O	O
virus-specific	NN	O	B-cell_type
CTL	NN	O	I-cell_type
in	NN	O	O
culture	NN	O	O
.	NN	O	O

The	NN	O	O
culture	NN	O	O
supernatants	NN	O	O
contained	NN	O	O
increased	NN	O	O
levels	NN	O	O
of	NN	O	O
prostaglandin	NN	O	B-protein
E	NN	O	I-protein
(	NN	O	I-protein
2	NN	O	I-protein
)	NN	O	I-protein
(	NN	O	I-protein
PGE	NN	O	I-protein
(	NN	O	I-protein
2	NN	O	I-protein
)	NN	O	I-protein
)	NN	O	I-protein
(	NN	O	O
shown	NN	O	O
to	NN	O	O
favor	NN	O	O
TH2	NN	O	O
cell	NN	O	O
development	NN	O	O
)	NN	O	O
and	NN	O	O
also	NN	O	O
inhibited	NN	O	O
EBV-CTL	NN	O	O
effector	NN	O	O
cell	NN	O	O
development	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
obtained	NN	O	O
PBL	NN	O	B-cell_type
from	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
(	NN	O	O
EBV	NN	O	O
)	NN	O	O
IgG	NN	O	B-protein
antibody	NN	O	I-protein
positive	NN	O	O
kidney	NN	O	O
transplant	NN	O	O
recipients	NN	O	O
(	NN	O	O
R	NN	O	O
)	NN	O	O
and	NN	O	O
their	NN	O	O
living-related	NN	O	O
donors	NN	O	O
(	NN	O	O
LRD	NN	O	O
)	NN	O	O
one	NN	O	O
year	NN	O	O
after	NN	O	O
renal	NN	O	O
transplantation	NN	O	O
.	NN	O	O

EBV-specific	NN	O	B-cell_type
CTL	NN	O	I-cell_type
were	NN	O	O
then	NN	O	O
generated	NN	O	O
in	NN	O	O
vitro	NN	O	O
by	NN	O	O
stimulating	NN	O	O
PBL	NN	O	B-cell_type
with	NN	O	O
autologous	NN	O	O
EBV-transformed	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
EBV-B	NN	O	B-cell_type
)	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
autologous	NN	O	B-cell_type
BMC	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
addition	NN	O	O
of	NN	O	O
BMC	NN	O	B-cell_type
to	NN	O	O
the	NN	O	O
EBV-CTL	NN	O	B-cell_type
generation	NN	O	I-cell_type
cultures	NN	O	I-cell_type
increased	NN	O	O
the	NN	O	O
intracellular	NN	O	O
expression	NN	O	O
in	NN	O	O
CD3+	NN	O	B-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	I-protein
-5	NN	O	I-protein
,	NN	O	I-protein
-6	NN	O	I-protein
,	NN	O	I-protein
-10	NN	O	I-protein
,	NN	O	I-protein
and	NN	O	I-protein
-13	NN	O	I-protein
.	NN	O	O

These	NN	O	O
CD3+	NN	O	B-cell_type
cells	NN	O	I-cell_type
also	NN	O	O
expressed	NN	O	O
increased	NN	O	O
levels	NN	O	O
of	NN	O	O
the	NN	O	O
TH2	NN	O	B-protein
associated	NN	O	I-protein
receptor	NN	O	I-protein
CCR3	NN	O	I-protein
.	NN	O	O

Inhibition	NN	O	O
was	NN	O	O
even	NN	O	O
observed	NN	O	O
by	NN	O	O
preparing	NN	O	O
EBV-CTL	NN	O	B-cell_type
generating	NN	O	I-cell_type
cultures	NN	O	I-cell_type
in	NN	O	O
trans-wells	NN	O	O
that	NN	O	O
separated	NN	O	O
the	NN	O	O
autologous	NN	O	B-cell_type
BMC	NN	O	I-cell_type
from	NN	O	O
the	NN	O	O
PBL	NN	O	B-cell_type
+	NN	O	O
EBV-B	NN	O	B-cell_type
.	NN	O	O

It	NN	O	O
was	NN	O	O
then	NN	O	O
observed	NN	O	O
that	NN	O	O
CD3+	NN	O	B-cell_type
cells	NN	O	I-cell_type
obtained	NN	O	O
after	NN	O	O
7	NN	O	O
days	NN	O	O
of	NN	O	O
culture	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
autologous	NN	O	B-cell_type
BMC	NN	O	I-cell_type
could	NN	O	O
be	NN	O	O
used	NN	O	O
as	NN	O	O
inhibitors	NN	O	O
of	NN	O	O
EBV-CTL	NN	O	O
generation	NN	O	O
.	NN	O	O

Protein	NN	O	B-protein
Kinase	NN	O	I-protein
A	NN	O	I-protein
(	NN	O	O
PKA	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
cAMP	NN	O	B-protein
kinase	NN	O	I-protein
that	NN	O	O
is	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
upregulation	NN	O	O
of	NN	O	O
TH2	NN	O	B-protein
cytokine	NN	O	I-protein
activity	NN	O	O
,	NN	O	O
was	NN	O	O
increased	NN	O	O
in	NN	O	O
EBV-CTL	NN	O	B-cell_type
cultures	NN	O	I-cell_type
by	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
BMC	NN	O	B-cell_type
.	NN	O	O

Additionally	NN	O	O
,	NN	O	O
IL-4-mediated	NN	O	O
signal	NN	O	O
transduction	NN	O	O
and	NN	O	O
activation	NN	O	O
of	NN	O	O
transcription	NN	O	O
(	NN	O	O
STAT-6	NN	O	B-protein
)	NN	O	O
phosphorylation	NN	O	O
was	NN	O	O
slightly	NN	O	O
increased	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
BMC	NN	O	B-cell_type
inhibition	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
soluble	NN	O	B-protein
factors	NN	O	I-protein
(	NN	O	O
cytokines	NN	O	B-protein
)	NN	O	O
and	NN	O	O
that	NN	O	O
cell-cell	NN	O	O
contact	NN	O	O
in	NN	O	O
this	NN	O	O
autologous	NN	O	O
system	NN	O	O
is	NN	O	O
not	NN	O	O
required	NN	O	O
,	NN	O	O
so	NN	O	O
that	NN	O	O
BMC	NN	O	B-cell_type
(	NN	O	O
at	NN	O	O
least	NN	O	O
partially	NN	O	O
,	NN	O	O
via	NN	O	O
cytokine	NN	O	O
production	NN	O	O
)	NN	O	O
promote	NN	O	O
TH2	NN	O	O
polarization	NN	O	O
in	NN	O	O
culture	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
TH2	NN	O	B-cell_type
cells	NN	O	I-cell_type
induced	NN	O	O
by	NN	O	O
culturing	NN	O	O
with	NN	O	O
autologous	NN	O	B-cell_type
BMC	NN	O	I-cell_type
directly	NN	O	O
inhibit	NN	O	O
EBV-CTL	NN	O	O
generation	NN	O	O
,	NN	O	O
and	NN	O	O
TH2	NN	O	B-protein
associated	NN	O	O
PKA	NN	O	B-protein
,	NN	O	O
CCR3	NN	O	B-protein
,	NN	O	O
and	NN	O	O
STAT-6	NN	O	B-protein
phosphorylation	NN	O	O
are	NN	O	O
enhanced	NN	O	O
by	NN	O	O
BMC	NN	O	B-cell_type
.	NN	O	O

-DOCSTART-	O

Reduction	NN	O	O
in	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
Pax-5a	NN	O	B-protein
in	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
of	NN	O	O
aged	NN	O	O
mice	NN	O	O
.	NN	O	O

Aging	NN	O	O
has	NN	O	O
been	NN	O	O
associated	NN	O	O
with	NN	O	O
intrinsic	NN	O	O
changes	NN	O	O
of	NN	O	O
the	NN	O	O
humoral	NN	O	O
immune	NN	O	O
response	NN	O	O
,	NN	O	O
which	NN	O	O
may	NN	O	O
lead	NN	O	O
to	NN	O	O
an	NN	O	O
increased	NN	O	O
occurrence	NN	O	O
of	NN	O	O
autoimmune	NN	O	O
disorders	NN	O	O
and	NN	O	O
pathogenic	NN	O	O
susceptibility	NN	O	O
.	NN	O	O

The	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
Pax-5	NN	O	B-protein
is	NN	O	O
a	NN	O	O
key	NN	O	O
regulator	NN	O	O
of	NN	O	O
B	NN	O	O
cell	NN	O	O
development	NN	O	O
.	NN	O	O

Pax-5a/B	NN	O	B-protein
cell-specific	NN	O	I-protein
activator	NN	O	I-protein
protein	NN	O	I-protein
and	NN	O	O
an	NN	O	O
alternatively	NN	O	B-protein
spliced	NN	O	I-protein
isoform	NN	O	I-protein
,	NN	O	I-protein
Pax-5d	NN	O	I-protein
,	NN	O	O
may	NN	O	O
have	NN	O	O
opposing	NN	O	O
functions	NN	O	O
in	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
due	NN	O	O
to	NN	O	O
the	NN	O	O
lack	NN	O	O
of	NN	O	O
a	NN	O	O
transactivation	NN	O	B-protein
domain	NN	O	I-protein
in	NN	O	O
Pax-5d	NN	O	B-protein
.	NN	O	O

To	NN	O	O
study	NN	O	O
B	NN	O	B-cell_type
cell	NN	O	I-cell_type
-specific	NN	O	O
changes	NN	O	O
that	NN	O	O
occur	NN	O	O
during	NN	O	O
the	NN	O	O
aging	NN	O	O
process	NN	O	O
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
expression	NN	O	O
patterns	NN	O	O
of	NN	O	O
Pax-5a	NN	O	B-protein
and	NN	O	I-protein
5d	NN	O	I-protein
in	NN	O	O
mature	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
young	NN	O	O
and	NN	O	O
aged	NN	O	O
mice	NN	O	O
.	NN	O	O

RNase	NN	O	O
protection	NN	O	O
assays	NN	O	O
showed	NN	O	O
a	NN	O	O
similar	NN	O	O
transcriptional	NN	O	O
pattern	NN	O	O
for	NN	O	O
both	NN	O	O
age	NN	O	O
groups	NN	O	O
that	NN	O	O
indicates	NN	O	O
that	NN	O	O
aging	NN	O	O
has	NN	O	O
no	NN	O	O
affect	NN	O	O
on	NN	O	O
transcription	NN	O	O
initiation	NN	O	O
or	NN	O	O
alternative	NN	O	O
splicing	NN	O	O
for	NN	O	O
either	NN	O	O
isoform	NN	O	B-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
a	NN	O	O
significant	NN	O	O
reduction	NN	O	O
in	NN	O	O
the	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
of	NN	O	O
Pax-5a	NN	O	B-protein
but	NN	O	O
not	NN	O	O
Pax-5d	NN	O	B-protein
protein	NN	O	O
was	NN	O	O
observed	NN	O	O
in	NN	O	O
aged	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
vitro	NN	O	O
,	NN	O	O
while	NN	O	O
Western	NN	O	O
blot	NN	O	O
analyses	NN	O	O
showed	NN	O	O
that	NN	O	O
similar	NN	O	O
levels	NN	O	O
of	NN	O	O
Pax-5a	NN	O	B-protein
and	NN	O	I-protein
5d	NN	O	I-protein
proteins	NN	O	O
were	NN	O	O
present	NN	O	O
in	NN	O	O
both	NN	O	O
age	NN	O	O
groups	NN	O	O
.	NN	O	O

The	NN	O	O
observed	NN	O	O
decrease	NN	O	O
in	NN	O	O
Pax-5a	NN	O	B-protein
binding	NN	O	O
activity	NN	O	O
correlated	NN	O	O
with	NN	O	O
changes	NN	O	O
in	NN	O	O
expression	NN	O	O
of	NN	O	O
two	NN	O	O
Pax-5	NN	O	B-DNA
target	NN	O	I-DNA
genes	NN	O	I-DNA
in	NN	O	O
aged	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
the	NN	O	O
Ig	NN	O	B-protein
J	NN	O	I-protein
chain	NN	O	I-protein
and	NN	O	O
the	NN	O	O
secreted	NN	O	O
form	NN	O	O
of	NN	O	O
Ig	NN	O	B-protein
mu	NN	O	I-protein
,	NN	O	O
which	NN	O	O
are	NN	O	O
both	NN	O	O
known	NN	O	O
to	NN	O	O
be	NN	O	O
suppressed	NN	O	O
by	NN	O	O
Pax-5a	NN	O	B-protein
in	NN	O	O
mature	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
were	NN	O	O
increased	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
aged	NN	O	O
mice	NN	O	O
.	NN	O	O

Together	NN	O	O
,	NN	O	O
our	NN	O	O
studies	NN	O	O
suggest	NN	O	O
that	NN	O	O
changes	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
aging	NN	O	O
phenotype	NN	O	O
cause	NN	O	O
posttranslational	NN	O	O
modification	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
of	NN	O	O
Pax-5a	NN	O	B-protein
but	NN	O	O
not	NN	O	O
Pax-5d	NN	O	B-protein
,	NN	O	O
which	NN	O	O
may	NN	O	O
lead	NN	O	O
to	NN	O	O
an	NN	O	O
abnormal	NN	O	O
B	NN	O	O
cell	NN	O	O
phenotype	NN	O	O
in	NN	O	O
aged	NN	O	O
mice	NN	O	O
,	NN	O	O
associated	NN	O	O
with	NN	O	O
elevated	NN	O	O
levels	NN	O	O
of	NN	O	O
J	NN	O	B-protein
chain	NN	O	I-protein
,	NN	O	O
and	NN	O	O
secretion	NN	O	O
of	NN	O	O
IgM	NN	O	B-protein

-DOCSTART-	O

NF-kappa	NN	O	B-protein
B/Rel	NN	O	I-protein
participation	NN	O	O
in	NN	O	O
the	NN	O	O
lymphokine-dependent	NN	O	O
proliferation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
lymphoid	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Proliferative	NN	O	O
responses	NN	O	O
of	NN	O	O
lymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
IL-4	NN	O	B-protein
depend	NN	O	O
on	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
cells	NN	O	O
,	NN	O	O
but	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
activation	NN	O	O
enhances	NN	O	O
cellular	NN	O	O
competence	NN	O	O
to	NN	O	O
respond	NN	O	O
to	NN	O	O
cytokines	NN	O	B-protein
is	NN	O	O
not	NN	O	O
fully	NN	O	O
understood	NN	O	O
.	NN	O	O

The	NN	O	O
NF-kappaB/Rel	NN	O	B-protein
family	NN	O	I-protein
represents	NN	O	O
one	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathway	NN	O	O
induced	NN	O	O
during	NN	O	O
such	NN	O	O
activation	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
in	NN	O	O
this	NN	O	O
study	NN	O	O
that	NN	O	O
inhibition	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
through	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
an	NN	O	O
IkappaBalpha	NN	O	B-protein
(	NN	O	O
inhibitory	NN	O	B-protein
protein	NN	O	I-protein
that	NN	O	O
dissociates	NN	O	O
from	NN	O	O
NF-kappaB	NN	O	B-protein
)	NN	O	O
mutant	NN	O	O
refractory	NN	O	O
to	NN	O	O
signal-induced	NN	O	O
degradation	NN	O	O
(	NN	O	O
IkappaBalpha	NN	O	B-protein
(	NN	O	I-protein
DeltaN	NN	O	I-protein
)	NN	O	I-protein
)	NN	O	O
interfered	NN	O	O
with	NN	O	O
the	NN	O	O
acquisition	NN	O	O
of	NN	O	O
competence	NN	O	O
to	NN	O	O
proliferate	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
IL-4	NN	O	B-protein
as	NN	O	O
well	NN	O	O
as	NN	O	O
IL-2	NN	O	B-protein
.	NN	O	O

Thymocytes	NN	O	B-cell_type
and	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
IkappaBalpha	NN	O	B-protein
(	NN	O	I-protein
DeltaN	NN	O	I-protein
)	NN	O	I-protein
transgenic	NN	O	O
mice	NN	O	O
expressed	NN	O	O
normal	NN	O	O
levels	NN	O	O
of	NN	O	O
IL-2R	NN	O	B-protein
subunits	NN	O	I-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
transgenic	NN	O	B-cell_type
cells	NN	O	I-cell_type
exhibited	NN	O	O
a	NN	O	O
dramatic	NN	O	O
defect	NN	O	O
in	NN	O	O
Stat5A	NN	O	O
activation	NN	O	O
treatment	NN	O	O
with	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
and	NN	O	O
a	NN	O	O
similar	NN	O	O
defect	NN	O	O
was	NN	O	O
observed	NN	O	O
for	NN	O	O
IL-4	NN	O	B-protein
-induced	NN	O	O
Stat5	NN	O	B-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
T	NN	O	B-cell_type
lymphoid	NN	O	I-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
inhibition	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
showed	NN	O	O
normal	NN	O	O
insulin	NN	O	O
receptor	NN	O	O
substrate-2	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
only	NN	O	O
a	NN	O	O
modest	NN	O	O
decrease	NN	O	O
in	NN	O	O
Stat6	NN	O	O
activation	NN	O	O
and	NN	O	O
insulin	NN	O	O
receptor	NN	O	O
substrate-1	NN	O	O
phosphorylation	NN	O	O
after	NN	O	O
IL-4	NN	O	O
stimulation	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
NF-kappaB/Rel/IkappaBalpha	NN	O	O
system	NN	O	O
can	NN	O	O
regulate	NN	O	O
cytokine	NN	O	B-protein
receptor	NN	O	I-protein
capacitation	NN	O	O
through	NN	O	O
effects	NN	O	O
on	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
downstream	NN	O	O
signaling	NN	O	O
by	NN	O	O
the	NN	O	O
Stat	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
family	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

BMS-189453	NN	O	O
,	NN	O	O
a	NN	O	O
novel	NN	O	O
retinoid	NN	O	O
receptor	NN	O	O
antagonist	NN	O	O
,	NN	O	O
is	NN	O	O
a	NN	O	O
potent	NN	O	O
testicular	NN	O	O
toxin	NN	O	O
.	NN	O	O

BMS-189453	NN	O	O
is	NN	O	O
a	NN	O	O
synthetic	NN	O	O
retinoid	NN	O	O
that	NN	O	O
acts	NN	O	O
as	NN	O	O
an	NN	O	O
antagonist	NN	O	O
at	NN	O	O
retinoic	NN	O	B-protein
acid	NN	O	I-protein
receptors	NN	O	I-protein
alpha	NN	O	I-protein
,	NN	O	I-protein
beta	NN	O	I-protein
,	NN	O	I-protein
and	NN	O	I-protein
gamma	NN	O	I-protein
.	NN	O	O

In	NN	O	O
Sprague	NN	O	O
Dawley	NN	O	O
rats	NN	O	O
at	NN	O	O
daily	NN	O	O
oral	NN	O	O
doses	NN	O	O
of	NN	O	O
15	NN	O	O
,	NN	O	O
60	NN	O	O
,	NN	O	O
or	NN	O	O
240	NN	O	O
mg/kg	NN	O	O
for	NN	O	O
1	NN	O	O
month	NN	O	O
,	NN	O	O
BMS-189453	NN	O	O
produced	NN	O	O
increases	NN	O	O
in	NN	O	O
leukocyte	NN	O	B-cell_type
counts	NN	O	O
,	NN	O	O
alkaline	NN	O	B-protein
phosphatase	NN	O	I-protein
and	NN	O	O
alanine	NN	O	B-protein
aminotransferase	NN	O	I-protein
levels	NN	O	O
,	NN	O	O
and	NN	O	O
marked	NN	O	O
testicular	NN	O	O
degeneration	NN	O	O
and	NN	O	O
atrophy	NN	O	O
at	NN	O	O
all	NN	O	O
doses	NN	O	O
.	NN	O	O

Significant	NN	O	O
overt	NN	O	O
signs	NN	O	O
of	NN	O	O
toxicity	NN	O	O
and	NN	O	O
deaths	NN	O	O
occurred	NN	O	O
at	NN	O	O
240	NN	O	O
mg/kg	NN	O	O
,	NN	O	O
whereas	NN	O	O
body-weight	NN	O	O
and	NN	O	O
food-consumption	NN	O	O
decreases	NN	O	O
occurred	NN	O	O
at	NN	O	O
60	NN	O	O
and	NN	O	O
240	NN	O	O
mg/kg	NN	O	O
.	NN	O	O

When	NN	O	O
BMS-189453	NN	O	O
was	NN	O	O
administered	NN	O	O
to	NN	O	O
male	NN	O	O
rats	NN	O	O
at	NN	O	O
daily	NN	O	O
doses	NN	O	O
ranging	NN	O	O
from	NN	O	O
12.5	NN	O	O
to	NN	O	O
100	NN	O	O
mg/kg	NN	O	O
for	NN	O	O
1	NN	O	O
week	NN	O	O
,	NN	O	O
only	NN	O	O
minimal	NN	O	O
testicular	NN	O	O
changes	NN	O	O
occurred	NN	O	O
at	NN	O	O
all	NN	O	O
doses	NN	O	O
,	NN	O	O
shortly	NN	O	O
after	NN	O	O
the	NN	O	O
dosing	NN	O	O
period	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
after	NN	O	O
a	NN	O	O
1-month	NN	O	O
drug-free	NN	O	O
observation	NN	O	O
period	NN	O	O
,	NN	O	O
marked	NN	O	O
testicular	NN	O	O
atrophy	NN	O	O
was	NN	O	O
evident	NN	O	O
at	NN	O	O
all	NN	O	O
doses	NN	O	O
.	NN	O	O

BMS-189453	NN	O	O
was	NN	O	O
then	NN	O	O
administered	NN	O	O
at	NN	O	O
doses	NN	O	O
of	NN	O	O
2	NN	O	O
,	NN	O	O
10	NN	O	O
,	NN	O	O
or	NN	O	O
50	NN	O	O
mg/kg	NN	O	O
to	NN	O	O
male	NN	O	O
rats	NN	O	O
for	NN	O	O
1	NN	O	O
,	NN	O	O
3	NN	O	O
,	NN	O	O
or	NN	O	O
7	NN	O	O
consecutive	NN	O	O
days	NN	O	O
.	NN	O	O

Dose-	NN	O	O
and	NN	O	O
duration-dependent	NN	O	O
testicular	NN	O	O
toxicity	NN	O	O
that	NN	O	O
occurred	NN	O	O
after	NN	O	O
a	NN	O	O
1-month	NN	O	O
observation	NN	O	O
period	NN	O	O
did	NN	O	O
not	NN	O	O
recover	NN	O	O
,	NN	O	O
and	NN	O	O
,	NN	O	O
in	NN	O	O
some	NN	O	O
cases	NN	O	O
,	NN	O	O
was	NN	O	O
more	NN	O	O
severe	NN	O	O
4	NN	O	O
months	NN	O	O
after	NN	O	O
the	NN	O	O
last	NN	O	O
dose	NN	O	O
.	NN	O	O

In	NN	O	O
rabbits	NN	O	O
administered	NN	O	O
BMS-189453	NN	O	O
at	NN	O	O
oral	NN	O	O
doses	NN	O	O
of	NN	O	O
2	NN	O	O
,	NN	O	O
10	NN	O	O
,	NN	O	O
or	NN	O	O
50	NN	O	O
mg/kg	NN	O	O
for	NN	O	O
1	NN	O	O
week	NN	O	O
,	NN	O	O
testicular	NN	O	O
degeneration	NN	O	O
and	NN	O	O
atrophy	NN	O	O
were	NN	O	O
evident	NN	O	O
in	NN	O	O
the	NN	O	O
high-dose	NN	O	O
group	NN	O	O
at	NN	O	O
1	NN	O	O
month	NN	O	O
following	NN	O	O
treatment	NN	O	O
.	NN	O	O

These	NN	O	O
studies	NN	O	O
indicate	NN	O	O
that	NN	O	O
retinoid	NN	O	O
antagonists	NN	O	O
can	NN	O	O
selectively	NN	O	O
produce	NN	O	O
progressive	NN	O	O
and	NN	O	O
prolonged	NN	O	O
testicular	NN	O	O
toxicity	NN	O	O
after	NN	O	O
single	NN	O	O
or	NN	O	O
repeated	NN	O	O
oral	NN	O	O
doses	NN	O	O
that	NN	O	O
are	NN	O	O
otherwise	NN	O	O
well	NN	O	O
tolerated	NN	O	O
.	NN	O	O

-DOCSTART-	O

Sequential	NN	O	O
involvement	NN	O	O
of	NN	O	O
NFAT	NN	O	B-protein
and	NN	O	O
Egr	NN	O	B-protein
transcription	NN	O	B-protein
factors	NN	O	I-protein
in	NN	O	O
FasL	NN	O	O
regulation	NN	O	O
.	NN	O	O

The	NN	O	O
critical	NN	O	O
function	NN	O	O
of	NN	O	O
NFAT	NN	O	B-protein
proteins	NN	O	I-protein
in	NN	O	O
maintaining	NN	O	O
lymphoid	NN	O	O
homeostasis	NN	O	O
was	NN	O	O
revealed	NN	O	O
in	NN	O	O
mice	NN	O	O
lacking	NN	O	O
both	NN	O	O
NFATp	NN	O	B-protein
and	NN	O	O
NFAT4	NN	O	B-protein
(	NN	O	O
DKO	NN	O	O
)	NN	O	O
.	NN	O	O

DKO	NN	O	O
mice	NN	O	O
exhibit	NN	O	O
increased	NN	O	O
lymphoproliferation	NN	O	O
,	NN	O	O
decreased	NN	O	O
activation-induced	NN	O	O
cell	NN	O	O
death	NN	O	O
,	NN	O	O
and	NN	O	O
impaired	NN	O	O
induction	NN	O	O
of	NN	O	O
FasL	NN	O	B-protein
.	NN	O	O

The	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
Egr2	NN	O	B-protein
and	NN	O	O
Egr3	NN	O	B-protein
are	NN	O	O
potent	NN	O	O
activators	NN	O	O
of	NN	O	O
FasL	NN	O	O
expression	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
find	NN	O	O
that	NN	O	O
Egr2	NN	O	B-DNA
and	NN	O	O
Egr3	NN	O	B-DNA
are	NN	O	O
NFAT	NN	O	B-DNA
target	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
FasL	NN	O	B-protein
occurs	NN	O	O
via	NN	O	O
the	NN	O	O
NFAT	NN	O	B-protein
-dependent	NN	O	O
induction	NN	O	O
of	NN	O	O
Egr3	NN	O	B-DNA
,	NN	O	O
as	NN	O	O
demonstrated	NN	O	O
by	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
exogenously	NN	O	O
provided	NN	O	O
NFATp	NN	O	B-protein
to	NN	O	O
restore	NN	O	O
Egr	NN	O	B-protein
-dependent	NN	O	O
FasL	NN	O	O
promoter	NN	O	O
activity	NN	O	O
in	NN	O	O
DKO	NN	O	B-cell_type
lymph	NN	O	I-cell_type
node	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Further	NN	O	O
,	NN	O	O
Egr3	NN	O	B-DNA
expression	NN	O	O
is	NN	O	O
enriched	NN	O	O
in	NN	O	O
Th1	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
suggesting	NN	O	O
a	NN	O	O
molecular	NN	O	O
basis	NN	O	O
for	NN	O	O
the	NN	O	O
known	NN	O	O
preferential	NN	O	O
expression	NN	O	O
of	NN	O	O
FasL	NN	O	B-protein
in	NN	O	O
the	NN	O	O
Th1	NN	O	O
versus	NN	O	O
Th2	NN	O	O
subset	NN	O	O
.	NN	O	O

-DOCSTART-	O

Mechanisms	NN	O	O
and	NN	O	O
clinical	NN	O	O
relevance	NN	O	O
of	NN	O	O
nongenomic	NN	O	O
glucocorticoid	NN	O	O
actions	NN	O	O
.	NN	O	O

Glucocorticoids	NN	O	O
have	NN	O	O
profound	NN	O	O
anti-inflammatory	NN	O	O
and	NN	O	O
immunosuppressive	NN	O	O
actions	NN	O	O
when	NN	O	O
used	NN	O	O
therapeutically	NN	O	O
.	NN	O	O

The	NN	O	O
therapeutic	NN	O	O
dose	NN	O	O
is	NN	O	O
quite	NN	O	O
variable	NN	O	O
and	NN	O	O
depends	NN	O	O
on	NN	O	O
the	NN	O	O
disease	NN	O	O
,	NN	O	O
but	NN	O	O
ranges	NN	O	O
from	NN	O	O
very	NN	O	O
low	NN	O	O
to	NN	O	O
extremely	NN	O	O
high	NN	O	O
.	NN	O	O

The	NN	O	O
rationale	NN	O	O
for	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
various	NN	O	O
dosage	NN	O	O
regimens	NN	O	O
for	NN	O	O
specific	NN	O	O
clinical	NN	O	O
indications	NN	O	O
is	NN	O	O
the	NN	O	O
existence	NN	O	O
of	NN	O	O
three	NN	O	O
distinct	NN	O	O
,	NN	O	O
therapeutically	NN	O	O
relevant	NN	O	O
effects	NN	O	O
:	NN	O	O
genomic	NN	O	O
,	NN	O	O
specific	NN	O	O
nongenomic	NN	O	O
and	NN	O	O
unspecific	NN	O	O
nongenomic	NN	O	O
.	NN	O	O

Genomic	NN	O	O
effects	NN	O	O
are	NN	O	O
mediated	NN	O	O
by	NN	O	O
cytosolic	NN	O	B-protein
receptors	NN	O	I-protein
that	NN	O	O
alter	NN	O	O
expression	NN	O	O
of	NN	O	O
specific	NN	O	O
genes	NN	O	O
.	NN	O	O

Specific	NN	O	O
nongenomic	NN	O	O
effects	NN	O	O
occur	NN	O	O
within	NN	O	O
a	NN	O	O
few	NN	O	O
minutes	NN	O	O
and	NN	O	O
are	NN	O	O
mediated	NN	O	O
by	NN	O	O
steroid-selective	NN	O	B-protein
membrane	NN	O	I-protein
receptors	NN	O	I-protein
.	NN	O	O

Unspecific	NN	O	O
nongenomic	NN	O	O
effects	NN	O	O
occur	NN	O	O
within	NN	O	O
seconds	NN	O	O
,	NN	O	O
but	NN	O	O
only	NN	O	O
at	NN	O	O
high	NN	O	O
glucocorticoid	NN	O	O
dosages	NN	O	O
,	NN	O	O
and	NN	O	O
seem	NN	O	O
to	NN	O	O
result	NN	O	O
from	NN	O	O
direct	NN	O	O
interactions	NN	O	O
with	NN	O	O
biological	NN	O	O
membranes	NN	O	O
.	NN	O	O

For	NN	O	O
unspecific	NN	O	O
nongenomic	NN	O	O
effects	NN	O	O
,	NN	O	O
methylprednisolone	NN	O	O
and	NN	O	O
other	NN	O	O
glucocorticoids	NN	O	O
have	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
inhibit	NN	O	O
cation	NN	O	O
cycling	NN	O	O
across	NN	O	O
the	NN	O	O
plasma	NN	O	O
membrane	NN	O	O
,	NN	O	O
but	NN	O	O
to	NN	O	O
have	NN	O	O
little	NN	O	O
effect	NN	O	O
on	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
glucocorticoids	NN	O	O
could	NN	O	O
diminish	NN	O	O
or	NN	O	O
prevent	NN	O	O
the	NN	O	O
acute	NN	O	O
immune	NN	O	O
response	NN	O	O
by	NN	O	O
interfering	NN	O	O
with	NN	O	O
processes	NN	O	O
such	NN	O	O
as	NN	O	O
the	NN	O	O
rise	NN	O	O
in	NN	O	O
intracellular	NN	O	O
Ca2+	NN	O	O
concentration	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
proposed	NN	O	O
that	NN	O	O
the	NN	O	O
additional	NN	O	O
therapeutic	NN	O	O
benefit	NN	O	O
of	NN	O	O
higher	NN	O	O
doses	NN	O	O
is	NN	O	O
obtained	NN	O	O
via	NN	O	O
these	NN	O	O
nongenomic	NN	O	O
effects	NN	O	O
.	NN	O	O

-DOCSTART-	O

High	NN	O	O
glucose-induced	NN	O	O
intercellular	NN	O	B-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
(	NN	O	O
ICAM-1	NN	O	B-protein
)	NN	O	O
expression	NN	O	O
through	NN	O	O
an	NN	O	O
osmotic	NN	O	O
effect	NN	O	O
in	NN	O	O
rat	NN	O	B-cell_type
mesangial	NN	O	I-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
PKC-NF-kappa	NN	O	O
B-dependent	NN	O	O
.	NN	O	O

AIMS/HYPOTHESIS	NN	O	O
:	NN	O	O
Infiltration	NN	O	O
of	NN	O	O
mononuclear	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
glomerular	NN	O	O
enlargement	NN	O	O
accompanied	NN	O	O
by	NN	O	O
glomerular	NN	O	B-cell_type
cell	NN	O	I-cell_type
proliferation	NN	O	O
are	NN	O	O
very	NN	O	O
early	NN	O	O
characteristics	NN	O	O
of	NN	O	O
the	NN	O	O
pathophysiology	NN	O	O
of	NN	O	O
diabetes	NN	O	O
.	NN	O	O

To	NN	O	O
clarify	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
early	NN	O	O
diabetic	NN	O	O
nephropathy	NN	O	O
,	NN	O	O
we	NN	O	O
measured	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
-thymidine	NN	O	O
incorporation	NN	O	O
and	NN	O	O
cell	NN	O	O
numbers	NN	O	O
to	NN	O	O
show	NN	O	O
the	NN	O	O
influence	NN	O	O
of	NN	O	O
a	NN	O	O
high	NN	O	O
ambient	NN	O	O
glucose	NN	O	O
concentration	NN	O	O
and	NN	O	O
the	NN	O	O
osmotic	NN	O	O
effect	NN	O	O
on	NN	O	O
rat	NN	O	B-cell_type
mesangial	NN	O	I-cell_type
cell	NN	O	I-cell_type
proliferation	NN	O	O
.	NN	O	O

We	NN	O	O
also	NN	O	O
measured	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
high	NN	O	O
glucose	NN	O	O
on	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
intercellular	NN	O	B-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
and	NN	O	O
vascular	NN	O	B-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
by	NN	O	O
flow	NN	O	O
cytometry	NN	O	O
and	NN	O	O
semiquantitative	NN	O	O
RT-PCR	NN	O	O
in	NN	O	O
mesangial	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
the	NN	O	O
adhesion	NN	O	O
of	NN	O	O
leukocytes	NN	O	B-cell_type
to	NN	O	O
mesangial	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

METHODS/RESULTS	NN	O	O
:	NN	O	O
Cells	NN	O	O
exposed	NN	O	O
to	NN	O	O
high	NN	O	O
D-glucose	NN	O	O
(	NN	O	O
30	NN	O	O
mmol/l	NN	O	O
)	NN	O	O
caused	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
-thymidine	NN	O	O
incorporation	NN	O	O
and	NN	O	O
cell	NN	O	O
numbers	NN	O	O
at	NN	O	O
24	NN	O	O
and	NN	O	O
48	NN	O	O
h	NN	O	O
and	NN	O	O
normalized	NN	O	O
at	NN	O	O
72	NN	O	O
h	NN	O	O
(	NN	O	O
p	NN	O	O
<	NN	O	O
0.05	NN	O	O
)	NN	O	O
,	NN	O	O
whereas	NN	O	O
these	NN	O	O
changes	NN	O	O
were	NN	O	O
not	NN	O	O
found	NN	O	O
in	NN	O	O
high	NN	O	O
mannitol	NN	O	O
(	NN	O	O
30	NN	O	O
mmol/l	NN	O	O
)	NN	O	O
,	NN	O	O
IL-1	NN	O	B-protein
beta	NN	O	I-protein
,	NN	O	O
or	NN	O	O
TNF	NN	O	B-protein
alpha	NN	O	I-protein
-stimulated	NN	O	O
mesangial	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Cells	NN	O	O
exposed	NN	O	O
to	NN	O	O
high-glucose	NN	O	O
(	NN	O	O
15	NN	O	O
,	NN	O	O
30	NN	O	O
,	NN	O	O
or	NN	O	O
60	NN	O	O
mmol/l	NN	O	O
)	NN	O	O
or	NN	O	O
osmotic	NN	O	O
agents	NN	O	O
(	NN	O	O
L-glucose	NN	O	O
,	NN	O	O
raffinose	NN	O	O
and	NN	O	O
mannitol	NN	O	O
)	NN	O	O
showed	NN	O	O
that	NN	O	O
intercellular	NN	O	B-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
expression	NN	O	O
began	NN	O	O
to	NN	O	O
increase	NN	O	O
after	NN	O	O
24	NN	O	O
h	NN	O	O
,	NN	O	O
reached	NN	O	O
its	NN	O	O
maximum	NN	O	O
at	NN	O	O
24	NN	O	O
and	NN	O	O
48	NN	O	O
h	NN	O	O
and	NN	O	O
gradually	NN	O	O
decreased	NN	O	O
afterwards	NN	O	O
.	NN	O	O

The	NN	O	O
stimulatory	NN	O	O
effects	NN	O	O
of	NN	O	O
high	NN	O	O
glucose	NN	O	O
and	NN	O	O
high	NN	O	O
mannitol	NN	O	O
on	NN	O	O
mRNA	NN	O	O
expression	NN	O	O
were	NN	O	O
observed	NN	O	O
as	NN	O	O
early	NN	O	O
as	NN	O	O
6	NN	O	O
h	NN	O	O
and	NN	O	O
reached	NN	O	O
its	NN	O	O
maximum	NN	O	O
at	NN	O	O
12	NN	O	O
h	NN	O	O
.	NN	O	O

Up-regulation	NN	O	O
of	NN	O	O
ICAM-1	NN	O	B-protein
protein	NN	O	O
and	NN	O	O
mRNA	NN	O	O
was	NN	O	O
also	NN	O	O
found	NN	O	O
in	NN	O	O
IL-1-beta	NN	O	B-protein
and	NN	O	O
TNF-alpha	NN	O	B-protein
-stimulated	NN	O	O
mesangial	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Neither	NN	O	O
vascular	NN	O	B-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
protein	NN	O	O
nor	NN	O	O
mRNA	NN	O	O
expression	NN	O	O
was	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
affected	NN	O	O
by	NN	O	O
high	NN	O	O
glucose	NN	O	O
and	NN	O	O
high	NN	O	O
mannitol	NN	O	O
.	NN	O	O

Notably	NN	O	O
,	NN	O	O
the	NN	O	O
protein	NN	O	O
kinase	NN	O	O
C	NN	O	O
inhibitors	NN	O	O
calphostin	NN	O	O
C	NN	O	O
and	NN	O	O
staurosporine	NN	O	O
reduced	NN	O	O
high	NN	O	O
glucose-	NN	O	O
or	NN	O	O
high	NN	O	O
mannitol-induced	NN	O	O
intercellular	NN	O	B-RNA
adhesion	NN	O	I-RNA
molecule-1	NN	O	I-RNA
mRNA	NN	O	O
expression	NN	O	O
and	NN	O	O
high	NN	O	O
glucose-induced	NN	O	O
proliferation	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
the	NN	O	O
NF-kappa	NN	O	O
B	NN	O	O
inhibitor	NN	O	O
N-tosyl-L-phenylalanine	NN	O	O
chloromethyl	NN	O	O
ketone	NN	O	O
reduced	NN	O	O
high	NN	O	O
glucose-	NN	O	O
or	NN	O	O
high	NN	O	O
mannitol-induced	NN	O	O
intercellular	NN	O	B-RNA
adhesion	NN	O	I-RNA
molecule-1	NN	O	I-RNA
mRNA	NN	O	O
expression	NN	O	O
and	NN	O	O
high	NN	O	O
glucose-induced	NN	O	O
proliferation	NN	O	O
.	NN	O	O

Results	NN	O	O
showed	NN	O	O
that	NN	O	O
high	NN	O	O
glucose	NN	O	O
(	NN	O	O
15	NN	O	O
,	NN	O	O
30	NN	O	O
mmol/l	NN	O	O
)	NN	O	O
or	NN	O	O
high	NN	O	O
concentrations	NN	O	O
of	NN	O	O
osmotic	NN	O	O
agents	NN	O	O
remarkably	NN	O	O
increased	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
adherent	NN	O	O
leukocytes	NN	O	B-cell_type
to	NN	O	O
mesangial	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
p	NN	O	O
<	NN	O	O
0.01	NN	O	O
)	NN	O	O
compared	NN	O	O
with	NN	O	O
control	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
5	NN	O	O
mmol/l	NN	O	O
D-glucose	NN	O	O
)	NN	O	O
.	NN	O	O

Functional	NN	O	O
blocking	NN	O	O
of	NN	O	O
intercellular	NN	O	B-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
on	NN	O	O
mesangial	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
rat	NN	O	B-protein
intercellular	NN	O	I-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
monoclonal	NN	O	I-protein
antibody	NN	O	I-protein
,	NN	O	O
calphostin	NN	O	O
C	NN	O	O
,	NN	O	O
staurosporine	NN	O	O
,	NN	O	O
or	NN	O	O
N-tosyl-L-phenylalanine	NN	O	O
chloromethyl	NN	O	O
ketone	NN	O	O
significantly	NN	O	O
inhibited	NN	O	O
high	NN	O	O
glucose-	NN	O	O
or	NN	O	O
high	NN	O	O
mannitol-induced	NN	O	O
increase	NN	O	O
in	NN	O	O
leukocyte	NN	O	O
adhesion	NN	O	O
(	NN	O	O
p	NN	O	O
<	NN	O	O
<	NN	O	O
0.05	NN	O	O
)	NN	O	O
.	NN	O	O

CONCLUSION/INTERPRETATION	NN	O	O
:	NN	O	O
These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
high	NN	O	O
glucose	NN	O	O
can	NN	O	O
upregulate	NN	O	O
intercellular	NN	O	B-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
protein	NN	O	O
and	NN	O	O
mRNA	NN	O	O
expression	NN	O	O
but	NN	O	O
not	NN	O	O
vascular	NN	O	O
adhesion	NN	O	O
molecule-1	NN	O	O
expression	NN	O	O
in	NN	O	O
mesangial	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
promote	NN	O	O
leukocyte	NN	O	O
adhesion	NN	O	O
through	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
intercellular	NN	O	B-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
through	NN	O	O
osmotic	NN	O	O
effect	NN	O	O
,	NN	O	O
possibly	NN	O	O
depending	NN	O	O
on	NN	O	O
the	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
PKC	NN	O	B-protein
-	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
pathway	NN	O	O
.	NN	O	O

High	NN	O	O
glucose	NN	O	O
itself	NN	O	O
can	NN	O	O
also	NN	O	O
promote	NN	O	O
mesangial	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
through	NN	O	O
the	NN	O	O
PKC-NF-kappa	NN	O	O
B	NN	O	O
pathways	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
hyperglycaemia	NN	O	O
in	NN	O	O
itself	NN	O	O
seems	NN	O	O
to	NN	O	O
be	NN	O	O
an	NN	O	O
important	NN	O	O
factor	NN	O	O
in	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
early	NN	O	O
diabetic	NN	O	O
nephropathy	NN	O	O
.	NN	O	O

-DOCSTART-	O

Combined	NN	O	O
corticosteroid/	NN	O	O
granulocyte	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
G-CSF	NN	O	B-protein
)	NN	O	O
therapy	NN	O	O
in	NN	O	O
the	NN	O	O
treatment	NN	O	O
of	NN	O	O
severe	NN	O	O
congenital	NN	O	O
neutropenia	NN	O	O
unresponsive	NN	O	O
to	NN	O	O
G-CSF	NN	O	B-protein
:	NN	O	O
Activated	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
synergize	NN	O	O
with	NN	O	O
G-CSF	NN	O	B-protein
signals	NN	O	O
.	NN	O	O

More	NN	O	O
than	NN	O	O
90	NN	O	O
%	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
severe	NN	O	O
congenital	NN	O	O
neutropenia	NN	O	O
(	NN	O	O
SCN	NN	O	O
)	NN	O	O
respond	NN	O	O
to	NN	O	O
granulocyte	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
G-CSF	NN	O	B-protein
)	NN	O	O
therapy	NN	O	O
.	NN	O	O

The	NN	O	O
basis	NN	O	O
for	NN	O	O
the	NN	O	O
refractory	NN	O	O
state	NN	O	O
in	NN	O	O
the	NN	O	O
remaining	NN	O	O
patients	NN	O	O
is	NN	O	O
unknown	NN	O	O
.	NN	O	O

To	NN	O	O
address	NN	O	O
this	NN	O	O
issue	NN	O	O
,	NN	O	O
we	NN	O	O
studied	NN	O	O
a	NN	O	O
child	NN	O	O
with	NN	O	O
SCN	NN	O	O
who	NN	O	O
was	NN	O	O
totally	NN	O	O
unresponsive	NN	O	O
to	NN	O	O
G-CSF	NN	O	B-protein
and	NN	O	O
had	NN	O	O
a	NN	O	O
novel	NN	O	O
point	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
extracellular	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
the	NN	O	O
G-CSF	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
GCSF-R	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Marrow	NN	O	O
stromal	NN	O	O
support	NN	O	O
of	NN	O	O
granulopoiesis	NN	O	O
was	NN	O	O
evaluated	NN	O	O
by	NN	O	O
plating	NN	O	O
CD34	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
on	NN	O	O
preformed	NN	O	O
stromal	NN	O	O
layers	NN	O	O
.	NN	O	O

Nonadherent	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
harvested	NN	O	O
and	NN	O	O
assayed	NN	O	O
in	NN	O	O
clonogenic	NN	O	O
assays	NN	O	O
for	NN	O	O
granulocytic	NN	O	O
colony	NN	O	O
production	NN	O	O
.	NN	O	O

The	NN	O	O
in	NN	O	O
vitro	NN	O	O
effect	NN	O	O
of	NN	O	O
G-CSF	NN	O	B-protein
and	NN	O	O
corticosteroids	NN	O	O
on	NN	O	O
granulopoiesis	NN	O	O
was	NN	O	O
evaluated	NN	O	O
in	NN	O	O
clonogenic	NN	O	O
assays	NN	O	O
of	NN	O	O
marrow	NN	O	B-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
by	NN	O	O
proliferation	NN	O	O
studies	NN	O	O
of	NN	O	O
the	NN	O	O
murine	NN	O	B-cell_line
myeloid	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
32D	NN	O	I-cell_line
expressing	NN	O	O
the	NN	O	O
patient	NN	O	O
's	NN	O	O
mutated	NN	O	B-protein
G-CSFR	NN	O	I-protein
,	NN	O	O
and	NN	O	O
by	NN	O	O
measuring	NN	O	O
STAT5	NN	O	O
activation	NN	O	O
in	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
from	NN	O	O
stimulated	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Patient	NN	O	O
's	NN	O	O
stroma	NN	O	O
supported	NN	O	O
granulopoiesis	NN	O	O
derived	NN	O	O
from	NN	O	O
control	NN	O	O
marrow	NN	O	B-cell_type
CD34	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
a	NN	O	O
normal	NN	O	O
manner	NN	O	O
.	NN	O	O

Normal	NN	O	O
stroma	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
failed	NN	O	O
to	NN	O	O
induce	NN	O	O
granulopoiesis	NN	O	O
from	NN	O	O
patient	NN	O	O
's	NN	O	O
CD34	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Clonogenic	NN	O	O
assays	NN	O	O
of	NN	O	O
the	NN	O	O
patient	NN	O	O
's	NN	O	O
marrow	NN	O	B-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
incorporating	NN	O	O
either	NN	O	O
G-CSF	NN	O	B-protein
or	NN	O	O
hydrocortisone	NN	O	O
produced	NN	O	O
little	NN	O	O
neutrophil	NN	O	O
growth	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
inclusion	NN	O	O
of	NN	O	O
both	NN	O	O
G-CSF	NN	O	B-protein
and	NN	O	O
hydrocortisone	NN	O	O
in	NN	O	O
the	NN	O	O
cytokine	NN	O	B-protein
``	NN	O	I-protein
cocktail	NN	O	I-protein
''	NN	O	I-protein
markedly	NN	O	O
increased	NN	O	O
the	NN	O	O
neutrophil	NN	O	O
numbers	NN	O	O
.	NN	O	O

Proliferation	NN	O	O
of	NN	O	O
32D	NN	O	B-cell_line
cells	NN	O	I-cell_line
expressing	NN	O	O
the	NN	O	O
mutated	NN	O	B-protein
receptor	NN	O	I-protein
and	NN	O	O
STAT5	NN	O	O
activation	NN	O	O
were	NN	O	O
improved	NN	O	O
by	NN	O	O
a	NN	O	O
combination	NN	O	O
of	NN	O	O
G-CSF	NN	O	B-protein
and	NN	O	O
dexamethasone	NN	O	O
.	NN	O	O

When	NN	O	O
small	NN	O	O
daily	NN	O	O
doses	NN	O	O
of	NN	O	O
oral	NN	O	O
prednisone	NN	O	O
were	NN	O	O
then	NN	O	O
administered	NN	O	O
to	NN	O	O
the	NN	O	O
patient	NN	O	O
with	NN	O	O
conventional	NN	O	O
doses	NN	O	O
of	NN	O	O
subcutaneous	NN	O	B-protein
G-CSF	NN	O	I-protein
,	NN	O	O
the	NN	O	O
patient	NN	O	O
responded	NN	O	O
with	NN	O	O
increased	NN	O	O
neutrophil	NN	O	O
numbers	NN	O	O
and	NN	O	O
with	NN	O	O
a	NN	O	O
complete	NN	O	O
reversal	NN	O	O
of	NN	O	O
the	NN	O	O
infectious	NN	O	O
problems	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
provide	NN	O	O
insight	NN	O	O
into	NN	O	O
SCN	NN	O	O
unresponsive	NN	O	O
to	NN	O	O
standard	NN	O	O
G-CSF	NN	O	B-protein
treatment	NN	O	O
and	NN	O	O
to	NN	O	O
the	NN	O	O
potential	NN	O	O
corrective	NN	O	O
action	NN	O	O
of	NN	O	O
combined	NN	O	O
treatment	NN	O	O
with	NN	O	O
G-CSF	NN	O	B-protein
and	NN	O	O
corticosteroids	NN	O	O
through	NN	O	O
synergistic	NN	O	O
activation	NN	O	O
of	NN	O	O
STAT5	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Human	NN	O	B-cell_type
renal	NN	O	I-cell_type
mesangial	NN	O	I-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
a	NN	O	O
target	NN	O	O
for	NN	O	O
the	NN	O	O
anti-inflammatory	NN	O	O
action	NN	O	O
of	NN	O	O
9-cis	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
.	NN	O	O

Mesangial	NN	O	B-cell_type
cells	NN	O	I-cell_type
play	NN	O	O
an	NN	O	O
active	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
inflammatory	NN	O	O
response	NN	O	O
to	NN	O	O
glomerular	NN	O	O
injury	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
studied	NN	O	O
in	NN	O	O
cultured	NN	O	B-cell_line
human	NN	O	I-cell_line
mesangial	NN	O	I-cell_line
cells	NN	O	I-cell_line
(	NN	O	O
CHMC	NN	O	B-cell_line
)	NN	O	O
several	NN	O	O
effects	NN	O	O
of	NN	O	O
9-cis	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
9-cRA	NN	O	O
)	NN	O	O
,	NN	O	O
an	NN	O	O
activator	NN	O	O
of	NN	O	O
both	NN	O	O
retinoic	NN	O	B-protein
acid	NN	O	I-protein
receptors	NN	O	I-protein
(	NN	O	O
RARs	NN	O	B-protein
)	NN	O	O
and	NN	O	O
retinoid	NN	O	B-protein
X	NN	O	I-protein
receptors	NN	O	I-protein
(	NN	O	O
RXRs	NN	O	B-protein
)	NN	O	O
.	NN	O	O

9-cRA	NN	O	O
inhibited	NN	O	O
foetal	NN	O	O
calf	NN	O	O
serum-induced	NN	O	O
CHMC	NN	O	B-cell_line
proliferation	NN	O	O
.	NN	O	O

It	NN	O	O
also	NN	O	O
prevented	NN	O	O
CHMC	NN	O	B-cell_line
death	NN	O	O
induced	NN	O	O
by	NN	O	O
the	NN	O	O
inflammatory	NN	O	O
mediator	NN	O	O
H	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
O	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
.	NN	O	O

This	NN	O	O
preventive	NN	O	O
effect	NN	O	O
was	NN	O	O
not	NN	O	O
due	NN	O	O
to	NN	O	O
any	NN	O	O
increase	NN	O	O
in	NN	O	O
H	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
O	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
catabolism	NN	O	O
and	NN	O	O
it	NN	O	O
persisted	NN	O	O
even	NN	O	O
when	NN	O	O
both	NN	O	O
catalase	NN	O	O
and	NN	O	O
glutathione	NN	O	O
synthesis	NN	O	O
were	NN	O	O
inhibited	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
9-cRA	NN	O	O
diminished	NN	O	O
monocyte	NN	O	O
adhesion	NN	O	O
to	NN	O	O
FCS-stimulated	NN	O	B-cell_line
CHMC	NN	O	I-cell_line
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
the	NN	O	O
retinoid	NN	O	O
also	NN	O	O
inhibited	NN	O	O
in	NN	O	O
FCS-stimulated	NN	O	B-cell_type
cells	NN	O	I-cell_type
the	NN	O	O
protein	NN	O	O
expression	NN	O	O
of	NN	O	O
two	NN	O	O
mesangial	NN	O	B-protein
adhesion	NN	O	I-protein
molecules	NN	O	I-protein
,	NN	O	O
fibronectin	NN	O	B-protein
and	NN	O	O
osteopontin	NN	O	B-protein
,	NN	O	O
but	NN	O	O
it	NN	O	O
did	NN	O	O
not	NN	O	O
modify	NN	O	O
the	NN	O	O
protein	NN	O	O
expression	NN	O	O
of	NN	O	O
intercellular	NN	O	B-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
and	NN	O	O
vascular	NN	O	B-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
.	NN	O	O

All	NN	O	O
major	NN	O	O
RARs	NN	O	B-protein
and	NN	O	I-protein
RXRs	NN	O	I-protein
isotypes	NN	O	I-protein
were	NN	O	O
expressed	NN	O	O
in	NN	O	O
CHMC	NN	O	B-cell_line
regardless	NN	O	O
of	NN	O	O
the	NN	O	O
presence	NN	O	O
or	NN	O	O
absence	NN	O	O
of	NN	O	O
9-cRA	NN	O	O
.	NN	O	O

Transcripts	NN	O	O
to	NN	O	O
RAR-alpha	NN	O	B-RNA
,	NN	O	O
RAR-beta	NN	O	B-RNA
and	NN	O	O
RXR-alpha	NN	O	B-RNA
increased	NN	O	O
after	NN	O	O
incubation	NN	O	O
with	NN	O	O
9-cRA	NN	O	O
whereas	NN	O	O
RXR-gamma	NN	O	B-RNA
was	NN	O	O
inhibited	NN	O	O
,	NN	O	O
suggesting	NN	O	O
a	NN	O	O
major	NN	O	O
role	NN	O	O
for	NN	O	O
RARs	NN	O	B-protein
and	NN	O	O
RXRs	NN	O	B-protein
in	NN	O	O
9-cRA-anti-inflammatory	NN	O	O
effects	NN	O	O
.	NN	O	O

9-cRA	NN	O	O
was	NN	O	O
toxic	NN	O	O
only	NN	O	O
at	NN	O	O
50	NN	O	O
microM	NN	O	O
(	NN	O	O
a	NN	O	O
concentration	NN	O	O
50	NN	O	O
-	NN	O	O
5000	NN	O	O
times	NN	O	O
higher	NN	O	O
than	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
effects	NN	O	O
above	NN	O	O
)	NN	O	O
.	NN	O	O

Cell	NN	O	O
death	NN	O	O
occurred	NN	O	O
by	NN	O	O
apoptosis	NN	O	O
,	NN	O	O
whose	NN	O	O
onset	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
pronounced	NN	O	O
increase	NN	O	O
in	NN	O	O
catalase	NN	O	O
activity	NN	O	O
and	NN	O	O
reduced	NN	O	O
glutathione	NN	O	O
content	NN	O	O
,	NN	O	O
being	NN	O	O
more	NN	O	O
effectively	NN	O	O
induced	NN	O	O
by	NN	O	O
all-trans	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
.	NN	O	O

Modulation	NN	O	O
of	NN	O	O
the	NN	O	O
oxidant/antioxidant	NN	O	O
balance	NN	O	O
failed	NN	O	O
to	NN	O	O
inhibit	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
mesangial	NN	O	B-cell_type
cells	NN	O	I-cell_type
might	NN	O	O
be	NN	O	O
a	NN	O	O
target	NN	O	O
for	NN	O	O
the	NN	O	O
treatment	NN	O	O
of	NN	O	O
inflammatory	NN	O	O
glomerulopathies	NN	O	O
with	NN	O	O
9-cRA	NN	O	O
.	NN	O	O

-DOCSTART-	O

Expression	NN	O	O
of	NN	O	O
mammalian	NN	O	B-DNA
defensin	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

Antimicrobial	NN	O	O
peptides	NN	O	O
are	NN	O	O
a	NN	O	O
prevalent	NN	O	O
mechanism	NN	O	O
of	NN	O	O
host	NN	O	O
defense	NN	O	O
found	NN	O	O
throughout	NN	O	O
nature	NN	O	O
.	NN	O	O

In	NN	O	O
mammals	NN	O	O
,	NN	O	O
defensins	NN	O	B-protein
are	NN	O	O
among	NN	O	O
the	NN	O	O
most	NN	O	O
abundant	NN	O	O
of	NN	O	O
these	NN	O	O
broad-spectrum	NN	O	O
antibiotics	NN	O	O
,	NN	O	O
and	NN	O	O
are	NN	O	O
expressed	NN	O	O
in	NN	O	O
epithelial	NN	O	B-cell_type
and	NN	O	I-cell_type
hematopoietic	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
defensin	NN	O	O
peptides	NN	O	O
are	NN	O	O
especially	NN	O	O
abundant	NN	O	O
in	NN	O	O
neutrophils	NN	O	B-cell_type
;	NN	O	O
however	NN	O	O
,	NN	O	O
gene	NN	O	O
expression	NN	O	O
is	NN	O	O
limited	NN	O	O
to	NN	O	O
the	NN	O	O
promyelocyte	NN	O	O
stage	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
defensin	NN	O	B-DNA
genes	NN	O	I-DNA
are	NN	O	O
found	NN	O	O
as	NN	O	O
both	NN	O	O
constitutively	NN	O	O
expressed	NN	O	O
and	NN	O	O
inducible	NN	O	O
.	NN	O	O

Induction	NN	O	O
has	NN	O	O
been	NN	O	O
observed	NN	O	O
in	NN	O	O
vitro	NN	O	O
by	NN	O	O
stimulation	NN	O	O
with	NN	O	O
bacterial	NN	O	O
lipopolysaccharide	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
inflammatory	NN	O	O
mediators	NN	O	O
.	NN	O	O

In	NN	O	O
vivo	NN	O	O
,	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
several	NN	O	O
defensin	NN	O	B-DNA
genes	NN	O	I-DNA
occurs	NN	O	O
in	NN	O	O
both	NN	O	O
infectious	NN	O	O
and	NN	O	O
inflammatory	NN	O	O
states	NN	O	O
.	NN	O	O

Gene	NN	O	O
regulation	NN	O	O
occurs	NN	O	O
via	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathways	NN	O	O
common	NN	O	O
to	NN	O	O
other	NN	O	O
innate	NN	O	O
immune	NN	O	O
responses	NN	O	O
,	NN	O	O
utilizing	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
such	NN	O	O
as	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	I-protein
NF	NN	O	I-protein
)	NN	O	I-protein
-kappaB	NN	O	I-protein
and	NN	O	O
NF	NN	O	B-protein
interleukin-6	NN	O	I-protein
.	NN	O	O

Together	NN	O	O
,	NN	O	O
the	NN	O	O
data	NN	O	O
suggest	NN	O	O
a	NN	O	O
broad-based	NN	O	O
innate	NN	O	O
host	NN	O	O
defense	NN	O	O
whereby	NN	O	O
potent	NN	O	O
antimicrobial	NN	O	O
peptides	NN	O	O
are	NN	O	O
present	NN	O	O
to	NN	O	O
prevent	NN	O	O
initial	NN	O	O
colonization	NN	O	O
by	NN	O	O
pathogenic	NN	O	O
microorganisms	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
the	NN	O	O
recognition	NN	O	O
of	NN	O	O
bacteria	NN	O	O
coupled	NN	O	O
with	NN	O	O
a	NN	O	O
nascent	NN	O	O
inflammatory	NN	O	O
response	NN	O	O
can	NN	O	O
bolster	NN	O	O
this	NN	O	O
defense	NN	O	O
by	NN	O	O
a	NN	O	O
coordinated	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
the	NN	O	O
peptides	NN	O	O
.	NN	O	O

-DOCSTART-	O

Multiple	NN	O	O
signals	NN	O	O
required	NN	O	O
for	NN	O	O
cyclic	NN	O	B-protein
AMP-responsive	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
binding	NN	O	O
protein	NN	O	O
interaction	NN	O	O
induced	NN	O	O
by	NN	O	O
CD3/CD28	NN	O	O
costimulation	NN	O	O
.	NN	O	O

The	NN	O	O
optimal	NN	O	O
activation	NN	O	O
of	NN	O	O
cAMP-responsive	NN	O	B-protein
element	NN	O	I-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
CREB	NN	O	B-protein
)	NN	O	O
,	NN	O	O
similar	NN	O	O
to	NN	O	O
the	NN	O	O
full	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
requires	NN	O	O
the	NN	O	O
stimulation	NN	O	O
of	NN	O	O
both	NN	O	O
CD3	NN	O	B-protein
and	NN	O	O
CD28	NN	O	B-protein
.	NN	O	O

Using	NN	O	O
a	NN	O	O
reporter	NN	O	O
system	NN	O	O
to	NN	O	O
detect	NN	O	O
interaction	NN	O	O
of	NN	O	O
CREB	NN	O	B-protein
and	NN	O	O
CREB-binding	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	O
CBP	NN	O	B-protein
)	NN	O	O
,	NN	O	O
in	NN	O	O
this	NN	O	O
study	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
CREB	NN	O	B-protein
binds	NN	O	O
to	NN	O	O
CBP	NN	O	B-protein
only	NN	O	O
by	NN	O	O
engagement	NN	O	O
of	NN	O	O
both	NN	O	O
CD3	NN	O	B-protein
and	NN	O	O
CD28	NN	O	B-protein
.	NN	O	O

CD3/CD28	NN	O	B-protein
-promoted	NN	O	O
CREB	NN	O	B-protein
-	NN	O	O
CBP	NN	O	B-protein
interaction	NN	O	O
was	NN	O	O
dependent	NN	O	O
on	NN	O	O
p38	NN	O	B-protein
mitogen-activated	NN	O	I-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	O
MAPK	NN	O	B-protein
)	NN	O	O
and	NN	O	O
calcium/calmodulin-dependent	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	I-protein
CaMK	NN	O	I-protein
)	NN	O	I-protein
IV	NN	O	I-protein
in	NN	O	O
addition	NN	O	O
to	NN	O	O
the	NN	O	O
previously	NN	O	O
identified	NN	O	O
extracellular	NN	O	O
signal-regulated	NN	O	O
kinase	NN	O	O
pathway	NN	O	O
.	NN	O	O

Extracellular	NN	O	B-protein
signal-regulated	NN	O	I-protein
kinase	NN	O	I-protein
,	NN	O	I-protein
CaMKIV	NN	O	I-protein
,	NN	O	O
and	NN	O	O
p38	NN	O	B-protein
MAPK	NN	O	I-protein
were	NN	O	O
also	NN	O	O
the	NN	O	O
kinases	NN	O	O
involved	NN	O	O
in	NN	O	O
CREB	NN	O	B-protein
Ser	NN	O	O
(	NN	O	O
133	NN	O	O
)	NN	O	O
phosphorylation	NN	O	O
induced	NN	O	O
by	NN	O	O
CD3/CD28	NN	O	B-protein
.	NN	O	O

A	NN	O	O
reconstitution	NN	O	O
experiment	NN	O	O
illustrated	NN	O	O
that	NN	O	O
optimum	NN	O	O
CREB	NN	O	B-protein
-	NN	O	O
CBP	NN	O	B-protein
interaction	NN	O	O
and	NN	O	O
CREB	NN	O	B-protein
trans-activation	NN	O	O
were	NN	O	O
attained	NN	O	O
when	NN	O	O
these	NN	O	O
three	NN	O	O
kinase	NN	O	O
pathways	NN	O	O
were	NN	O	O
simultaneously	NN	O	O
activated	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Our	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
coordinated	NN	O	O
activation	NN	O	O
of	NN	O	O
different	NN	O	O
kinases	NN	O	B-protein
leads	NN	O	O
to	NN	O	O
full	NN	O	O
activation	NN	O	O
of	NN	O	O
CREB	NN	O	B-protein
.	NN	O	O

Notably	NN	O	O
,	NN	O	O
CD28	NN	O	O
ligation	NN	O	O
activated	NN	O	O
p38	NN	O	B-protein
MAPK	NN	O	I-protein
and	NN	O	O
CaMKIV	NN	O	B-protein
,	NN	O	O
the	NN	O	O
kinases	NN	O	B-protein
stimulated	NN	O	O
by	NN	O	O
CD3	NN	O	O
engagement	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
CD28	NN	O	B-protein
acts	NN	O	O
by	NN	O	O
increasing	NN	O	O
the	NN	O	O
activation	NN	O	O
extent	NN	O	O
of	NN	O	O
p38	NN	O	B-protein
MAPK	NN	O	I-protein
and	NN	O	O
CaMKIV	NN	O	B-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
support	NN	O	O
the	NN	O	O
model	NN	O	O
of	NN	O	O
a	NN	O	O
minimum	NN	O	O
activation	NN	O	O
threshold	NN	O	O
for	NN	O	O
CREB	NN	O	B-protein
-	NN	O	O
CBP	NN	O	B-protein
interaction	NN	O	O
that	NN	O	O
can	NN	O	O
be	NN	O	O
reached	NN	O	O
only	NN	O	O
when	NN	O	O
both	NN	O	O
CD3	NN	O	B-protein
and	NN	O	O
CD28	NN	O	B-protein
are	NN	O	O
stimulated	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
murine	NN	O	B-DNA
IL-2	NN	O	I-DNA
promoter	NN	O	I-DNA
contains	NN	O	O
distal	NN	O	B-DNA
regulatory	NN	O	I-DNA
elements	NN	O	I-DNA
responsive	NN	O	O
to	NN	O	O
the	NN	O	O
Ah	NN	O	B-protein
receptor	NN	O	I-protein
,	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
evolutionarily	NN	O	O
conserved	NN	O	O
bHLH-PAS	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
family	NN	O	I-protein
.	NN	O	O

Signaling	NN	O	O
through	NN	O	O
the	NN	O	O
TCR	NN	O	B-protein
and	NN	O	O
costimulatory	NN	O	O
signals	NN	O	O
primarily	NN	O	O
control	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
naive	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
minimal	NN	O	B-DNA
promoter	NN	O	I-DNA
necessary	NN	O	O
for	NN	O	O
this	NN	O	O
expression	NN	O	O
lies	NN	O	O
proximal	NN	O	O
,	NN	O	O
between	NN	O	O
-300	NN	O	O
and	NN	O	O
the	NN	O	O
transcription	NN	O	B-DNA
start	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
had	NN	O	O
previously	NN	O	O
shown	NN	O	O
that	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
arylhydrocarbon	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
AHR	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
bHLH-PAS	NN	O	B-protein
family	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
,	NN	O	O
leads	NN	O	O
to	NN	O	O
increased	NN	O	O
mRNA	NN	O	O
expression	NN	O	O
of	NN	O	O
IL-2	NN	O	B-RNA
in	NN	O	O
murine	NN	O	B-cell_type
fetal	NN	O	I-cell_type
thymocytes	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
AHR	NN	O	B-protein
is	NN	O	O
abundant	NN	O	O
in	NN	O	O
the	NN	O	O
thymus	NN	O	O
and	NN	O	O
may	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
its	NN	O	O
overactivation	NN	O	O
by	NN	O	O
chemicals	NN	O	O
such	NN	O	O
as	NN	O	O
dioxins	NN	O	O
leads	NN	O	O
to	NN	O	O
immunosuppression	NN	O	O
and	NN	O	O
thymic	NN	O	O
involution	NN	O	O
.	NN	O	O

Binding	NN	O	O
motifs	NN	O	O
for	NN	O	O
the	NN	O	O
liganded	NN	O	B-protein
AHR	NN	O	I-protein
can	NN	O	O
be	NN	O	O
identified	NN	O	O
in	NN	O	O
the	NN	O	O
distal	NN	O	B-DNA
region	NN	O	I-DNA
-1300	NN	O	I-DNA
to	NN	O	I-DNA
-800	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
mouse	NN	O	B-DNA
IL-2	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
show	NN	O	O
here	NN	O	O
that	NN	O	O
these	NN	O	O
DNA	NN	O	B-DNA
motifs	NN	O	I-DNA
,	NN	O	O
the	NN	O	O
so-called	NN	O	O
dioxin	NN	O	B-DNA
response	NN	O	I-DNA
elements	NN	O	I-DNA
,	NN	O	O
after	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
liganded	NN	O	B-protein
AHR	NN	O	I-protein
are	NN	O	O
sufficient	NN	O	O
to	NN	O	O
transactivate	NN	O	O
luciferase	NN	O	O
expression	NN	O	O
in	NN	O	O
a	NN	O	O
reporter	NN	O	O
gene	NN	O	O
system	NN	O	O
.	NN	O	O

The	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
can	NN	O	O
be	NN	O	O
induced	NN	O	O
by	NN	O	O
the	NN	O	O
AHR	NN	O	B-protein
also	NN	O	O
in	NN	O	O
thymocytes	NN	O	B-cell_type
in	NN	O	O
vivo	NN	O	O
after	NN	O	O
injection	NN	O	O
of	NN	O	O
2	NN	O	O
,	NN	O	O
3	NN	O	O
,	NN	O	O
7	NN	O	O
,	NN	O	O
8-tetrachlorodibenzo-p-dioxin	NN	O	O
,	NN	O	O
a	NN	O	O
potent	NN	O	O
ligand	NN	O	O
of	NN	O	O
the	NN	O	O
AHR	NN	O	B-protein
.	NN	O	O

The	NN	O	O
AHR	NN	O	B-protein
mediates	NN	O	O
the	NN	O	O
IL-2	NN	O	O
induction	NN	O	O
as	NN	O	O
shown	NN	O	O
with	NN	O	O
AHR	NN	O	B-protein
-deficient	NN	O	O
mice	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
in	NN	O	O
spleen	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
vitro	NN	O	O
costimulation	NN	O	O
via	NN	O	O
the	NN	O	O
TCR	NN	O	B-protein
is	NN	O	O
necessary	NN	O	O
for	NN	O	O
optimal	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
induction	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
region	NN	O	I-DNA
contains	NN	O	O
novel	NN	O	O
distal	NN	O	B-DNA
regulatory	NN	O	I-DNA
elements	NN	O	I-DNA
that	NN	O	O
can	NN	O	O
be	NN	O	O
addressed	NN	O	O
by	NN	O	O
the	NN	O	O
AHR	NN	O	B-protein
to	NN	O	O
induce	NN	O	O
IL-2	NN	O	B-DNA
and	NN	O	O
can	NN	O	O
cooperate	NN	O	O
with	NN	O	O
the	NN	O	O
proximal	NN	O	B-DNA
promoter	NN	O	I-DNA

-DOCSTART-	O

Visualization	NN	O	O
of	NN	O	O
Syk-antigen	NN	O	B-protein
receptor	NN	O	I-protein
interactions	NN	O	O
using	NN	O	O
green	NN	O	B-protein
fluorescent	NN	O	I-protein
protein	NN	O	I-protein
:	NN	O	O
differential	NN	O	O
roles	NN	O	O
for	NN	O	O
Syk	NN	O	B-protein
and	NN	O	O
Lyn	NN	O	B-protein
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
receptor	NN	O	O
capping	NN	O	O
and	NN	O	O
internalization	NN	O	O
.	NN	O	O

The	NN	O	O
cross-linking	NN	O	O
of	NN	O	O
the	NN	O	O
B	NN	O	B-protein
cell	NN	O	I-protein
Ag	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
BCR	NN	O	B-protein
)	NN	O	O
is	NN	O	O
coupled	NN	O	O
to	NN	O	O
the	NN	O	O
stimulation	NN	O	O
of	NN	O	O
multiple	NN	O	O
intracellular	NN	O	O
signal	NN	O	O
transduction	NN	O	O
cascades	NN	O	O
via	NN	O	O
receptor-associated	NN	O	O
,	NN	O	O
protein	NN	O	B-protein
tyrosine	NN	O	I-protein
kinases	NN	O	I-protein
of	NN	O	O
both	NN	O	O
the	NN	O	O
Src	NN	O	B-protein
and	NN	O	I-protein
Syk	NN	O	I-protein
families	NN	O	I-protein
.	NN	O	O

To	NN	O	O
monitor	NN	O	O
changes	NN	O	O
in	NN	O	O
the	NN	O	O
subcellular	NN	O	O
distribution	NN	O	O
of	NN	O	O
Syk	NN	O	B-protein
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
responding	NN	O	O
to	NN	O	O
BCR	NN	O	B-protein
cross-linking	NN	O	O
,	NN	O	O
we	NN	O	O
expressed	NN	O	O
in	NN	O	O
Syk	NN	O	B-protein
-deficient	NN	O	O
DT40	NN	O	B-cell_line
B	NN	O	I-cell_line
cells	NN	O	I-cell_line
a	NN	O	O
fusion	NN	O	B-protein
protein	NN	O	I-protein
consisting	NN	O	O
of	NN	O	O
Syk	NN	O	B-protein
coupled	NN	O	O
to	NN	O	O
green	NN	O	B-protein
fluorescent	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
these	NN	O	O
cells	NN	O	O
with	NN	O	O
anti-IgM	NN	O	B-protein
Abs	NN	O	I-protein
leads	NN	O	O
to	NN	O	O
the	NN	O	O
recruitment	NN	O	O
of	NN	O	O
the	NN	O	O
kinase	NN	O	B-protein
from	NN	O	O
cytoplasmic	NN	O	O
and	NN	O	O
nuclear	NN	O	O
compartments	NN	O	O
to	NN	O	O
the	NN	O	O
site	NN	O	O
of	NN	O	O
the	NN	O	O
cross-linked	NN	O	B-protein
receptor	NN	O	I-protein
at	NN	O	O
the	NN	O	O
plasma	NN	O	O
membrane	NN	O	O
.	NN	O	O

The	NN	O	O
Syk-receptor	NN	O	B-protein
complexes	NN	O	I-protein
aggregate	NN	O	O
into	NN	O	O
membrane	NN	O	O
patches	NN	O	O
that	NN	O	O
redistribute	NN	O	O
to	NN	O	O
form	NN	O	O
a	NN	O	O
cap	NN	O	O
at	NN	O	O
one	NN	O	O
pole	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	O
.	NN	O	O

Syk	NN	O	B-protein
is	NN	O	O
not	NN	O	O
demonstrably	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
internalized	NN	O	B-protein
receptor	NN	O	I-protein
.	NN	O	O

Catalytically	NN	O	O
active	NN	O	O
Syk	NN	O	B-protein
promotes	NN	O	O
and	NN	O	O
stabilizes	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
tightly	NN	O	B-protein
capped	NN	O	I-protein
BCR	NN	O	I-protein
complexes	NN	O	I-protein
at	NN	O	O
the	NN	O	O
plasma	NN	O	O
membrane	NN	O	O
.	NN	O	O

Lyn	NN	O	B-protein
is	NN	O	O
not	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
recruitment	NN	O	O
of	NN	O	O
Syk	NN	O	B-protein
to	NN	O	O
the	NN	O	O
cross-linked	NN	O	B-protein
receptor	NN	O	I-protein
,	NN	O	O
but	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
internalization	NN	O	O
of	NN	O	O
the	NN	O	O
clustered	NN	O	O
BCR	NN	O	B-protein
complexes	NN	O	I-protein
.	NN	O	O

In	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
Lyn	NN	O	B-protein
,	NN	O	O
receptor-Syk	NN	O	B-protein
complexes	NN	O	I-protein
at	NN	O	O
the	NN	O	O
plasma	NN	O	O
membrane	NN	O	O
are	NN	O	O
long	NN	O	O
lived	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
receptor-mediated	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
NF-AT	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
is	NN	O	O
enhanced	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
Lyn	NN	O	B-protein
appears	NN	O	O
to	NN	O	O
function	NN	O	O
to	NN	O	O
negatively	NN	O	O
regulate	NN	O	O
aspects	NN	O	O
of	NN	O	O
BCR	NN	O	B-protein
-dependent	NN	O	O
signaling	NN	O	O
by	NN	O	O
stimulating	NN	O	O
receptor	NN	O	O
internalization	NN	O	O
and	NN	O	O
down-regulation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Renal	NN	O	O
interstitial	NN	O	O
fibrosis	NN	O	O
is	NN	O	O
reduced	NN	O	O
in	NN	O	O
angiotensin	NN	O	B-protein
II	NN	O	I-protein
type	NN	O	I-protein
1a	NN	O	I-protein
receptor	NN	O	I-protein
-deficient	NN	O	O
mice	NN	O	O
.	NN	O	O

Unilateral	NN	O	O
ureteral	NN	O	O
obstruction	NN	O	O
(	NN	O	O
UUO	NN	O	O
)	NN	O	O
results	NN	O	O
in	NN	O	O
tubulointerstitial	NN	O	O
fibrosis	NN	O	O
of	NN	O	O
the	NN	O	O
affected	NN	O	O
kidney	NN	O	O
by	NN	O	O
stimulating	NN	O	O
the	NN	O	O
renin-angiotensin	NN	O	O
system	NN	O	O
.	NN	O	O

This	NN	O	O
study	NN	O	O
established	NN	O	O
a	NN	O	O
UUO	NN	O	O
model	NN	O	O
in	NN	O	O
angiotensin	NN	O	B-protein
type	NN	O	I-protein
1a	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
AT1a	NN	O	B-protein
)	NN	O	O
deficient	NN	O	O
(	NN	O	O
mutant	NN	O	O
)	NN	O	O
mice	NN	O	O
to	NN	O	O
elucidate	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
angiotensin	NN	O	B-protein
II	NN	O	I-protein
through	NN	O	O
AT1a	NN	O	B-protein
on	NN	O	O
the	NN	O	O
fibrosis	NN	O	O
of	NN	O	O
the	NN	O	O
obstructed	NN	O	O
kidney	NN	O	O
(	NN	O	O
OBK	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
relative	NN	O	O
volume	NN	O	O
of	NN	O	O
the	NN	O	O
tubulointerstitium	NN	O	O
was	NN	O	O
measured	NN	O	O
by	NN	O	O
an	NN	O	O
image	NN	O	O
analyzer	NN	O	O
;	NN	O	O
deposition	NN	O	O
of	NN	O	O
collagen	NN	O	B-protein
types	NN	O	I-protein
III	NN	O	I-protein
and	NN	O	I-protein
IV	NN	O	I-protein
and	NN	O	O
monocyte	NN	O	B-cell_type
/	NN	O	O
macrophage	NN	O	B-cell_type
infiltration	NN	O	O
were	NN	O	O
histologically	NN	O	O
examined	NN	O	O
using	NN	O	O
specific	NN	O	B-protein
antibodies	NN	O	I-protein
.	NN	O	O

Also	NN	O	O
determined	NN	O	O
were	NN	O	O
the	NN	O	O
mRNA	NN	O	O
levels	NN	O	O
of	NN	O	O
transforming	NN	O	B-protein
growth	NN	O	I-protein
factor-beta	NN	O	I-protein
by	NN	O	O
Northern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
.	NN	O	O

Nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
activity	NN	O	O
was	NN	O	O
assessed	NN	O	O
by	NN	O	O
gel	NN	O	O
shift	NN	O	O
assay	NN	O	O
.	NN	O	O

UUO	NN	O	O
in	NN	O	O
wild	NN	O	O
mice	NN	O	O
resulted	NN	O	O
in	NN	O	O
a	NN	O	O
marked	NN	O	O
expansion	NN	O	O
of	NN	O	O
relative	NN	O	O
volume	NN	O	O
of	NN	O	O
the	NN	O	O
tubulointerstitium	NN	O	O
,	NN	O	O
together	NN	O	O
with	NN	O	O
increased	NN	O	O
deposition	NN	O	O
of	NN	O	O
collagen	NN	O	B-protein
types	NN	O	I-protein
III	NN	O	I-protein
and	NN	O	I-protein
IV	NN	O	I-protein
and	NN	O	O
number	NN	O	O
of	NN	O	O
infiltrated	NN	O	O
monocytes	NN	O	B-cell_type
/	NN	O	O
macrophages	NN	O	B-cell_type
in	NN	O	O
the	NN	O	O
interstitium	NN	O	O
,	NN	O	O
relative	NN	O	O
to	NN	O	O
sham-operated	NN	O	O
mice	NN	O	O
.	NN	O	O

In	NN	O	O
comparison	NN	O	O
,	NN	O	O
these	NN	O	O
changes	NN	O	O
were	NN	O	O
significantly	NN	O	O
lower	NN	O	O
in	NN	O	O
mutant	NN	O	O
mice	NN	O	O
with	NN	O	O
UUO	NN	O	O
.	NN	O	O

The	NN	O	O
mRNA	NN	O	O
level	NN	O	O
of	NN	O	O
transforming	NN	O	B-protein
growth	NN	O	I-protein
factor-beta	NN	O	I-protein
was	NN	O	O
significantly	NN	O	O
higher	NN	O	O
in	NN	O	O
the	NN	O	O
OBK	NN	O	O
of	NN	O	O
wild	NN	O	O
mice	NN	O	O
with	NN	O	O
UUO	NN	O	O
compared	NN	O	O
with	NN	O	O
sham-operated	NN	O	O
mice	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
the	NN	O	O
increase	NN	O	O
in	NN	O	O
mRNA	NN	O	O
level	NN	O	O
in	NN	O	O
the	NN	O	O
OBK	NN	O	O
of	NN	O	O
mutant	NN	O	O
mice	NN	O	O
was	NN	O	O
significantly	NN	O	O
less	NN	O	O
than	NN	O	O
in	NN	O	O
wild	NN	O	O
mice	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
UUO	NN	O	O
resulted	NN	O	O
in	NN	O	O
activation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
in	NN	O	O
wild	NN	O	O
mice	NN	O	O
but	NN	O	O
was	NN	O	O
inhibited	NN	O	O
in	NN	O	O
the	NN	O	O
OBK	NN	O	O
of	NN	O	O
mutant	NN	O	O
mice	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
provide	NN	O	O
direct	NN	O	O
evidence	NN	O	O
that	NN	O	O
angiotensin	NN	O	B-protein
II	NN	O	I-protein
acting	NN	O	O
via	NN	O	O
the	NN	O	O
AT1a	NN	O	B-protein
plays	NN	O	O
a	NN	O	O
pivotal	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
tubulointerstitial	NN	O	O
fibrosis	NN	O	O
in	NN	O	O
UUO	NN	O	O
.	NN	O	O

-DOCSTART-	O

HLA-DQ	NN	O	B-protein
tetramers	NN	O	I-protein
identify	NN	O	O
epitope-specific	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
of	NN	O	O
herpes	NN	O	O
simplex	NN	O	O
virus	NN	O	O
type	NN	O	O
2-infected	NN	O	O
individuals	NN	O	O
:	NN	O	O
direct	NN	O	O
detection	NN	O	O
of	NN	O	O
immunodominant	NN	O	B-cell_type
antigen-responsive	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Ag-specific	NN	O	B-cell_type
CD4+	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
present	NN	O	O
in	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
in	NN	O	O
low	NN	O	O
frequency	NN	O	O
,	NN	O	O
where	NN	O	O
they	NN	O	O
undergo	NN	O	O
recruitment	NN	O	O
and	NN	O	O
expansion	NN	O	O
during	NN	O	O
immune	NN	O	O
responses	NN	O	O
and	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
numerous	NN	O	O
autoimmune	NN	O	O
diseases	NN	O	O
.	NN	O	O

MHC	NN	O	B-protein
tetramers	NN	O	I-protein
,	NN	O	O
which	NN	O	O
constitute	NN	O	O
a	NN	O	O
labeled	NN	O	B-protein
MHC-peptide	NN	O	I-protein
ligand	NN	O	I-protein
suitable	NN	O	O
for	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
Ag-specific	NN	O	B-protein
receptor	NN	O	I-protein
on	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
provide	NN	O	O
a	NN	O	O
novel	NN	O	O
approach	NN	O	O
for	NN	O	O
the	NN	O	O
detection	NN	O	O
and	NN	O	O
characterization	NN	O	O
of	NN	O	O
such	NN	O	O
rare	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
utilized	NN	O	O
this	NN	O	O
technology	NN	O	O
to	NN	O	O
identify	NN	O	O
HLA	NN	O	B-protein
DQ	NN	O	I-protein
-restricted	NN	O	O
Ag-specific	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
of	NN	O	O
human	NN	O	O
subjects	NN	O	O
and	NN	O	O
to	NN	O	O
identify	NN	O	O
immunodominant	NN	O	O
epitopes	NN	O	O
associated	NN	O	O
with	NN	O	O
viral	NN	O	O
infection	NN	O	O
.	NN	O	O

Peptides	NN	O	O
representing	NN	O	O
potential	NN	O	O
epitope	NN	O	B-protein
regions	NN	O	I-protein
of	NN	O	O
the	NN	O	O
VP16	NN	O	B-protein
protein	NN	O	I-protein
from	NN	O	O
HSV-2	NN	O	O
were	NN	O	O
loaded	NN	O	O
onto	NN	O	O
recombinant	NN	O	B-protein
DQ0602	NN	O	I-protein
molecules	NN	O	I-protein
to	NN	O	O
generate	NN	O	O
a	NN	O	O
panel	NN	O	O
of	NN	O	O
Ag-specific	NN	O	B-protein
DQ0602	NN	O	I-protein
tetramers	NN	O	I-protein
.	NN	O	O

VP16	NN	O	B-protein
Ag	NN	O	I-protein
-specific	NN	O	O
DQ-restricted	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
identified	NN	O	O
and	NN	O	O
expanded	NN	O	O
from	NN	O	O
the	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
of	NN	O	O
HSV-2-infected	NN	O	O
individuals	NN	O	O
,	NN	O	O
representing	NN	O	O
two	NN	O	O
predominant	NN	O	O
epitope	NN	O	O
specificities	NN	O	O
.	NN	O	O

Although	NN	O	O
the	NN	O	O
VP16	NN	O	O
369-380	NN	O	O
peptide	NN	O	O
has	NN	O	O
a	NN	O	O
lower	NN	O	O
binding	NN	O	O
affinity	NN	O	O
for	NN	O	O
DQ0602	NN	O	B-protein
molecules	NN	O	I-protein
than	NN	O	O
the	NN	O	O
VP16	NN	O	O
33-52	NN	O	O
peptide	NN	O	O
,	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
that	NN	O	O
recognized	NN	O	O
the	NN	O	O
VP16	NN	O	O
369-380	NN	O	O
peptide	NN	O	O
occurred	NN	O	O
at	NN	O	O
a	NN	O	O
much	NN	O	O
higher	NN	O	O
frequency	NN	O	O
than	NN	O	O
those	NN	O	O
that	NN	O	O
were	NN	O	O
specific	NN	O	O
for	NN	O	O
the	NN	O	O
VP16	NN	O	O
33-52	NN	O	O
peptide	NN	O	O
.	NN	O	O

-DOCSTART-	O

Down-regulation	NN	O	O
of	NN	O	O
BOB.1/OBF.1	NN	O	B-protein
and	NN	O	O
Oct2	NN	O	B-protein
in	NN	O	O
classical	NN	O	O
Hodgkin	NN	O	O
disease	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
lymphocyte	NN	O	O
predominant	NN	O	O
Hodgkin	NN	O	O
disease	NN	O	O
correlates	NN	O	O
with	NN	O	O
immunoglobulin	NN	O	O
transcription	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
the	NN	O	O
tumor	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
L	NN	O	B-cell_type
&	NN	O	I-cell_type
H	NN	O	I-cell_type
cells	NN	O	I-cell_type
)	NN	O	O
of	NN	O	O
lymphocyte	NN	O	O
predominant	NN	O	O
Hodgkin	NN	O	O
disease	NN	O	O
(	NN	O	O
LPHD	NN	O	O
)	NN	O	O
,	NN	O	O
Hodgkin	NN	O	B-cell_type
and	NN	O	I-cell_type
Reed-Sternberg	NN	O	I-cell_type
(	NN	O	I-cell_type
HRS	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
classical	NN	O	O
Hodgkin	NN	O	O
disease	NN	O	O
(	NN	O	O
cHD	NN	O	O
)	NN	O	O
are	NN	O	O
unable	NN	O	O
to	NN	O	O
transcribe	NN	O	O
immunoglobulin	NN	O	B-protein
,	NN	O	O
despite	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
rearranged	NN	O	B-DNA
immunoglobulin	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

Although	NN	O	O
initial	NN	O	O
studies	NN	O	O
have	NN	O	O
suggested	NN	O	O
crippling	NN	O	O
immunoglobulin	NN	O	O
gene	NN	O	O
mutations	NN	O	O
to	NN	O	O
be	NN	O	O
the	NN	O	O
cause	NN	O	O
of	NN	O	O
absent	NN	O	O
immunoglobulin	NN	O	O
expression	NN	O	O
in	NN	O	O
cHD	NN	O	O
,	NN	O	O
recent	NN	O	O
work	NN	O	O
of	NN	O	O
our	NN	O	O
group	NN	O	O
has	NN	O	O
demonstrated	NN	O	O
an	NN	O	O
impaired	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
immunoglobulin	NN	O	B-DNA
promoter	NN	O	I-DNA
as	NN	O	O
a	NN	O	O
superior	NN	O	O
mechanism	NN	O	O
.	NN	O	O

As	NN	O	O
immunoglobulin	NN	O	O
transcription	NN	O	O
is	NN	O	O
mainly	NN	O	O
regulated	NN	O	O
by	NN	O	O
the	NN	O	O
B-cell	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
Oct2	NN	O	B-protein
and	NN	O	O
BOB.1/OBF.1	NN	O	B-protein
,	NN	O	O
we	NN	O	O
analyzed	NN	O	O
35	NN	O	O
cases	NN	O	O
of	NN	O	O
LPHD	NN	O	O
,	NN	O	O
32	NN	O	O
cases	NN	O	O
of	NN	O	O
cHD	NN	O	O
,	NN	O	O
and	NN	O	O
2	NN	O	O
Hodgkin	NN	O	B-cell_line
disease	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
for	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
these	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
and	NN	O	O
also	NN	O	O
in	NN	O	O
parallel	NN	O	O
for	NN	O	O
immunoglobulin	NN	O	O
expression	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
an	NN	O	O
absence	NN	O	O
of	NN	O	O
Oct2	NN	O	B-protein
and/or	NN	O	O
BOB.1/OBF.1	NN	O	B-protein
in	NN	O	O
cHD	NN	O	O
and	NN	O	O
a	NN	O	O
striking	NN	O	O
overexpression	NN	O	O
of	NN	O	O
Oct2	NN	O	B-protein
in	NN	O	O
LPHD	NN	O	O
.	NN	O	O

Immunoglobulin	NN	O	O
expression	NN	O	O
was	NN	O	O
lacking	NN	O	O
in	NN	O	O
cHD	NN	O	O
but	NN	O	O
present	NN	O	O
in	NN	O	O
LPHD	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
the	NN	O	O
reintroduction	NN	O	O
of	NN	O	O
BOB.1/OBF.1	NN	O	B-protein
and	NN	O	O
Oct2	NN	O	B-protein
into	NN	O	O
cultured	NN	O	O
HRS	NN	O	B-cell_type
cells	NN	O	I-cell_type
restored	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
cotransduced	NN	O	O
immunoglobulin	NN	O	B-DNA
promoter	NN	O	I-DNA
constructs	NN	O	I-DNA
.	NN	O	O

Our	NN	O	O
findings	NN	O	O
dismiss	NN	O	O
the	NN	O	O
concept	NN	O	O
that	NN	O	O
the	NN	O	O
different	NN	O	O
immunoglobulin	NN	O	O
expression	NN	O	O
in	NN	O	O
cHD	NN	O	O
and	NN	O	O
LPHD	NN	O	O
is	NN	O	O
due	NN	O	O
to	NN	O	O
disrupting	NN	O	O
mutations	NN	O	O
of	NN	O	O
immunoglobulin	NN	O	B-DNA
V	NN	O	I-DNA
genes	NN	O	I-DNA
in	NN	O	O
cHD	NN	O	O
but	NN	O	O
is	NN	O	O
most	NN	O	O
likely	NN	O	O
due	NN	O	O
to	NN	O	O
a	NN	O	O
down-regulation	NN	O	O
of	NN	O	O
Oct2	NN	O	B-protein
and/or	NN	O	O
BOB.1/OBF.1	NN	O	B-protein
.	NN	O	O

This	NN	O	O
study	NN	O	O
further	NN	O	O
revealed	NN	O	O
Oct2	NN	O	B-protein
as	NN	O	O
a	NN	O	O
new	NN	O	O
and	NN	O	O
valuable	NN	O	O
marker	NN	O	O
for	NN	O	O
the	NN	O	O
identification	NN	O	O
of	NN	O	O
L	NN	O	B-cell_type
&	NN	O	I-cell_type
H	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
their	NN	O	O
distinction	NN	O	O
from	NN	O	O
HRS	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
impairment	NN	O	O
of	NN	O	O
immunoglobulin	NN	O	O
transcription	NN	O	O
with	NN	O	O
a	NN	O	O
down-regulated	NN	O	O
synthesis	NN	O	O
of	NN	O	O
Oct2	NN	O	B-protein
and	NN	O	O
BOB.1/OBF.1	NN	O	B-protein
is	NN	O	O
the	NN	O	O
first	NN	O	O
established	NN	O	O
general	NN	O	O
recurrent	NN	O	O
defect	NN	O	O
found	NN	O	O
in	NN	O	O
HRS	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Constitutive	NN	O	O
expression	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
is	NN	O	O
a	NN	O	O
characteristic	NN	O	O
feature	NN	O	O
of	NN	O	O
mycosis	NN	O	O
fungoides	NN	O	O
:	NN	O	O
implications	NN	O	O
for	NN	O	O
apoptosis	NN	O	O
resistance	NN	O	O
and	NN	O	O
pathogenesis	NN	O	O
.	NN	O	O

The	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
family	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
is	NN	O	O
an	NN	O	O
important	NN	O	O
regulator	NN	O	O
of	NN	O	O
genes	NN	O	O
expressed	NN	O	O
during	NN	O	O
inflammatory	NN	O	O
responses	NN	O	O
,	NN	O	O
immunoglobulin	NN	O	O
(	NN	O	O
Ig	NN	O	O
)	NN	O	O
class	NN	O	O
switching	NN	O	O
,	NN	O	O
cellular	NN	O	O
differentiation	NN	O	O
,	NN	O	O
and	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-protein
family	NN	O	I-protein
,	NN	O	O
including	NN	O	O
p65	NN	O	B-protein
(	NN	O	I-protein
Rel	NN	O	I-protein
A	NN	O	I-protein
)	NN	O	I-protein
,	NN	O	O
have	NN	O	O
been	NN	O	O
implicated	NN	O	O
in	NN	O	O
promoting	NN	O	O
survival	NN	O	O
of	NN	O	O
various	NN	O	O
hematopoeitic	NN	O	O
neoplasms	NN	O	O
,	NN	O	O
including	NN	O	O
T	NN	O	O
cell	NN	O	O
malignancies	NN	O	O
such	NN	O	O
as	NN	O	O
adult	NN	O	O
T	NN	O	O
cell	NN	O	O
leukemia-lymphoma	NN	O	O
.	NN	O	O

We	NN	O	O
investigated	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
active	NN	O	B-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
p65	NN	O	I-protein
(	NN	O	I-protein
Rel	NN	O	I-protein
A	NN	O	I-protein
)	NN	O	I-protein
in	NN	O	O
cases	NN	O	O
of	NN	O	O
mycosis	NN	O	O
fungoides	NN	O	O
(	NN	O	O
MF	NN	O	O
)	NN	O	O
and	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
chemical	NN	O	O
inhibitors	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
on	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
cutaneous	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
lymphoma	NN	O	I-cell_line
(	NN	O	I-cell_line
CTCL	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

Paraffin-embedded	NN	O	O
tissues	NN	O	O
from	NN	O	O
23	NN	O	O
cutaneous	NN	O	O
lesions	NN	O	O
and	NN	O	O
a	NN	O	O
single	NN	O	O
lymph	NN	O	O
node	NN	O	O
biopsy	NN	O	O
from	NN	O	O
patients	NN	O	O
diagnosed	NN	O	O
with	NN	O	O
MF	NN	O	O
were	NN	O	O
evaluated	NN	O	O
for	NN	O	O
p65	NN	O	O
(	NN	O	O
Rel	NN	O	O
A	NN	O	O
)	NN	O	O
expression	NN	O	O
by	NN	O	O
using	NN	O	O
a	NN	O	O
monoclonal	NN	O	B-protein
mouse	NN	O	I-protein
antibody	NN	O	I-protein
that	NN	O	O
detects	NN	O	O
the	NN	O	O
activated	NN	O	O
form	NN	O	O
of	NN	O	O
p65	NN	O	B-protein
(	NN	O	I-protein
Rel	NN	O	I-protein
A	NN	O	I-protein
)	NN	O	I-protein
.	NN	O	O

Apoptosis	NN	O	O
after	NN	O	O
treatment	NN	O	O
with	NN	O	O
the	NN	O	O
NF-kappa	NN	O	O
B	NN	O	O
inhibitors	NN	O	O
gliotoxin	NN	O	O
,	NN	O	O
MG132	NN	O	O
,	NN	O	O
BAY	NN	O	O
11-7082	NN	O	O
,	NN	O	O
and	NN	O	O
BAY	NN	O	O
11-7085	NN	O	O
was	NN	O	O
quantitatively	NN	O	O
measured	NN	O	O
in	NN	O	O
the	NN	O	O
CTCL	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
HuT-78	NN	O	I-cell_line
and	NN	O	I-cell_line
HH	NN	O	I-cell_line
by	NN	O	O
propidium	NN	O	O
iodide	NN	O	O
(	NN	O	O
PI	NN	O	O
)	NN	O	O
/cell	NN	O	O
cycle	NN	O	O
analysis	NN	O	O
for	NN	O	O
detection	NN	O	O
of	NN	O	O
a	NN	O	O
hypodiploid	NN	O	O
(	NN	O	O
sub-G	NN	O	O
(	NN	O	O
0	NN	O	O
)	NN	O	O
)	NN	O	O
population	NN	O	O
and	NN	O	O
by	NN	O	O
determination	NN	O	O
of	NN	O	O
increased	NN	O	O
Annexin	NN	O	O
V/7-amino-actinomycin	NN	O	O
D	NN	O	O
(	NN	O	O
7-AAD	NN	O	O
)	NN	O	O
expression	NN	O	O
.	NN	O	O

Nuclear	NN	O	O
extracts	NN	O	O
from	NN	O	O
CTCL	NN	O	B-cell_type
cells	NN	O	I-cell_type
before	NN	O	O
and	NN	O	O
after	NN	O	O
chemical	NN	O	O
inhibition	NN	O	O
were	NN	O	O
analyzed	NN	O	O
for	NN	O	O
NF-kappa	NN	O	O
B	NN	O	O
nuclear	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
by	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
(	NN	O	O
EMSA	NN	O	O
)	NN	O	O
with	NN	O	O
quantitative	NN	O	O
densitometry	NN	O	O
.	NN	O	O

Nuclear	NN	O	O
expression	NN	O	O
of	NN	O	O
p65	NN	O	B-protein
(	NN	O	I-protein
Rel	NN	O	I-protein
A	NN	O	I-protein
)	NN	O	I-protein
before	NN	O	O
and	NN	O	O
after	NN	O	O
treatment	NN	O	O
with	NN	O	O
the	NN	O	O
various	NN	O	O
inhibitory	NN	O	O
compounds	NN	O	O
was	NN	O	O
measured	NN	O	O
by	NN	O	O
immunofluorescence	NN	O	O
staining	NN	O	O
in	NN	O	O
each	NN	O	O
CTCL	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

Neoplastic	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
from	NN	O	O
22	NN	O	O
of	NN	O	O
24	NN	O	O
cases	NN	O	O
of	NN	O	O
MF	NN	O	O
showed	NN	O	O
strong	NN	O	O
nuclear	NN	O	O
and	NN	O	O
cytoplasmic	NN	O	O
expression	NN	O	O
of	NN	O	O
active	NN	O	B-protein
p65	NN	O	I-protein
(	NN	O	I-protein
Rel	NN	O	I-protein
A	NN	O	I-protein
)	NN	O	I-protein
.	NN	O	O

Compared	NN	O	O
with	NN	O	O
untreated	NN	O	O
control	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
a	NN	O	O
marked	NN	O	O
increase	NN	O	O
in	NN	O	O
apoptosis	NN	O	O
,	NN	O	O
a	NN	O	O
significant	NN	O	O
decrease	NN	O	O
in	NN	O	O
NF-kappa	NN	O	O
B	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
,	NN	O	O
and	NN	O	O
a	NN	O	O
marked	NN	O	O
decrease	NN	O	O
in	NN	O	O
nuclear	NN	O	O
p65	NN	O	O
(	NN	O	O
Rel	NN	O	O
A	NN	O	O
)	NN	O	O
expression	NN	O	O
were	NN	O	O
seen	NN	O	O
in	NN	O	O
cells	NN	O	O
from	NN	O	O
both	NN	O	O
CTCL	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
after	NN	O	O
chemical	NN	O	O
NF-kappa	NN	O	O
B	NN	O	O
inhibition	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
active	NN	O	O
form	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
p65	NN	O	I-protein
(	NN	O	I-protein
Rel	NN	O	I-protein
A	NN	O	I-protein
)	NN	O	I-protein
is	NN	O	O
commonly	NN	O	O
expressed	NN	O	O
in	NN	O	O
neoplastic	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
in	NN	O	O
patients	NN	O	O
with	NN	O	O
MF	NN	O	O
.	NN	O	O

In	NN	O	O
CTCL	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
the	NN	O	O
significant	NN	O	O
decrease	NN	O	O
in	NN	O	O
nuclear	NN	O	O
NF-kappa	NN	O	O
B	NN	O	O
expression	NN	O	O
and	NN	O	O
the	NN	O	O
marked	NN	O	O
increase	NN	O	O
in	NN	O	O
spontaneous	NN	O	O
apoptosis	NN	O	O
caused	NN	O	O
by	NN	O	O
chemical	NN	O	O
NF-kappa	NN	O	O
B	NN	O	O
inhibition	NN	O	O
suggest	NN	O	O
a	NN	O	O
critical	NN	O	O
role	NN	O	O
for	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
and	NN	O	O
tumor	NN	O	O
cell	NN	O	O
maintenance	NN	O	O
of	NN	O	O
CTCLs	NN	O	O
.	NN	O	O

HUM	NN	O	O
PATHOL	NN	O	O
31	NN	O	O
:	NN	O	O
1482-1490	NN	O	O
.	NN	O	O

-DOCSTART-	O

Human	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
tax	NN	O	B-protein
protein	NN	O	I-protein
activates	NN	O	O
transcription	NN	O	O
through	NN	O	O
AP-1	NN	O	B-DNA
site	NN	O	I-DNA
by	NN	O	O
inducing	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Human	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
(	NN	O	O
HTLV-1	NN	O	O
)	NN	O	O
Tax	NN	O	B-protein
protein	NN	O	I-protein
induces	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
various	NN	O	O
family	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
AP-1	NN	O	I-protein
,	NN	O	O
such	NN	O	O
as	NN	O	O
c-Jun	NN	O	B-protein
,	NN	O	O
JunD	NN	O	B-protein
,	NN	O	O
c-Fos	NN	O	B-protein
,	NN	O	O
and	NN	O	O
Fra-1	NN	O	B-protein
,	NN	O	O
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
RNA	NN	O	O
expression	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
examined	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
Tax	NN	O	B-protein
in	NN	O	O
transcription	NN	O	O
through	NN	O	O
AP-1-binding	NN	O	B-DNA
sites	NN	O	I-DNA
(	NN	O	O
AP-1	NN	O	B-DNA
site	NN	O	I-DNA
)	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Transient	NN	O	O
transfection	NN	O	O
studies	NN	O	O
showed	NN	O	O
that	NN	O	O
Tax	NN	O	B-protein
activated	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
luciferase	NN	O	B-DNA
gene	NN	O	I-DNA
regulated	NN	O	O
by	NN	O	O
two	NN	O	O
copies	NN	O	O
of	NN	O	O
an	NN	O	O
AP-1	NN	O	B-DNA
site	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
Jurkat	NN	O	I-cell_line
T-cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

Tax	NN	O	B-protein
activates	NN	O	O
the	NN	O	O
expression	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
through	NN	O	O
two	NN	O	O
different	NN	O	O
enhancers	NN	O	O
:	NN	O	O
a	NN	O	O
cAMP-responsive	NN	O	B-DNA
(	NN	O	I-DNA
CRE	NN	O	I-DNA
)	NN	O	I-DNA
-like	NN	O	I-DNA
element	NN	O	I-DNA
and	NN	O	O
a	NN	O	O
kappaB	NN	O	B-DNA
element	NN	O	I-DNA
.	NN	O	O

Two	NN	O	O
Tax	NN	O	B-protein
mutants	NN	O	I-protein
differentially	NN	O	O
activated	NN	O	O
expression	NN	O	O
of	NN	O	O
these	NN	O	O
two	NN	O	O
elements	NN	O	O
.	NN	O	O

Tax703	NN	O	B-protein
preferentially	NN	O	O
activated	NN	O	O
the	NN	O	O
kappaB	NN	O	B-DNA
element	NN	O	I-DNA
but	NN	O	O
not	NN	O	O
the	NN	O	O
CRE-like	NN	O	O
one	NN	O	O
,	NN	O	O
whereas	NN	O	O
TaxM22	NN	O	B-protein
showed	NN	O	O
the	NN	O	O
reverse	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
Tax703	NN	O	B-protein
and	NN	O	O
Tax	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
TaxM22	NN	O	B-protein
,	NN	O	O
converted	NN	O	O
cell	NN	O	O
growth	NN	O	O
of	NN	O	O
a	NN	O	O
mouse	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
line	NN	O	I-cell_line
from	NN	O	O
being	NN	O	O
interleukin	NN	O	O
(	NN	O	O
IL	NN	O	O
)	NN	O	O
-2-dependent	NN	O	O
to	NN	O	O
being	NN	O	O
IL-2-independent	NN	O	O
.	NN	O	O

Unlike	NN	O	O
the	NN	O	O
wild-type	NN	O	B-protein
Tax	NN	O	I-protein
,	NN	O	O
Tax703	NN	O	B-protein
and	NN	O	O
TaxM22	NN	O	B-protein
only	NN	O	O
weakly	NN	O	O
activated	NN	O	O
the	NN	O	O
AP-1	NN	O	B-DNA
site	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
T-cell	NN	O	B-cell_line
line	NN	O	I-cell_line
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
Tax	NN	O	B-protein
seems	NN	O	O
to	NN	O	O
activate	NN	O	O
the	NN	O	O
AP-1	NN	O	B-DNA
site	NN	O	I-DNA
via	NN	O	O
mechanisms	NN	O	O
distinct	NN	O	O
from	NN	O	O
those	NN	O	O
of	NN	O	O
kappaB	NN	O	B-DNA
or	NN	O	I-DNA
CRE-like	NN	O	I-DNA
elements	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
AP-1	NN	O	B-DNA
site	NN	O	I-DNA
is	NN	O	O
dispensable	NN	O	O
for	NN	O	O
IL-2-independent	NN	O	O
growth	NN	O	O
of	NN	O	O
CTLL-2	NN	O	O
.	NN	O	O

Electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
showed	NN	O	O
that	NN	O	O
Tax	NN	O	B-protein
induced	NN	O	O
strong	NN	O	O
binding	NN	O	O
activity	NN	O	O
to	NN	O	O
an	NN	O	O
AP-1	NN	O	B-DNA
site	NN	O	I-DNA
in	NN	O	O
CTLL-2	NN	O	O
,	NN	O	O
whereas	NN	O	O
Tax703	NN	O	B-protein
did	NN	O	O
not	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
binding	NN	O	O
activity	NN	O	O
to	NN	O	O
the	NN	O	O
AP-1	NN	O	B-DNA
site	NN	O	I-DNA
is	NN	O	O
essential	NN	O	O
for	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
by	NN	O	O
Tax	NN	O	B-protein
.	NN	O	O

The	NN	O	O
binding	NN	O	B-protein
complex	NN	O	I-protein
induced	NN	O	O
by	NN	O	O
Tax	NN	O	B-protein
in	NN	O	O
CTLL-2	NN	O	O
contained	NN	O	O
JunD	NN	O	B-protein
and	NN	O	O
Fra-2	NN	O	B-protein
.	NN	O	O

Other	NN	O	O
AP-1	NN	O	B-protein
proteins	NN	O	I-protein
were	NN	O	O
undetectable	NN	O	O
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
transcription	NN	O	O
through	NN	O	O
the	NN	O	O
AP-1	NN	O	B-DNA
site	NN	O	I-DNA
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
by	NN	O	O
JunD	NN	O	B-protein
and/or	NN	O	O
Fra-2	NN	O	B-protein
was	NN	O	O
weak	NN	O	O
.	NN	O	O

c-Jun	NN	O	B-protein
,	NN	O	O
JunB	NN	O	B-protein
,	NN	O	O
and	NN	O	O
c-Fos	NN	O	B-protein
activation	NN	O	O
was	NN	O	O
greater	NN	O	O
,	NN	O	O
although	NN	O	O
the	NN	O	O
level	NN	O	O
was	NN	O	O
still	NN	O	O
less	NN	O	O
than	NN	O	O
that	NN	O	O
with	NN	O	O
Tax	NN	O	B-protein
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
AP-1	NN	O	B-RNA
mRNA	NN	O	I-RNA
by	NN	O	O
Tax	NN	O	B-protein
may	NN	O	O
not	NN	O	O
be	NN	O	O
sufficient	NN	O	O
for	NN	O	O
a	NN	O	O
complete	NN	O	O
activation	NN	O	O
of	NN	O	O
AP-1	NN	O	B-DNA
site	NN	O	I-DNA
by	NN	O	O
Tax	NN	O	B-protein
.	NN	O	O

Our	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
Tax	NN	O	B-protein
activates	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
cellular	NN	O	B-DNA
genes	NN	O	I-DNA
with	NN	O	O
AP-1	NN	O	B-DNA
sites	NN	O	I-DNA
by	NN	O	O
inducing	NN	O	O
the	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
proteins	NN	O	I-protein
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
a	NN	O	O
mechanism	NN	O	O
distinct	NN	O	O
from	NN	O	O
those	NN	O	O
of	NN	O	O
CRE-like	NN	O	B-DNA
and	NN	O	I-DNA
kappaB	NN	O	I-DNA
elements	NN	O	I-DNA
.	NN	O	O

Copyright	NN	O	O
2001	NN	O	O
Academic	NN	O	O
Press	NN	O	O
.	NN	O	O

-DOCSTART-	O

Strict	NN	O	O
control	NN	O	O
of	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
replication	NN	O	O
by	NN	O	O
a	NN	O	O
genetic	NN	O	O
switch	NN	O	O
:	NN	O	O
Tet	NN	O	O
for	NN	O	O
Tat	NN	O	O
.	NN	O	O

Live-attenuated	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
variants	NN	O	O
have	NN	O	O
shown	NN	O	O
great	NN	O	O
promise	NN	O	O
as	NN	O	O
AIDS	NN	O	O
vaccines	NN	O	O
,	NN	O	O
but	NN	O	O
continued	NN	O	O
replication	NN	O	O
can	NN	O	O
lead	NN	O	O
to	NN	O	O
the	NN	O	O
selection	NN	O	O
of	NN	O	O
faster-replicating	NN	O	O
variants	NN	O	O
that	NN	O	O
are	NN	O	O
pathogenic	NN	O	O
.	NN	O	O

We	NN	O	O
therefore	NN	O	O
designed	NN	O	O
HIV-1	NN	O	B-DNA
genomes	NN	O	I-DNA
that	NN	O	O
replicate	NN	O	O
exclusively	NN	O	O
upon	NN	O	O
addition	NN	O	O
of	NN	O	O
the	NN	O	O
nontoxic	NN	O	O
effector	NN	O	O
doxycycline	NN	O	O
(	NN	O	O
dox	NN	O	O
)	NN	O	O
.	NN	O	O

This	NN	O	O
was	NN	O	O
achieved	NN	O	O
by	NN	O	O
replacement	NN	O	O
of	NN	O	O
the	NN	O	O
viral	NN	O	O
TAR-Tat	NN	O	O
system	NN	O	O
for	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
by	NN	O	O
the	NN	O	O
Escherichia	NN	O	O
coli-derived	NN	O	O
Tet	NN	O	O
system	NN	O	O
for	NN	O	O
inducible	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

These	NN	O	O
designer	NN	O	O
``	NN	O	O
HIV-rtTA	NN	O	O
''	NN	O	O
viruses	NN	O	O
replicate	NN	O	O
in	NN	O	O
a	NN	O	O
strictly	NN	O	O
dox-dependent	NN	O	O
manner	NN	O	O
both	NN	O	O
in	NN	O	O
a	NN	O	O
T-cell	NN	O	B-cell_line
line	NN	O	I-cell_line
and	NN	O	O
in	NN	O	O
primary	NN	O	B-cell_type
blood	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
the	NN	O	O
rate	NN	O	O
of	NN	O	O
replication	NN	O	O
can	NN	O	O
be	NN	O	O
fine-tuned	NN	O	O
by	NN	O	O
simple	NN	O	O
variation	NN	O	O
of	NN	O	O
the	NN	O	O
dox	NN	O	O
concentration	NN	O	O
.	NN	O	O

These	NN	O	O
HIV-rtTA	NN	O	O
viruses	NN	O	O
provide	NN	O	O
a	NN	O	O
tool	NN	O	O
to	NN	O	O
perform	NN	O	O
genetics	NN	O	O
,	NN	O	O
e.g.	NN	O	O
,	NN	O	O
selection	NN	O	O
and	NN	O	O
optimization	NN	O	O
experiments	NN	O	O
,	NN	O	O
with	NN	O	O
the	NN	O	O
E.	NN	O	O
coli-derived	NN	O	O
Tet	NN	O	O
reagents	NN	O	O
in	NN	O	O
a	NN	O	O
eukaryotic	NN	O	O
background	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
such	NN	O	O
viruses	NN	O	O
may	NN	O	O
represent	NN	O	O
improved	NN	O	O
vaccine	NN	O	O
candidates	NN	O	O
because	NN	O	O
their	NN	O	O
replication	NN	O	O
can	NN	O	O
be	NN	O	O
turned	NN	O	O
on	NN	O	O
and	NN	O	O
off	NN	O	O
at	NN	O	O
will	NN	O	O
.	NN	O	O

-DOCSTART-	O

Synovial	NN	O	O
fluid	NN	O	O
induced	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
DNA	NN	O	O
binding	NN	O	O
in	NN	O	O
a	NN	O	O
monocytic	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

OBJECTIVE	NN	O	O
:	NN	O	O
To	NN	O	O
determine	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
synovial	NN	O	O
fluids	NN	O	O
(	NN	O	O
SF	NN	O	O
)	NN	O	O
on	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
(	NN	O	O
NF-kappaB	NN	O	B-protein
)	NN	O	O
in	NN	O	O
the	NN	O	O
Mono	NN	O	B-cell_line
Mac	NN	O	I-cell_line
6	NN	O	I-cell_line
monocytic/macrophage	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
as	NN	O	O
a	NN	O	O
model	NN	O	O
for	NN	O	O
the	NN	O	O
interaction	NN	O	O
between	NN	O	O
SF	NN	O	O
and	NN	O	O
synovial	NN	O	B-cell_type
tissue	NN	O	I-cell_type
macrophages	NN	O	I-cell_type
in	NN	O	O
arthritis	NN	O	O
.	NN	O	O

METHODS	NN	O	O
:	NN	O	O
Mono	NN	O	B-cell_line
Mac	NN	O	I-cell_line
6	NN	O	I-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
incubated	NN	O	O
with	NN	O	O
SF	NN	O	O
from	NN	O	O
the	NN	O	O
knee	NN	O	O
joints	NN	O	O
of	NN	O	O
human	NN	O	O
subjects	NN	O	O
with	NN	O	O
rheumatoid	NN	O	O
arthritis	NN	O	O
(	NN	O	O
RA	NN	O	O
)	NN	O	O
,	NN	O	O
undifferentiated	NN	O	O
seronegative	NN	O	O
oligoarthritis	NN	O	O
,	NN	O	O
and	NN	O	O
osteoarthritis	NN	O	O
(	NN	O	O
OA	NN	O	O
)	NN	O	O
.	NN	O	O

Nuclear	NN	O	O
extracts	NN	O	O
prepared	NN	O	O
from	NN	O	O
the	NN	O	O
Mono	NN	O	B-cell_line
Mac	NN	O	I-cell_line
6	NN	O	I-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
RA	NN	O	O
synovial	NN	O	O
tissue	NN	O	O
were	NN	O	O
analyzed	NN	O	O
by	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
analysis	NN	O	O
(	NN	O	O
EMSA	NN	O	O
)	NN	O	O
for	NN	O	O
NF-kappaB	NN	O	B-protein
DNA	NN	O	I-protein
binding	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
Induction	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
by	NN	O	O
the	NN	O	O
p65	NN	O	B-protein
(	NN	O	I-protein
RelA	NN	O	I-protein
)	NN	O	I-protein
/p50	NN	O	I-protein
heterodimer	NN	O	I-protein
was	NN	O	O
observed	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
incubation	NN	O	O
of	NN	O	O
Mono	NN	O	B-cell_line
Mac	NN	O	I-cell_line
6	NN	O	I-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
SF	NN	O	O
(	NN	O	O
20	NN	O	O
%	NN	O	O
in	NN	O	O
culture	NN	O	O
medium	NN	O	O
)	NN	O	O
from	NN	O	O
5	NN	O	O
of	NN	O	O
8	NN	O	O
subjects	NN	O	O
with	NN	O	O
RA	NN	O	O
,	NN	O	O
4	NN	O	O
of	NN	O	O
5	NN	O	O
with	NN	O	O
OA	NN	O	O
,	NN	O	O
and	NN	O	O
none	NN	O	O
of	NN	O	O
3	NN	O	O
with	NN	O	O
undifferentiated	NN	O	O
seronegative	NN	O	O
oligoarthritis	NN	O	O
.	NN	O	O

Incubation	NN	O	O
of	NN	O	O
SF	NN	O	O
with	NN	O	O
neutralizing	NN	O	B-protein
antibodies	NN	O	I-protein
against	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
(	NN	O	I-protein
TNF-alpha	NN	O	I-protein
)	NN	O	I-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
antibodies	NN	O	B-protein
against	NN	O	O
interleukin	NN	O	B-protein
6	NN	O	I-protein
(	NN	O	I-protein
IL-6	NN	O	I-protein
)	NN	O	I-protein
,	NN	O	O
significantly	NN	O	O
reduced	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
p65/p50	NN	O	O
binding	NN	O	O
activity	NN	O	O
in	NN	O	O
SF	NN	O	O
from	NN	O	O
subjects	NN	O	O
with	NN	O	O
RA	NN	O	O
and	NN	O	O
OA	NN	O	O
.	NN	O	O

Unexpectedly	NN	O	O
,	NN	O	O
a	NN	O	O
slowly	NN	O	O
migrating	NN	O	O
SF	NN	O	O
inducible	NN	O	O
NF-kappaB	NN	O	B-protein
-binding	NN	O	O
complex	NN	O	O
was	NN	O	O
observed	NN	O	O
in	NN	O	O
EMSA	NN	O	O
of	NN	O	O
Mono	NN	O	B-cell_line
Mac	NN	O	I-cell_line
6	NN	O	I-cell_line
cells	NN	O	I-cell_line
after	NN	O	O
incubation	NN	O	O
with	NN	O	O
SF	NN	O	O
from	NN	O	O
5	NN	O	O
of	NN	O	O
8	NN	O	O
RA	NN	O	O
and	NN	O	O
2	NN	O	O
of	NN	O	O
5	NN	O	O
OA	NN	O	O
subjects	NN	O	O
.	NN	O	O

The	NN	O	O
induction	NN	O	O
of	NN	O	O
this	NN	O	O
complex	NN	O	O
by	NN	O	O
SF	NN	O	O
was	NN	O	O
not	NN	O	O
affected	NN	O	O
by	NN	O	O
neutralization	NN	O	O
of	NN	O	O
TNF-alpha	NN	O	B-protein
or	NN	O	O
IL-6	NN	O	B-protein
in	NN	O	O
SF	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
complex	NN	O	O
was	NN	O	O
not	NN	O	O
inducible	NN	O	O
by	NN	O	O
TNF-alpha	NN	O	B-protein
,	NN	O	O
IL-1beta	NN	O	O
,	NN	O	O
TNF-alpha	NN	O	B-protein
/	NN	O	O
IL-1beta	NN	O	B-protein
,	NN	O	O
IL-6	NN	O	B-protein
,	NN	O	O
platelet	NN	O	B-protein
derived	NN	O	I-protein
growth	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
lipopolysaccharide	NN	O	O
,	NN	O	O
or	NN	O	O
tetradecanoyl	NN	O	O
phorbol	NN	O	O
acetate	NN	O	O
.	NN	O	O

The	NN	O	O
slowly	NN	O	O
migrating	NN	O	B-protein
complex	NN	O	I-protein
could	NN	O	O
not	NN	O	O
be	NN	O	O
supershifted	NN	O	O
with	NN	O	O
antibodies	NN	O	B-protein
against	NN	O	O
NF-kappaB	NN	O	B-protein
,	NN	O	O
Jun	NN	O	B-protein
,	NN	O	O
or	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
coactivators	NN	O	O
p300	NN	O	B-protein
or	NN	O	O
CBP	NN	O	B-protein
.	NN	O	O

A	NN	O	O
NF-kappaB-binding	NN	O	B-protein
complex	NN	O	I-protein
with	NN	O	O
similar	NN	O	O
slow	NN	O	O
mobility	NN	O	O
was	NN	O	O
observed	NN	O	O
in	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
prepared	NN	O	O
from	NN	O	O
fresh	NN	O	O
human	NN	O	O
RA	NN	O	O
synovial	NN	O	O
tissue	NN	O	O
.	NN	O	O

CONCLUSION	NN	O	O
:	NN	O	O
Biological	NN	O	O
activity	NN	O	O
of	NN	O	O
TNF-alpha	NN	O	B-protein
in	NN	O	O
SF	NN	O	O
from	NN	O	O
RA	NN	O	O
and	NN	O	O
OA	NN	O	O
subjects	NN	O	O
is	NN	O	O
capable	NN	O	O
of	NN	O	O
inducing	NN	O	O
p65/p50	NN	O	B-protein
NF-kappaB	NN	O	I-protein
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
in	NN	O	O
macrophages	NN	O	B-cell_type
.	NN	O	O

A	NN	O	O
property	NN	O	O
of	NN	O	O
SF	NN	O	O
that	NN	O	O
is	NN	O	O
independent	NN	O	O
of	NN	O	O
TNF-alpha	NN	O	B-protein
and	NN	O	O
other	NN	O	O
cytokines	NN	O	B-protein
is	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
a	NN	O	O
novel	NN	O	O
slowly	NN	O	O
migrating	NN	O	O
NF-kappaB-binding	NN	O	B-protein
complex	NN	O	I-protein
.	NN	O	O

Soluble	NN	O	O
mediators	NN	O	O
in	NN	O	O
SF	NN	O	O
of	NN	O	O
subjects	NN	O	O
with	NN	O	O
RA	NN	O	O
and	NN	O	O
OA	NN	O	O
can	NN	O	O
therefore	NN	O	O
modulate	NN	O	O
binding	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
proteins	NN	O	I-protein
to	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-protein
binding	NN	O	I-protein
site	NN	O	I-protein
in	NN	O	O
macrophages	NN	O	B-cell_type
and	NN	O	O
may	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
inflammatory	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
arthritis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Peroxisome	NN	O	B-protein
proliferator	NN	O	I-protein
activator	NN	O	I-protein
receptor-gamma	NN	O	I-protein
agonists	NN	O	O
and	NN	O	O
15-deoxy-Delta	NN	O	O
(	NN	O	O
12	NN	O	O
,	NN	O	O
14	NN	O	O
)	NN	O	O
(	NN	O	O
12	NN	O	O
,	NN	O	O
14	NN	O	O
)	NN	O	O
-PGJ	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
induce	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
normal	NN	O	O
and	NN	O	O
malignant	NN	O	O
B-lineage	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
research	NN	O	O
described	NN	O	O
herein	NN	O	O
evaluates	NN	O	O
the	NN	O	O
expression	NN	O	O
and	NN	O	O
functional	NN	O	O
significance	NN	O	O
of	NN	O	O
peroxisome	NN	O	B-protein
proliferator	NN	O	I-protein
activator	NN	O	I-protein
receptor-gamma	NN	O	I-protein
(	NN	O	I-protein
PPAR-gamma	NN	O	I-protein
)	NN	O	I-protein
on	NN	O	O
B-lineage	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Normal	NN	O	B-cell_type
mouse	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
B	NN	O	B-cell_type
lymphoma	NN	O	I-cell_type
cells	NN	O	I-cell_type
reflective	NN	O	O
of	NN	O	O
stages	NN	O	O
of	NN	O	O
B	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
(	NN	O	O
e.g.	NN	O	O
,	NN	O	O
70Z/3	NN	O	O
,	NN	O	O
CH31	NN	O	O
,	NN	O	O
WEHI-231	NN	O	O
,	NN	O	O
CH12	NN	O	O
,	NN	O	O
and	NN	O	O
J558	NN	O	O
)	NN	O	O
express	NN	O	O
PPAR-gamma	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
,	NN	O	O
by	NN	O	O
Western	NN	O	O
blot	NN	O	O
analysis	NN	O	O
,	NN	O	O
the	NN	O	O
67-kDa	NN	O	B-protein
PPAR-gamma	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

15-Deoxy-Delta	NN	O	O
(	NN	O	O
12	NN	O	O
,	NN	O	O
14	NN	O	O
)	NN	O	O
-PGJ	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
(	NN	O	O
15d-PGJ	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
)	NN	O	O
,	NN	O	O
a	NN	O	O
PPAR-gamma	NN	O	O
agonist	NN	O	O
,	NN	O	O
has	NN	O	O
a	NN	O	O
dose-dependent	NN	O	O
antiproliferative	NN	O	O
and	NN	O	O
cytotoxic	NN	O	O
effect	NN	O	O
on	NN	O	O
normal	NN	O	B-cell_type
and	NN	O	I-cell_type
malignant	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
as	NN	O	O
shown	NN	O	O
by	NN	O	O
[	NN	O	O
(	NN	O	O
3	NN	O	O
)	NN	O	O
H	NN	O	O
]	NN	O	O
thymidine	NN	O	O
and	NN	O	O
3-	NN	O	O
[	NN	O	O
4	NN	O	O
,	NN	O	O
5-dimethylthiazol-2-yl	NN	O	O
]	NN	O	O
-2	NN	O	O
,	NN	O	O
5-diphenyltetrazolium	NN	O	O
bromide	NN	O	O
assays	NN	O	O
.	NN	O	O

Only	NN	O	O
PPAR-gamma	NN	O	O
agonists	NN	O	O
(	NN	O	O
thiazolidinediones	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
not	NN	O	O
PPAR-alpha	NN	O	O
agonists	NN	O	O
,	NN	O	O
mimicked	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
15d-PGJ	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
on	NN	O	O
B-lineage	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
the	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
15d-PGJ	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
negatively	NN	O	O
affects	NN	O	O
B-lineage	NN	O	B-cell_type
cells	NN	O	I-cell_type
involves	NN	O	O
in	NN	O	O
part	NN	O	O
PPAR-gamma	NN	O	B-protein
.	NN	O	O

The	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
PPAR-gamma	NN	O	O
agonists	NN	O	O
induce	NN	O	O
cytotoxicity	NN	O	O
is	NN	O	O
via	NN	O	O
apoptosis	NN	O	O
,	NN	O	O
as	NN	O	O
shown	NN	O	O
by	NN	O	O
annexin	NN	O	O
V	NN	O	O
staining	NN	O	O
and	NN	O	O
as	NN	O	O
confirmed	NN	O	O
by	NN	O	O
DNA	NN	O	O
fragmentation	NN	O	O
detected	NN	O	O
using	NN	O	O
the	NN	O	O
TUNEL	NN	O	O
assay	NN	O	O
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
addition	NN	O	O
of	NN	O	O
PGF	NN	O	O
(	NN	O	O
2alpha	NN	O	O
)	NN	O	O
,	NN	O	O
which	NN	O	O
was	NN	O	O
not	NN	O	O
known	NN	O	O
to	NN	O	O
affect	NN	O	O
lymphocytes	NN	O	B-cell_type
,	NN	O	O
dramatically	NN	O	O
attenuated	NN	O	O
the	NN	O	O
deleterious	NN	O	O
effects	NN	O	O
of	NN	O	O
PPAR-gamma	NN	O	O
agonists	NN	O	O
on	NN	O	O
B	NN	O	B-cell_type
lymphomas	NN	O	I-cell_type
.	NN	O	O

Surprisingly	NN	O	O
,	NN	O	O
15d-PGJ	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
induced	NN	O	O
a	NN	O	O
massive	NN	O	O
increase	NN	O	O
in	NN	O	O
nuclear	NN	O	O
mitogen-activated	NN	O	O
protein	NN	O	O
kinase	NN	O	O
activation	NN	O	O
,	NN	O	O
and	NN	O	O
pretreatment	NN	O	O
with	NN	O	O
PGF	NN	O	O
(	NN	O	O
2alpha	NN	O	O
)	NN	O	O
blunted	NN	O	O
the	NN	O	O
mitogen-activated	NN	O	O
protein	NN	O	O
kinase	NN	O	O
activation	NN	O	O
.	NN	O	O

This	NN	O	O
is	NN	O	O
the	NN	O	O
first	NN	O	O
study	NN	O	O
evaluating	NN	O	O
PPAR-gamma	NN	O	O
expression	NN	O	O
and	NN	O	O
its	NN	O	O
significance	NN	O	O
on	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

PPAR-gamma	NN	O	O
agonists	NN	O	O
may	NN	O	O
serve	NN	O	O
as	NN	O	O
a	NN	O	O
counterbalance	NN	O	O
to	NN	O	O
the	NN	O	O
stimulating	NN	O	O
effects	NN	O	O
of	NN	O	O
other	NN	O	O
PGs	NN	O	O
,	NN	O	O
namely	NN	O	O
PGE	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
,	NN	O	O
which	NN	O	O
promotes	NN	O	O
B	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
PGs	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
15d-PGJ	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
synthetic	NN	O	O
PPAR-gamma	NN	O	O
agonists	NN	O	O
to	NN	O	O
induce	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
B-lineage	NN	O	B-cell_type
cells	NN	O	I-cell_type
may	NN	O	O
lead	NN	O	O
to	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
novel	NN	O	O
therapies	NN	O	O
for	NN	O	O
fatal	NN	O	B-cell_type
B	NN	O	I-cell_type
lymphomas	NN	O	I-cell_type

-DOCSTART-	O

Inhibition	NN	O	O
of	NN	O	O
STAT3	NN	O	O
signaling	NN	O	O
leads	NN	O	O
to	NN	O	O
apoptosis	NN	O	O
of	NN	O	O
leukemic	NN	O	B-cell_type
large	NN	O	I-cell_type
granular	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
decreased	NN	O	O
Mcl-1	NN	O	O
expression	NN	O	O
.	NN	O	O

Large	NN	O	O
granular	NN	O	O
lymphocyte	NN	O	O
(	NN	O	O
LGL	NN	O	O
)	NN	O	O
leukemia	NN	O	O
is	NN	O	O
characterized	NN	O	O
by	NN	O	O
the	NN	O	O
expansion	NN	O	O
of	NN	O	O
antigen-activated	NN	O	B-cell_type
cytotoxic	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
leukemic	NN	O	B-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
resistant	NN	O	O
to	NN	O	O
Fas-mediated	NN	O	O
apoptosis	NN	O	O
despite	NN	O	O
expressing	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
Fas	NN	O	B-protein
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
leukemic	NN	O	B-cell_type
LGL	NN	O	I-cell_type
from	NN	O	O
19	NN	O	O
patients	NN	O	O
displayed	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
activated	NN	O	B-protein
STAT3	NN	O	I-protein
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
leukemic	NN	O	B-cell_type
LGL	NN	O	I-cell_type
with	NN	O	O
the	NN	O	O
JAK-selective	NN	O	O
tyrosine	NN	O	O
kinase	NN	O	O
inhibitor	NN	O	O
AG-490	NN	O	O
induced	NN	O	O
apoptosis	NN	O	O
with	NN	O	O
a	NN	O	O
corresponding	NN	O	O
decrease	NN	O	O
in	NN	O	O
STAT	NN	O	B-protein
-DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
using	NN	O	O
an	NN	O	O
antisense	NN	O	O
oligonucleotide	NN	O	O
approach	NN	O	O
to	NN	O	O
diminish	NN	O	O
STAT3	NN	O	O
expression	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
Fas	NN	O	O
sensitivity	NN	O	O
was	NN	O	O
restored	NN	O	O
in	NN	O	O
leukemic	NN	O	B-cell_type
LGL	NN	O	I-cell_type
.	NN	O	O

AG-490-induced	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
leukemic	NN	O	B-cell_type
LGL	NN	O	I-cell_type
was	NN	O	O
independent	NN	O	O
of	NN	O	O
Bcl-xL	NN	O	O
or	NN	O	O
Bcl-2	NN	O	O
expression	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
Bcl-2-family	NN	O	B-protein
protein	NN	O	I-protein
Mcl-1	NN	O	I-protein
was	NN	O	O
significantly	NN	O	O
reduced	NN	O	O
by	NN	O	O
AG-490	NN	O	O
treatment	NN	O	O
.	NN	O	O

Activated	NN	O	B-protein
STAT3	NN	O	I-protein
was	NN	O	O
shown	NN	O	O
to	NN	O	O
bind	NN	O	O
an	NN	O	O
SIE-related	NN	O	B-DNA
element	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
murine	NN	O	B-DNA
mcl-1	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Using	NN	O	O
a	NN	O	O
luciferase	NN	O	O
reporter	NN	O	O
assay	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
v-src	NN	O	O
overexpression	NN	O	O
in	NN	O	O
NIH3T3	NN	O	O
induced	NN	O	O
STAT3	NN	O	B-protein
-dependent	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
from	NN	O	O
the	NN	O	O
mcl-1	NN	O	B-DNA
promoter	NN	O	I-DNA
and	NN	O	O
increased	NN	O	O
endogenous	NN	O	O
Mcl-1	NN	O	O
protein	NN	O	O
levels	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
STAT3	NN	O	B-protein
activation	NN	O	O
contributed	NN	O	O
to	NN	O	O
accumulation	NN	O	O
of	NN	O	O
the	NN	O	O
leukemic	NN	O	B-cell_type
LGL	NN	O	I-cell_type
clones	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
investigation	NN	O	O
should	NN	O	O
focus	NN	O	O
on	NN	O	O
novel	NN	O	O
strategies	NN	O	O
targeting	NN	O	O
STAT3	NN	O	B-protein
in	NN	O	O
the	NN	O	O
treatment	NN	O	O
of	NN	O	O
LGL	NN	O	O
leukemia	NN	O	O
.	NN	O	O

-DOCSTART-	O

Decreased	NN	O	O
immediate	NN	O	O
inflammatory	NN	O	B-DNA
gene	NN	O	I-DNA
induction	NN	O	O
in	NN	O	O
activating	NN	O	B-protein
transcription	NN	O	I-protein
factor-2	NN	O	I-protein
mutant	NN	O	O
mice	NN	O	O
.	NN	O	O

Transcription	NN	O	B-protein
factor	NN	O	I-protein
activating	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	I-protein
ATF	NN	O	I-protein
)	NN	O	I-protein
-2	NN	O	I-protein
is	NN	O	O
activated	NN	O	O
by	NN	O	O
inflammatory	NN	O	O
signals	NN	O	O
transduced	NN	O	O
by	NN	O	O
the	NN	O	O
JNK	NN	O	O
and	NN	O	O
p38	NN	O	O
MAP	NN	O	O
kinase	NN	O	O
pathways	NN	O	O
.	NN	O	O

To	NN	O	O
better	NN	O	O
define	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
ATF-2	NN	O	B-protein
in	NN	O	O
inflammation	NN	O	O
,	NN	O	O
adult	NN	O	O
mice	NN	O	O
expressing	NN	O	O
small	NN	O	O
amounts	NN	O	O
of	NN	O	O
a	NN	O	O
mutant	NN	O	B-protein
ATF-2	NN	O	I-protein
protein	NN	O	I-protein
were	NN	O	O
challenged	NN	O	O
with	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
,	NN	O	O
anti-CD3	NN	O	B-protein
antibody	NN	O	I-protein
or	NN	O	O
virus	NN	O	O
.	NN	O	O

Within	NN	O	O
3	NN	O	O
h	NN	O	O
of	NN	O	O
challenge	NN	O	O
by	NN	O	O
LPS	NN	O	O
,	NN	O	O
ATF-2	NN	O	O
mutant	NN	O	O
mice	NN	O	O
had	NN	O	O
decreased	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
adhesion	NN	O	B-protein
molecules	NN	O	I-protein
E-selectin	NN	O	B-protein
,	NN	O	O
P-selectin	NN	O	B-protein
and	NN	O	O
VCAM-1	NN	O	B-protein
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
cytokines	NN	O	B-protein
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
,	NN	O	O
IL-1beta	NN	O	B-protein
and	NN	O	O
IL-6	NN	O	B-protein
compared	NN	O	O
with	NN	O	O
control	NN	O	O
mice	NN	O	O
.	NN	O	O

Stimulation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
by	NN	O	O
anti-CD3	NN	O	B-protein
antibody	NN	O	I-protein
also	NN	O	O
showed	NN	O	O
less	NN	O	O
induction	NN	O	O
of	NN	O	O
IL-1	NN	O	B-protein
and	NN	O	O
IL-6	NN	O	B-protein
in	NN	O	O
ATF-2	NN	O	O
mutant	NN	O	O
tissues	NN	O	O
.	NN	O	O

ATF-2	NN	O	B-cell_type
mutant	NN	O	I-cell_type
thymocytes	NN	O	I-cell_type
treated	NN	O	O
with	NN	O	O
anti-CD3	NN	O	B-protein
antibody	NN	O	I-protein
in	NN	O	O
vitro	NN	O	O
demonstrated	NN	O	O
reduced	NN	O	O
induction	NN	O	O
of	NN	O	O
c-Jun	NN	O	B-protein
,	NN	O	O
JunB	NN	O	B-protein
,	NN	O	O
JunD	NN	O	B-protein
and	NN	O	O
Fra-2	NN	O	B-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
similar	NN	O	O
to	NN	O	O
what	NN	O	O
was	NN	O	O
observed	NN	O	O
after	NN	O	O
p38	NN	O	O
kinase	NN	O	O
inhibition	NN	O	O
in	NN	O	O
normal	NN	O	O
mice	NN	O	O
,	NN	O	O
relative	NN	O	O
ATF-2	NN	O	O
deficiency	NN	O	O
did	NN	O	O
not	NN	O	O
prevent	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
a	NN	O	O
mononuclear	NN	O	B-cell_type
cell	NN	O	I-cell_type
infiltrate	NN	O	O
in	NN	O	O
the	NN	O	O
week	NN	O	O
following	NN	O	O
an	NN	O	O
inflammatory	NN	O	O
stimulus	NN	O	O
.	NN	O	O

ATF-2	NN	O	O
mutant	NN	O	O
mice	NN	O	O
proved	NN	O	O
more	NN	O	O
susceptible	NN	O	O
to	NN	O	O
death	NN	O	O
than	NN	O	O
control	NN	O	O
mice	NN	O	O
from	NN	O	O
LPS	NN	O	O
plus	NN	O	O
D-galactosamine	NN	O	O
injection	NN	O	O
or	NN	O	O
Coxsackievirus	NN	O	O
B3	NN	O	O
infection	NN	O	O
and	NN	O	O
had	NN	O	O
a	NN	O	O
higher	NN	O	O
incidence	NN	O	O
of	NN	O	O
mononuclear	NN	O	O
pulmonary	NN	O	O
infiltrates	NN	O	O
after	NN	O	O
exposure	NN	O	O
to	NN	O	O
Herpes	NN	O	O
simplex	NN	O	O
virus-1	NN	O	O
.	NN	O	O

ATF-2	NN	O	B-protein
is	NN	O	O
essential	NN	O	O
for	NN	O	O
maximal	NN	O	O
immediate	NN	O	O
induction	NN	O	O
of	NN	O	O
adhesion	NN	O	B-protein
molecules	NN	O	I-protein
and	NN	O	O
cytokine	NN	O	B-DNA
genes	NN	O	I-DNA
,	NN	O	O
but	NN	O	O
at	NN	O	O
later	NN	O	O
time	NN	O	O
points	NN	O	O
may	NN	O	O
even	NN	O	O
protect	NN	O	O
against	NN	O	O
overactive	NN	O	O
immune	NN	O	O
responses	NN	O	O
.	NN	O	O

-DOCSTART-	O

LIGHT	NN	O	B-protein
,	NN	O	O
a	NN	O	O
TNF-like	NN	O	B-protein
molecule	NN	O	I-protein
,	NN	O	O
costimulates	NN	O	O
T	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
and	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
dendritic	NN	O	B-cell_type
cell	NN	O	I-cell_type
-mediated	NN	O	O
allogeneic	NN	O	O
T	NN	O	O
cell	NN	O	O
response	NN	O	O
.	NN	O	O

LIGHT	NN	O	B-protein
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
TNF	NN	O	B-protein
superfamily	NN	O	I-protein
and	NN	O	O
its	NN	O	O
receptors	NN	O	O
,	NN	O	O
herpesvirus	NN	O	B-protein
entry	NN	O	I-protein
mediator	NN	O	I-protein
and	NN	O	O
lymphotoxin	NN	O	B-protein
beta	NN	O	I-protein
receptor	NN	O	I-protein
,	NN	O	O
are	NN	O	O
found	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
stromal	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
LIGHT	NN	O	B-protein
is	NN	O	O
selectively	NN	O	O
expressed	NN	O	O
on	NN	O	O
immature	NN	O	B-cell_type
dendritic	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
DCs	NN	O	B-cell_type
)	NN	O	O
generated	NN	O	O
from	NN	O	O
human	NN	O	B-cell_type
PBMCs	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
LIGHT	NN	O	B-protein
is	NN	O	O
not	NN	O	O
detectable	NN	O	O
in	NN	O	O
DCs	NN	O	B-cell_type
either	NN	O	O
freshly	NN	O	O
isolated	NN	O	O
from	NN	O	O
PBMCs	NN	O	B-cell_type
or	NN	O	O
rendered	NN	O	O
mature	NN	O	O
in	NN	O	O
vitro	NN	O	O
by	NN	O	O
LPS	NN	O	O
treatment	NN	O	O
.	NN	O	O

Blockade	NN	O	O
of	NN	O	O
LIGHT	NN	O	B-protein
by	NN	O	O
its	NN	O	O
soluble	NN	O	B-protein
receptors	NN	O	I-protein
,	NN	O	O
lymphotoxin	NN	O	B-protein
beta	NN	O	I-protein
receptor-Ig	NN	O	I-protein
or	NN	O	O
HVEM-Ig	NN	O	B-protein
,	NN	O	O
inhibits	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
DC	NN	O	B-cell_type
-mediated	NN	O	O
primary	NN	O	O
allogeneic	NN	O	O
T	NN	O	O
cell	NN	O	O
response	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
engagement	NN	O	O
of	NN	O	O
LIGHT	NN	O	B-protein
costimulates	NN	O	O
human	NN	O	O
T	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
,	NN	O	O
amplifies	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
,	NN	O	O
and	NN	O	O
preferentially	NN	O	O
induces	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
IFN-gamma	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
an	NN	O	O
antigenic	NN	O	O
signal	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
LIGHT	NN	O	B-protein
is	NN	O	O
a	NN	O	O
costimulatory	NN	O	B-protein
molecule	NN	O	I-protein
involved	NN	O	O
in	NN	O	O
DC	NN	O	B-cell_type
-mediated	NN	O	O
cellular	NN	O	O
immune	NN	O	O
responses	NN	O	O
.	NN	O	O

-DOCSTART-	O

Suppression	NN	O	O
of	NN	O	O
HIV	NN	O	O
type	NN	O	O
1	NN	O	O
replication	NN	O	O
by	NN	O	O
a	NN	O	O
dominant-negative	NN	O	B-protein
Ets-1	NN	O	I-protein
mutant	NN	O	I-protein
.	NN	O	O

Activity	NN	O	O
of	NN	O	O
the	NN	O	O
distal	NN	O	B-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
(	NN	O	O
HIV-1	NN	O	O
)	NN	O	O
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
which	NN	O	O
contains	NN	O	O
binding	NN	O	B-DNA
sites	NN	O	I-DNA
for	NN	O	O
the	NN	O	O
Ets-1	NN	O	B-protein
and	NN	O	I-protein
USF-1	NN	O	I-protein
proteins	NN	O	I-protein
,	NN	O	O
is	NN	O	O
integral	NN	O	O
for	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
.	NN	O	O

The	NN	O	O
Ets-1	NN	O	B-protein
and	NN	O	I-protein
USF-1	NN	O	I-protein
proteins	NN	O	I-protein
play	NN	O	O
a	NN	O	O
critical	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
LTR	NN	O	I-DNA
distal	NN	O	I-DNA
enhancer	NN	O	I-DNA
region	NN	O	I-DNA
,	NN	O	O
as	NN	O	O
indicated	NN	O	O
by	NN	O	O
the	NN	O	O
potent	NN	O	O
dominant	NN	O	O
negative	NN	O	O
effect	NN	O	O
of	NN	O	O
a	NN	O	O
mutant	NN	O	B-DNA
Ets-1	NN	O	I-DNA
lacking	NN	O	I-DNA
trans-activation	NN	O	I-DNA
domains	NN	O	I-DNA
on	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
LTR	NN	O	B-DNA
.	NN	O	O

To	NN	O	O
determine	NN	O	O
the	NN	O	O
biological	NN	O	O
relevance	NN	O	O
of	NN	O	O
the	NN	O	O
Ets-1	NN	O	B-protein
and	NN	O	I-protein
USF-1	NN	O	I-protein
proteins	NN	O	I-protein
in	NN	O	O
HIV-1	NN	O	O
replication	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
expression	NN	O	O
of	NN	O	O
the	NN	O	O
dominant-negative	NN	O	O
mutant	NN	O	O
of	NN	O	O
Ets-1	NN	O	B-DNA
(	NN	O	O
dnEts-1	NN	O	B-DNA
)	NN	O	O
on	NN	O	O
HIV-1	NN	O	O
infection	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
expression	NN	O	O
of	NN	O	O
dnEts	NN	O	B-DNA
markedly	NN	O	O
suppressed	NN	O	O
HIV-1	NN	O	O
infection	NN	O	O
of	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
finding	NN	O	O
indicates	NN	O	O
that	NN	O	O
formation	NN	O	O
of	NN	O	O
a	NN	O	O
transcriptionaly	NN	O	B-protein
active	NN	O	I-protein
USF-1/Ets-1	NN	O	I-protein
complex	NN	O	I-protein
is	NN	O	O
important	NN	O	O
in	NN	O	O
the	NN	O	O
productive	NN	O	O
infection	NN	O	O
of	NN	O	O
cells	NN	O	O
by	NN	O	O
HIV-1	NN	O	O
,	NN	O	O
and	NN	O	O
suggests	NN	O	O
that	NN	O	O
inhibition	NN	O	O
of	NN	O	O
the	NN	O	O
interaction	NN	O	O
between	NN	O	O
USF-1	NN	O	B-protein
and	NN	O	O
Ets-1	NN	O	B-protein
with	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
LTR	NN	O	I-DNA
may	NN	O	O
provide	NN	O	O
a	NN	O	O
new	NN	O	O
target	NN	O	O
for	NN	O	O
anti-HIV-1	NN	O	O
gene	NN	O	O
therapy	NN	O	O
.	NN	O	O

-DOCSTART-	O

Human	NN	O	B-cell_type
eosinophils	NN	O	I-cell_type
constitutively	NN	O	O
express	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
p	NN	O	I-protein
and	NN	O	I-protein
c	NN	O	I-protein
.	NN	O	O

BACKGROUND	NN	O	O
:	NN	O	O
Eosinophils	NN	O	B-cell_type
are	NN	O	O
now	NN	O	O
known	NN	O	O
to	NN	O	O
produce	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
proinflammatory	NN	O	B-protein
cytokines	NN	O	I-protein
,	NN	O	O
although	NN	O	O
the	NN	O	O
molecular	NN	O	O
factors	NN	O	O
that	NN	O	O
regulate	NN	O	O
their	NN	O	O
production	NN	O	O
are	NN	O	O
poorly	NN	O	O
understood	NN	O	O
.	NN	O	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
almost	NN	O	O
all	NN	O	O
of	NN	O	O
the	NN	O	O
cytokines	NN	O	B-protein
produced	NN	O	O
by	NN	O	O
eosinophils	NN	O	B-cell_type
,	NN	O	O
including	NN	O	O
the	NN	O	O
proallergic	NN	O	B-protein
cytokine	NN	O	I-protein
IL-4	NN	O	I-protein
,	NN	O	O
is	NN	O	O
now	NN	O	O
known	NN	O	O
to	NN	O	O
be	NN	O	O
regulated	NN	O	O
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
transcription	NN	O	O
by	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
(	NN	O	O
NFAT	NN	O	B-protein
)	NN	O	O
family	NN	O	O
of	NN	O	O
transcription	NN	O	O
factors	NN	O	O
.	NN	O	O

OBJECTIVE	NN	O	O
:	NN	O	O
We	NN	O	O
sought	NN	O	O
to	NN	O	O
characterize	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
different	NN	O	O
NFAT	NN	O	B-protein
proteins	NN	O	I-protein
in	NN	O	O
resting	NN	O	B-cell_type
and	NN	O	I-cell_type
activated	NN	O	I-cell_type
eosinophils	NN	O	I-cell_type
.	NN	O	O

METHODS	NN	O	O
:	NN	O	O
Nuclear	NN	O	O
and	NN	O	O
whole	NN	O	O
cell	NN	O	O
extracts	NN	O	O
were	NN	O	O
obtained	NN	O	O
from	NN	O	O
both	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
eosinophils	NN	O	I-cell_type
and	NN	O	O
those	NN	O	O
obtained	NN	O	O
from	NN	O	O
bronchoalveolar	NN	O	O
lavage	NN	O	O
fluid	NN	O	O
of	NN	O	O
asthmatic	NN	O	O
subjects	NN	O	O
after	NN	O	O
endobronchial	NN	O	O
allergen	NN	O	O
challenge	NN	O	O
.	NN	O	O

NFAT	NN	O	B-protein
expression	NN	O	O
was	NN	O	O
determined	NN	O	O
by	NN	O	O
using	NN	O	O
immunoprecipitation	NN	O	O
and	NN	O	O
Western	NN	O	O
blot	NN	O	O
analysis	NN	O	O
,	NN	O	O
DNA-binding	NN	O	O
assays	NN	O	O
,	NN	O	O
and	NN	O	O
RT-PCR	NN	O	O
analysis	NN	O	O
of	NN	O	O
eosinophil	NN	O	B-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
Both	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
and	NN	O	I-cell_type
bronchoalveolar	NN	O	I-cell_type
lavage	NN	O	I-cell_type
fluid	NN	O	I-cell_type
eosinophils	NN	O	I-cell_type
expressed	NN	O	O
NFATp	NN	O	B-protein
and	NN	O	I-protein
NFATc	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

Unlike	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
which	NN	O	O
express	NN	O	O
multiple	NN	O	O
NFATc	NN	O	O
isoforms	NN	O	O
,	NN	O	O
eosinophils	NN	O	B-cell_type
preferentially	NN	O	O
express	NN	O	O
the	NN	O	O
approximately	NN	O	O
85-kd	NN	O	O
isoform	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
eosinophils	NN	O	B-cell_type
were	NN	O	O
found	NN	O	O
to	NN	O	O
constitutively	NN	O	O
express	NN	O	O
NFATc	NN	O	B-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

A	NN	O	O
brief	NN	O	O
incubation	NN	O	O
with	NN	O	O
the	NN	O	O
T	NN	O	B-protein
(	NN	O	I-protein
H	NN	O	I-protein
)	NN	O	I-protein
2	NN	O	I-protein
cytokines	NN	O	I-protein
IL-4	NN	O	B-protein
and	NN	O	O
IL-5	NN	O	B-protein
was	NN	O	O
sufficient	NN	O	O
to	NN	O	O
induce	NN	O	O
the	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
NFATc	NN	O	B-protein
.	NN	O	O

Eosinophil	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
contain	NN	O	O
multiple	NN	O	O
factors	NN	O	O
that	NN	O	O
can	NN	O	O
specifically	NN	O	O
recognize	NN	O	O
the	NN	O	O
IL-4	NN	O	B-DNA
promoter	NN	O	I-DNA
P1	NN	O	I-DNA
NFAT	NN	O	I-DNA
site	NN	O	I-DNA
in	NN	O	O
DNA-binding	NN	O	O
assays	NN	O	O
,	NN	O	O
including	NN	O	O
NFATp	NN	O	B-protein
.	NN	O	O

CONCLUSION	NN	O	O
:	NN	O	O
NFATp	NN	O	B-protein
and	NN	O	O
NFATc	NN	O	B-protein
can	NN	O	O
regulate	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
and	NN	O	O
other	NN	O	O
genes	NN	O	O
in	NN	O	O
eosinophils	NN	O	B-cell_type
but	NN	O	O
appear	NN	O	O
to	NN	O	O
be	NN	O	O
regulated	NN	O	O
by	NN	O	O
a	NN	O	O
novel	NN	O	O
signal	NN	O	O
transduction	NN	O	O
mechanism	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

-DOCSTART-	O

BLyS	NN	O	B-protein
BINDS	NN	O	O
TO	NN	O	O
B	NN	O	B-cell_type
CELLS	NN	O	I-cell_type
WITH	NN	O	O
HIGH	NN	O	O
AFFINITY	NN	O	O
AND	NN	O	O
INDUCES	NN	O	O
ACTIVATION	NN	O	O
OF	NN	O	O
THE	NN	O	O
TRANSCRIPTION	NN	O	B-protein
FACTORS	NN	O	I-protein
NF-kappaB	NN	O	B-protein
AND	NN	O	O
ELF-1	NN	O	B-protein
.	NN	O	O

B	NN	O	B-protein
lymphocyte	NN	O	I-protein
stimulator	NN	O	I-protein
(	NN	O	O
BLyS	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
novel	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
TNF	NN	O	B-protein
family	NN	O	I-protein
of	NN	O	I-protein
proteins	NN	O	I-protein
expressed	NN	O	O
by	NN	O	O
myeloid	NN	O	B-cell_type
cells	NN	O	I-cell_type
as	NN	O	O
membrane-bound	NN	O	O
and	NN	O	O
soluble	NN	O	O
forms	NN	O	O
.	NN	O	O

BLyS	NN	O	B-protein
was	NN	O	O
shown	NN	O	O
to	NN	O	O
act	NN	O	O
specifically	NN	O	O
on	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
inducing	NN	O	O
proliferation	NN	O	O
and	NN	O	O
immunoglobulin	NN	O	O
production	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

The	NN	O	O
present	NN	O	O
study	NN	O	O
was	NN	O	O
undertaken	NN	O	O
to	NN	O	O
characterize	NN	O	O
binding	NN	O	O
of	NN	O	O
radiolabeled	NN	O	B-protein
BLyS	NN	O	I-protein
to	NN	O	O
its	NN	O	O
cognate	NN	O	B-protein
receptor	NN	O	I-protein
on	NN	O	O
human	NN	O	B-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
examine	NN	O	O
intracellular	NN	O	O
events	NN	O	O
initiated	NN	O	O
by	NN	O	O
BLyS	NN	O	B-protein
binding	NN	O	O
.	NN	O	O

Similar	NN	O	O
to	NN	O	O
other	NN	O	O
TNF	NN	O	B-protein
family	NN	O	I-protein
members	NN	O	I-protein
,	NN	O	O
BLyS	NN	O	B-protein
is	NN	O	O
present	NN	O	O
in	NN	O	O
solution	NN	O	O
as	NN	O	O
a	NN	O	O
homotrimer	NN	O	B-protein
as	NN	O	O
determined	NN	O	O
by	NN	O	O
gel	NN	O	O
filtration	NN	O	O
chromatography	NN	O	O
and	NN	O	O
light	NN	O	O
scattering	NN	O	O
analysis	NN	O	O
.	NN	O	O

BLyS	NN	O	B-protein
binding	NN	O	O
to	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
specific	NN	O	O
as	NN	O	O
other	NN	O	O
TNF	NN	O	B-protein
family	NN	O	I-protein
members	NN	O	I-protein
tested	NN	O	O
did	NN	O	O
not	NN	O	O
compete	NN	O	O
for	NN	O	O
(	NN	O	O
125	NN	O	O
)	NN	O	O
I-	NN	O	O
BLyS	NN	O	B-protein
binding	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
equilibrium	NN	O	O
binding	NN	O	O
of	NN	O	O
(	NN	O	B-protein
125	NN	O	I-protein
)	NN	O	I-protein
I-labeled	NN	O	I-protein
BLyS	NN	O	I-protein
to	NN	O	O
purified	NN	O	O
human	NN	O	B-cell_type
tonsillar	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
demonstrated	NN	O	O
saturable	NN	O	O
binding	NN	O	O
.	NN	O	O

Scatchard	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
binding	NN	O	O
data	NN	O	O
revealed	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
on	NN	O	O
human	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
approximately	NN	O	O
2600	NN	O	O
binding	NN	O	O
sites	NN	O	O
per	NN	O	O
cell	NN	O	O
and	NN	O	O
an	NN	O	O
apparent	NN	O	O
dissociation	NN	O	O
constant	NN	O	O
(	NN	O	O
K	NN	O	O
(	NN	O	O
D	NN	O	O
)	NN	O	O
)	NN	O	O
of	NN	O	O
about	NN	O	O
0.1	NN	O	O
nM	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
BLyS	NN	O	B-protein
binding	NN	O	O
to	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
results	NN	O	O
in	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
and	NN	O	O
the	NN	O	O
Ets	NN	O	B-protein
family	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
ELF-1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
in	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
mRNA	NN	O	O
for	NN	O	O
Polo-like	NN	O	B-protein
kinase	NN	O	I-protein
(	NN	O	O
PLK	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Copyright	NN	O	O
2001	NN	O	O
Academic	NN	O	O
Press	NN	O	O
.	NN	O	O

-DOCSTART-	O

Design	NN	O	O
and	NN	O	O
use	NN	O	O
of	NN	O	O
an	NN	O	O
inducibly	NN	O	O
activated	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
Nef	NN	O	B-protein
to	NN	O	O
study	NN	O	O
immune	NN	O	O
modulation	NN	O	O
.	NN	O	O

The	NN	O	O
Nef	NN	O	B-protein
protein	NN	O	I-protein
of	NN	O	O
the	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
(	NN	O	O
HIV-1	NN	O	O
)	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
infectivity	NN	O	O
of	NN	O	O
virus	NN	O	O
particles	NN	O	O
,	NN	O	O
downmodulate	NN	O	O
cell	NN	O	B-protein
surface	NN	O	I-protein
proteins	NN	O	I-protein
,	NN	O	O
and	NN	O	O
associate	NN	O	O
with	NN	O	O
many	NN	O	O
intracellular	NN	O	B-protein
proteins	NN	O	I-protein
that	NN	O	O
are	NN	O	O
thought	NN	O	O
to	NN	O	O
facilitate	NN	O	O
HIV	NN	O	O
infection	NN	O	O
.	NN	O	O

One	NN	O	O
of	NN	O	O
the	NN	O	O
challenges	NN	O	O
in	NN	O	O
defining	NN	O	O
the	NN	O	O
molecular	NN	O	O
events	NN	O	O
regulated	NN	O	O
by	NN	O	O
Nef	NN	O	B-protein
has	NN	O	O
been	NN	O	O
obtaining	NN	O	O
good	NN	O	O
expression	NN	O	O
of	NN	O	O
Nef	NN	O	B-protein
protein	NN	O	I-protein
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

This	NN	O	O
has	NN	O	O
been	NN	O	O
attributed	NN	O	O
to	NN	O	O
effects	NN	O	O
of	NN	O	O
Nef	NN	O	B-protein
on	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
and	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
designed	NN	O	O
a	NN	O	O
Nef	NN	O	B-protein
protein	NN	O	I-protein
that	NN	O	O
is	NN	O	O
readily	NN	O	O
expressed	NN	O	O
in	NN	O	O
T-cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
and	NN	O	O
whose	NN	O	O
function	NN	O	O
is	NN	O	O
inducibly	NN	O	O
activated	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
composed	NN	O	O
of	NN	O	O
a	NN	O	O
fusion	NN	O	O
between	NN	O	O
full-length	NN	O	B-protein
Nef	NN	O	I-protein
and	NN	O	O
the	NN	O	O
estrogen	NN	O	B-protein
receptor	NN	O	I-protein
hormone-binding	NN	O	I-protein
domain	NN	O	I-protein
(	NN	O	O
Nef-ER	NN	O	B-protein
)	NN	O	O
.	NN	O	O

The	NN	O	O
Nef-ER	NN	O	B-protein
is	NN	O	O
kept	NN	O	O
in	NN	O	O
an	NN	O	O
inactive	NN	O	O
state	NN	O	O
due	NN	O	O
to	NN	O	O
steric	NN	O	O
hindrance	NN	O	O
,	NN	O	O
and	NN	O	O
addition	NN	O	O
of	NN	O	O
the	NN	O	O
membrane-permeable	NN	O	O
drug	NN	O	O
4-hydroxytamoxifen	NN	O	O
(	NN	O	O
4-HT	NN	O	O
)	NN	O	O
,	NN	O	O
which	NN	O	O
binds	NN	O	O
to	NN	O	O
the	NN	O	O
ER	NN	O	B-protein
domain	NN	O	I-protein
,	NN	O	O
leads	NN	O	O
to	NN	O	O
inducible	NN	O	O
activation	NN	O	O
of	NN	O	O
Nef-ER	NN	O	B-protein
within	NN	O	O
cells	NN	O	O
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
Nef-ER	NN	O	B-protein
inducibly	NN	O	O
associates	NN	O	O
with	NN	O	O
the	NN	O	O
62-kDa	NN	O	B-protein
Ser/Thr	NN	O	I-protein
kinase	NN	O	I-protein
and	NN	O	O
is	NN	O	O
localized	NN	O	O
to	NN	O	O
specific	NN	O	O
membrane	NN	O	O
microdomains	NN	O	O
(	NN	O	O
lipid	NN	O	O
rafts	NN	O	O
)	NN	O	O
only	NN	O	O
after	NN	O	O
activation	NN	O	O
.	NN	O	O

Using	NN	O	O
this	NN	O	O
inducible	NN	O	O
Nef	NN	O	B-protein
,	NN	O	O
we	NN	O	O
also	NN	O	O
compared	NN	O	O
the	NN	O	O
specific	NN	O	O
requirements	NN	O	O
for	NN	O	O
CD4	NN	O	B-protein
and	NN	O	O
HLA-A2	NN	O	B-protein
downmodulation	NN	O	O
in	NN	O	O
a	NN	O	O
SupT1	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

Half-maximal	NN	O	O
downmodulation	NN	O	O
of	NN	O	O
cell	NN	O	B-protein
surface	NN	O	I-protein
CD4	NN	O	I-protein
required	NN	O	O
very	NN	O	O
little	NN	O	O
active	NN	O	O
Nef-ER	NN	O	B-protein
and	NN	O	O
occurred	NN	O	O
as	NN	O	O
early	NN	O	O
as	NN	O	O
4	NN	O	O
h	NN	O	O
after	NN	O	O
addition	NN	O	O
of	NN	O	O
4-HT	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
50	NN	O	O
%	NN	O	O
downmodulation	NN	O	O
of	NN	O	O
HLA-A2	NN	O	B-protein
by	NN	O	O
Nef	NN	O	B-protein
required	NN	O	O
16	NN	O	O
to	NN	O	O
24	NN	O	O
h	NN	O	O
and	NN	O	O
about	NN	O	O
50-	NN	O	O
to	NN	O	O
100-fold-greater	NN	O	O
concentrations	NN	O	O
of	NN	O	O
4-HT	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
HLA-A2	NN	O	B-protein
downmodulation	NN	O	O
may	NN	O	O
require	NN	O	O
certain	NN	O	O
threshold	NN	O	O
levels	NN	O	O
of	NN	O	O
active	NN	O	O
Nef	NN	O	B-protein
.	NN	O	O

The	NN	O	O
differential	NN	O	O
timing	NN	O	O
of	NN	O	O
CD4	NN	O	B-protein
and	NN	O	O
HLA-A2	NN	O	B-protein
downmodulation	NN	O	O
may	NN	O	O
have	NN	O	O
implications	NN	O	O
for	NN	O	O
HIV	NN	O	O
pathogenesis	NN	O	O
and	NN	O	O
immune	NN	O	O
evasion	NN	O	O
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
chemokine	NN	O	O
mRNA	NN	O	O
expression	NN	O	O
in	NN	O	O
a	NN	O	O
rat	NN	O	O
model	NN	O	O
of	NN	O	O
vanadium-induced	NN	O	O
pulmonary	NN	O	O
inflammation	NN	O	O
.	NN	O	O

Environmental	NN	O	O
and	NN	O	O
occupational	NN	O	O
exposure	NN	O	O
to	NN	O	O
vanadium	NN	O	O
dusts	NN	O	O
results	NN	O	O
in	NN	O	O
toxic	NN	O	O
effects	NN	O	O
mainly	NN	O	O
confined	NN	O	O
to	NN	O	O
the	NN	O	O
respiratory	NN	O	O
system	NN	O	O
.	NN	O	O

Using	NN	O	O
a	NN	O	O
rat	NN	O	O
model	NN	O	O
of	NN	O	O
acute	NN	O	O
lung	NN	O	O
inflammation	NN	O	O
induced	NN	O	O
by	NN	O	O
intratracheal	NN	O	O
instillation	NN	O	O
of	NN	O	O
sodium	NN	O	O
metavanadate	NN	O	O
(	NN	O	O
NaVO3	NN	O	O
)	NN	O	O
at	NN	O	O
the	NN	O	O
dose	NN	O	O
of	NN	O	O
200	NN	O	O
microg	NN	O	O
V/kg	NN	O	O
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
the	NN	O	O
relationship	NN	O	O
between	NN	O	O
the	NN	O	O
cytologic	NN	O	O
characterization	NN	O	O
of	NN	O	O
pulmonary	NN	O	O
inflammation	NN	O	O
and	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
chemokine	NN	O	B-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

Significant	NN	O	O
polymorphonuclear	NN	O	B-cell_type
leukocyte	NN	O	I-cell_type
(	NN	O	O
PMN	NN	O	B-cell_type
)	NN	O	O
influx	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.01	NN	O	O
)	NN	O	O
into	NN	O	O
the	NN	O	O
lung	NN	O	O
was	NN	O	O
noted	NN	O	O
4	NN	O	O
h	NN	O	O
after	NN	O	O
NaVO3	NN	O	O
instillation	NN	O	O
,	NN	O	O
whereas	NN	O	O
alveolar	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
(	NN	O	O
AMs	NN	O	B-cell_type
)	NN	O	O
in	NN	O	O
bronchoalveolar	NN	O	B-cell_type
lavage	NN	O	I-cell_type
(	NN	O	I-cell_type
BAL	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
appeared	NN	O	O
to	NN	O	O
decrease	NN	O	O
significantly	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
neither	NN	O	O
PMNs	NN	O	B-cell_type
nor	NN	O	O
AMs	NN	O	B-cell_type
changed	NN	O	O
substantially	NN	O	O
1	NN	O	O
h	NN	O	O
after	NN	O	O
NaVO3	NN	O	O
instillation	NN	O	O
.	NN	O	O

By	NN	O	O
Northern	NN	O	O
analysis	NN	O	O
,	NN	O	O
macrophage	NN	O	B-RNA
inflammatory	NN	O	I-RNA
protein	NN	O	I-RNA
(	NN	O	I-RNA
MIP	NN	O	I-RNA
)	NN	O	I-RNA
-2	NN	O	I-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
BAL	NN	O	B-cell_type
cells	NN	O	I-cell_type
increased	NN	O	O
markedly	NN	O	O
1	NN	O	O
h	NN	O	O
after	NN	O	O
NaVO3	NN	O	O
instillation	NN	O	O
and	NN	O	O
reduced	NN	O	O
a	NN	O	O
little	NN	O	O
bit	NN	O	O
at	NN	O	O
4	NN	O	O
h	NN	O	O
,	NN	O	O
whereas	NN	O	O
MIP-1alpha	NN	O	B-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
BAL	NN	O	B-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
expressed	NN	O	O
relatively	NN	O	O
high	NN	O	O
1	NN	O	O
h	NN	O	O
after	NN	O	O
NaVO3	NN	O	O
instillation	NN	O	O
,	NN	O	O
although	NN	O	O
a	NN	O	O
basal	NN	O	O
expression	NN	O	O
was	NN	O	O
detected	NN	O	O
in	NN	O	O
control	NN	O	O
group	NN	O	O
,	NN	O	O
and	NN	O	O
returned	NN	O	O
rapidly	NN	O	O
nearly	NN	O	O
to	NN	O	O
control	NN	O	O
level	NN	O	O
at	NN	O	O
4	NN	O	O
h	NN	O	O
.	NN	O	O

Since	NN	O	O
MIP-2	NN	O	B-protein
is	NN	O	O
a	NN	O	O
potent	NN	O	O
PMN	NN	O	B-cell_type
chemoattractant	NN	O	O
and	NN	O	O
MIP-1alpha	NN	O	B-protein
is	NN	O	O
a	NN	O	O
potent	NN	O	O
macrophage/monocyte	NN	O	O
chemoattractant	NN	O	O
has	NN	O	O
been	NN	O	O
well	NN	O	O
known	NN	O	O
.	NN	O	O

The	NN	O	O
facts	NN	O	O
that	NN	O	O
PMN	NN	O	B-cell_type
influx	NN	O	O
was	NN	O	O
preceded	NN	O	O
by	NN	O	O
increased	NN	O	O
MIP-2	NN	O	O
mRNA	NN	O	O
expression	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
MIP-2	NN	O	B-protein
is	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
NaVO3-induced	NN	O	O
pulmonary	NN	O	O
inflammation	NN	O	O
,	NN	O	O
whereas	NN	O	O
increased	NN	O	O
MIP-1alpha	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
was	NN	O	O
followed	NN	O	O
by	NN	O	O
decreased	NN	O	O
AMs	NN	O	B-cell_type
in	NN	O	O
BAL	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
suggesting	NN	O	O
AMs	NN	O	B-cell_type
might	NN	O	O
be	NN	O	O
activated	NN	O	O
by	NN	O	O
MIP-1alpha	NN	O	B-protein
,	NN	O	O
adherent	NN	O	O
to	NN	O	O
the	NN	O	O
lining	NN	O	O
surface	NN	O	O
of	NN	O	O
the	NN	O	O
airways	NN	O	O
and	NN	O	O
then	NN	O	O
resistant	NN	O	O
to	NN	O	O
be	NN	O	O
washed	NN	O	O
out	NN	O	O
.	NN	O	O

To	NN	O	O
delineate	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
,	NN	O	O
we	NN	O	O
recently	NN	O	O
cloned	NN	O	O
the	NN	O	O
5'-flanking	NN	O	B-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
MIP-2	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
promotor	NN	O	B-DNA
region	NN	O	I-DNA
contains	NN	O	O
consensus	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
for	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
nuclear	NN	O	I-protein
factor	NN	O	I-protein
kappaB	NN	O	I-protein
(	NN	O	O
NF-kappaB	NN	O	B-protein
)	NN	O	O
and	NN	O	O
activator	NN	O	B-protein
protein-1	NN	O	I-protein
(	NN	O	O
AP-1	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Using	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
,	NN	O	O
increased	NN	O	O
nuclear	NN	O	O
NF-kappaB	NN	O	B-protein
,	NN	O	O
not	NN	O	O
AP-1	NN	O	B-protein
,	NN	O	O
binding	NN	O	O
activity	NN	O	O
was	NN	O	O
detected	NN	O	O
1	NN	O	O
h	NN	O	O
after	NN	O	O
NaVO3	NN	O	O
instillation	NN	O	O
,	NN	O	O
which	NN	O	O
correlated	NN	O	O
with	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
MIP-2	NN	O	B-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

p65	NN	O	B-protein
(	NN	O	I-protein
Rel	NN	O	I-protein
A	NN	O	I-protein
)	NN	O	I-protein
and	NN	O	O
p50	NN	O	B-protein
protein	NN	O	I-protein
appears	NN	O	O
to	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
MIP-2	NN	O	O
NF-kappaB	NN	O	B-protein
binding	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
suggest	NN	O	O
that	NN	O	O
MIP-2	NN	O	B-protein
is	NN	O	O
an	NN	O	O
important	NN	O	O
mediator	NN	O	O
of	NN	O	O
NaVO3-induced	NN	O	O
pulmonary	NN	O	O
inflammation	NN	O	O
in	NN	O	O
the	NN	O	O
rat	NN	O	O
model	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
elevated	NN	O	O
MIP-2	NN	O	B-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
are	NN	O	O
accompanied	NN	O	O
by	NN	O	O
increased	NN	O	O
NF-kappaB	NN	O	B-protein
binding	NN	O	O
activity	NN	O	O
in	NN	O	O
BAL	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
suggesting	NN	O	O
possible	NN	O	O
MIP-2	NN	O	B-protein
transcriptional	NN	O	O
regulation	NN	O	O
through	NN	O	O
NF-kappaB	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
of	NN	O	O
signal	NN	O	O
transduction	NN	O	O
and	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
healthy	NN	O	B-cell_type
lymphomonocytes	NN	O	I-cell_type
exposed	NN	O	O
to	NN	O	O
bystander	NN	O	O
HIV-1-infected	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Persistent	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
is	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
hallmarks	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
infection	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
we	NN	O	O
analysed	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
factors	NN	O	O
involved	NN	O	O
in	NN	O	O
cytokine	NN	O	O
signal	NN	O	O
transduction	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
STAT	NN	O	B-protein
1	NN	O	I-protein
proteins	NN	O	I-protein
and	NN	O	O
IRF-1	NN	O	B-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
in	NN	O	O
normal	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
PBMC	NN	O	B-cell_type
)	NN	O	O
exposed	NN	O	O
to	NN	O	O
HIV-infected	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

Western	NN	O	O
blot	NN	O	O
analyses	NN	O	O
and	NN	O	O
reverse	NN	O	O
transcriptase-polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
both	NN	O	O
cells	NN	O	O
infected	NN	O	O
with	NN	O	O
a	NN	O	O
X4	NN	O	O
strain	NN	O	O
and	NN	O	O
cells	NN	O	O
infected	NN	O	O
with	NN	O	O
a	NN	O	O
R5	NN	O	O
strain	NN	O	O
are	NN	O	O
able	NN	O	O
to	NN	O	O
increase	NN	O	O
intracellular	NN	O	O
levels	NN	O	O
of	NN	O	O
STAT	NN	O	B-protein
1alpha	NN	O	I-protein
and	NN	O	I-protein
beta	NN	O	I-protein
proteins	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
IRF-1	NN	O	B-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

This	NN	O	O
effect	NN	O	O
was	NN	O	O
prevented	NN	O	O
by	NN	O	O
neutralizing	NN	O	O
antibodies	NN	O	O
against	NN	O	O
interferon-alpha	NN	O	B-protein
(	NN	O	O
IFN-alpha	NN	O	B-protein
)	NN	O	O
.	NN	O	O

HIV-1-infected	NN	O	B-cell_type
cells	NN	O	I-cell_type
dose-dependently	NN	O	O
induced	NN	O	O
apoptotic	NN	O	O
commitment	NN	O	O
in	NN	O	O
normal	NN	O	B-cell_type
PBMC	NN	O	I-cell_type
,	NN	O	O
as	NN	O	O
revealed	NN	O	O
by	NN	O	O
DNA	NN	O	O
fragmentation	NN	O	O
analysis	NN	O	O
,	NN	O	O
but	NN	O	O
this	NN	O	O
was	NN	O	O
not	NN	O	O
accompanied	NN	O	O
by	NN	O	O
an	NN	O	O
increase	NN	O	O
of	NN	O	O
caspase-3	NN	O	O
activity	NN	O	O
,	NN	O	O
even	NN	O	O
if	NN	O	O
a	NN	O	O
slight	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
IL-1beta-converting	NN	O	B-RNA
enzyme	NN	O	I-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
detected	NN	O	O
.	NN	O	O

Apoptosis	NN	O	O
induction	NN	O	O
could	NN	O	O
be	NN	O	O
abrogated	NN	O	O
mainly	NN	O	O
by	NN	O	O
antibodies	NN	O	B-protein
against	NN	O	O
tumour	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
and	NN	O	O
,	NN	O	O
to	NN	O	O
a	NN	O	O
lesser	NN	O	O
extent	NN	O	O
,	NN	O	O
by	NN	O	O
antibodies	NN	O	B-protein
against	NN	O	O
IFN-gamma	NN	O	B-protein
.	NN	O	O

All	NN	O	O
these	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
uninfected	NN	O	B-cell_type
PBMC	NN	O	I-cell_type
can	NN	O	O
undergo	NN	O	O
activation	NN	O	O
of	NN	O	O
signal	NN	O	O
transduction	NN	O	O
and	NN	O	O
apoptosis	NN	O	O
after	NN	O	O
exposure	NN	O	O
to	NN	O	O
bystander	NN	O	O
HIV-infected	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
subsequent	NN	O	O
to	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
such	NN	O	O
as	NN	O	O
IFNs	NN	O	B-protein
and	NN	O	O
TNF-alpha	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
physical	NN	O	O
association	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
theta	NN	O	I-protein
with	NN	O	O
a	NN	O	O
lipid	NN	O	O
raft-associated	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
kappa	NN	O	B-protein
B	NN	O	I-protein
factor	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	I-protein
IKK	NN	O	I-protein
)	NN	O	I-protein
complex	NN	O	I-protein
plays	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappa	NN	O	O
B	NN	O	O
cascade	NN	O	O
by	NN	O	O
TCR	NN	O	B-protein
and	NN	O	O
CD28	NN	O	B-protein
.	NN	O	O

We	NN	O	O
investigated	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
theta	NN	O	I-protein
(	NN	O	O
PKCtheta	NN	O	B-protein
)	NN	O	O
in	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-protein
cascade	NN	O	O
in	NN	O	O
primary	NN	O	B-cell_type
human	NN	O	I-cell_type
CD4	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Among	NN	O	O
six	NN	O	O
or	NN	O	O
so	NN	O	O
PKC	NN	O	B-protein
isoforms	NN	O	I-protein
expressed	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
only	NN	O	O
PKCtheta	NN	O	B-protein
participates	NN	O	O
in	NN	O	O
the	NN	O	O
assembly	NN	O	O
of	NN	O	O
the	NN	O	O
supramolecular	NN	O	O
activation	NN	O	O
clusters	NN	O	O
at	NN	O	O
the	NN	O	O
contact	NN	O	O
site	NN	O	O
of	NN	O	O
the	NN	O	O
TCR	NN	O	B-protein
with	NN	O	O
Ag	NN	O	B-protein
.	NN	O	O

Signaling	NN	O	O
via	NN	O	O
both	NN	O	O
the	NN	O	O
TCR	NN	O	B-protein
and	NN	O	O
CD28	NN	O	B-protein
is	NN	O	O
required	NN	O	O
for	NN	O	O
optimal	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
multisubunit	NN	O	B-protein
IkappaB	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	I-protein
IKK	NN	O	I-protein
)	NN	O	I-protein
complex	NN	O	I-protein
in	NN	O	O
primary	NN	O	B-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
;	NN	O	O
this	NN	O	O
activation	NN	O	O
could	NN	O	O
be	NN	O	O
inhibited	NN	O	O
by	NN	O	O
a	NN	O	O
Ca	NN	O	O
(	NN	O	O
2+	NN	O	O
)	NN	O	O
-independent	NN	O	O
PKC	NN	O	O
isoform	NN	O	O
inhibitor	NN	O	O
,	NN	O	O
rottlerin	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
endogenous	NN	O	O
PKCtheta	NN	O	B-protein
physically	NN	O	O
associates	NN	O	O
with	NN	O	O
activated	NN	O	O
IKK	NN	O	B-protein
complexes	NN	O	I-protein
in	NN	O	O
CD3/CD28-costimulated	NN	O	B-cell_type
primary	NN	O	I-cell_type
CD4	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
same	NN	O	O
set	NN	O	O
of	NN	O	O
stimuli	NN	O	O
also	NN	O	O
induced	NN	O	O
relocation	NN	O	O
of	NN	O	O
endogenous	NN	O	O
PKCtheta	NN	O	B-protein
and	NN	O	O
IKKs	NN	O	B-protein
to	NN	O	O
a	NN	O	O
GM1	NN	O	O
ganglioside-enriched	NN	O	O
,	NN	O	O
detergent-insoluble	NN	O	O
membrane	NN	O	O
compartment	NN	O	O
in	NN	O	O
primary	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

IKKs	NN	O	B-protein
recruited	NN	O	O
to	NN	O	O
these	NN	O	O
lipid	NN	O	O
rafts	NN	O	O
were	NN	O	O
capable	NN	O	O
of	NN	O	O
phosphorylating	NN	O	O
a	NN	O	O
recombinant	NN	O	O
IkappaBalpha	NN	O	O
sustrate	NN	O	O
.	NN	O	O

Confocal	NN	O	O
microscopy	NN	O	O
further	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
exogenously	NN	O	O
expressed	NN	O	O
PKCtheta	NN	O	B-protein
and	NN	O	O
IKKss	NN	O	B-protein
colocalize	NN	O	O
in	NN	O	O
the	NN	O	O
membrane	NN	O	O
of	NN	O	O
CD3/CD28-costimulated	NN	O	B-cell_line
Jurkat	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Constitutively	NN	O	O
active	NN	O	O
but	NN	O	O
not	NN	O	O
kinase-inactive	NN	O	B-protein
PKCtheta	NN	O	I-protein
activated	NN	O	O
IKKbeta	NN	O	B-protein
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
dominant-active	NN	O	B-protein
PKCtheta	NN	O	I-protein
also	NN	O	O
had	NN	O	O
stimulatory	NN	O	O
effects	NN	O	O
on	NN	O	O
the	NN	O	O
CD28	NN	O	B-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
these	NN	O	O
data	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
PKCtheta	NN	O	B-protein
by	NN	O	O
the	NN	O	O
TCR	NN	O	B-protein
and	NN	O	O
CD28	NN	O	B-protein
plays	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
assembly	NN	O	O
and	NN	O	O
activation	NN	O	O
of	NN	O	O
IKK	NN	O	B-protein
complexes	NN	O	I-protein
in	NN	O	O
the	NN	O	O
T	NN	O	O
cell	NN	O	O
membrane	NN	O	O

-DOCSTART-	O

T-cell	NN	O	B-cell_type
-mediated	NN	O	O
regulation	NN	O	O
of	NN	O	O
osteoclastogenesis	NN	O	O
by	NN	O	O
signalling	NN	O	O
cross-talk	NN	O	O
between	NN	O	O
RANKL	NN	O	B-protein
and	NN	O	O
IFN-gamma	NN	O	B-protein
.	NN	O	O

Bone	NN	O	O
resorption	NN	O	O
is	NN	O	O
regulated	NN	O	O
by	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
,	NN	O	O
where	NN	O	O
T-cell	NN	O	B-cell_type
expression	NN	O	O
of	NN	O	O
RANKL	NN	O	B-protein
(	NN	O	O
receptor	NN	O	B-protein
activator	NN	O	I-protein
of	NN	O	I-protein
nuclear	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	I-protein
NF	NN	O	I-protein
)	NN	O	I-protein
-kappaB	NN	O	I-protein
ligand	NN	O	I-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
tumour-necrosis	NN	O	B-protein
factor	NN	O	I-protein
family	NN	O	I-protein
that	NN	O	O
is	NN	O	O
essential	NN	O	O
for	NN	O	O
osteoclastogenesis	NN	O	O
,	NN	O	O
may	NN	O	O
contribute	NN	O	O
to	NN	O	O
pathological	NN	O	O
conditions	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
autoimmune	NN	O	O
arthritis	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
whether	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
maintain	NN	O	O
bone	NN	O	O
homeostasis	NN	O	O
by	NN	O	O
counterbalancing	NN	O	O
the	NN	O	O
action	NN	O	O
of	NN	O	O
RANKL	NN	O	B-protein
remains	NN	O	O
unknown	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
T-cell	NN	O	B-cell_type
production	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
strongly	NN	O	O
suppresses	NN	O	O
osteoclastogenesis	NN	O	O
by	NN	O	O
interfering	NN	O	O
with	NN	O	O
the	NN	O	O
RANKL	NN	O	B-protein
-RANK	NN	O	O
signalling	NN	O	O
pathway	NN	O	O
.	NN	O	O

IFN-gamma	NN	O	B-protein
induces	NN	O	O
rapid	NN	O	O
degradation	NN	O	O
of	NN	O	O
the	NN	O	O
RANK	NN	O	B-protein
adapter	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
TRAF6	NN	O	B-protein
(	NN	O	O
tumour	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
receptor-associated	NN	O	I-protein
factor	NN	O	I-protein
6	NN	O	I-protein
)	NN	O	O
,	NN	O	O
which	NN	O	O
results	NN	O	O
in	NN	O	O
strong	NN	O	O
inhibition	NN	O	O
of	NN	O	O
the	NN	O	O
RANKL	NN	O	B-protein
-induced	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappaB	NN	O	B-protein
and	NN	O	O
JNK	NN	O	B-protein
.	NN	O	O

This	NN	O	O
inhibition	NN	O	O
of	NN	O	O
osteoclastogenesis	NN	O	O
is	NN	O	O
rescued	NN	O	O
by	NN	O	O
overexpressing	NN	O	O
TRAF6	NN	O	B-protein
in	NN	O	O
precursor	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
which	NN	O	O
indicates	NN	O	O
that	NN	O	O
TRAF6	NN	O	B-protein
is	NN	O	O
the	NN	O	O
target	NN	O	O
critical	NN	O	O
for	NN	O	O
the	NN	O	O
IFN-gamma	NN	O	B-protein
action	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
we	NN	O	O
provide	NN	O	O
evidence	NN	O	O
that	NN	O	O
the	NN	O	O
accelerated	NN	O	O
degradation	NN	O	O
of	NN	O	O
TRAF6	NN	O	B-protein
requires	NN	O	O
both	NN	O	O
its	NN	O	O
ubiquitination	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
initiated	NN	O	O
by	NN	O	O
RANKL	NN	O	B-protein
,	NN	O	O
and	NN	O	O
IFN-gamma	NN	O	B-protein
-induced	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
ubiquitin-proteasome	NN	O	O
system	NN	O	O
.	NN	O	O

Our	NN	O	O
study	NN	O	O
shows	NN	O	O
that	NN	O	O
there	NN	O	O
is	NN	O	O
cross-talk	NN	O	O
between	NN	O	O
the	NN	O	O
tumour	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
and	NN	O	O
IFN	NN	O	B-protein
families	NN	O	I-protein
of	NN	O	O
cytokines	NN	O	B-protein
,	NN	O	O
through	NN	O	O
which	NN	O	O
IFN-gamma	NN	O	B-protein
provides	NN	O	O
a	NN	O	O
negative	NN	O	O
link	NN	O	O
between	NN	O	O
T-cell	NN	O	B-cell_type
activation	NN	O	O
and	NN	O	O
bone	NN	O	O
resorption	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
may	NN	O	O
offer	NN	O	O
a	NN	O	O
therapeutic	NN	O	O
approach	NN	O	O
to	NN	O	O
treat	NN	O	O
the	NN	O	O
inflammation-induced	NN	O	O
tissue	NN	O	O
breakdown	NN	O	O
.	NN	O	O

-DOCSTART-	O

Stromal-derived	NN	O	B-protein
factor	NN	O	I-protein
1	NN	O	I-protein
and	NN	O	O
thrombopoietin	NN	O	B-protein
regulate	NN	O	O
distinct	NN	O	O
aspects	NN	O	O
of	NN	O	O
human	NN	O	O
megakaryopoiesis	NN	O	O
.	NN	O	O

The	NN	O	O
role	NN	O	O
of	NN	O	O
the	NN	O	O
chemokine	NN	O	B-protein
binding	NN	O	O
stromal-derived	NN	O	B-protein
factor	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	O
SDF-1	NN	O	B-protein
)	NN	O	O
in	NN	O	O
normal	NN	O	O
human	NN	O	O
megakaryopoiesis	NN	O	O
at	NN	O	O
the	NN	O	O
cellular	NN	O	O
and	NN	O	O
molecular	NN	O	O
levels	NN	O	O
and	NN	O	O
its	NN	O	O
comparison	NN	O	O
with	NN	O	O
that	NN	O	O
of	NN	O	O
thrombopoietin	NN	O	B-protein
(	NN	O	O
TPO	NN	O	B-protein
)	NN	O	O
have	NN	O	O
not	NN	O	O
been	NN	O	O
determined	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
it	NN	O	O
was	NN	O	O
found	NN	O	O
that	NN	O	O
SDF-1	NN	O	B-protein
,	NN	O	O
unlike	NN	O	O
TPO	NN	O	B-protein
,	NN	O	O
does	NN	O	O
not	NN	O	O
stimulate	NN	O	O
alpha	NN	O	O
(	NN	O	O
IIb	NN	O	O
)	NN	O	O
beta	NN	O	O
(	NN	O	O
3	NN	O	O
)	NN	O	O
(	NN	O	O
+	NN	O	O
)	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
or	NN	O	O
differentiation	NN	O	O
or	NN	O	O
have	NN	O	O
an	NN	O	O
antiapoptotic	NN	O	O
effect	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
it	NN	O	O
does	NN	O	O
induce	NN	O	O
chemotaxis	NN	O	O
,	NN	O	O
trans-Matrigel	NN	O	O
migration	NN	O	O
,	NN	O	O
and	NN	O	O
secretion	NN	O	O
of	NN	O	O
matrix	NN	O	O
metalloproteinase	NN	O	B-protein
9	NN	O	I-protein
(	NN	O	O
MMP-9	NN	O	B-protein
)	NN	O	O
and	NN	O	O
vascular	NN	O	B-protein
endothelial	NN	O	I-protein
growth	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
VEGF	NN	O	B-protein
)	NN	O	O
by	NN	O	O
these	NN	O	O
cells	NN	O	O
,	NN	O	O
and	NN	O	O
both	NN	O	O
SDF-1	NN	O	B-protein
and	NN	O	O
TPO	NN	O	B-protein
increase	NN	O	O
the	NN	O	O
adhesion	NN	O	O
of	NN	O	O
alpha	NN	O	B-cell_type
(	NN	O	I-cell_type
IIb	NN	O	I-cell_type
)	NN	O	I-cell_type
beta	NN	O	I-cell_type
(	NN	O	I-cell_type
3	NN	O	I-cell_type
)	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
fibrinogen	NN	O	B-protein
and	NN	O	O
vitronectin	NN	O	B-protein
.	NN	O	O

Investigating	NN	O	O
the	NN	O	O
intracellular	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
induced	NN	O	O
by	NN	O	O
SDF-1	NN	O	B-protein
and	NN	O	O
TPO	NN	O	B-protein
revealed	NN	O	O
some	NN	O	O
overlapping	NN	O	O
patterns	NN	O	O
of	NN	O	O
protein	NN	O	O
phosphorylation/activation	NN	O	O
(	NN	O	O
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
[	NN	O	I-protein
MAPK	NN	O	I-protein
]	NN	O	I-protein
p42/44	NN	O	I-protein
,	NN	O	O
MAPK	NN	O	B-protein
p38	NN	O	I-protein
,	NN	O	O
and	NN	O	O
AKT	NN	O	B-protein
[	NN	O	I-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
B	NN	O	I-protein
]	NN	O	I-protein
)	NN	O	O
and	NN	O	O
some	NN	O	O
that	NN	O	O
were	NN	O	O
distinct	NN	O	O
for	NN	O	O
TPO	NN	O	B-protein
(	NN	O	O
eg	NN	O	O
,	NN	O	O
JAK-STAT	NN	O	B-protein
)	NN	O	O
and	NN	O	O
for	NN	O	O
SDF-1	NN	O	B-protein
(	NN	O	O
eg	NN	O	O
,	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
.	NN	O	O

It	NN	O	O
was	NN	O	O
also	NN	O	O
found	NN	O	O
that	NN	O	O
though	NN	O	O
inhibition	NN	O	O
of	NN	O	O
phosphatidyl-inositol	NN	O	B-protein
3-kinase	NN	O	I-protein
(	NN	O	O
PI-3K	NN	O	B-protein
)	NN	O	O
by	NN	O	O
LY294002	NN	O	O
in	NN	O	O
alpha	NN	O	B-cell_type
(	NN	O	I-cell_type
IIb	NN	O	I-cell_type
)	NN	O	I-cell_type
beta	NN	O	I-cell_type
(	NN	O	I-cell_type
3	NN	O	I-cell_type
)	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
induced	NN	O	O
apoptosis	NN	O	O
and	NN	O	O
inhibited	NN	O	O
chemotaxis	NN	O	O
adhesion	NN	O	O
and	NN	O	O
the	NN	O	O
secretion	NN	O	O
of	NN	O	O
MMP-9	NN	O	B-protein
and	NN	O	O
VEGF	NN	O	B-protein
,	NN	O	O
the	NN	O	O
inhibition	NN	O	O
of	NN	O	O
MAPK	NN	O	B-protein
p42/44	NN	O	I-protein
(	NN	O	O
by	NN	O	O
the	NN	O	O
MEK	NN	O	O
inhibitor	NN	O	O
U0126	NN	O	O
)	NN	O	O
had	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
survival	NN	O	O
,	NN	O	O
proliferation	NN	O	O
,	NN	O	O
and	NN	O	O
migration	NN	O	O
of	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

Hence	NN	O	O
,	NN	O	O
it	NN	O	O
is	NN	O	O
suggested	NN	O	O
that	NN	O	O
the	NN	O	O
proliferative	NN	O	O
effect	NN	O	O
of	NN	O	O
TPO	NN	O	B-protein
is	NN	O	O
more	NN	O	O
related	NN	O	O
to	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
JAK-STAT	NN	O	O
pathway	NN	O	O
(	NN	O	O
unique	NN	O	O
to	NN	O	O
TPO	NN	O	B-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
PI-3K-AKT	NN	O	O
axis	NN	O	O
is	NN	O	O
differentially	NN	O	O
involved	NN	O	O
in	NN	O	O
TPO-	NN	O	O
and	NN	O	O
SDF-1-dependent	NN	O	O
signaling	NN	O	O
.	NN	O	O

Accordingly	NN	O	O
,	NN	O	O
PI-3K	NN	O	B-protein
is	NN	O	O
involved	NN	O	O
in	NN	O	O
TPO	NN	O	B-protein
-mediated	NN	O	O
inhibition	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
,	NN	O	O
TPO-	NN	O	O
and	NN	O	O
SDF-1-regulated	NN	O	O
adhesion	NN	O	O
to	NN	O	O
fibrinogen	NN	O	B-protein
and	NN	O	O
vitronectin	NN	O	B-protein
,	NN	O	O
and	NN	O	O
SDF-1	NN	O	B-protein
-mediated	NN	O	O
migration	NN	O	O
.	NN	O	O

This	NN	O	O
study	NN	O	O
expands	NN	O	O
the	NN	O	O
understanding	NN	O	O
of	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
SDF-1	NN	O	B-protein
and	NN	O	O
TPO	NN	O	B-protein
in	NN	O	O
normal	NN	O	O
human	NN	O	O
megakaryopoiesis	NN	O	O
and	NN	O	O
indicates	NN	O	O
the	NN	O	O
molecular	NN	O	O
basis	NN	O	O
of	NN	O	O
the	NN	O	O
observed	NN	O	O
differences	NN	O	O
in	NN	O	O
cellular	NN	O	O
responses	NN	O	O
.	NN	O	O

(	NN	O	O
Blood.	NN	O	O
2000	NN	O	O
;	NN	O	O
96	NN	O	O
:	NN	O	O
4142-4151	NN	O	O
)	NN	O	O

-DOCSTART-	O

Adhesion	NN	O	O
of	NN	O	O
immature	NN	O	B-cell_type
and	NN	O	I-cell_type
mature	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
induces	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
thymic	NN	O	I-cell_type
epithelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
TEC	NN	O	B-cell_type
)	NN	O	O
activation	NN	O	O
of	NN	O	O
IL-6	NN	O	B-protein
gene	NN	O	I-protein
trascription	NN	O	I-protein
factors	NN	O	I-protein
(	NN	O	O
NF-kappaB	NN	O	B-protein
and	NN	O	O
NF-IL6	NN	O	B-protein
)	NN	O	O
and	NN	O	O
IL-6	NN	O	O
gene	NN	O	O
expression	NN	O	O
:	NN	O	O
role	NN	O	O
of	NN	O	O
alpha3beta1	NN	O	B-protein
and	NN	O	I-protein
alpha6beta4	NN	O	I-protein
integrins	NN	O	I-protein
.	NN	O	O

T	NN	O	B-cell_type
cell	NN	O	I-cell_type
precursors	NN	O	I-cell_type
homed	NN	O	O
to	NN	O	O
thymus	NN	O	O
develop	NN	O	O
in	NN	O	O
close	NN	O	O
contact	NN	O	O
with	NN	O	O
stromal	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Among	NN	O	O
them	NN	O	O
,	NN	O	O
thymic	NN	O	B-cell_type
epithelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
TEC	NN	O	B-cell_type
)	NN	O	O
are	NN	O	O
known	NN	O	O
to	NN	O	O
exert	NN	O	O
dominant	NN	O	O
roles	NN	O	O
in	NN	O	O
their	NN	O	O
survival	NN	O	O
and	NN	O	O
functional	NN	O	O
shaping	NN	O	O
.	NN	O	O

Key	NN	O	O
molecules	NN	O	O
mediating	NN	O	O
TEC	NN	O	B-cell_type
/	NN	O	O
thymocytes	NN	O	B-cell_type
interactions	NN	O	O
include	NN	O	O
cytokines	NN	O	B-protein
and	NN	O	O
growth	NN	O	B-protein
factors	NN	O	I-protein
secreted	NN	O	O
by	NN	O	O
the	NN	O	O
two	NN	O	O
cell	NN	O	O
types	NN	O	O
and	NN	O	O
adhesion	NN	O	B-protein
receptors	NN	O	I-protein
mediating	NN	O	O
cell	NN	O	O
contact	NN	O	O
.	NN	O	O

Signaling	NN	O	O
events	NN	O	O
triggered	NN	O	O
in	NN	O	O
thymocytes	NN	O	B-cell_type
by	NN	O	O
adhesion	NN	O	O
to	NN	O	O
epithelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
have	NN	O	O
been	NN	O	O
extensively	NN	O	O
investigated	NN	O	O
,	NN	O	O
whereas	NN	O	O
little	NN	O	O
is	NN	O	O
known	NN	O	O
on	NN	O	O
the	NN	O	O
opposite	NN	O	O
phenomenon	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
investigated	NN	O	O
this	NN	O	O
issue	NN	O	O
in	NN	O	O
a	NN	O	O
co-culture	NN	O	O
system	NN	O	O
composed	NN	O	O
of	NN	O	O
TEC	NN	O	B-cell_type
cultures	NN	O	I-cell_type
derived	NN	O	O
from	NN	O	O
human	NN	O	O
normal	NN	O	O
thymus	NN	O	O
and	NN	O	O
heterologous	NN	O	B-cell_type
thymocytes	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
thymocytes	NN	O	B-cell_type
adhere	NN	O	O
to	NN	O	O
TEC	NN	O	B-cell_type
involving	NN	O	O
beta1	NN	O	B-protein
and	NN	O	I-protein
beta4	NN	O	I-protein
integrins	NN	O	I-protein
and	NN	O	O
induce	NN	O	O
the	NN	O	O
clustering	NN	O	O
of	NN	O	O
alpha3beta1	NN	O	B-protein
and	NN	O	I-protein
alpha6beta4	NN	O	I-protein
heterodimers	NN	O	I-protein
at	NN	O	O
the	NN	O	O
TEC	NN	O	O
surface	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
thymocyte	NN	O	O
adhesion	NN	O	O
was	NN	O	O
followed	NN	O	O
by	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
and	NN	O	O
NF-IL6	NN	O	B-protein
gene	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
and	NN	O	O
enhanced	NN	O	O
IL-6	NN	O	O
production	NN	O	O
.	NN	O	O

The	NN	O	O
two	NN	O	O
latter	NN	O	O
phenomena	NN	O	O
were	NN	O	O
reproduced	NN	O	O
by	NN	O	O
the	NN	O	O
cross-linking	NN	O	O
of	NN	O	O
the	NN	O	O
alpha3	NN	O	B-protein
,	NN	O	O
alpha6	NN	O	B-protein
,	NN	O	O
beta1	NN	O	B-protein
and	NN	O	I-protein
beta4	NN	O	I-protein
integrins	NN	O	I-protein
,	NN	O	O
thus	NN	O	O
implying	NN	O	O
that	NN	O	O
the	NN	O	O
alpha3beta1	NN	O	B-protein
and	NN	O	I-protein
alpha6beta4	NN	O	I-protein
heterodimers	NN	O	I-protein
can	NN	O	O
signal	NN	O	O
during	NN	O	O
thymocyte	NN	O	O
adhesion	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
extended	NN	O	O
our	NN	O	O
previous	NN	O	O
work	NN	O	O
investigating	NN	O	O
in	NN	O	O
the	NN	O	O
same	NN	O	O
experimental	NN	O	O
setting	NN	O	O
the	NN	O	O
inducing	NN	O	O
activity	NN	O	O
of	NN	O	O
non	NN	O	O
stimulated	NN	O	O
or	NN	O	O
activated	NN	O	O
policlonal	NN	O	B-cell_type
or	NN	O	I-cell_type
clonal	NN	O	I-cell_type
mature	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
as	NN	O	O
representative	NN	O	O
of	NN	O	O
the	NN	O	O
more	NN	O	O
mature	NN	O	B-cell_type
thymocyte	NN	O	I-cell_type
subset	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
adhesion	NN	O	O
of	NN	O	O
unstimulated	NN	O	B-cell_type
T	NN	O	I-cell_type
cell	NN	O	I-cell_type
i	NN	O	O
)	NN	O	O
involved	NN	O	O
beta1	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
beta4	NN	O	O
integrin	NN	O	O
functions	NN	O	O
at	NN	O	O
the	NN	O	O
surface	NN	O	O
ii	NN	O	O
)	NN	O	O
induced	NN	O	O
the	NN	O	O
clustering	NN	O	O
of	NN	O	O
alpha3beta1	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
alpha2beta1	NN	O	B-protein
heterodimers	NN	O	I-protein
at	NN	O	O
the	NN	O	O
TEC	NN	O	O
surface	NN	O	O
and	NN	O	O
iii	NN	O	O
)	NN	O	O
up-regulated	NN	O	O
the	NN	O	O
nuclear	NN	O	O
binding	NN	O	O
activity	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
and	NN	O	O
the	NN	O	O
IL-6	NN	O	O
secretion	NN	O	O
.	NN	O	O

We	NN	O	O
propose	NN	O	O
that	NN	O	O
alpha3beta1	NN	O	B-protein
and	NN	O	I-protein
alpha6beta4	NN	O	I-protein
heterodimers	NN	O	I-protein
are	NN	O	O
induced	NN	O	O
to	NN	O	O
cluster	NN	O	O
at	NN	O	O
the	NN	O	O
TEC	NN	O	O
surface	NN	O	O
recognizing	NN	O	O
yet	NN	O	O
unknown	NN	O	O
cellular	NN	O	B-protein
ligands	NN	O	I-protein
differentially	NN	O	O
expressed	NN	O	O
during	NN	O	O
T	NN	O	O
cell	NN	O	O
development	NN	O	O
.	NN	O	O

-DOCSTART-	O

Identification	NN	O	O
and	NN	O	O
characterization	NN	O	O
of	NN	O	O
SKAT-2	NN	O	B-DNA
,	NN	O	O
a	NN	O	O
novel	NN	O	O
Th2-specific	NN	O	B-DNA
zinc	NN	O	I-DNA
finger	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
have	NN	O	O
identified	NN	O	O
a	NN	O	O
novel	NN	O	O
Kruppel-type	NN	O	B-DNA
zinc	NN	O	I-DNA
finger	NN	O	I-DNA
(	NN	O	I-DNA
ZF	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
SKAT-2	NN	O	B-DNA
,	NN	O	O
which	NN	O	O
is	NN	O	O
selectively	NN	O	O
expressed	NN	O	O
by	NN	O	O
murine	NN	O	B-cell_type
Th2	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
protein	NN	O	O
encoded	NN	O	O
by	NN	O	O
this	NN	O	O
gene	NN	O	O
has	NN	O	O
14	NN	O	O
C2H2-type	NN	O	O
ZF	NN	O	O
tandemly	NN	O	O
arrayed	NN	O	O
at	NN	O	O
its	NN	O	O
C	NN	O	B-DNA
terminus	NN	O	I-DNA
and	NN	O	O
N-terminal	NN	O	B-DNA
SCAN	NN	O	B-DNA
box	NN	O	I-DNA
and	NN	O	O
KRAB	NN	O	B-DNA
domains	NN	O	I-DNA
.	NN	O	O

SKAT-2	NN	O	B-DNA
is	NN	O	O
tissue	NN	O	O
restricted	NN	O	O
in	NN	O	O
expression	NN	O	O
at	NN	O	O
the	NN	O	O
RNA	NN	O	O
level	NN	O	O
,	NN	O	O
detectable	NN	O	O
only	NN	O	O
in	NN	O	O
brain	NN	O	O
and	NN	O	O
at	NN	O	O
low	NN	O	O
levels	NN	O	O
in	NN	O	O
kidney	NN	O	O
and	NN	O	O
spleen	NN	O	O
and	NN	O	O
few	NN	O	O
hematopoietic	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

By	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
,	NN	O	O
SKAT-2	NN	O	B-DNA
expression	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
peak	NN	O	O
in	NN	O	O
antigen-stimulated	NN	O	O
CD4	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
after	NN	O	O
2-3	NN	O	O
days	NN	O	O
of	NN	O	O
culture	NN	O	O
under	NN	O	O
Th2	NN	O	O
but	NN	O	O
not	NN	O	O
Th1	NN	O	O
biasing	NN	O	O
conditions	NN	O	O
.	NN	O	O

This	NN	O	O
pattern	NN	O	O
of	NN	O	O
expression	NN	O	O
closely	NN	O	O
mirrored	NN	O	O
that	NN	O	O
of	NN	O	O
GATA-3	NN	O	B-protein
in	NN	O	O
the	NN	O	O
same	NN	O	O
cells	NN	O	O
.	NN	O	O

In	NN	O	O
transient	NN	O	O
transfection	NN	O	O
experiments	NN	O	O
in	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate/ionomycin-stimulated	NN	O	O
EL4	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
SKAT-2	NN	O	B-DNA
was	NN	O	O
found	NN	O	O
to	NN	O	O
up-regulate	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
IL-4	NN	O	B-DNA
but	NN	O	I-DNA
not	NN	O	I-DNA
the	NN	O	I-DNA
IL-5	NN	O	I-DNA
promoter	NN	O	I-DNA
,	NN	O	O
contrasting	NN	O	O
with	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
GATA-3	NN	O	B-protein
to	NN	O	O
activate	NN	O	O
both	NN	O	O
promoters	NN	O	B-DNA
.	NN	O	O

This	NN	O	O
result	NN	O	O
was	NN	O	O
confirmed	NN	O	O
using	NN	O	O
clones	NN	O	O
of	NN	O	O
EL4	NN	O	B-cell_line
cells	NN	O	I-cell_line
stably	NN	O	O
expressing	NN	O	O
an	NN	O	O
inducible	NN	O	O
form	NN	O	O
of	NN	O	O
SKAT-2	NN	O	B-DNA
,	NN	O	O
thus	NN	O	O
SKAT-2	NN	O	B-DNA
is	NN	O	O
a	NN	O	O
novel	NN	O	O
Th2-specific	NN	O	B-DNA
gene	NN	O	I-DNA
that	NN	O	O
may	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
selective	NN	O	O
regulation	NN	O	O
of	NN	O	O
cytokine	NN	O	B-DNA
genes	NN	O	I-DNA
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

hsp70	NN	O	B-protein
interacting	NN	O	I-protein
protein	NN	O	I-protein
Hip	NN	O	I-protein
does	NN	O	O
not	NN	O	O
affect	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
folding	NN	O	O
by	NN	O	O
the	NN	O	O
hsp90-based	NN	O	O
chaperone	NN	O	O
machinery	NN	O	O
except	NN	O	O
to	NN	O	O
oppose	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
BAG-1	NN	O	B-protein
.	NN	O	O

Reticulocyte	NN	O	O
lysate	NN	O	O
contains	NN	O	O
a	NN	O	O
chaperone	NN	O	O
system	NN	O	O
that	NN	O	O
assembles	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	I-protein
GR	NN	O	I-protein
)	NN	O	I-protein
.hsp90	NN	O	I-protein
heterocomplexes	NN	O	I-protein
.	NN	O	O

Using	NN	O	O
purified	NN	O	B-protein
proteins	NN	O	I-protein
,	NN	O	O
we	NN	O	O
have	NN	O	O
prepared	NN	O	O
a	NN	O	O
five-protein	NN	O	O
heterocomplex	NN	O	O
assembly	NN	O	O
system	NN	O	O
consisting	NN	O	O
of	NN	O	O
two	NN	O	O
proteins	NN	O	O
essential	NN	O	O
for	NN	O	O
heterocomplex	NN	O	O
assembly-	NN	O	O
hsp90	NN	O	B-protein
and	NN	O	O
hsp70	NN	O	B-protein
-and	NN	O	O
three	NN	O	O
proteins	NN	O	O
that	NN	O	O
act	NN	O	O
as	NN	O	O
co-chaperones	NN	O	O
to	NN	O	O
enhance	NN	O	O
assembly-	NN	O	O
Hop	NN	O	B-protein
,	NN	O	O
hsp40	NN	O	B-protein
,	NN	O	O
p23	NN	O	B-protein
[	NN	O	O
Morishima	NN	O	O
,	NN	O	O
Y.	NN	O	O
,	NN	O	O
Kanelakis	NN	O	O
,	NN	O	O
K.	NN	O	O
C.	NN	O	O
,	NN	O	O
Silverstein	NN	O	O
,	NN	O	O
A.	NN	O	O
M.	NN	O	O
,	NN	O	O
Dittmar	NN	O	O
,	NN	O	O
K.	NN	O	O
D.	NN	O	O
,	NN	O	O
Estrada	NN	O	O
,	NN	O	O
L.	NN	O	O
,	NN	O	O
and	NN	O	O
Pratt	NN	O	O
,	NN	O	O
W.	NN	O	O
B.	NN	O	O
(	NN	O	O
2000	NN	O	O
)	NN	O	O
J.	NN	O	O
Biol.	NN	O	O
Chem.	NN	O	O
275	NN	O	O
,	NN	O	O
6894-6900	NN	O	O
]	NN	O	O
.	NN	O	O

The	NN	O	O
hsp70	NN	O	B-protein
co-chaperone	NN	O	I-protein
Hip	NN	O	I-protein
has	NN	O	O
been	NN	O	O
recovered	NN	O	O
in	NN	O	O
receptor.hsp90	NN	O	B-protein
heterocomplexes	NN	O	I-protein
at	NN	O	O
an	NN	O	O
intermediate	NN	O	O
stage	NN	O	O
of	NN	O	O
assembly	NN	O	O
in	NN	O	O
reticulocyte	NN	O	O
lysate	NN	O	O
,	NN	O	O
and	NN	O	O
Hip	NN	O	B-protein
is	NN	O	O
also	NN	O	O
thought	NN	O	O
to	NN	O	O
be	NN	O	O
an	NN	O	O
intrinsic	NN	O	O
component	NN	O	O
of	NN	O	O
the	NN	O	O
assembly	NN	O	O
machinery	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
immunodepletion	NN	O	O
of	NN	O	O
Hip	NN	O	B-protein
from	NN	O	O
reticulocyte	NN	O	O
lysate	NN	O	O
or	NN	O	O
addition	NN	O	O
of	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
Hip	NN	O	B-protein
to	NN	O	O
the	NN	O	O
purified	NN	O	O
five-protein	NN	O	O
system	NN	O	O
does	NN	O	O
not	NN	O	O
affect	NN	O	O
GR.hsp90	NN	O	O
heterocomplex	NN	O	O
assembly	NN	O	O
or	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
steroid	NN	O	O
binding	NN	O	O
activity	NN	O	O
that	NN	O	O
occurs	NN	O	O
with	NN	O	O
assembly	NN	O	O
.	NN	O	O

Despite	NN	O	O
the	NN	O	O
fact	NN	O	O
that	NN	O	O
Hip	NN	O	B-protein
does	NN	O	O
not	NN	O	O
affect	NN	O	O
assembly	NN	O	O
,	NN	O	O
it	NN	O	O
is	NN	O	O
recovered	NN	O	O
in	NN	O	O
GR.hsp90	NN	O	B-protein
heterocomplexes	NN	O	I-protein
assembled	NN	O	O
by	NN	O	O
both	NN	O	O
systems	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
five-protein	NN	O	O
system	NN	O	O
,	NN	O	O
Hip	NN	O	B-protein
prevents	NN	O	O
inhibition	NN	O	O
of	NN	O	O
assembly	NN	O	O
by	NN	O	O
the	NN	O	O
hsp70	NN	O	B-protein
co-chaperone	NN	O	I-protein
BAG-1	NN	O	I-protein
,	NN	O	O
and	NN	O	O
cotransfection	NN	O	O
of	NN	O	O
Hip	NN	O	B-protein
with	NN	O	O
BAG-1	NN	O	B-protein
opposes	NN	O	O
BAG-1	NN	O	B-protein
reduction	NN	O	O
of	NN	O	O
steroid	NN	O	O
binding	NN	O	O
activity	NN	O	O
in	NN	O	O
COS	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
Hip	NN	O	B-protein
is	NN	O	O
not	NN	O	O
a	NN	O	O
component	NN	O	O
of	NN	O	O
the	NN	O	O
assembly	NN	O	O
machinery	NN	O	O
but	NN	O	O
that	NN	O	O
it	NN	O	O
could	NN	O	O
play	NN	O	O
a	NN	O	O
regulatory	NN	O	O
role	NN	O	O
in	NN	O	O
opposition	NN	O	O
to	NN	O	O
BAG-1	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Cutting	NN	O	O
edge	NN	O	O
:	NN	O	O
STAT6-deficient	NN	O	O
mice	NN	O	O
have	NN	O	O
enhanced	NN	O	O
tumor	NN	O	O
immunity	NN	O	O
to	NN	O	O
primary	NN	O	O
and	NN	O	O
metastatic	NN	O	O
mammary	NN	O	O
carcinoma	NN	O	O
.	NN	O	O

STAT4	NN	O	B-protein
and	NN	O	O
STAT6	NN	O	B-protein
are	NN	O	O
essential	NN	O	O
for	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
CD4	NN	O	O
(	NN	O	O
+	NN	O	O
)	NN	O	O
Th1	NN	O	O
and	NN	O	O
Th2	NN	O	O
development	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

Tumor	NN	O	O
immunologists	NN	O	O
have	NN	O	O
hypothesized	NN	O	O
that	NN	O	O
Th1	NN	O	B-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
critical	NN	O	O
in	NN	O	O
tumor	NN	O	O
immunity	NN	O	O
because	NN	O	O
they	NN	O	O
facilitate	NN	O	O
differentiation	NN	O	O
of	NN	O	O
CD8	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
which	NN	O	O
are	NN	O	O
potent	NN	O	O
anti-tumor	NN	O	B-cell_type
effectors	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
have	NN	O	O
used	NN	O	O
STAT4	NN	O	O
(	NN	O	O
-/-	NN	O	O
)	NN	O	O
and	NN	O	O
STAT6	NN	O	O
(	NN	O	O
-/-	NN	O	O
)	NN	O	O
mice	NN	O	O
to	NN	O	O
test	NN	O	O
this	NN	O	O
hypothesis	NN	O	O
.	NN	O	O

BALB/c	NN	O	O
and	NN	O	O
knockout	NN	O	O
mice	NN	O	O
were	NN	O	O
challenged	NN	O	O
in	NN	O	O
the	NN	O	O
mammary	NN	O	O
gland	NN	O	O
with	NN	O	O
the	NN	O	O
highly	NN	O	O
malignant	NN	O	O
and	NN	O	O
spontaneously	NN	O	O
metastatic	NN	O	O
BALB/c-derived	NN	O	O
4T1	NN	O	O
mammary	NN	O	O
carcinoma	NN	O	O
.	NN	O	O

Primary	NN	O	O
tumor	NN	O	O
growth	NN	O	O
and	NN	O	O
metastatic	NN	O	O
disease	NN	O	O
are	NN	O	O
reduced	NN	O	O
in	NN	O	O
STAT6	NN	O	O
(	NN	O	O
-/-	NN	O	O
)	NN	O	O
mice	NN	O	O
relative	NN	O	O
to	NN	O	O
BALB/c	NN	O	O
and	NN	O	O
STAT4	NN	O	O
(	NN	O	O
-/-	NN	O	O
)	NN	O	O
mice	NN	O	O
.	NN	O	O

Ab	NN	O	O
depletions	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
effect	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
CD8	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
immunized	NN	O	O
STAT6	NN	O	O
(	NN	O	O
-/-	NN	O	O
)	NN	O	O
mice	NN	O	O
have	NN	O	O
higher	NN	O	O
levels	NN	O	O
of	NN	O	O
4T1-specific	NN	O	B-cell_type
CTL	NN	O	I-cell_type
than	NN	O	O
BALB/c	NN	O	O
or	NN	O	O
STAT4	NN	O	O
(	NN	O	O
-/-	NN	O	O
)	NN	O	O
mice	NN	O	O
.	NN	O	O

Surprisingly	NN	O	O
,	NN	O	O
Th1	NN	O	B-cell_type
or	NN	O	I-cell_type
Th2	NN	O	I-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
not	NN	O	O
involved	NN	O	O
,	NN	O	O
because	NN	O	O
CD4	NN	O	O
depletion	NN	O	O
does	NN	O	O
not	NN	O	O
diminish	NN	O	O
the	NN	O	O
anti-tumor	NN	O	O
effect	NN	O	O
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
deletion	NN	O	O
of	NN	O	O
the	NN	O	O
STAT6	NN	O	B-DNA
gene	NN	O	I-DNA
facilitates	NN	O	O
development	NN	O	O
of	NN	O	O
potent	NN	O	O
anti-tumor	NN	O	O
immunity	NN	O	O
via	NN	O	O
a	NN	O	O
CD4	NN	O	O
(	NN	O	O
+	NN	O	O
)	NN	O	O
-independent	NN	O	O
pathway	NN	O	O
.	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
of	NN	O	O
oncogenic	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
AP-1	NN	O	I-protein
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
infected	NN	O	O
with	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
1	NN	O	O
.	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
1	NN	O	O
(	NN	O	O
HTLV-1	NN	O	O
)	NN	O	O
Tax	NN	O	B-protein
protein	NN	O	I-protein
transforms	NN	O	O
primary	NN	O	B-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

We	NN	O	O
previously	NN	O	O
showed	NN	O	O
that	NN	O	O
Tax	NN	O	B-protein
induces	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
various	NN	O	O
family	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
AP-1	NN	O	I-protein
such	NN	O	O
as	NN	O	O
c-Jun	NN	O	B-protein
,	NN	O	O
JunD	NN	O	B-protein
,	NN	O	O
c-Fos	NN	O	B-protein
,	NN	O	O
and	NN	O	O
Fra-1	NN	O	B-protein
at	NN	O	O
the	NN	O	O
mRNA	NN	O	O
level	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
study	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
examined	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
Tax	NN	O	B-protein
to	NN	O	O
activate	NN	O	O
transcription	NN	O	O
through	NN	O	O
the	NN	O	O
AP-1-binding	NN	O	B-DNA
site	NN	O	I-DNA
(	NN	O	O
AP-1	NN	O	B-DNA
site	NN	O	I-DNA
)	NN	O	O
.	NN	O	O

A	NN	O	O
transient	NN	O	O
transfection	NN	O	O
study	NN	O	O
showed	NN	O	O
that	NN	O	O
Tax	NN	O	B-protein
can	NN	O	O
activate	NN	O	O
transcription	NN	O	O
through	NN	O	O
the	NN	O	O
AP-1-binding	NN	O	B-DNA
site	NN	O	I-DNA
in	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
whereas	NN	O	O
any	NN	O	O
combination	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
proteins	NN	O	I-protein
did	NN	O	O
so	NN	O	O
much	NN	O	O
less	NN	O	O
than	NN	O	O
Tax	NN	O	B-protein
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
AP-1	NN	O	B-DNA
site	NN	O	I-DNA
by	NN	O	O
Tax	NN	O	B-protein
may	NN	O	O
require	NN	O	O
a	NN	O	O
mechanism	NN	O	O
other	NN	O	O
than	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
AP-1	NN	O	B-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

Fresh	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
leukemia	NN	O	I-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
all	NN	O	O
surveyed	NN	O	O
ATL	NN	O	O
patients	NN	O	O
displayed	NN	O	O
constitutive	NN	O	O
AP-1	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
,	NN	O	O
whereas	NN	O	O
no	NN	O	O
normal	NN	O	O
individuals	NN	O	O
did	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
HTLV-1	NN	O	B-DNA
genes	NN	O	I-DNA
,	NN	O	O
including	NN	O	O
tax	NN	O	B-DNA
,	NN	O	O
are	NN	O	O
not	NN	O	O
significantly	NN	O	O
expressed	NN	O	O
in	NN	O	O
fresh	NN	O	O
leukemia	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
ATL	NN	O	O
patients	NN	O	O
.	NN	O	O

Our	NN	O	O
present	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
activation	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
occurs	NN	O	O
through	NN	O	O
Tax-dependent	NN	O	O
and	NN	O	O
-independent	NN	O	O
mechanisms	NN	O	O
in	NN	O	O
HTLV-1-infected	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
which	NN	O	O
may	NN	O	O
play	NN	O	O
some	NN	O	O
roles	NN	O	O
in	NN	O	O
dysregulated	NN	O	O
phenotypes	NN	O	O
of	NN	O	O
HTLV-1-infected	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

2	NN	O	O
,	NN	O	O
2	NN	O	O
'	NN	O	O
,	NN	O	O
4	NN	O	O
,	NN	O	O
6	NN	O	O
,	NN	O	O
6'-pentachlorobiphenyl	NN	O	O
induces	NN	O	O
apoptosis	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

Polychlorinatedbiphenyls	NN	O	O
(	NN	O	O
PCBs	NN	O	O
)	NN	O	O
are	NN	O	O
a	NN	O	O
group	NN	O	O
of	NN	O	O
persistent	NN	O	O
and	NN	O	O
widely	NN	O	O
dispersed	NN	O	O
environmental	NN	O	O
pollutants	NN	O	O
,	NN	O	O
some	NN	O	O
of	NN	O	O
which	NN	O	O
may	NN	O	O
be	NN	O	O
immunotoxic	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
PCBs	NN	O	O
on	NN	O	O
immune	NN	O	O
system	NN	O	O
by	NN	O	O
assessing	NN	O	O
apoptotic	NN	O	O
cell	NN	O	O
death	NN	O	O
in	NN	O	O
human	NN	O	B-cell_line
monocytic	NN	O	I-cell_line
U937	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Among	NN	O	O
the	NN	O	O
various	NN	O	O
congeners	NN	O	O
tested	NN	O	O
,	NN	O	O
2	NN	O	O
,	NN	O	O
2	NN	O	O
'	NN	O	O
,	NN	O	O
4	NN	O	O
,	NN	O	O
6	NN	O	O
,	NN	O	O
6'-pentachlorobiphenyl	NN	O	O
(	NN	O	O
PeCB	NN	O	O
)	NN	O	O
,	NN	O	O
a	NN	O	O
highly	NN	O	O
ortho-substituted	NN	O	O
congener	NN	O	O
,	NN	O	O
specifically	NN	O	O
induced	NN	O	O
DNA	NN	O	O
fragmentation	NN	O	O
,	NN	O	O
a	NN	O	O
hallmark	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
,	NN	O	O
while	NN	O	O
the	NN	O	O
other	NN	O	O
examined	NN	O	O
di-	NN	O	O
,	NN	O	O
tri-	NN	O	O
,	NN	O	O
tetra-	NN	O	O
,	NN	O	O
and	NN	O	O
pentachlorobiphenyls	NN	O	O
did	NN	O	O
not	NN	O	O
.	NN	O	O

To	NN	O	O
further	NN	O	O
study	NN	O	O
the	NN	O	O
2	NN	O	O
,	NN	O	O
2	NN	O	O
'	NN	O	O
,	NN	O	O
4	NN	O	O
,	NN	O	O
6	NN	O	O
,	NN	O	O
6'-PeCB-induced	NN	O	O
cell	NN	O	O
death	NN	O	O
,	NN	O	O
various	NN	O	O
features	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
were	NN	O	O
examined	NN	O	O
.	NN	O	O

2	NN	O	O
,	NN	O	O
2	NN	O	O
'	NN	O	O
,	NN	O	O
4	NN	O	O
,	NN	O	O
6	NN	O	O
,	NN	O	O
6'-PeCB	NN	O	O
caused	NN	O	O
a	NN	O	O
decrease	NN	O	O
in	NN	O	O
cell	NN	O	O
viability	NN	O	O
and	NN	O	O
induced	NN	O	O
cellular	NN	O	O
morphologic	NN	O	O
features	NN	O	O
characteristic	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
such	NN	O	O
as	NN	O	O
chromatin	NN	O	O
aggregation	NN	O	O
and	NN	O	O
apoptotic	NN	O	O
bodies	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
caspase-3	NN	O	B-protein
,	NN	O	O
an	NN	O	O
executioner	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
,	NN	O	O
was	NN	O	O
activated	NN	O	O
and	NN	O	O
its	NN	O	O
substrate	NN	O	O
,	NN	O	O
poly	NN	O	B-protein
(	NN	O	I-protein
ADP-ribose	NN	O	I-protein
)	NN	O	I-protein
polymerase	NN	O	I-protein
(	NN	O	O
PARP	NN	O	B-protein
)	NN	O	O
,	NN	O	O
was	NN	O	O
cleaved	NN	O	O
during	NN	O	O
2	NN	O	O
,	NN	O	O
2	NN	O	O
'	NN	O	O
,	NN	O	O
4	NN	O	O
,	NN	O	O
6	NN	O	O
,	NN	O	O
6'-PeCB-induced	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
3	NN	O	O
,	NN	O	O
3	NN	O	O
'	NN	O	O
,	NN	O	O
4	NN	O	O
,	NN	O	O
4	NN	O	O
'	NN	O	O
,	NN	O	O
5-PeCB	NN	O	O
,	NN	O	O
a	NN	O	O
congener	NN	O	O
of	NN	O	O
coplanar	NN	O	O
structure	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
2	NN	O	O
,	NN	O	O
3	NN	O	O
,	NN	O	O
7	NN	O	O
,	NN	O	O
8-TCDD	NN	O	O
did	NN	O	O
not	NN	O	O
induce	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
these	NN	O	O
human	NN	O	B-cell_type
monocytic	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
although	NN	O	O
they	NN	O	O
potently	NN	O	O
induced	NN	O	O
CYP	NN	O	B-protein
1A1	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_line
hepatoma	NN	O	I-cell_line
Hep	NN	O	I-cell_line
G2	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
the	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
2	NN	O	O
,	NN	O	O
2	NN	O	O
'	NN	O	O
,	NN	O	O
4	NN	O	O
,	NN	O	O
6	NN	O	O
,	NN	O	O
6'-PeCB	NN	O	O
induces	NN	O	O
apoptosis	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
through	NN	O	O
a	NN	O	O
mechanism	NN	O	O
that	NN	O	O
is	NN	O	O
independent	NN	O	O
of	NN	O	O
the	NN	O	O
arylhydrocarbon	NN	O	B-protein
receptor	NN	O	I-protein
.	NN	O	O

This	NN	O	O
suggests	NN	O	O
a	NN	O	O
possibly	NN	O	O
separate	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
PCBs	NN	O	O
cause	NN	O	O
immunosuppression	NN	O	O
.	NN	O	O

-DOCSTART-	O

Effects	NN	O	O
of	NN	O	O
deregulated	NN	O	O
Raf	NN	O	O
activation	NN	O	O
on	NN	O	O
integrin	NN	O	B-protein
,	NN	O	O
cytokine-receptor	NN	O	O
expression	NN	O	O
and	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
hematopoietic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
effects	NN	O	O
of	NN	O	O
deregulated	NN	O	O
Raf	NN	O	O
activation	NN	O	O
on	NN	O	O
the	NN	O	O
growth	NN	O	O
and	NN	O	O
differentiation	NN	O	O
of	NN	O	O
hematopoietic	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
investigated	NN	O	O
.	NN	O	O

The	NN	O	O
cytokine-dependent	NN	O	B-cell_line
murine	NN	O	I-cell_line
myeloid	NN	O	I-cell_line
FDC-P1	NN	O	I-cell_line
and	NN	O	O
human	NN	O	B-cell_line
erythroleukemic	NN	O	I-cell_line
TF-1	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
were	NN	O	O
transformed	NN	O	O
to	NN	O	O
grow	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
deregulated	NN	O	O
Raf	NN	O	O
expression	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
exogenous	NN	O	B-protein
cytokines	NN	O	I-protein
.	NN	O	O

The	NN	O	O
conditionally	NN	O	O
active	NN	O	O
Raf	NN	O	B-protein
proteins	NN	O	I-protein
were	NN	O	O
regulated	NN	O	O
by	NN	O	O
beta-estradiol	NN	O	O
as	NN	O	O
cDNAs	NN	O	B-DNA
containing	NN	O	O
the	NN	O	O
Raf	NN	O	B-protein
catalytic	NN	O	I-protein
,	NN	O	O
but	NN	O	O
lacking	NN	O	O
negative-regulatory	NN	O	B-protein
domains	NN	O	I-protein
,	NN	O	O
were	NN	O	O
ligated	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
estrogen	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
deltaRaf	NN	O	B-protein
:	NN	O	I-protein
ER	NN	O	I-protein
)	NN	O	O
.	NN	O	O

Continuous	NN	O	O
deltaRaf	NN	O	B-protein
expression	NN	O	O
prevented	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
exogenous	NN	O	B-protein
cytokines	NN	O	I-protein
and	NN	O	O
altered	NN	O	O
the	NN	O	O
morphology	NN	O	O
of	NN	O	O
the	NN	O	O
FD/deltaRaf	NN	O	B-cell_type
:	NN	O	I-cell_type
ER	NN	O	I-cell_type
cells	NN	O	I-cell_type
as	NN	O	O
they	NN	O	O
grew	NN	O	O
in	NN	O	O
large	NN	O	O
aggregated	NN	O	O
masses	NN	O	O
(	NN	O	O
>	NN	O	O
100	NN	O	O
cells	NN	O	O
)	NN	O	O
whereas	NN	O	O
the	NN	O	O
parental	NN	O	B-cell_line
cytokine-dependent	NN	O	I-cell_line
FDC-P1	NN	O	I-cell_line
cells	NN	O	I-cell_line
grew	NN	O	O
in	NN	O	O
smaller	NN	O	O
grape-like	NN	O	O
clusters	NN	O	O
(	NN	O	O
<	NN	O	O
10	NN	O	O
cells	NN	O	O
)	NN	O	O
.	NN	O	O

FD/deltaRaf-1	NN	O	B-cell_line
:	NN	O	I-cell_line
ER	NN	O	I-cell_line
cells	NN	O	I-cell_line
growing	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
Raf	NN	O	O
activation	NN	O	O
displayed	NN	O	O
decreased	NN	O	O
levels	NN	O	O
of	NN	O	O
the	NN	O	O
Mac-2	NN	O	B-protein
and	NN	O	I-protein
Mac-3	NN	O	I-protein
molecules	NN	O	I-protein
on	NN	O	O
their	NN	O	O
cell	NN	O	B-cell_type
surface	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
when	NN	O	O
these	NN	O	O
cells	NN	O	O
were	NN	O	O
cultured	NN	O	O
in	NN	O	O
IL-3	NN	O	B-protein
,	NN	O	O
higher	NN	O	O
levels	NN	O	O
of	NN	O	O
these	NN	O	O
adhesion	NN	O	B-protein
molecules	NN	O	I-protein
were	NN	O	O
detected	NN	O	O
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
activated	NN	O	O
Raf	NN	O	B-protein
oncoproteins	NN	O	I-protein
also	NN	O	O
abrogated	NN	O	O
cytokine	NN	O	O
dependency	NN	O	O
and	NN	O	O
prevented	NN	O	O
apoptosis	NN	O	O
of	NN	O	O
TF-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
the	NN	O	O
differentiation	NN	O	O
status	NN	O	O
of	NN	O	O
these	NN	O	O
Raf-responsive	NN	O	B-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
more	NN	O	O
immature	NN	O	O
upon	NN	O	O
Raf	NN	O	O
activation	NN	O	O
as	NN	O	O
culture	NN	O	O
with	NN	O	O
the	NN	O	O
differentiation-inducing	NN	O	O
agent	NN	O	O
phorbol	NN	O	O
12	NN	O	O
myristate	NN	O	O
13-acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
and	NN	O	O
beta-estradiol	NN	O	O
resulted	NN	O	O
in	NN	O	O
decreased	NN	O	O
levels	NN	O	O
of	NN	O	O
the	NN	O	O
CD11b	NN	O	B-protein
and	NN	O	I-protein
CD18	NN	O	I-protein
integrin	NN	O	I-protein
molecules	NN	O	I-protein
on	NN	O	O
the	NN	O	O
cell	NN	O	B-cell_type
surface	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
when	NN	O	O
the	NN	O	O
Raf-responsive	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
induced	NN	O	O
to	NN	O	O
differentiate	NN	O	O
with	NN	O	O
PMA	NN	O	O
and	NN	O	O
GM-CSF	NN	O	B-protein
,	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
deltaRaf	NN	O	B-protein
:	NN	O	I-protein
ER	NN	O	I-protein
activation	NN	O	O
,	NN	O	O
increased	NN	O	O
levels	NN	O	O
of	NN	O	O
the	NN	O	O
CD11b	NN	O	B-protein
and	NN	O	I-protein
CD18	NN	O	I-protein
molecules	NN	O	I-protein
were	NN	O	O
detected	NN	O	O
.	NN	O	O

Retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
RA	NN	O	O
)	NN	O	O
inhibited	NN	O	O
3H-thymidine	NN	O	O
incorporation	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
GM-CSF	NN	O	B-protein
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
Raf	NN	O	O
activation	NN	O	O
counterbalanced	NN	O	O
the	NN	O	O
inhibition	NN	O	O
of	NN	O	O
DNA	NN	O	O
synthesis	NN	O	O
caused	NN	O	O
by	NN	O	O
RA	NN	O	O
but	NN	O	O
not	NN	O	O
PMA	NN	O	O
.	NN	O	O

Thus	NN	O	O
deregulated	NN	O	O
Raf	NN	O	O
expression	NN	O	O
can	NN	O	O
alter	NN	O	O
cytokine	NN	O	O
dependency	NN	O	O
,	NN	O	O
integrin	NN	O	O
expression	NN	O	O
and	NN	O	O
the	NN	O	O
stage	NN	O	O
of	NN	O	O
differentiation	NN	O	O
.	NN	O	O

These	NN	O	O
Raf-responsive	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
will	NN	O	O
be	NN	O	O
useful	NN	O	O
in	NN	O	O
elucidating	NN	O	O
the	NN	O	O
roles	NN	O	O
of	NN	O	O
the	NN	O	O
MAP	NN	O	O
kinase	NN	O	O
cascade	NN	O	O
on	NN	O	O
hematopoietic	NN	O	B-cell_type
cell	NN	O	I-cell_type
differentiation	NN	O	O
and	NN	O	O
malignant	NN	O	O
transformation	NN	O	O

-DOCSTART-	O

Cyclic	NN	O	O
AMP	NN	O	O
activates	NN	O	O
p38	NN	O	B-protein
mitogen-activated	NN	O	I-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
in	NN	O	O
Th2	NN	O	B-cell_type
cells	NN	O	I-cell_type
:	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
GATA-3	NN	O	B-protein
and	NN	O	O
stimulation	NN	O	O
of	NN	O	O
Th2	NN	O	B-cell_type
cytokine	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

cAMP	NN	O	O
is	NN	O	O
an	NN	O	O
important	NN	O	O
second	NN	O	O
messenger	NN	O	O
with	NN	O	O
immunomodulatory	NN	O	O
properties	NN	O	O
.	NN	O	O

Elevation	NN	O	O
of	NN	O	O
intracellular	NN	O	O
cAMP	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
induced	NN	O	O
by	NN	O	O
agents	NN	O	O
such	NN	O	O
as	NN	O	O
IL-1alpha	NN	O	B-protein
or	NN	O	O
PGs	NN	O	B-protein
,	NN	O	O
inhibits	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
.	NN	O	O

In	NN	O	O
effector	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	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
intracellular	NN	O	O
cAMP	NN	O	O
inhibits	NN	O	O
cytokine	NN	O	B-protein
production	NN	O	O
in	NN	O	O
Th1	NN	O	B-cell_type
cells	NN	O	I-cell_type
but	NN	O	O
stimulates	NN	O	O
cytokine	NN	O	B-protein
production	NN	O	O
in	NN	O	O
Th2	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
cAMP-induced	NN	O	O
effects	NN	O	O
in	NN	O	O
Th2	NN	O	B-cell_type
cells	NN	O	I-cell_type
occur	NN	O	O
independently	NN	O	O
of	NN	O	O
the	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
A	NN	O	I-protein
pathway	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
the	NN	O	O
major	NN	O	O
mediator	NN	O	O
of	NN	O	O
cAMP-induced	NN	O	O
signaling	NN	O	O
events	NN	O	O
in	NN	O	O
most	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

Instead	NN	O	O
,	NN	O	O
cAMP	NN	O	O
stimulates	NN	O	O
activation	NN	O	O
of	NN	O	O
p38	NN	O	B-protein
mitogen-activated	NN	O	I-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
in	NN	O	O
Th2	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

This	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
Th2	NN	O	B-cell_type
-selective	NN	O	O
event	NN	O	O
because	NN	O	O
cAMP	NN	O	O
barely	NN	O	O
increased	NN	O	O
p38	NN	O	B-protein
phosphorylation	NN	O	O
in	NN	O	O
Th1	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
in	NN	O	O
Th2	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
cAMP	NN	O	O
promotes	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
both	NN	O	O
IL-5	NN	O	B-protein
and	NN	O	O
IL-13	NN	O	B-protein
,	NN	O	O
which	NN	O	O
play	NN	O	O
distinct	NN	O	O
but	NN	O	O
critical	NN	O	O
roles	NN	O	O
in	NN	O	O
asthma	NN	O	O
pathogenesis	NN	O	O
.	NN	O	O

Our	NN	O	O
data	NN	O	O
also	NN	O	O
show	NN	O	O
that	NN	O	O
cAMP	NN	O	O
causes	NN	O	O
increased	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
GATA-3	NN	O	I-protein
,	NN	O	O
which	NN	O	O
we	NN	O	O
have	NN	O	O
shown	NN	O	O
is	NN	O	O
a	NN	O	O
critical	NN	O	O
regulator	NN	O	O
of	NN	O	O
Th2	NN	O	B-cell_type
cytokine	NN	O	O
gene	NN	O	O
expression	NN	O	O
and	NN	O	O
,	NN	O	O
in	NN	O	O
turn	NN	O	O
,	NN	O	O
of	NN	O	O
airway	NN	O	O
inflammation	NN	O	O
in	NN	O	O
mice	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
Th2	NN	O	B-cell_type
-specific	NN	O	O
GATA-3	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
p38	NN	O	B-protein
mitogen-activated	NN	O	I-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
activation	NN	O	O
together	NN	O	O
provide	NN	O	O
a	NN	O	O
molecular	NN	O	O
basis	NN	O	O
for	NN	O	O
the	NN	O	O
differential	NN	O	O
effects	NN	O	O
of	NN	O	O
cAMP	NN	O	O
in	NN	O	O
the	NN	O	O
two	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

-DOCSTART-	O

Characterization	NN	O	O
of	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
IL-13	NN	O	B-protein
signals	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
human	NN	O	B-protein
IL-13	NN	O	I-protein
receptor	NN	O	I-protein
alpha	NN	O	I-protein
chain	NN	O	I-protein
1	NN	O	I-protein
:	NN	O	O
redundancy	NN	O	O
of	NN	O	O
requirement	NN	O	O
of	NN	O	O
tyrosine	NN	O	O
residue	NN	O	O
for	NN	O	O
STAT3	NN	O	O
activation	NN	O	O
.	NN	O	O

IL-4	NN	O	B-protein
and	NN	O	O
IL-13	NN	O	B-protein
are	NN	O	O
pleiotropic	NN	O	B-protein
cytokines	NN	O	I-protein
whose	NN	O	O
biological	NN	O	O
activities	NN	O	O
overlap	NN	O	O
with	NN	O	O
each	NN	O	O
other	NN	O	O
.	NN	O	O

IL-13	NN	O	B-protein
receptor	NN	O	I-protein
alpha	NN	O	I-protein
chain	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	O
IL-13R	NN	O	B-protein
alpha	NN	O	I-protein
1	NN	O	I-protein
)	NN	O	O
is	NN	O	O
necessary	NN	O	O
for	NN	O	O
binding	NN	O	O
to	NN	O	O
IL-13	NN	O	B-protein
,	NN	O	O
and	NN	O	O
the	NN	O	O
heterodimer	NN	O	O
composed	NN	O	O
of	NN	O	O
IL-13R	NN	O	B-protein
alpha	NN	O	I-protein
1	NN	O	I-protein
and	NN	O	O
IL-4R	NN	O	B-protein
alpha	NN	O	I-protein
chain	NN	O	I-protein
transduces	NN	O	O
IL-13	NN	O	B-protein
and	NN	O	O
IL-4	NN	O	B-protein
signals	NN	O	O
;	NN	O	O
however	NN	O	O
,	NN	O	O
the	NN	O	O
functional	NN	O	O
mapping	NN	O	O
of	NN	O	O
the	NN	O	O
intracellular	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
IL-13R	NN	O	B-protein
alpha	NN	O	I-protein
1	NN	O	I-protein
is	NN	O	O
not	NN	O	O
fully	NN	O	O
understood	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
constructed	NN	O	O
wild	NN	O	O
and	NN	O	O
mutated	NN	O	O
types	NN	O	O
of	NN	O	O
human	NN	O	B-protein
IL-13R	NN	O	I-protein
alpha	NN	O	I-protein
1	NN	O	I-protein
,	NN	O	O
and	NN	O	O
analyzed	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
IL-13	NN	O	B-protein
signals	NN	O	O
using	NN	O	O
an	NN	O	O
IL-13R	NN	O	B-protein
alpha	NN	O	I-protein
1	NN	O	I-protein
-transfected	NN	O	O
human	NN	O	B-cell_line
B	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
IL-13R	NN	O	B-protein
alpha	NN	O	I-protein
1	NN	O	I-protein
evoked	NN	O	O
STAT3	NN	O	B-protein
activation	NN	O	O
by	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
IL-13	NN	O	B-protein
,	NN	O	O
and	NN	O	O
in	NN	O	O
stimulated	NN	O	O
human	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
on	NN	O	O
which	NN	O	O
IL-13R	NN	O	B-protein
alpha	NN	O	I-protein
1	NN	O	I-protein
was	NN	O	O
highly	NN	O	O
expressed	NN	O	O
,	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
IL-13	NN	O	B-protein
induced	NN	O	O
STAT3	NN	O	B-protein
activation	NN	O	O
.	NN	O	O

Replacement	NN	O	O
of	NN	O	O
the	NN	O	O
two	NN	O	O
tyrosine	NN	O	O
residues	NN	O	O
completely	NN	O	O
abolished	NN	O	O
STAT3	NN	O	B-protein
activation	NN	O	O
,	NN	O	O
although	NN	O	O
replacing	NN	O	O
either	NN	O	O
tyrosine	NN	O	O
residue	NN	O	O
alone	NN	O	O
retained	NN	O	O
it	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
Box1	NN	O	B-protein
region	NN	O	I-protein
and	NN	O	O
the	NN	O	O
C-terminal	NN	O	B-protein
tail	NN	O	I-protein
of	NN	O	O
IL-13R	NN	O	B-protein
alpha	NN	O	I-protein
1	NN	O	I-protein
were	NN	O	O
critical	NN	O	O
for	NN	O	O
binding	NN	O	O
to	NN	O	O
Tyk2	NN	O	B-protein
,	NN	O	O
and	NN	O	O
activation	NN	O	O
of	NN	O	O
Jak1	NN	O	B-protein
,	NN	O	O
Tyk2	NN	O	B-protein
,	NN	O	O
the	NN	O	O
insulin	NN	O	B-protein
receptor	NN	O	I-protein
substrate-1	NN	O	I-protein
and	NN	O	O
STAT6	NN	O	B-protein
respectively	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
STAT3	NN	O	B-protein
activation	NN	O	O
is	NN	O	O
involved	NN	O	O
with	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
IL-13	NN	O	B-protein
signals	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
along	NN	O	O
with	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
STAT6	NN	O	B-protein
,	NN	O	O
and	NN	O	O
that	NN	O	O
there	NN	O	O
is	NN	O	O
a	NN	O	O
unique	NN	O	O
sequence	NN	O	O
in	NN	O	O
IL-13R	NN	O	B-protein
alpha	NN	O	I-protein
1	NN	O	I-protein
to	NN	O	O
activate	NN	O	O
STAT3	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Functional	NN	O	O
uncoupling	NN	O	O
of	NN	O	O
the	NN	O	O
Janus	NN	O	O
kinase	NN	O	O
3-Stat5	NN	O	O
pathway	NN	O	O
in	NN	O	O
malignant	NN	O	O
growth	NN	O	O
of	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
1-transformed	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Human	NN	O	O
T	NN	O	O
cell	NN	O	O
leukemia	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
(	NN	O	O
HTLV-1	NN	O	O
)	NN	O	O
transforms	NN	O	O
cytokine	NN	O	B-protein
-dependent	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
causes	NN	O	O
adult	NN	O	O
T	NN	O	O
cell	NN	O	O
leukemia	NN	O	O
.	NN	O	O

Janus	NN	O	B-protein
tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	I-protein
Jak	NN	O	I-protein
)	NN	O	I-protein
3	NN	O	I-protein
and	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
Stat5a	NN	O	B-protein
and	NN	O	O
Stat5b	NN	O	B-protein
are	NN	O	O
essential	NN	O	O
for	NN	O	O
the	NN	O	O
proliferation	NN	O	O
of	NN	O	O
normal	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
are	NN	O	O
constitutively	NN	O	O
hyperactivated	NN	O	O
in	NN	O	O
both	NN	O	O
HTLV-1-transformed	NN	O	B-cell_line
human	NN	O	I-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
and	NN	O	O
lymphocytes	NN	O	B-cell_type
isolated	NN	O	O
from	NN	O	O
HTLV-1-infected	NN	O	O
patients	NN	O	O
;	NN	O	O
therefore	NN	O	O
,	NN	O	O
a	NN	O	O
critical	NN	O	O
role	NN	O	O
for	NN	O	O
the	NN	O	O
Jak3-Stat5	NN	O	O
pathway	NN	O	O
in	NN	O	O
the	NN	O	O
progression	NN	O	O
of	NN	O	O
this	NN	O	O
disease	NN	O	O
has	NN	O	O
been	NN	O	O
postulated	NN	O	O
.	NN	O	O

We	NN	O	O
recently	NN	O	O
reported	NN	O	O
that	NN	O	O
tyrphostin	NN	O	O
AG-490	NN	O	O
selectively	NN	O	O
blocked	NN	O	O
IL-2	NN	O	O
activation	NN	O	O
of	NN	O	O
Jak3/Stat5	NN	O	B-protein
and	NN	O	O
growth	NN	O	O
of	NN	O	O
murine	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

Here	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
disruption	NN	O	O
of	NN	O	O
Jak3/Stat5a/b	NN	O	O
signaling	NN	O	O
with	NN	O	O
AG-490	NN	O	O
(	NN	O	O
50	NN	O	O
&	NN	O	O
mgr	NN	O	O
;	NN	O	O
M	NN	O	O
)	NN	O	O
blocked	NN	O	O
the	NN	O	O
proliferation	NN	O	O
of	NN	O	O
primary	NN	O	B-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
but	NN	O	O
paradoxically	NN	O	O
failed	NN	O	O
to	NN	O	O
inhibit	NN	O	O
the	NN	O	O
proliferation	NN	O	O
of	NN	O	O
HTLV-1-transformed	NN	O	B-cell_line
human	NN	O	I-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
HuT-102	NN	O	B-cell_line
and	NN	O	O
MT-2	NN	O	B-cell_line
.	NN	O	O

Structural	NN	O	O
homologues	NN	O	O
of	NN	O	O
AG-490	NN	O	O
also	NN	O	O
inhibited	NN	O	O
the	NN	O	O
proliferation	NN	O	O
of	NN	O	O
primary	NN	O	B-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
but	NN	O	O
not	NN	O	O
HTLV-1-infected	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Disruption	NN	O	O
of	NN	O	O
constitutive	NN	O	O
Jak3/Stat5	NN	O	O
activation	NN	O	O
by	NN	O	O
AG-490	NN	O	O
was	NN	O	O
demonstrated	NN	O	O
by	NN	O	O
inhibition	NN	O	O
of	NN	O	O
1	NN	O	O
)	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
Jak3	NN	O	B-protein
,	NN	O	O
Stat5a	NN	O	B-protein
(	NN	O	O
Tyr	NN	O	O
(	NN	O	O
694	NN	O	O
)	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
Stat5b	NN	O	B-protein
(	NN	O	O
Tyr	NN	O	O
(	NN	O	O
699	NN	O	O
)	NN	O	O
)	NN	O	O
;	NN	O	O
2	NN	O	O
)	NN	O	O
serine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
Stat5a	NN	O	B-protein
(	NN	O	O
Ser	NN	O	O
(	NN	O	O
726	NN	O	O
)	NN	O	O
)	NN	O	O
as	NN	O	O
determined	NN	O	O
by	NN	O	O
a	NN	O	O
novel	NN	O	O
phosphospecific	NN	O	B-protein
Ab	NN	O	I-protein
;	NN	O	O
and	NN	O	O
3	NN	O	O
)	NN	O	O
Stat5a/b	NN	O	O
DNA	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
Stat5-responsive	NN	O	B-DNA
beta-casein	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
AG-490	NN	O	O
had	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
DNA	NN	O	O
binding	NN	O	O
by	NN	O	O
p50/p65	NN	O	B-protein
components	NN	O	I-protein
of	NN	O	O
NF-kappaB	NN	O	B-protein
,	NN	O	O
a	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
activated	NN	O	O
by	NN	O	O
the	NN	O	O
HTLV-1-encoded	NN	O	B-protein
phosphoprotein	NN	O	I-protein
,	NN	O	I-protein
Tax	NN	O	I-protein
.	NN	O	O

Collectively	NN	O	O
,	NN	O	O
these	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
Jak3-Stat5	NN	O	O
pathway	NN	O	O
in	NN	O	O
HTLV-1-transformed	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
has	NN	O	O
become	NN	O	O
functionally	NN	O	O
redundant	NN	O	O
for	NN	O	O
proliferation	NN	O	O
.	NN	O	O

Reversal	NN	O	O
of	NN	O	O
this	NN	O	O
functional	NN	O	O
uncoupling	NN	O	O
may	NN	O	O
be	NN	O	O
required	NN	O	O
before	NN	O	O
Jak3/Stat5	NN	O	O
inhibitors	NN	O	O
will	NN	O	O
be	NN	O	O
useful	NN	O	O
in	NN	O	O
the	NN	O	O
treatment	NN	O	O
of	NN	O	O
this	NN	O	O
malignancy	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
Epstein-Barr	NN	O	B-DNA
virus	NN	O	I-DNA
promoter	NN	O	I-DNA
initiating	NN	O	O
B-cell	NN	O	O
transformation	NN	O	O
is	NN	O	O
activated	NN	O	O
by	NN	O	O
RFX	NN	O	B-protein
proteins	NN	O	I-protein
and	NN	O	O
the	NN	O	O
B-cell-specific	NN	O	B-protein
activator	NN	O	I-protein
protein	NN	O	I-protein
BSAP/Pax5	NN	O	I-protein
.	NN	O	O

Epstein-Barr	NN	O	O
virus	NN	O	O
(	NN	O	O
EBV	NN	O	O
)	NN	O	O
-induced	NN	O	O
B-cell	NN	O	B-cell_type
growth	NN	O	O
transformation	NN	O	O
,	NN	O	O
a	NN	O	O
central	NN	O	O
feature	NN	O	O
of	NN	O	O
the	NN	O	O
virus	NN	O	O
'	NN	O	O
strategy	NN	O	O
for	NN	O	O
colonizing	NN	O	O
the	NN	O	O
human	NN	O	O
B-cell	NN	O	O
system	NN	O	O
,	NN	O	O
requires	NN	O	O
full	NN	O	O
virus	NN	O	O
latent	NN	O	O
gene	NN	O	O
expression	NN	O	O
and	NN	O	O
is	NN	O	O
initiated	NN	O	O
by	NN	O	O
transcription	NN	O	O
from	NN	O	O
the	NN	O	O
viral	NN	O	B-DNA
promoter	NN	O	I-DNA
Wp	NN	O	I-DNA
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
when	NN	O	O
EBV	NN	O	O
accesses	NN	O	O
other	NN	O	O
cell	NN	O	O
types	NN	O	O
,	NN	O	O
this	NN	O	O
growth-transforming	NN	O	O
program	NN	O	O
is	NN	O	O
not	NN	O	O
activated	NN	O	O
.	NN	O	O

The	NN	O	O
present	NN	O	O
work	NN	O	O
focuses	NN	O	O
on	NN	O	O
a	NN	O	O
region	NN	O	O
of	NN	O	O
Wp	NN	O	B-DNA
which	NN	O	O
in	NN	O	O
reporter	NN	O	O
assays	NN	O	O
confers	NN	O	O
B-cell	NN	O	B-cell_type
-specific	NN	O	O
activity	NN	O	O
.	NN	O	O

Bandshift	NN	O	O
studies	NN	O	O
indicate	NN	O	O
that	NN	O	O
this	NN	O	O
region	NN	O	O
contains	NN	O	O
three	NN	O	O
factor	NN	O	O
binding	NN	O	O
sites	NN	O	O
,	NN	O	O
termed	NN	O	O
sites	NN	O	B-DNA
B	NN	O	I-DNA
,	NN	O	I-DNA
C	NN	O	I-DNA
,	NN	O	I-DNA
and	NN	O	I-DNA
D	NN	O	I-DNA
,	NN	O	O
in	NN	O	O
addition	NN	O	O
to	NN	O	O
a	NN	O	O
previously	NN	O	O
characterized	NN	O	O
CREB	NN	O	B-DNA
site	NN	O	I-DNA
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
site	NN	O	B-DNA
C	NN	O	I-DNA
binds	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
ubiquitously	NN	O	O
expressed	NN	O	O
RFX	NN	O	B-protein
family	NN	O	I-protein
of	NN	O	I-protein
proteins	NN	O	I-protein
,	NN	O	O
notably	NN	O	O
RFX1	NN	O	B-protein
,	NN	O	O
RFX3	NN	O	B-protein
,	NN	O	O
and	NN	O	O
the	NN	O	O
associated	NN	O	O
factor	NN	O	O
MIBP1	NN	O	B-protein
,	NN	O	O
whereas	NN	O	O
sites	NN	O	B-DNA
B	NN	O	I-DNA
and	NN	O	I-DNA
D	NN	O	I-DNA
both	NN	O	O
bind	NN	O	O
the	NN	O	O
B-cell-specific	NN	O	B-protein
activator	NN	O	I-protein
protein	NN	O	I-protein
BSAP/Pax5	NN	O	I-protein
.	NN	O	O

In	NN	O	O
reporter	NN	O	O
assays	NN	O	O
with	NN	O	O
mutant	NN	O	B-DNA
Wp	NN	O	I-DNA
constructs	NN	O	I-DNA
,	NN	O	O
the	NN	O	O
loss	NN	O	O
of	NN	O	O
factor	NN	O	O
binding	NN	O	O
to	NN	O	O
any	NN	O	O
one	NN	O	O
of	NN	O	O
these	NN	O	O
sites	NN	O	O
severely	NN	O	O
impaired	NN	O	O
promoter	NN	O	O
activity	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
while	NN	O	O
the	NN	O	O
wild-type	NN	O	B-DNA
promoter	NN	O	I-DNA
could	NN	O	O
be	NN	O	O
activated	NN	O	O
in	NN	O	O
non-B	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
ectopic	NN	O	O
BSAP	NN	O	O
expression	NN	O	O
.	NN	O	O

We	NN	O	O
suggest	NN	O	O
that	NN	O	O
Wp	NN	O	B-DNA
regulation	NN	O	O
by	NN	O	O
BSAP	NN	O	B-protein
helps	NN	O	O
to	NN	O	O
ensure	NN	O	O
the	NN	O	O
B-cell	NN	O	B-cell_type
specificity	NN	O	O
of	NN	O	O
EBV	NN	O	O
's	NN	O	O
growth-transforming	NN	O	O
function	NN	O	O
.	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
of	NN	O	O
the	NN	O	O
Lck	NN	O	B-protein
tyrosine	NN	O	I-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
by	NN	O	O
the	NN	O	O
Herpesvirus	NN	O	O
saimiri	NN	O	O
tip	NN	O	B-protein
protein	NN	O	I-protein
involves	NN	O	O
two	NN	O	O
binding	NN	O	O
interactions	NN	O	O
.	NN	O	O

The	NN	O	O
Tip	NN	O	B-protein
protein	NN	O	I-protein
of	NN	O	O
Herpesvirus	NN	O	O
saimiri	NN	O	O
strain	NN	O	O
484C	NN	O	O
binds	NN	O	O
to	NN	O	O
and	NN	O	O
activates	NN	O	O
the	NN	O	O
Lck	NN	O	B-protein
tyrosine	NN	O	I-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
.	NN	O	O

Two	NN	O	O
sequences	NN	O	O
in	NN	O	O
the	NN	O	O
Tip	NN	O	B-protein
protein	NN	O	I-protein
were	NN	O	O
previously	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
binding	NN	O	O
to	NN	O	O
Lck	NN	O	B-protein
.	NN	O	O

A	NN	O	O
proline-rich	NN	O	B-protein
region	NN	O	I-protein
,	NN	O	O
residues	NN	O	O
132-141	NN	O	O
,	NN	O	O
binds	NN	O	O
to	NN	O	O
the	NN	O	O
SH3	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
the	NN	O	O
Lck	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

We	NN	O	O
show	NN	O	O
here	NN	O	O
that	NN	O	O
the	NN	O	O
other	NN	O	O
Lck-binding	NN	O	B-protein
domain	NN	O	I-protein
,	NN	O	O
residues	NN	O	O
104-113	NN	O	O
,	NN	O	O
binds	NN	O	O
to	NN	O	O
the	NN	O	O
carboxyl-terminal	NN	O	B-protein
half	NN	O	I-protein
of	NN	O	O
Lck	NN	O	B-protein
and	NN	O	O
that	NN	O	O
this	NN	O	O
binding	NN	O	O
does	NN	O	O
not	NN	O	O
require	NN	O	O
the	NN	O	O
Lck	NN	O	B-protein
SH3	NN	O	I-protein
domain	NN	O	I-protein
.	NN	O	O

Mutated	NN	O	O
Tip	NN	O	B-protein
containing	NN	O	O
only	NN	O	O
one	NN	O	O
functional	NN	O	O
Lck-binding	NN	O	B-protein
domain	NN	O	I-protein
can	NN	O	O
bind	NN	O	O
stably	NN	O	O
to	NN	O	O
Lck	NN	O	B-protein
,	NN	O	O
although	NN	O	O
not	NN	O	O
as	NN	O	O
strongly	NN	O	O
as	NN	O	O
wild-type	NN	O	B-protein
Tip	NN	O	I-protein
.	NN	O	O

Interaction	NN	O	O
of	NN	O	O
Tip	NN	O	B-protein
with	NN	O	O
Lck	NN	O	B-protein
through	NN	O	O
either	NN	O	O
Lck-binding	NN	O	B-protein
domain	NN	O	I-protein
increases	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
Lck	NN	O	B-protein
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

Simultaneous	NN	O	O
binding	NN	O	O
of	NN	O	O
both	NN	O	O
domains	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
maximal	NN	O	O
activation	NN	O	O
of	NN	O	O
Lck	NN	O	B-protein
.	NN	O	O

The	NN	O	O
transient	NN	O	O
expression	NN	O	O
of	NN	O	O
Tip	NN	O	B-protein
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
found	NN	O	O
to	NN	O	O
stimulate	NN	O	O
both	NN	O	O
Stat3	NN	O	B-protein
-dependent	NN	O	O
and	NN	O	O
NF-AT	NN	O	B-protein
-dependent	NN	O	O
transcription	NN	O	O
.	NN	O	O

Mutant	NN	O	O
forms	NN	O	O
of	NN	O	O
Tip	NN	O	B-protein
lacking	NN	O	O
one	NN	O	O
or	NN	O	O
the	NN	O	O
other	NN	O	O
of	NN	O	O
the	NN	O	O
two	NN	O	O
Lck-binding	NN	O	B-protein
domains	NN	O	I-protein
retained	NN	O	O
the	NN	O	O
ability	NN	O	O
to	NN	O	O
stimulate	NN	O	O
Stat3	NN	O	B-protein
-dependent	NN	O	O
transcription	NN	O	O
.	NN	O	O

Tip	NN	O	B-protein
lacking	NN	O	O
the	NN	O	O
proline-rich	NN	O	O
Lck-binding	NN	O	B-protein
domain	NN	O	I-protein
exhibited	NN	O	O
almost	NN	O	O
wild-type	NN	O	O
activity	NN	O	O
in	NN	O	O
this	NN	O	O
assay	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
ablation	NN	O	O
of	NN	O	O
either	NN	O	O
Lck-binding	NN	O	B-protein
domain	NN	O	I-protein
abolished	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
Tip	NN	O	B-protein
to	NN	O	O
stimulate	NN	O	O
NF-AT	NN	O	B-protein
-dependent	NN	O	O
transcription	NN	O	O
.	NN	O	O

Full	NN	O	O
biological	NN	O	O
activity	NN	O	O
of	NN	O	O
Tip	NN	O	B-protein
,	NN	O	O
therefore	NN	O	O
,	NN	O	O
appears	NN	O	O
to	NN	O	O
require	NN	O	O
both	NN	O	O
Lck-binding	NN	O	B-protein
domains	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Accumulation	NN	O	O
of	NN	O	O
RXR	NN	O	B-protein
alpha	NN	O	I-protein
during	NN	O	O
activation	NN	O	O
of	NN	O	O
cycling	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
:	NN	O	O
modulation	NN	O	O
of	NN	O	O
RXRE	NN	O	O
transactivation	NN	O	O
function	NN	O	O
by	NN	O	O
mitogen-activated	NN	O	O
protein	NN	O	O
kinase	NN	O	O
pathways	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
reported	NN	O	O
that	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
resting	NN	O	O
human	NN	O	B-cell_type
immature	NN	O	I-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
T	NN	O	I-cell_type
(	NN	O	I-cell_type
PBT	NN	O	I-cell_type
)	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
is	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
loss	NN	O	O
of	NN	O	O
retinoid	NN	O	B-protein
X	NN	O	I-protein
receptor	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
RXRalpha	NN	O	B-protein
)	NN	O	O
expression	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
,	NN	O	O
unlike	NN	O	O
resting	NN	O	O
cells	NN	O	O
,	NN	O	O
activation	NN	O	O
of	NN	O	O
cycling	NN	O	B-cell_type
human	NN	O	I-cell_type
mature	NN	O	I-cell_type
PBT	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
T	NN	O	B-cell_line
lymphocyte	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
is	NN	O	O
accompanied	NN	O	O
by	NN	O	O
the	NN	O	O
accumulation	NN	O	O
of	NN	O	O
RXRalpha	NN	O	B-protein
mRNA	NN	O	O
and	NN	O	O
protein	NN	O	O
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
further	NN	O	O
augmented	NN	O	O
RXRalpha	NN	O	B-protein
expression	NN	O	O
,	NN	O	O
indicating	NN	O	O
the	NN	O	O
involvement	NN	O	O
of	NN	O	O
calcineurin	NN	O	O
pathways	NN	O	O
in	NN	O	O
the	NN	O	O
process	NN	O	O
.	NN	O	O

9-cis	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
inhibited	NN	O	O
the	NN	O	O
accumulation	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
retinoids	NN	O	O
can	NN	O	O
regulate	NN	O	O
the	NN	O	O
synthesis	NN	O	O
of	NN	O	O
their	NN	O	O
own	NN	O	O
receptors	NN	O	O
during	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
.	NN	O	O

Transfection	NN	O	O
analysis	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
using	NN	O	O
RXRE-dependent	NN	O	O
reporter	NN	O	O
assays	NN	O	O
,	NN	O	O
showed	NN	O	O
that	NN	O	O
RXRalpha	NN	O	B-protein
accumulated	NN	O	O
during	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
was	NN	O	O
transcriptionally	NN	O	O
inactive	NN	O	O
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
such	NN	O	O
inhibition	NN	O	O
,	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
two	NN	O	O
mitogen-activated	NN	O	O
protein	NN	O	O
kinase	NN	O	O
pathways	NN	O	O
,	NN	O	O
c-Jun	NN	O	B-protein
N-terminal	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	O
JNK	NN	O	B-protein
)	NN	O	O
and	NN	O	O
extracellular	NN	O	B-protein
signal-regulated	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	O
ERK	NN	O	B-protein
)	NN	O	O
,	NN	O	O
in	NN	O	O
modulating	NN	O	O
RXRE-dependent	NN	O	O
transcription	NN	O	O
,	NN	O	O
was	NN	O	O
explored	NN	O	O
.	NN	O	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
constitutively	NN	O	O
active	NN	O	O
MAP/ERK	NN	O	B-protein
kinase	NN	O	I-protein
kinase	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	O
MEKK1	NN	O	B-protein
)	NN	O	O
inhibited	NN	O	O
RXRE-dependent	NN	O	O
transcription	NN	O	O
,	NN	O	O
whereas	NN	O	O
dominant	NN	O	B-protein
negative	NN	O	I-protein
MEKK1	NN	O	I-protein
increased	NN	O	O
the	NN	O	O
transcription	NN	O	O
,	NN	O	O
indicating	NN	O	O
the	NN	O	O
involvement	NN	O	O
of	NN	O	O
JNK	NN	O	B-protein
signaling	NN	O	O
pathways	NN	O	O
in	NN	O	O
the	NN	O	O
process	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
expression	NN	O	O
of	NN	O	O
constitutively	NN	O	B-protein
active	NN	O	I-protein
MEK1	NN	O	I-protein
,	NN	O	O
which	NN	O	O
activates	NN	O	O
ERK	NN	O	B-protein
pathway	NN	O	O
,	NN	O	O
enhanced	NN	O	O
RXRE-dependent	NN	O	O
activation	NN	O	O
.	NN	O	O

When	NN	O	O
both	NN	O	O
were	NN	O	O
activated	NN	O	O
simultaneously	NN	O	O
,	NN	O	O
JNK	NN	O	B-protein
pathway	NN	O	O
was	NN	O	O
dominant	NN	O	O
over	NN	O	O
ERK	NN	O	B-protein
pathway	NN	O	O
and	NN	O	O
resulted	NN	O	O
in	NN	O	O
inhibition	NN	O	O
of	NN	O	O
RXRE-mediated	NN	O	O
transcription	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
demonstrate	NN	O	O
a	NN	O	O
dual	NN	O	O
regulatory	NN	O	O
control	NN	O	O
of	NN	O	O
RXRalpha	NN	O	B-protein
expression	NN	O	O
during	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
resting	NN	O	B-cell_type
and	NN	O	I-cell_type
cycling	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
indicate	NN	O	O
a	NN	O	O
dynamic	NN	O	O
balance	NN	O	O
between	NN	O	O
JNK	NN	O	O
and	NN	O	O
ERK	NN	O	O
pathways	NN	O	O
in	NN	O	O
modulating	NN	O	O
RXRE-mediated	NN	O	O
transactivation	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
proteasome	NN	O	B-protein
regulates	NN	O	O
receptor-mediated	NN	O	O
endocytosis	NN	O	O
of	NN	O	O
interleukin-2	NN	O	B-protein
.	NN	O	O

Recent	NN	O	O
studies	NN	O	O
have	NN	O	O
increasingly	NN	O	O
implicated	NN	O	O
the	NN	O	O
proteasome	NN	O	B-protein
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
cell	NN	O	B-protein
surface	NN	O	I-protein
receptors	NN	O	I-protein
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
the	NN	O	O
proteasome	NN	O	B-protein
for	NN	O	O
ligand-dependent	NN	O	O
endocytosis	NN	O	O
and	NN	O	O
degradation	NN	O	O
of	NN	O	O
the	NN	O	O
interleukin-2	NN	O	B-protein
(	NN	O	I-protein
IL-2	NN	O	I-protein
)	NN	O	I-protein
-interleukin-2	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	I-protein
IL-2R	NN	O	I-protein
)	NN	O	I-protein
complex	NN	O	I-protein
.	NN	O	O

Proteasome	NN	O	O
inhibitors	NN	O	O
impaired	NN	O	O
internalization	NN	O	O
of	NN	O	O
IL-2.IL-2R	NN	O	B-protein
and	NN	O	O
prevented	NN	O	O
the	NN	O	O
lysosomal	NN	O	O
degradation	NN	O	O
of	NN	O	O
this	NN	O	O
cytokine	NN	O	B-protein
.	NN	O	O

Based	NN	O	O
on	NN	O	O
time-course	NN	O	O
studies	NN	O	O
,	NN	O	O
proteasome	NN	O	B-protein
activity	NN	O	O
is	NN	O	O
primarily	NN	O	O
required	NN	O	O
after	NN	O	O
initial	NN	O	O
endocytosis	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2.IL-2R	NN	O	B-protein
.	NN	O	O

Proteasome	NN	O	O
function	NN	O	O
was	NN	O	O
also	NN	O	O
necessary	NN	O	O
for	NN	O	O
the	NN	O	O
lysosomal	NN	O	O
degradation	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
internalized	NN	O	O
by	NN	O	O
IL-2R	NN	O	B-protein
that	NN	O	O
were	NN	O	O
comprised	NN	O	O
of	NN	O	O
cytoplasmic	NN	O	B-protein
tailless	NN	O	I-protein
beta-	NN	O	B-protein
or	NN	O	I-protein
gamma	NN	O	I-protein
c-subunits	NN	O	I-protein
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
the	NN	O	O
target	NN	O	O
protein	NN	O	O
for	NN	O	O
the	NN	O	O
proteasome	NN	O	B-protein
is	NN	O	O
independent	NN	O	O
of	NN	O	O
either	NN	O	O
the	NN	O	O
cytoplasmic	NN	O	B-protein
tail	NN	O	I-protein
of	NN	O	O
the	NN	O	O
IL-2R	NN	O	B-protein
beta-	NN	O	B-protein
or	NN	O	I-protein
gamma	NN	O	I-protein
c-subunits	NN	O	I-protein
and	NN	O	O
their	NN	O	O
associated	NN	O	O
signaling	NN	O	O
components	NN	O	O
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
a	NN	O	O
functional	NN	O	O
proteasome	NN	O	B-protein
is	NN	O	O
required	NN	O	O
for	NN	O	O
optimal	NN	O	O
endocytosis	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2R/ligand	NN	O	B-protein
complex	NN	O	I-protein
and	NN	O	O
is	NN	O	O
essential	NN	O	O
for	NN	O	O
the	NN	O	O
subsequent	NN	O	O
lysosomal	NN	O	O
degradation	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
possibly	NN	O	O
by	NN	O	O
regulating	NN	O	O
trafficking	NN	O	O
to	NN	O	O
the	NN	O	O
lysosome	NN	O	O
.	NN	O	O

-DOCSTART-	O

Functional	NN	O	O
characterization	NN	O	O
of	NN	O	O
the	NN	O	O
two	NN	O	O
alternative	NN	O	O
promoters	NN	O	O
of	NN	O	O
human	NN	O	B-DNA
p45	NN	O	I-DNA
NF-E2	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

OBJECTIVE	NN	O	O
:	NN	O	O
The	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-E2	NN	O	I-protein
,	NN	O	O
a	NN	O	O
heterodimeric	NN	O	B-protein
protein	NN	O	I-protein
complex	NN	O	I-protein
composed	NN	O	O
of	NN	O	O
p45	NN	O	B-protein
and	NN	O	O
small	NN	O	O
Maf	NN	O	B-protein
family	NN	O	I-protein
proteins	NN	O	I-protein
,	NN	O	O
is	NN	O	O
considered	NN	O	O
crucial	NN	O	O
for	NN	O	O
the	NN	O	O
proper	NN	O	O
differentiation	NN	O	O
of	NN	O	O
erythrocytes	NN	O	B-cell_type
and	NN	O	O
megakaryocytes	NN	O	B-cell_type
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
the	NN	O	O
results	NN	O	O
of	NN	O	O
studies	NN	O	O
aimed	NN	O	O
at	NN	O	O
understanding	NN	O	O
the	NN	O	O
regulatory	NN	O	O
mechanisms	NN	O	O
controlling	NN	O	O
p45	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
erythroid	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

MATERIALS	NN	O	O
AND	NN	O	O
METHODS	NN	O	O
:	NN	O	O
Human	NN	O	B-RNA
p45	NN	O	I-RNA
mRNAs	NN	O	I-RNA
have	NN	O	O
two	NN	O	O
alternative	NN	O	O
isoforms	NN	O	O
,	NN	O	O
aNF-E2	NN	O	B-RNA
and	NN	O	O
fNF-E2	NN	O	B-RNA
,	NN	O	O
and	NN	O	O
these	NN	O	O
isoforms	NN	O	O
are	NN	O	O
transcribed	NN	O	O
from	NN	O	O
the	NN	O	O
alternative	NN	O	B-DNA
promoters	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
investigated	NN	O	O
lineage-specific	NN	O	O
expression	NN	O	O
of	NN	O	O
both	NN	O	O
isomers	NN	O	O
in	NN	O	O
human	NN	O	O
erythroid	NN	O	B-cell_type
and	NN	O	I-cell_type
megakaryocytic	NN	O	I-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
reverse	NN	O	O
transcriptase	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
or	NN	O	O
Northern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
.	NN	O	O

For	NN	O	O
functional	NN	O	O
characterization	NN	O	O
of	NN	O	O
both	NN	O	O
promoters	NN	O	O
,	NN	O	O
plasmids	NN	O	O
in	NN	O	O
which	NN	O	O
reporter	NN	O	B-DNA
genes	NN	O	I-DNA
were	NN	O	O
placed	NN	O	O
under	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
a	NN	O	O
series	NN	O	O
of	NN	O	O
truncated	NN	O	O
or	NN	O	O
mutated	NN	O	O
promoter	NN	O	B-DNA
fragments	NN	O	I-DNA
were	NN	O	O
transfected	NN	O	O
to	NN	O	O
human	NN	O	B-cell_line
hematopoietic	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
When	NN	O	O
CD34	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
isolated	NN	O	O
from	NN	O	O
human	NN	O	O
cord	NN	O	O
blood	NN	O	O
were	NN	O	O
induced	NN	O	O
to	NN	O	O
unilineage	NN	O	O
erythroid	NN	O	O
or	NN	O	O
megakaryocytic	NN	O	O
differentiation	NN	O	O
in	NN	O	O
liquid	NN	O	O
suspension	NN	O	O
culture	NN	O	O
,	NN	O	O
both	NN	O	O
transcripts	NN	O	O
,	NN	O	O
although	NN	O	O
barely	NN	O	O
detected	NN	O	O
at	NN	O	O
day	NN	O	O
0	NN	O	O
,	NN	O	O
were	NN	O	O
induced	NN	O	O
in	NN	O	O
both	NN	O	O
erythroid	NN	O	O
and	NN	O	O
megakaryocytic	NN	O	O
cultures	NN	O	O
.	NN	O	O

fNF-E2	NN	O	B-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
more	NN	O	O
abundant	NN	O	O
in	NN	O	O
erythroid	NN	O	B-cell_type
cells	NN	O	I-cell_type
than	NN	O	O
megakaryocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
at	NN	O	O
day	NN	O	O
7	NN	O	O
of	NN	O	O
culture	NN	O	O
.	NN	O	O

Although	NN	O	O
both	NN	O	O
isomers	NN	O	O
were	NN	O	O
expressed	NN	O	O
in	NN	O	O
human	NN	O	B-cell_line
erythroid-megakaryocytic	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
megakaryocytic	NN	O	O
maturation	NN	O	O
with	NN	O	O
loss	NN	O	O
of	NN	O	O
erythroid	NN	O	O
phenotype	NN	O	O
induced	NN	O	O
by	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
resulted	NN	O	O
in	NN	O	O
exclusive	NN	O	O
downregulation	NN	O	O
of	NN	O	O
fNF-E2	NN	O	B-RNA
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
fNF-E2	NN	O	B-DNA
promoter	NN	O	I-DNA
is	NN	O	O
more	NN	O	O
erythroid	NN	O	O
specific	NN	O	O
.	NN	O	O

Functional	NN	O	O
analysis	NN	O	O
of	NN	O	O
fNF-E2	NN	O	B-DNA
promoter	NN	O	I-DNA
showed	NN	O	O
that	NN	O	O
the	NN	O	O
promoter	NN	O	O
is	NN	O	O
active	NN	O	O
only	NN	O	O
in	NN	O	O
erythroid-megakaryocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
that	NN	O	O
the	NN	O	O
double	NN	O	B-DNA
GATA	NN	O	I-DNA
sit	NN	O	I-DNA
e	NN	O	O
in	NN	O	O
the	NN	O	O
proximal	NN	O	O
region	NN	O	O
is	NN	O	O
necessary	NN	O	O
for	NN	O	O
its	NN	O	O
efficient	NN	O	O
activity	NN	O	O
.	NN	O	O

CONCLUSION	NN	O	O
:	NN	O	O
These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
GATA	NN	O	B-protein
proteins	NN	O	I-protein
,	NN	O	O
which	NN	O	O
govern	NN	O	O
the	NN	O	O
differentiation	NN	O	O
of	NN	O	O
erythroid	NN	O	B-cell_type
lineage	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
are	NN	O	O
required	NN	O	O
for	NN	O	O
full	NN	O	O
promoter	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
p45	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
heme	NN	O	B-DNA
oxygenase-1	NN	O	I-DNA
and	NN	O	O
its	NN	O	O
functional	NN	O	O
significance	NN	O	O
in	NN	O	O
acetaminophen-induced	NN	O	O
hepatitis	NN	O	O
and	NN	O	O
hepatocellular	NN	O	O
injury	NN	O	O
in	NN	O	O
the	NN	O	O
rat	NN	O	O
.	NN	O	O

BACKGROUND/AIM	NN	O	O
:	NN	O	O
Glutathione	NN	O	O
depletion	NN	O	O
contributes	NN	O	O
to	NN	O	O
acetaminophen	NN	O	O
hepatotoxicity	NN	O	O
and	NN	O	O
is	NN	O	O
known	NN	O	O
to	NN	O	O
induce	NN	O	O
the	NN	O	O
oxidative	NN	O	O
stress	NN	O	O
reactant	NN	O	O
heme	NN	O	B-protein
oxygenase-1	NN	O	I-protein
.	NN	O	O

The	NN	O	O
metabolites	NN	O	O
of	NN	O	O
the	NN	O	O
heme	NN	O	O
oxygenase	NN	O	O
pathway	NN	O	O
,	NN	O	O
biliverdin	NN	O	O
,	NN	O	O
carbon	NN	O	O
monoxide	NN	O	O
,	NN	O	O
and	NN	O	O
iron	NN	O	O
may	NN	O	O
modulate	NN	O	O
acetaminophen	NN	O	O
toxicity	NN	O	O
.	NN	O	O

The	NN	O	O
aim	NN	O	O
of	NN	O	O
this	NN	O	O
study	NN	O	O
was	NN	O	O
to	NN	O	O
assess	NN	O	O
cell-type	NN	O	O
specific	NN	O	O
expression	NN	O	O
of	NN	O	O
heme	NN	O	B-DNA
oxygenase-1	NN	O	I-DNA
and	NN	O	O
its	NN	O	O
impact	NN	O	O
on	NN	O	O
liver	NN	O	O
injury	NN	O	O
and	NN	O	O
microcirculatory	NN	O	O
disturbances	NN	O	O
in	NN	O	O
a	NN	O	O
model	NN	O	O
of	NN	O	O
acetaminophen-induced	NN	O	O
hepatitis	NN	O	O
.	NN	O	O

METHODS	NN	O	O
:	NN	O	O
Gene	NN	O	O
expression	NN	O	O
of	NN	O	O
heme	NN	O	B-DNA
oxygenase-1	NN	O	I-DNA
was	NN	O	O
studied	NN	O	O
by	NN	O	O
Northern-	NN	O	O
and	NN	O	O
Western	NN	O	O
analysis	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
immunohistochemistry	NN	O	O
.	NN	O	O

The	NN	O	O
time	NN	O	O
course	NN	O	O
of	NN	O	O
heme	NN	O	B-protein
oxygenase-1	NN	O	I-protein
and	NN	O	I-protein
-2	NN	O	I-protein
,	NN	O	O
cytokine-induced	NN	O	B-protein
neutrophil	NN	O	I-protein
chemoattractant-1	NN	O	I-protein
,	NN	O	O
and	NN	O	O
intercellular	NN	O	B-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
was	NN	O	O
studied	NN	O	O
by	NN	O	O
Northern	NN	O	O
analysis	NN	O	O
.	NN	O	O

DNA-binding	NN	O	O
activity	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
was	NN	O	O
determined	NN	O	O
by	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
.	NN	O	O

Sinusoidal	NN	O	O
perfusion	NN	O	O
and	NN	O	O
leukocyte-endothelial	NN	O	O
interactions	NN	O	O
were	NN	O	O
assessed	NN	O	O
by	NN	O	O
intravital	NN	O	O
microscopy	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
Acetaminophen	NN	O	O
caused	NN	O	O
a	NN	O	O
moderate	NN	O	O
sinusoidal	NN	O	O
perfusion	NN	O	O
failure	NN	O	O
(	NN	O	O
-15	NN	O	O
%	NN	O	O
)	NN	O	O
and	NN	O	O
infiltration	NN	O	O
of	NN	O	O
neutrophils	NN	O	B-cell_type
along	NN	O	O
with	NN	O	O
activation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
and	NN	O	O
intercellular	NN	O	B-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
and	NN	O	O
cytokine-induced	NN	O	B-protein
neutrophil	NN	O	I-protein
chemoattractant-1	NN	O	I-protein
mRNAs	NN	O	O
.	NN	O	O

Induction	NN	O	O
of	NN	O	O
heme	NN	O	B-protein
oxygenase-1	NN	O	I-protein
mRNA	NN	O	O
and	NN	O	O
protein	NN	O	O
(	NN	O	O
approximately	NN	O	O
30-fold	NN	O	O
)	NN	O	O
in	NN	O	O
hepatocytes	NN	O	B-cell_type
and	NN	O	O
non-parenchymal	NN	O	B-cell_type
cells	NN	O	I-cell_type
paralleled	NN	O	O
the	NN	O	O
inflammatory	NN	O	O
response	NN	O	O
.	NN	O	O

Blockade	NN	O	O
of	NN	O	O
heme	NN	O	O
oxygenase	NN	O	O
activity	NN	O	O
with	NN	O	O
tin-protoporphyrin-IX	NN	O	O
abrogated	NN	O	O
acetaminophen-induced	NN	O	O
hepatic	NN	O	B-cell_type
neutrophil	NN	O	I-cell_type
accumulation	NN	O	O
and	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
activation	NN	O	O
,	NN	O	O
but	NN	O	O
failed	NN	O	O
to	NN	O	O
affect	NN	O	O
sinusoidal	NN	O	O
perfusion	NN	O	O
and	NN	O	O
liver	NN	O	O
injury	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
:	NN	O	O
The	NN	O	O
inflammatory	NN	O	O
response	NN	O	O
associated	NN	O	O
with	NN	O	O
acetaminophen	NN	O	O
hepatotoxicity	NN	O	O
is	NN	O	O
modulated	NN	O	O
by	NN	O	O
the	NN	O	O
parallel	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
heme	NN	O	B-DNA
oxygenase-1	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

However	NN	O	O
,	NN	O	O
heme	NN	O	B-DNA
oxygenase-1	NN	O	I-DNA
has	NN	O	O
no	NN	O	O
permissive	NN	O	O
effect	NN	O	O
on	NN	O	O
sinusoidal	NN	O	O
perfusion	NN	O	O
and	NN	O	O
does	NN	O	O
not	NN	O	O
affect	NN	O	O
liver	NN	O	O
injury	NN	O	O
in	NN	O	O
this	NN	O	O
model	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
argue	NN	O	O
against	NN	O	O
a	NN	O	O
central	NN	O	O
role	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
activation	NN	O	O
and	NN	O	O
neutrophil	NN	O	O
infiltration	NN	O	O
as	NN	O	O
perpetuating	NN	O	O
factors	NN	O	O
of	NN	O	O
liver	NN	O	O
injury	NN	O	O
in	NN	O	O
acetaminophen	NN	O	O
toxicity	NN	O	O
.	NN	O	O

-DOCSTART-	O

Tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
-induced	NN	O	O
proliferation	NN	O	O
requires	NN	O	O
synthesis	NN	O	O
of	NN	O	O
granulocyte-macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

OBJECTIVE	NN	O	O
:	NN	O	O
Tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
induces	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
cellular	NN	O	O
responses	NN	O	O
,	NN	O	O
some	NN	O	O
of	NN	O	O
them	NN	O	O
being	NN	O	O
at	NN	O	O
least	NN	O	O
seemingly	NN	O	O
contradictory	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
we	NN	O	O
set	NN	O	O
out	NN	O	O
to	NN	O	O
find	NN	O	O
differences	NN	O	O
in	NN	O	O
the	NN	O	O
modes	NN	O	O
of	NN	O	O
proliferative	NN	O	O
and	NN	O	O
apoptotic	NN	O	O
responses	NN	O	O
to	NN	O	O
TNF-	NN	O	B-protein
alpha	NN	O	I-protein
.	NN	O	O

MATERIALS	NN	O	O
AND	NN	O	O
METHODS	NN	O	O
:	NN	O	O
We	NN	O	O
screened	NN	O	O
a	NN	O	O
panel	NN	O	O
of	NN	O	O
acute	NN	O	B-cell_line
myeloid	NN	O	I-cell_line
leukemia-derived	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
for	NN	O	O
TNF-	NN	O	B-protein
alpha	NN	O	I-protein
-responsiveness	NN	O	O
.	NN	O	O

In	NN	O	O
two	NN	O	O
lines	NN	O	O
(	NN	O	O
OCI-AML-1	NN	O	B-cell_line
,	NN	O	O
OCI-AML-11	NN	O	B-cell_line
)	NN	O	O
,	NN	O	O
TNF-	NN	O	B-protein
alpha	NN	O	I-protein
acted	NN	O	O
as	NN	O	O
an	NN	O	O
apoptotic	NN	O	O
agent	NN	O	O
;	NN	O	O
in	NN	O	O
others	NN	O	O
(	NN	O	O
HU-3	NN	O	B-cell_line
,	NN	O	O
M-07e	NN	O	B-cell_line
,	NN	O	O
TF-1	NN	O	B-cell_line
)	NN	O	O
,	NN	O	O
it	NN	O	O
had	NN	O	O
the	NN	O	O
opposite	NN	O	O
effect	NN	O	O
,	NN	O	O
preventing	NN	O	O
apoptosis	NN	O	O
and	NN	O	O
inducing	NN	O	O
proliferation	NN	O	O
.	NN	O	O

Direct	NN	O	O
and	NN	O	O
indirect	NN	O	O
signaling	NN	O	O
mechanisms	NN	O	O
,	NN	O	O
including	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
and	NN	O	O
cytokine	NN	O	O
synthesis	NN	O	O
,	NN	O	O
were	NN	O	O
analyzed	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
All	NN	O	O
cell	NN	O	O
lines	NN	O	O
tested	NN	O	O
expressed	NN	O	O
TNF-	NN	O	B-protein
alpha	NN	O	I-protein
receptors	NN	O	I-protein
I	NN	O	I-protein
and	NN	O	I-protein
II	NN	O	I-protein
and	NN	O	O
responded	NN	O	O
to	NN	O	O
TNF-	NN	O	B-protein
alpha	NN	O	I-protein
by	NN	O	O
upregulation	NN	O	O
of	NN	O	O
intercellular	NN	O	B-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
granulocyte-macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
GM-CSF	NN	O	B-protein
)	NN	O	O
,	NN	O	O
TNF-	NN	O	B-protein
alpha	NN	O	I-protein
did	NN	O	O
not	NN	O	O
activate	NN	O	O
the	NN	O	O
MAP	NN	O	B-protein
kinase	NN	O	I-protein
and	NN	O	O
p70S6	NN	O	O
kinase	NN	O	O
pathways	NN	O	O
.	NN	O	O

Nevertheless	NN	O	O
,	NN	O	O
inhibitors	NN	O	O
of	NN	O	O
these	NN	O	O
pathways	NN	O	O
clearly	NN	O	O
reduced	NN	O	O
the	NN	O	O
TNF-alpha	NN	O	B-protein
-induced	NN	O	O
cell	NN	O	O
growth	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
TNF-	NN	O	B-cell_type
alpha-proliferative	NN	O	I-cell_type
cells	NN	O	I-cell_type
produced	NN	O	O
a	NN	O	O
growth	NN	O	O
factor	NN	O	O
that	NN	O	O
induced	NN	O	O
proliferation	NN	O	O
upon	NN	O	O
stimulation	NN	O	O
of	NN	O	O
the	NN	O	O
above	NN	O	O
pathways	NN	O	O
.	NN	O	O

Anti-GM-CSF	NN	O	B-protein
antibodies	NN	O	I-protein
inhibited	NN	O	O
the	NN	O	O
TNF-alpha	NN	O	B-protein
-induced	NN	O	O
growth	NN	O	O
,	NN	O	O
suggesting	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
an	NN	O	O
autocrine	NN	O	O
loop	NN	O	O
for	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
mediated	NN	O	O
by	NN	O	O
GM-CSF	NN	O	B-protein
.	NN	O	O

Supporting	NN	O	O
this	NN	O	O
notion	NN	O	O
,	NN	O	O
TNF-alpha	NN	O	B-protein
-induced	NN	O	O
upregulation	NN	O	O
of	NN	O	O
GM-CSF	NN	O	B-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
and	NN	O	O
protein	NN	O	O
secretion	NN	O	O
in	NN	O	O
the	NN	O	O
TNF-alpha	NN	O	B-protein
-proliferative	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
the	NN	O	O
TNF-alpha-apoptotic	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

CONCLUSION	NN	O	O
:	NN	O	O
These	NN	O	O
data	NN	O	O
identify	NN	O	O
GM-CSF	NN	O	B-protein
synthesis	NN	O	O
as	NN	O	O
an	NN	O	O
early	NN	O	O
and	NN	O	O
essential	NN	O	O
step	NN	O	O
in	NN	O	O
TNF-	NN	O	O
alpha-induced	NN	O	O
proliferation	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
for	NN	O	O
the	NN	O	O
first	NN	O	O
time	NN	O	O
that	NN	O	O
TNF-alpha-treated	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
producing	NN	O	O
no	NN	O	O
or	NN	O	O
only	NN	O	O
minimal	NN	O	O
amounts	NN	O	O
of	NN	O	O
GM-CSF	NN	O	B-protein
demonstrate	NN	O	O
an	NN	O	O
apoptotic	NN	O	O
phenotype	NN	O	O
,	NN	O	O
while	NN	O	O
cell	NN	O	O
lines	NN	O	O
with	NN	O	O
high	NN	O	O
GM-CSF	NN	O	B-protein
expression	NN	O	O
rates	NN	O	O
can	NN	O	O
escape	NN	O	O
from	NN	O	O
growth	NN	O	O
arrest	NN	O	O
or	NN	O	O
even	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
context	NN	O	O
,	NN	O	O
we	NN	O	O
discuss	NN	O	O
arguments	NN	O	O
pointing	NN	O	O
at	NN	O	O
NF-kappaB	NN	O	B-protein
as	NN	O	O
regulator	NN	O	O
of	NN	O	O
GM-CSF	NN	O	B-protein
synthesis	NN	O	O
and	NN	O	O
thus	NN	O	O
indirectly	NN	O	O
as	NN	O	O
regulator	NN	O	O
for	NN	O	O
the	NN	O	O
escape	NN	O	O
of	NN	O	O
TNF-alpha	NN	O	B-protein
-induced	NN	O	O
apoptosis	NN	O	O

-DOCSTART-	O

Glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
content	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
:	NN	O	O
evidence	NN	O	O
for	NN	O	O
heterogeneity	NN	O	O
.	NN	O	O

Glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
were	NN	O	O
measured	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
that	NN	O	O
were	NN	O	O
isolated	NN	O	O
from	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
by	NN	O	O
either	NN	O	O
nylon	NN	O	O
wool	NN	O	O
filtration	NN	O	O
or	NN	O	O
E-rosette	NN	O	O
sedimentation	NN	O	O
.	NN	O	O

T	NN	O	B-cell_type
cells	NN	O	I-cell_type
isolated	NN	O	O
by	NN	O	O
nylon	NN	O	O
wool	NN	O	O
filtration	NN	O	O
specifically	NN	O	O
bind	NN	O	O
6.7	NN	O	O
+/-	NN	O	O
0.2	NN	O	O
fmol	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
per	NN	O	O
million	NN	O	O
cells	NN	O	O
(	NN	O	O
equivalent	NN	O	O
to	NN	O	O
4000	NN	O	O
+/-	NN	O	O
200	NN	O	O
receptors	NN	O	O
per	NN	O	O
cell	NN	O	O
)	NN	O	O
,	NN	O	O
whereas	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
isolated	NN	O	O
by	NN	O	O
E-rosette	NN	O	O
sedimentation	NN	O	O
bind	NN	O	O
12.0	NN	O	O
+/-	NN	O	O
0.7	NN	O	O
fmol	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
per	NN	O	O
million	NN	O	O
cells	NN	O	O
(	NN	O	O
equivalent	NN	O	O
to	NN	O	O
7200	NN	O	O
+/-	NN	O	O
400	NN	O	O
receptors	NN	O	O
per	NN	O	O
cell	NN	O	O
)	NN	O	O
.	NN	O	O

This	NN	O	O
difference	NN	O	O
in	NN	O	O
the	NN	O	O
amount	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
bound	NN	O	O
by	NN	O	O
the	NN	O	O
two	NN	O	O
T	NN	O	O
cell	NN	O	O
preparations	NN	O	O
was	NN	O	O
significant	NN	O	O
(	NN	O	O
p	NN	O	O
less	NN	O	O
than	NN	O	O
.001	NN	O	O
)	NN	O	O
and	NN	O	O
was	NN	O	O
present	NN	O	O
immediately	NN	O	O
after	NN	O	O
cell	NN	O	O
isolation	NN	O	O
.	NN	O	O

The	NN	O	O
binding	NN	O	O
affinities	NN	O	O
of	NN	O	O
the	NN	O	O
different	NN	O	O
T	NN	O	B-cell_line
cell	NN	O	I-cell_line
preparations	NN	O	I-cell_line
for	NN	O	O
dexamethasone	NN	O	O
were	NN	O	O
similar	NN	O	O
.	NN	O	O

T	NN	O	O
cells	NN	O	O
that	NN	O	O
are	NN	O	O
isolated	NN	O	O
by	NN	O	O
a	NN	O	O
combination	NN	O	O
of	NN	O	O
nylon	NN	O	O
wool	NN	O	O
filtration	NN	O	O
followed	NN	O	O
by	NN	O	O
E-rosette	NN	O	O
sedimentation	NN	O	O
bind	NN	O	O
the	NN	O	O
same	NN	O	O
amount	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
as	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
isolated	NN	O	O
by	NN	O	O
nylon	NN	O	O
wool	NN	O	O
filtration	NN	O	O
alone	NN	O	O
.	NN	O	O

T	NN	O	B-cell_type
cells	NN	O	I-cell_type
isolated	NN	O	O
by	NN	O	O
a	NN	O	O
combination	NN	O	O
of	NN	O	O
E-rosette	NN	O	O
sedimentation	NN	O	O
following	NN	O	O
by	NN	O	O
nylon	NN	O	O
wool	NN	O	O
filtration	NN	O	O
bind	NN	O	O
less	NN	O	O
dexamethasone	NN	O	O
than	NN	O	O
do	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
isolated	NN	O	O
by	NN	O	O
E-rosette	NN	O	O
sedimentation	NN	O	O
alone	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
heterogeneous	NN	O	O
with	NN	O	O
respect	NN	O	O
to	NN	O	O
their	NN	O	O
quantity	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
.	NN	O	O

Isolation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
E-rosette	NN	O	O
sedimentation	NN	O	O
enriches	NN	O	O
for	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
that	NN	O	O
have	NN	O	O
a	NN	O	O
greater	NN	O	O
number	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
,	NN	O	O
and	NN	O	O
isolation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
nylon	NN	O	O
wool	NN	O	O
filtration	NN	O	O
enriches	NN	O	O
for	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
that	NN	O	O
have	NN	O	O
a	NN	O	O
lesser	NN	O	O
number	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
and	NN	O	O
glucocorticoid	NN	O	O
sensitivity	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
leukemic	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
have	NN	O	O
established	NN	O	O
optimal	NN	O	O
conditions	NN	O	O
for	NN	O	O
the	NN	O	O
measurement	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
(	NN	O	O
GR	NN	O	B-protein
)	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
white	NN	O	I-cell_type
cells	NN	O	I-cell_type
using	NN	O	O
a	NN	O	O
whole-cell	NN	O	O
binding	NN	O	O
assay	NN	O	O
with	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
dexamethasone	NN	O	O
as	NN	O	O
the	NN	O	O
ligand	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
subsequent	NN	O	O
determination	NN	O	O
of	NN	O	O
the	NN	O	O
GR	NN	O	B-protein
content	NN	O	O
in	NN	O	O
normal	NN	O	B-cell_type
human	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
in	NN	O	O
leukemic	NN	O	B-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
patients	NN	O	O
with	NN	O	O
various	NN	O	O
forms	NN	O	O
of	NN	O	O
acute	NN	O	O
and	NN	O	O
chronic	NN	O	O
leukemia	NN	O	O
.	NN	O	O

A	NN	O	O
number	NN	O	O
of	NN	O	O
leukemia	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
in	NN	O	O
continuous	NN	O	O
culture	NN	O	O
were	NN	O	O
also	NN	O	O
subjected	NN	O	O
to	NN	O	O
the	NN	O	O
GR	NN	O	B-protein
assay	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
results	NN	O	O
were	NN	O	O
correlated	NN	O	O
with	NN	O	O
the	NN	O	O
sensitivity	NN	O	O
of	NN	O	O
these	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
to	NN	O	O
glucocorticoid	NN	O	O
steroids	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

The	NN	O	O
GR	NN	O	B-protein
content	NN	O	O
of	NN	O	O
normal	NN	O	B-cell_type
human	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
amounted	NN	O	O
to	NN	O	O
4	NN	O	O
,	NN	O	O
850	NN	O	O
+/-	NN	O	O
1	NN	O	O
,	NN	O	O
340	NN	O	O
(	NN	O	O
mean	NN	O	O
+/-	NN	O	O
SD	NN	O	O
)	NN	O	O
receptors/cell	NN	O	O
.	NN	O	O

The	NN	O	O
mean	NN	O	O
equilibrium	NN	O	O
dissociation	NN	O	O
constant	NN	O	O
(	NN	O	O
KD	NN	O	O
)	NN	O	O
of	NN	O	O
the	NN	O	O
interaction	NN	O	O
of	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
dexamethasone	NN	O	O
with	NN	O	O
the	NN	O	O
GR	NN	O	B-protein
was	NN	O	O
1.2	NN	O	O
x	NN	O	O
10	NN	O	O
(	NN	O	O
-8	NN	O	O
)	NN	O	O
M	NN	O	O
.	NN	O	O

Steroidal	NN	O	O
compounds	NN	O	O
with	NN	O	O
a	NN	O	O
known	NN	O	O
glucocorticoid	NN	O	O
potency	NN	O	O
effectively	NN	O	O
competed	NN	O	O
for	NN	O	O
the	NN	O	O
binding	NN	O	O
,	NN	O	O
whereas	NN	O	O
steroids	NN	O	O
devoid	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	O
activity	NN	O	O
(	NN	O	O
e.g.	NN	O	O
estradiol-17	NN	O	O
beta	NN	O	O
and	NN	O	O
testosterone	NN	O	O
)	NN	O	O
were	NN	O	O
ineffective	NN	O	O
.	NN	O	O

The	NN	O	O
GR	NN	O	B-protein
content	NN	O	O
of	NN	O	O
the	NN	O	O
blast	NN	O	B-cell_type
cells	NN	O	I-cell_type
obtained	NN	O	O
from	NN	O	O
eight	NN	O	O
patients	NN	O	O
suffering	NN	O	O
from	NN	O	O
acute	NN	O	O
leukemia	NN	O	O
and	NN	O	O
four	NN	O	O
patients	NN	O	O
with	NN	O	O
a	NN	O	O
blast	NN	O	O
crisis	NN	O	O
of	NN	O	O
chronic	NN	O	O
myelocytic	NN	O	O
leukemia	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
highly	NN	O	O
variable	NN	O	O
(	NN	O	O
3	NN	O	O
,	NN	O	O
230-29	NN	O	O
,	NN	O	O
900	NN	O	O
receptors/cell	NN	O	O
)	NN	O	O
,	NN	O	O
while	NN	O	O
the	NN	O	O
lymphocytes	NN	O	B-cell_type
of	NN	O	O
six	NN	O	O
patients	NN	O	O
with	NN	O	O
chronic	NN	O	O
lymphatic	NN	O	O
leukemia	NN	O	O
contained	NN	O	O
a	NN	O	O
rather	NN	O	O
stable	NN	O	O
GR	NN	O	B-protein
content	NN	O	O
(	NN	O	O
2	NN	O	O
,	NN	O	O
930-5	NN	O	O
,	NN	O	O
120	NN	O	O
receptors/cell	NN	O	O
)	NN	O	O
,	NN	O	O
which	NN	O	O
was	NN	O	O
comparable	NN	O	O
with	NN	O	O
that	NN	O	O
of	NN	O	O
normal	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

GR	NN	O	B-protein
was	NN	O	O
identified	NN	O	O
in	NN	O	O
all	NN	O	O
the	NN	O	O
12	NN	O	O
malignant	NN	O	B-cell_line
continuous	NN	O	I-cell_line
white	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
studied	NN	O	O
.	NN	O	O

Large	NN	O	O
cells	NN	O	O
contained	NN	O	O
more	NN	O	O
GR	NN	O	B-protein
than	NN	O	O
the	NN	O	O
smaller	NN	O	O
ones	NN	O	O
.	NN	O	O

There	NN	O	O
was	NN	O	O
no	NN	O	O
apparent	NN	O	O
correlation	NN	O	O
between	NN	O	O
the	NN	O	O
GR	NN	O	B-protein
concentration	NN	O	O
and	NN	O	O
the	NN	O	O
sensitivity	NN	O	O
of	NN	O	O
the	NN	O	O
cells	NN	O	O
in	NN	O	O
vitro	NN	O	O
to	NN	O	O
glucocorticoids	NN	O	O
as	NN	O	O
judged	NN	O	O
by	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
thymidine	NN	O	O
incorporation	NN	O	O
studies	NN	O	O
.	NN	O	O

Distribution	NN	O	O
of	NN	O	O
the	NN	O	O
surface	NN	O	O
markers	NN	O	O
in	NN	O	O
the	NN	O	O
leukemic	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
did	NN	O	O
not	NN	O	O
relate	NN	O	O
to	NN	O	O
the	NN	O	O
GR	NN	O	B-protein
concentration	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
GR	NN	O	B-protein
is	NN	O	O
probably	NN	O	O
a	NN	O	O
universal	NN	O	O
feature	NN	O	O
of	NN	O	O
the	NN	O	O
leukemic	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
,	NN	O	O
from	NN	O	O
a	NN	O	O
clinical	NN	O	O
standpoint	NN	O	O
,	NN	O	O
probably	NN	O	O
does	NN	O	O
not	NN	O	O
alone	NN	O	O
imply	NN	O	O
steroid	NN	O	O
responsiveness	NN	O	O
.	NN	O	O

-DOCSTART-	O

Evidence	NN	O	O
for	NN	O	O
a	NN	O	O
steroid	NN	O	B-protein
receptor	NN	O	I-protein
in	NN	O	O
rheumatoid	NN	O	B-cell_type
synovial	NN	O	I-cell_type
tissue	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

One	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
glucocorticoids	NN	O	O
could	NN	O	O
exert	NN	O	O
their	NN	O	O
anti-inflammatory	NN	O	O
action	NN	O	O
is	NN	O	O
via	NN	O	O
rapidly	NN	O	O
saturable	NN	O	O
,	NN	O	O
stereo-specific	NN	O	B-protein
cytoplasmic	NN	O	I-protein
protein	NN	O	I-protein
receptors	NN	O	I-protein
.	NN	O	O

This	NN	O	O
report	NN	O	O
is	NN	O	O
of	NN	O	O
an	NN	O	O
investigation	NN	O	O
into	NN	O	O
such	NN	O	O
a	NN	O	O
possibility	NN	O	O
in	NN	O	O
synovial	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Synovium	NN	O	O
,	NN	O	O
obtained	NN	O	O
from	NN	O	O
knee	NN	O	O
joints	NN	O	O
of	NN	O	O
rheumatoid	NN	O	O
patients	NN	O	O
undergoing	NN	O	O
surgery	NN	O	O
,	NN	O	O
was	NN	O	O
incubated	NN	O	O
with	NN	O	O
clostridiopeptidase	NN	O	B-protein
A	NN	O	I-protein
and	NN	O	O
trypsin-EDTA	NN	O	B-protein
to	NN	O	O
obtain	NN	O	O
cell	NN	O	O
suspensions	NN	O	O
.	NN	O	O

These	NN	O	O
,	NN	O	O
together	NN	O	O
with	NN	O	O
cells	NN	O	O
obtained	NN	O	O
from	NN	O	O
synovial	NN	O	O
fluid	NN	O	O
aspirated	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
rheumatoid	NN	O	O
arthritis	NN	O	O
,	NN	O	O
were	NN	O	O
identified	NN	O	O
by	NN	O	O
electron	NN	O	O
microscopy	NN	O	O
.	NN	O	O

Duplicate	NN	O	O
samples	NN	O	O
of	NN	O	O
these	NN	O	O
cell	NN	O	O
suspensions	NN	O	O
were	NN	O	O
incubated	NN	O	O
with	NN	O	O
increasing	NN	O	O
concentrations	NN	O	O
of	NN	O	O
H3Dexamethasone	NN	O	O
(	NN	O	O
1	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
x	NN	O	O
10	NN	O	O
(	NN	O	O
-9	NN	O	O
)	NN	O	O
M	NN	O	O
)	NN	O	O
for	NN	O	O
30	NN	O	O
minutes	NN	O	O
at	NN	O	O
37	NN	O	O
degrees	NN	O	O
C	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
the	NN	O	O
proportion	NN	O	O
of	NN	O	O
steroid	NN	O	O
bound	NN	O	O
to	NN	O	O
whole	NN	O	B-cell_type
cells	NN	O	I-cell_type
showed	NN	O	O
evidence	NN	O	O
for	NN	O	O
specific	NN	O	O
,	NN	O	O
rapidly	NN	O	O
saturable	NN	O	O
,	NN	O	O
receptors	NN	O	O
in	NN	O	O
the	NN	O	O
cells	NN	O	O
obtained	NN	O	O
from	NN	O	O
synovial	NN	O	O
tissue	NN	O	O
,	NN	O	O
but	NN	O	O
this	NN	O	O
was	NN	O	O
not	NN	O	O
found	NN	O	O
in	NN	O	O
synovial	NN	O	B-cell_type
fluid	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Electron	NN	O	O
micrographs	NN	O	O
showed	NN	O	O
that	NN	O	O
cells	NN	O	O
obtained	NN	O	O
from	NN	O	O
synovial	NN	O	O
tissue	NN	O	O
consisted	NN	O	O
of	NN	O	O
synovial	NN	O	B-cell_type
fibroblast	NN	O	I-cell_type
-	NN	O	O
and	NN	O	O
macrophage-types	NN	O	B-cell_type
,	NN	O	O
lymphocytes	NN	O	B-cell_type
,	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
macrophages	NN	O	B-cell_type
.	NN	O	O

Polymorphonuclear	NN	O	B-cell_type
leucocytes	NN	O	I-cell_type
appeared	NN	O	O
to	NN	O	O
be	NN	O	O
absent	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
in	NN	O	O
synovial	NN	O	B-cell_type
fluid	NN	O	I-cell_type
cell	NN	O	I-cell_type
type	NN	O	I-cell_type
polymorphonuclear	NN	O	I-cell_type
leucocytes	NN	O	I-cell_type
were	NN	O	O
the	NN	O	O
predominant	NN	O	O
cell	NN	O	O
type	NN	O	O
.	NN	O	O

We	NN	O	O
concluded	NN	O	O
from	NN	O	O
this	NN	O	O
,	NN	O	O
that	NN	O	O
one	NN	O	O
or	NN	O	O
more	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	O
types	NN	O	O
present	NN	O	O
in	NN	O	O
synovial	NN	O	O
tissue	NN	O	O
contain	NN	O	O
a	NN	O	O
specific	NN	O	B-protein
steroid	NN	O	I-protein
receptor	NN	O	I-protein
,	NN	O	O
but	NN	O	O
that	NN	O	O
this	NN	O	O
is	NN	O	O
lacking	NN	O	O
in	NN	O	O
synovial	NN	O	B-cell_type
fluid	NN	O	I-cell_type
polymorphonuclear	NN	O	I-cell_type
leucocytes	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Clinical	NN	O	O
implications	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
human	NN	O	O
leukemia	NN	O	O
.	NN	O	O

Glucorticoid	NN	O	B-protein
receptors	NN	O	I-protein
were	NN	O	O
studied	NN	O	O
in	NN	O	O
various	NN	O	O
populations	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
and	NN	O	O
leukemic	NN	O	B-cell_type
lymphoblasts	NN	O	I-cell_type
.	NN	O	O

Normal	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
contain	NN	O	O
low	NN	O	O
levels	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
approximately	NN	O	O
2	NN	O	O
,	NN	O	O
500	NN	O	O
sites/cell	NN	O	O
)	NN	O	O
which	NN	O	O
are	NN	O	O
identical	NN	O	O
in	NN	O	O
T-	NN	O	O
and	NN	O	O
non-T-fractions	NN	O	O
.	NN	O	O

Phytohemagglutinin	NN	O	B-protein
treatment	NN	O	O
increases	NN	O	O
levels	NN	O	O
about	NN	O	O
3-fold	NN	O	O
.	NN	O	O

Leukemic	NN	O	B-cell_type
lymphoblasts	NN	O	I-cell_type
contain	NN	O	O
larger	NN	O	O
numbers	NN	O	O
of	NN	O	O
receptor	NN	O	O
sites	NN	O	O
.	NN	O	O

Presence	NN	O	O
of	NN	O	O
receptor	NN	O	O
is	NN	O	O
correlated	NN	O	O
with	NN	O	O
in	NN	O	O
vitro	NN	O	O
sensitivitiy	NN	O	O
to	NN	O	O
glucocorticoids	NN	O	O
and	NN	O	O
in	NN	O	O
vivo	NN	O	O
response	NN	O	O
to	NN	O	O
therapy	NN	O	O
.	NN	O	O

Quantity	NN	O	O
of	NN	O	O
receptor	NN	O	O
is	NN	O	O
also	NN	O	O
correlated	NN	O	O
with	NN	O	O
complete	NN	O	O
remission	NN	O	O
duration	NN	O	O
independently	NN	O	O
of	NN	O	O
leukemic	NN	O	B-cell_type
cell	NN	O	I-cell_type
type	NN	O	I-cell_type
(	NN	O	O
T	NN	O	O
or	NN	O	O
null	NN	O	O
)	NN	O	O
,	NN	O	O
initial	NN	O	O
WBC	NN	O	B-cell_type
,	NN	O	O
or	NN	O	O
age	NN	O	O
of	NN	O	O
patient	NN	O	O
.	NN	O	O

Quantitative	NN	O	O
determination	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
levels	NN	O	O
in	NN	O	O
acute	NN	O	O
lymphoblastic	NN	O	O
leukemia	NN	O	O
may	NN	O	O
be	NN	O	O
of	NN	O	O
value	NN	O	O
both	NN	O	O
as	NN	O	O
an	NN	O	O
independent	NN	O	O
prognostic	NN	O	O
variable	NN	O	O
and	NN	O	O
in	NN	O	O
suggesting	NN	O	O
which	NN	O	O
patients	NN	O	O
should	NN	O	O
receive	NN	O	O
glucocorticoid	NN	O	O
therapy	NN	O	O
.	NN	O	O

-DOCSTART-	O

Functional	NN	O	O
and	NN	O	O
physical	NN	O	O
interaction	NN	O	O
of	NN	O	O
protein-tyrosine	NN	O	B-protein
kinases	NN	O	I-protein
Fyn	NN	O	I-protein
and	NN	O	I-protein
Csk	NN	O	I-protein
in	NN	O	O
the	NN	O	O
T-cell	NN	O	O
signaling	NN	O	O
system	NN	O	O
.	NN	O	O

The	NN	O	O
Src-like	NN	O	B-protein
protein-tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
Fyn	NN	O	B-protein
is	NN	O	O
associated	NN	O	O
with	NN	O	O
T-cell	NN	O	B-protein
antigen	NN	O	I-protein
receptor	NN	O	I-protein
.	NN	O	O

Transient	NN	O	O
expression	NN	O	O
of	NN	O	O
actively	NN	O	O
mutated	NN	O	O
Fyn	NN	O	B-protein
,	NN	O	O
having	NN	O	O
Phe-528	NN	O	O
instead	NN	O	O
of	NN	O	O
Tyr-528	NN	O	O
or	NN	O	O
Thr-338	NN	O	O
instead	NN	O	O
of	NN	O	O
Ile-338	NN	O	O
,	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_type
T-cells	NN	O	I-cell_type
stimulated	NN	O	B-DNA
the	NN	O	I-DNA
serum	NN	O	I-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
SRE	NN	O	B-DNA
)	NN	O	O
,	NN	O	O
12-O-tetradecanoyl-phorbol-13-acetate	NN	O	B-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
,	NN	O	O
cyclic	NN	O	B-DNA
AMP	NN	O	I-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
c-fos	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
stimulation	NN	O	O
of	NN	O	O
SRE	NN	O	B-DNA
was	NN	O	O
particularly	NN	O	O
prominent	NN	O	O
not	NN	O	O
only	NN	O	O
with	NN	O	O
active	NN	O	O
Fyn	NN	O	B-protein
but	NN	O	O
also	NN	O	O
with	NN	O	O
normal	NN	O	B-protein
(	NN	O	I-protein
wild-type	NN	O	I-protein
)	NN	O	I-protein
Fyn	NN	O	I-protein
.	NN	O	O

SRE	NN	O	B-DNA
was	NN	O	O
also	NN	O	O
stimulated	NN	O	O
by	NN	O	O
both	NN	O	O
normal	NN	O	B-protein
and	NN	O	I-protein
active	NN	O	I-protein
Lck	NN	O	I-protein
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
normal	NN	O	B-protein
and	NN	O	I-protein
active	NN	O	I-protein
Fyn	NN	O	I-protein
stimulated	NN	O	O
transcription	NN	O	O
from	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
when	NN	O	O
transfected	NN	O	O
cells	NN	O	O
were	NN	O	O
stimulated	NN	O	O
by	NN	O	O
concanavalin	NN	O	B-protein
A	NN	O	I-protein
plus	NN	O	O
12-O-tetradecanoylphorbol-13-acetate	NN	O	O
.	NN	O	O

Under	NN	O	O
the	NN	O	O
same	NN	O	O
conditions	NN	O	O
,	NN	O	O
Lck	NN	O	B-protein
did	NN	O	O
not	NN	O	O
stimulate	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
unless	NN	O	O
it	NN	O	O
was	NN	O	O
activated	NN	O	O
by	NN	O	O
mutation	NN	O	O
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
a	NN	O	O
mutant	NN	O	B-protein
Fyn	NN	O	I-protein
,	NN	O	O
which	NN	O	O
has	NN	O	O
deletions	NN	O	O
within	NN	O	O
the	NN	O	O
SH2	NN	O	B-protein
region	NN	O	I-protein
and	NN	O	O
so	NN	O	O
is	NN	O	O
able	NN	O	O
to	NN	O	O
transform	NN	O	O
chicken	NN	O	B-cell_type
embryo	NN	O	I-cell_type
fibroblasts	NN	O	I-cell_type
,	NN	O	O
did	NN	O	O
not	NN	O	O
stimulate	NN	O	O
either	NN	O	O
the	NN	O	O
c-fos	NN	O	B-DNA
or	NN	O	I-DNA
IL-2	NN	O	I-DNA
promoter	NN	O	I-DNA
,	NN	O	O
suggesting	NN	O	O
the	NN	O	O
importance	NN	O	O
of	NN	O	O
this	NN	O	O
region	NN	O	O
in	NN	O	O
T-cell	NN	O	B-cell_type
signaling	NN	O	O
.	NN	O	O

Csk	NN	O	B-protein
,	NN	O	O
which	NN	O	O
phosphorylates	NN	O	O
tyrosine	NN	O	O
residues	NN	O	O
in	NN	O	O
the	NN	O	O
negative	NN	O	O
regulatory	NN	O	O
sites	NN	O	O
of	NN	O	O
Src	NN	O	B-protein
family	NN	O	I-protein
kinases	NN	O	I-protein
,	NN	O	O
down-regulated	NN	O	O
Fyn-	NN	O	O
and	NN	O	O
Lck-mediated	NN	O	O
stimulation	NN	O	O
of	NN	O	O
the	NN	O	O
serum	NN	O	B-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
and	NN	O	O
Fyn-mediated	NN	O	O
enhancement	NN	O	O
of	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
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
Fyn	NN	O	B-protein
and	NN	O	O
Lck	NN	O	B-protein
,	NN	O	O
whose	NN	O	O
activities	NN	O	O
are	NN	O	O
regulated	NN	O	O
by	NN	O	O
Csk	NN	O	B-protein
,	NN	O	O
are	NN	O	O
involved	NN	O	O
in	NN	O	O
different	NN	O	O
phases	NN	O	O
of	NN	O	O
T-cell	NN	O	B-cell_type
activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
novel	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
complex	NN	O	I-protein
containing	NN	O	O
p65	NN	O	B-protein
homodimers	NN	O	I-protein
:	NN	O	O
implications	NN	O	O
for	NN	O	O
transcriptional	NN	O	O
control	NN	O	O
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
subunit	NN	O	O
dimerization	NN	O	O
.	NN	O	O

The	NN	O	O
predominant	NN	O	O
inducible	NN	O	O
form	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
is	NN	O	O
a	NN	O	O
heteromeric	NN	O	B-protein
complex	NN	O	I-protein
containing	NN	O	O
two	NN	O	O
Rel-related	NN	O	B-protein
DNA-binding	NN	O	I-protein
subunits	NN	O	I-protein
,	NN	O	O
termed	NN	O	B-protein
p65	NN	O	I-protein
and	NN	O	O
p50	NN	O	B-protein
.	NN	O	O

Prior	NN	O	O
transfection	NN	O	O
studies	NN	O	O
have	NN	O	O
shown	NN	O	O
that	NN	O	O
when	NN	O	O
these	NN	O	O
p65	NN	O	B-protein
and	NN	O	I-protein
p50	NN	O	I-protein
subunits	NN	O	I-protein
are	NN	O	O
expressed	NN	O	O
independently	NN	O	O
as	NN	O	O
stable	NN	O	B-protein
homodimers	NN	O	I-protein
,	NN	O	O
p65	NN	O	B-protein
stimulates	NN	O	O
kappa	NN	O	O
B-directed	NN	O	O
transcription	NN	O	O
,	NN	O	O
whereas	NN	O	O
p50	NN	O	B-protein
functions	NN	O	O
as	NN	O	O
a	NN	O	O
kappa	NN	O	O
B-specific	NN	O	O
repressor	NN	O	O
.	NN	O	O

While	NN	O	O
authentic	NN	O	O
p50	NN	O	B-protein
homodimers	NN	O	I-protein
(	NN	O	O
previously	NN	O	O
termed	NN	O	O
KBF1	NN	O	B-protein
)	NN	O	O
have	NN	O	O
been	NN	O	O
detected	NN	O	O
in	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
from	NN	O	O
nontransfected	NN	O	O
cells	NN	O	O
,	NN	O	O
experimental	NN	O	O
evidence	NN	O	O
supporting	NN	O	O
the	NN	O	O
existence	NN	O	O
of	NN	O	O
p65	NN	O	B-protein
homodimers	NN	O	I-protein
in	NN	O	O
vivo	NN	O	O
was	NN	O	O
lacking	NN	O	O
.	NN	O	O

We	NN	O	O
now	NN	O	O
provide	NN	O	O
direct	NN	O	O
biochemical	NN	O	O
evidence	NN	O	O
for	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
an	NN	O	O
endogenous	NN	O	O
pool	NN	O	O
of	NN	O	O
inducible	NN	O	O
p65	NN	O	B-protein
homodimers	NN	O	I-protein
in	NN	O	O
intact	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

As	NN	O	O
with	NN	O	O
the	NN	O	O
prototypical	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
p50-p65	NN	O	I-protein
heterodimer	NN	O	I-protein
,	NN	O	O
this	NN	O	O
novel	NN	O	O
p65	NN	O	B-protein
homodimeric	NN	O	I-protein
form	NN	O	I-protein
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
is	NN	O	O
functionally	NN	O	O
sequestered	NN	O	O
in	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
but	NN	O	O
rapidly	NN	O	O
appears	NN	O	O
in	NN	O	O
the	NN	O	O
nuclear	NN	O	O
compartment	NN	O	O
following	NN	O	O
cellular	NN	O	O
stimulation	NN	O	O
.	NN	O	O

Site-directed	NN	O	O
mutagenesis	NN	O	O
studies	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
homodimerization	NN	O	O
function	NN	O	O
of	NN	O	O
p65	NN	O	B-protein
is	NN	O	O
dependent	NN	O	O
upon	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
cysteine	NN	O	O
216	NN	O	O
and	NN	O	O
a	NN	O	O
conserved	NN	O	O
recognition	NN	O	O
motif	NN	O	O
for	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
A	NN	O	I-protein
(	NN	O	O
RRPS	NN	O	B-protein
;	NN	O	O
amino	NN	O	O
acids	NN	O	O
273	NN	O	O
to	NN	O	O
276	NN	O	O
)	NN	O	O
,	NN	O	O
both	NN	O	O
of	NN	O	O
which	NN	O	O
reside	NN	O	O
within	NN	O	O
a	NN	O	O
91-amino-acid	NN	O	O
segment	NN	O	O
of	NN	O	O
the	NN	O	O
Rel	NN	O	B-protein
homology	NN	O	I-protein
domain	NN	O	I-protein
that	NN	O	O
mediates	NN	O	O
self-association	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
mutations	NN	O	O
at	NN	O	O
these	NN	O	O
two	NN	O	O
sites	NN	O	O
do	NN	O	O
not	NN	O	O
affect	NN	O	O
heterodimerization	NN	O	O
of	NN	O	O
p65	NN	O	B-protein
with	NN	O	O
p50	NN	O	B-protein
or	NN	O	O
its	NN	O	O
functional	NN	O	O
interaction	NN	O	O
with	NN	O	O
I	NN	O	O
kappa	NN	O	O
B	NN	O	O
alpha	NN	O	O
.	NN	O	O

These	NN	O	O
later	NN	O	O
findings	NN	O	O
indicate	NN	O	O
that	NN	O	O
neither	NN	O	O
homo-	NN	O	O
nor	NN	O	O
heterodimer	NN	O	O
formation	NN	O	O
is	NN	O	O
an	NN	O	O
absolute	NN	O	O
prerequisite	NN	O	O
for	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
recognition	NN	O	O
of	NN	O	O
p65	NN	O	B-protein
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
with	NN	O	O
prior	NN	O	O
in	NN	O	O
vivo	NN	O	O
transcription	NN	O	O
studies	NN	O	O
,	NN	O	O
these	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
biological	NN	O	O
activities	NN	O	O
of	NN	O	O
p65	NN	O	B-protein
and	NN	O	I-protein
p50	NN	O	I-protein
homodimers	NN	O	I-protein
are	NN	O	O
independently	NN	O	O
regulated	NN	O	O
,	NN	O	O
thereby	NN	O	O
providing	NN	O	O
an	NN	O	O
integrated	NN	O	O
and	NN	O	O
flexible	NN	O	O
control	NN	O	O
mechanism	NN	O	O
for	NN	O	O
the	NN	O	O
rapid	NN	O	O
activation	NN	O	O
and	NN	O	O
repression	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
/	NN	O	O
Rel	NN	O	B-protein
-directed	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

Carrier	NN	O	O
determination	NN	O	O
for	NN	O	O
X-linked	NN	O	O
agammaglobulinemia	NN	O	O
using	NN	O	O
X	NN	O	O
inactivation	NN	O	O
analysis	NN	O	O
of	NN	O	O
purified	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
report	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
a	NN	O	O
relatively	NN	O	O
quick	NN	O	O
and	NN	O	O
simple	NN	O	O
method	NN	O	O
for	NN	O	O
the	NN	O	O
assessment	NN	O	O
of	NN	O	O
X	NN	O	O
inactivation	NN	O	O
status	NN	O	O
for	NN	O	O
carrier	NN	O	O
determination	NN	O	O
in	NN	O	O
families	NN	O	O
affected	NN	O	O
by	NN	O	O
X-linked	NN	O	O
agammaglobulinemia	NN	O	O
(	NN	O	O
XLA	NN	O	O
)	NN	O	O
.	NN	O	O

This	NN	O	O
method	NN	O	O
utilises	NN	O	O
an	NN	O	O
immunomagnetic	NN	O	O
separation	NN	O	O
technique	NN	O	O
for	NN	O	O
B	NN	O	B-cell_type
cell	NN	O	I-cell_type
purification	NN	O	O
and	NN	O	O
a	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
(	NN	O	O
PCR	NN	O	O
)	NN	O	O
based	NN	O	O
assay	NN	O	O
for	NN	O	O
the	NN	O	O
determination	NN	O	O
of	NN	O	O
methylation	NN	O	O
status	NN	O	O
at	NN	O	O
the	NN	O	O
androgen	NN	O	B-DNA
receptor	NN	O	I-DNA
(	NN	O	I-DNA
AR	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
locus	NN	O	I-DNA
to	NN	O	O
assess	NN	O	O
whether	NN	O	O
X	NN	O	O
inactivation	NN	O	O
is	NN	O	O
random	NN	O	O
or	NN	O	O
non-random	NN	O	O
at	NN	O	O
this	NN	O	O
locus	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
the	NN	O	O
results	NN	O	O
we	NN	O	O
have	NN	O	O
obtained	NN	O	O
using	NN	O	O
this	NN	O	O
assay	NN	O	O
to	NN	O	O
investigate	NN	O	O
females	NN	O	O
known	NN	O	O
to	NN	O	O
be	NN	O	O
carriers	NN	O	O
of	NN	O	O
various	NN	O	O
X-linked	NN	O	O
immunodeficiency	NN	O	O
disorders	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
four	NN	O	O
females	NN	O	O
from	NN	O	O
different	NN	O	O
families	NN	O	O
affected	NN	O	O
by	NN	O	O
XLA	NN	O	O
,	NN	O	O
two	NN	O	O
of	NN	O	O
whom	NN	O	O
were	NN	O	O
of	NN	O	O
unknown	NN	O	O
carrier	NN	O	O
status	NN	O	O
,	NN	O	O
and	NN	O	O
we	NN	O	O
discuss	NN	O	O
the	NN	O	O
results	NN	O	O
obtained	NN	O	O
with	NN	O	O
this	NN	O	O
and	NN	O	O
other	NN	O	O
X-inactivation	NN	O	O
assays	NN	O	O
.	NN	O	O

A	NN	O	O
similar	NN	O	O
assay	NN	O	O
has	NN	O	O
recently	NN	O	O
been	NN	O	O
described	NN	O	O
by	NN	O	O
Allen	NN	O	O
et	NN	O	O
al.	NN	O	O
(	NN	O	O
1992	NN	O	O
)	NN	O	O
and	NN	O	O
applied	NN	O	O
to	NN	O	O
members	NN	O	O
of	NN	O	O
one	NN	O	O
family	NN	O	O
affected	NN	O	O
by	NN	O	O
XLA	NN	O	O
.	NN	O	O

-DOCSTART-	O

Effects	NN	O	O
of	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
Fc	NN	O	B-protein
gamma	NN	O	I-protein
receptor	NN	O	I-protein
II	NN	O	I-protein
engagement	NN	O	O
on	NN	O	O
Egr-1	NN	O	B-protein
expression	NN	O	O
during	NN	O	O
stimulation	NN	O	O
of	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
by	NN	O	O
membrane	NN	O	O
immunoglobulin	NN	O	O
crosslinking	NN	O	O
.	NN	O	O

Egr-1	NN	O	B-DNA
is	NN	O	O
an	NN	O	O
immediate	NN	O	B-DNA
early	NN	O	I-DNA
gene	NN	O	I-DNA
that	NN	O	O
is	NN	O	O
rapidly	NN	O	O
upregulated	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
mitogenic	NN	O	O
signals	NN	O	O
induced	NN	O	O
by	NN	O	O
antigen	NN	O	O
receptor	NN	O	O
crosslinking	NN	O	O
on	NN	O	O
murine	NN	O	B-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

It	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
that	NN	O	O
levels	NN	O	O
of	NN	O	O
Egr-1	NN	O	B-DNA
expression	NN	O	O
are	NN	O	O
closely	NN	O	O
correlated	NN	O	O
with	NN	O	O
B	NN	O	B-cell_type
cell	NN	O	I-cell_type
proliferation	NN	O	O
in	NN	O	O
several	NN	O	O
models	NN	O	O
of	NN	O	O
B	NN	O	O
cell	NN	O	O
activation	NN	O	O
and	NN	O	O
tolerance	NN	O	O
.	NN	O	O

We	NN	O	O
compared	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
Egr-1	NN	O	B-DNA
during	NN	O	O
B	NN	O	O
cell	NN	O	O
stimulation	NN	O	O
with	NN	O	O
Fab'2	NN	O	B-protein
and	NN	O	O
IgG	NN	O	B-protein
anti-immunoglobulin	NN	O	I-protein
(	NN	O	O
anti-Ig	NN	O	B-protein
)	NN	O	O
,	NN	O	O
since	NN	O	O
it	NN	O	O
is	NN	O	O
known	NN	O	O
that	NN	O	O
Fab'2	NN	O	B-protein
anti-Ig	NN	O	I-protein
is	NN	O	O
mitogenic	NN	O	O
while	NN	O	O
IgG	NN	O	B-protein
anti-Ig	NN	O	I-protein
is	NN	O	O
not	NN	O	O
,	NN	O	O
owing	NN	O	O
to	NN	O	O
a	NN	O	O
dominant	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
of	NN	O	O
crosslinking	NN	O	O
the	NN	O	O
B	NN	O	B-protein
cell	NN	O	I-protein
Fc	NN	O	I-protein
gamma	NN	O	I-protein
RII	NN	O	I-protein
to	NN	O	O
membrane	NN	O	B-protein
Ig	NN	O	I-protein
.	NN	O	O

While	NN	O	O
mitogenic	NN	O	O
doses	NN	O	O
of	NN	O	O
Fab'2	NN	O	B-protein
anti-Ig	NN	O	I-protein
induce	NN	O	O
large	NN	O	O
and	NN	O	O
rapid	NN	O	O
increases	NN	O	O
in	NN	O	O
Egr-1	NN	O	O
expression	NN	O	O
,	NN	O	O
IgG	NN	O	B-protein
anti-Ig	NN	O	I-protein
results	NN	O	O
in	NN	O	O
smaller	NN	O	O
increases	NN	O	O
in	NN	O	O
Egr-1	NN	O	B-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
comparable	NN	O	O
to	NN	O	O
that	NN	O	O
seen	NN	O	O
with	NN	O	O
submitogenic	NN	O	O
concentrations	NN	O	O
of	NN	O	O
Fab'2	NN	O	B-protein
anti-Ig	NN	O	I-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
correlation	NN	O	O
between	NN	O	O
Egr-1	NN	O	O
expression	NN	O	O
and	NN	O	O
B	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
breaks	NN	O	O
down	NN	O	O
when	NN	O	O
IL-4	NN	O	B-protein
is	NN	O	O
added	NN	O	O
as	NN	O	O
a	NN	O	O
co-mitogen	NN	O	O
to	NN	O	O
induce	NN	O	O
B	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
with	NN	O	O
IgG	NN	O	B-protein
anti-Ig	NN	O	I-protein
or	NN	O	O
submitogenic	NN	O	O
concentrations	NN	O	O
of	NN	O	O
Fab'2	NN	O	B-protein
anti-Ig	NN	O	I-protein
.	NN	O	O

No	NN	O	O
corresponding	NN	O	O
increases	NN	O	O
in	NN	O	O
Egr-1	NN	O	B-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
are	NN	O	O
observed	NN	O	O
when	NN	O	O
IL-4	NN	O	B-protein
is	NN	O	O
added	NN	O	O
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
IL-4	NN	O	B-protein
overcomes	NN	O	O
Fc	NN	O	O
receptor-mediated	NN	O	O
inhibition	NN	O	O
of	NN	O	O
B	NN	O	B-cell_type
cell	NN	O	I-cell_type
proliferation	NN	O	O
without	NN	O	O
affecting	NN	O	O
inhibition	NN	O	O
of	NN	O	O
Egr-1	NN	O	B-RNA
mRNA	NN	O	I-RNA
induction	NN	O	O
,	NN	O	O
as	NN	O	O
demonstrated	NN	O	O
earlier	NN	O	O
for	NN	O	O
c-myc	NN	O	B-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
this	NN	O	O
system	NN	O	O
.	NN	O	O

-DOCSTART-	O

Identification	NN	O	O
of	NN	O	O
a	NN	O	O
novel	NN	O	O
cyclosporin-sensitive	NN	O	B-DNA
element	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
tumor	NN	O	I-DNA
necrosis	NN	O	I-DNA
factor	NN	O	I-DNA
alpha	NN	O	I-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
cytokine	NN	O	B-protein
with	NN	O	O
pleiotropic	NN	O	O
biological	NN	O	O
effects	NN	O	O
,	NN	O	O
is	NN	O	O
produced	NN	O	O
by	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
cell	NN	O	O
types	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
induction	NN	O	O
by	NN	O	O
diverse	NN	O	O
stimuli	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
paper	NN	O	O
,	NN	O	O
TNF-alpha	NN	O	B-RNA
mRNA	NN	O	I-RNA
is	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
highly	NN	O	O
induced	NN	O	O
in	NN	O	O
a	NN	O	O
murine	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
clone	NN	O	I-cell_line
by	NN	O	O
stimulation	NN	O	O
with	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
TCR	NN	O	B-protein
)	NN	O	O
ligands	NN	O	O
or	NN	O	O
by	NN	O	O
calcium	NN	O	O
ionophores	NN	O	O
alone	NN	O	O
.	NN	O	O

Induction	NN	O	O
is	NN	O	O
rapid	NN	O	O
,	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
and	NN	O	O
is	NN	O	O
completely	NN	O	O
blocked	NN	O	O
by	NN	O	O
the	NN	O	O
immunosuppressant	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
(	NN	O	O
CsA	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
identified	NN	O	O
a	NN	O	O
human	NN	O	B-DNA
TNF-alpha	NN	O	I-DNA
promoter	NN	O	I-DNA
element	NN	O	I-DNA
,	NN	O	O
kappa	NN	O	B-DNA
3	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
plays	NN	O	O
a	NN	O	O
key	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
calcium-mediated	NN	O	O
inducibility	NN	O	O
and	NN	O	O
CsA	NN	O	O
sensitivity	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
.	NN	O	O

In	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
,	NN	O	O
an	NN	O	O
oligonucleotide	NN	O	O
containing	NN	O	O
kappa	NN	O	B-DNA
3	NN	O	I-DNA
forms	NN	O	O
two	NN	O	O
DNA	NN	O	O
protein	NN	O	O
complexes	NN	O	O
with	NN	O	O
proteins	NN	O	O
that	NN	O	O
are	NN	O	O
present	NN	O	O
in	NN	O	O
extracts	NN	O	O
from	NN	O	O
unstimulated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
complexes	NN	O	O
appear	NN	O	O
in	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
only	NN	O	O
after	NN	O	O
T	NN	O	O
cell	NN	O	O
stimulation	NN	O	O
.	NN	O	O

Induction	NN	O	O
of	NN	O	O
the	NN	O	O
inducible	NN	O	O
nuclear	NN	O	B-protein
complexes	NN	O	I-protein
is	NN	O	O
rapid	NN	O	O
,	NN	O	O
independent	NN	O	O
of	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
,	NN	O	O
and	NN	O	O
blocked	NN	O	O
by	NN	O	O
CsA	NN	O	O
,	NN	O	O
and	NN	O	O
thus	NN	O	O
,	NN	O	O
exactly	NN	O	O
parallels	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
TNF-alpha	NN	O	B-RNA
mRNA	NN	O	I-RNA
by	NN	O	O
TCR	NN	O	O
ligands	NN	O	O
or	NN	O	O
by	NN	O	O
calcium	NN	O	O
ionophore	NN	O	O
.	NN	O	O

Our	NN	O	O
studies	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
kappa	NN	O	B-protein
3	NN	O	I-protein
binding	NN	O	I-protein
factor	NN	O	I-protein
resembles	NN	O	O
the	NN	O	O
preexisting	NN	O	O
component	NN	O	O
of	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

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
TNF-alpha	NN	O	B-DNA
gene	NN	O	I-DNA
is	NN	O	O
an	NN	O	O
immediate	NN	O	O
early	NN	O	O
gene	NN	O	O
in	NN	O	O
activated	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
provides	NN	O	O
a	NN	O	O
new	NN	O	O
model	NN	O	O
system	NN	O	O
in	NN	O	O
which	NN	O	O
to	NN	O	O
study	NN	O	O
CsA-sensitive	NN	O	B-DNA
gene	NN	O	I-DNA
induction	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Differences	NN	O	O
in	NN	O	O
expression	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
AP-1	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_line
promyelocytic	NN	O	I-cell_line
HL-60	NN	O	I-cell_line
cells	NN	O	I-cell_line
during	NN	O	O
differentiation	NN	O	O
towards	NN	O	O
macrophages	NN	O	B-cell_type
versus	NN	O	O
granulocytes	NN	O	B-cell_type
.	NN	O	O

Commitment	NN	O	O
of	NN	O	O
HL-60	NN	O	B-cell_line
cells	NN	O	I-cell_line
to	NN	O	O
macrophage	NN	O	O
or	NN	O	O
granulocytic	NN	O	O
differentiation	NN	O	O
was	NN	O	O
achieved	NN	O	O
by	NN	O	O
incubation	NN	O	O
with	NN	O	O
4	NN	O	O
beta-phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
for	NN	O	O
30-60	NN	O	O
min	NN	O	O
or	NN	O	O
with	NN	O	O
dimethyl	NN	O	O
sulphoxide	NN	O	O
(	NN	O	O
DMSO	NN	O	O
)	NN	O	O
for	NN	O	O
24	NN	O	O
h	NN	O	O
respectively	NN	O	O
.	NN	O	O

The	NN	O	O
commitment	NN	O	O
stage	NN	O	O
towards	NN	O	O
PMA-induced	NN	O	O
macrophage	NN	O	O
differentiation	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
increases	NN	O	O
in	NN	O	O
jun	NN	O	O
B	NN	O	O
and	NN	O	O
c-fos	NN	O	O
mRNA	NN	O	O
levels	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
with	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
binding	NN	O	O
activity	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
AP-1	NN	O	I-protein
.	NN	O	O

Nevertheless	NN	O	O
,	NN	O	O
gel	NN	O	O
retardation	NN	O	O
analysis	NN	O	O
indicated	NN	O	O
that	NN	O	O
the	NN	O	O
AP-1	NN	O	B-protein
activity	NN	O	O
detected	NN	O	O
in	NN	O	O
untreated	NN	O	O
cells	NN	O	O
was	NN	O	O
drastically	NN	O	O
reduced	NN	O	O
during	NN	O	O
the	NN	O	O
commitment	NN	O	O
stage	NN	O	O
of	NN	O	O
DMSO-induced	NN	O	O
HL-60	NN	O	O
differentiation	NN	O	O
towards	NN	O	O
granulocytes	NN	O	B-cell_type
.	NN	O	O

When	NN	O	O
HL-60	NN	O	B-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
treated	NN	O	O
with	NN	O	O
sodium	NN	O	O
butyrate	NN	O	O
,	NN	O	O
which	NN	O	O
induced	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
,	NN	O	O
a	NN	O	O
remarkable	NN	O	O
increase	NN	O	O
in	NN	O	O
AP-1	NN	O	O
binding	NN	O	O
activity	NN	O	O
was	NN	O	O
detected	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
1	NN	O	O
alpha	NN	O	O
,	NN	O	O
25-dihydroxyvitamin	NN	O	O
D3	NN	O	O
,	NN	O	O
another	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
agent	NN	O	O
,	NN	O	O
induced	NN	O	O
a	NN	O	O
weak	NN	O	O
,	NN	O	O
but	NN	O	O
appreciable	NN	O	O
,	NN	O	O
increase	NN	O	O
in	NN	O	O
AP-1	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
addition	NN	O	O
of	NN	O	O
sodium	NN	O	O
butyrate	NN	O	O
or	NN	O	O
1	NN	O	O
alpha	NN	O	O
,	NN	O	O
25-dihydroxyvitamin	NN	O	O
D3	NN	O	O
to	NN	O	O
HL-60	NN	O	B-cell_line
cells	NN	O	I-cell_line
induced	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
c-fos	NN	O	B-DNA
,	NN	O	I-DNA
c-jun	NN	O	I-DNA
,	NN	O	I-DNA
jun	NN	O	I-DNA
B	NN	O	I-DNA
and	NN	O	I-DNA
jun	NN	O	I-DNA
D	NN	O	I-DNA
proto-oncogenes	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
when	NN	O	O
HL-60	NN	O	B-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
treated	NN	O	O
with	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
,	NN	O	O
a	NN	O	O
granulocytic	NN	O	O
differentiation	NN	O	O
inducer	NN	O	O
,	NN	O	O
no	NN	O	O
enhanced	NN	O	O
AP-1	NN	O	O
binding	NN	O	O
activity	NN	O	O
was	NN	O	O
observed	NN	O	O
,	NN	O	O
and	NN	O	O
only	NN	O	O
a	NN	O	O
weak	NN	O	O
increase	NN	O	O
in	NN	O	O
jun	NN	O	B-RNA
D	NN	O	I-RNA
mRNA	NN	O	I-RNA
level	NN	O	O
was	NN	O	O
detected	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
formation	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
is	NN	O	O
not	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
HL-60	NN	O	B-cell_line
differentiation	NN	O	O
towards	NN	O	O
granulocytes	NN	O	B-cell_type
,	NN	O	O
whereas	NN	O	O
induction	NN	O	O
of	NN	O	O
monocytic	NN	O	O
differentiation	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
AP-1	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

The	NN	O	O
differential	NN	O	O
expression	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
activity	NN	O	O
may	NN	O	O
be	NN	O	O
critical	NN	O	O
in	NN	O	O
the	NN	O	O
differentiation	NN	O	O
of	NN	O	O
HL-60	NN	O	B-cell_line
cells	NN	O	I-cell_line
towards	NN	O	O
monocytic	NN	O	B-cell_type
or	NN	O	I-cell_type
granulocytic	NN	O	I-cell_type
lineages	NN	O	I-cell_type

-DOCSTART-	O

Glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
and	NN	O	O
sensitivity	NN	O	O
in	NN	O	O
leukemias	NN	O	O
.	NN	O	O

In	NN	O	O
an	NN	O	O
attempt	NN	O	O
to	NN	O	O
investigate	NN	O	O
the	NN	O	O
utility	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
determination	NN	O	O
to	NN	O	O
predict	NN	O	O
clinical	NN	O	O
responsiveness	NN	O	O
in	NN	O	O
human	NN	O	O
leukemias	NN	O	O
we	NN	O	O
have	NN	O	O
studied	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
the	NN	O	O
leukemic	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
46	NN	O	O
patients	NN	O	O
and	NN	O	O
in	NN	O	O
the	NN	O	O
lymphocytes	NN	O	B-cell_type
from	NN	O	O
18	NN	O	O
normal	NN	O	O
donors	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
normal	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
there	NN	O	O
were	NN	O	O
3	NN	O	O
,	NN	O	O
875	NN	O	O
(	NN	O	O
Median	NN	O	O
)	NN	O	O
specific	NN	O	O
binding	NN	O	O
sites	NN	O	O
per	NN	O	O
cell	NN	O	O
.	NN	O	O

The	NN	O	O
blasts	NN	O	B-cell_type
from	NN	O	O
17	NN	O	O
patients	NN	O	O
with	NN	O	O
ANLL	NN	O	O
had	NN	O	O
on	NN	O	O
average	NN	O	O
higher	NN	O	O
levels	NN	O	O
of	NN	O	O
binding	NN	O	B-protein
sites	NN	O	I-protein
per	NN	O	O
cell	NN	O	O
(	NN	O	O
Median	NN	O	O
=	NN	O	O
7	NN	O	O
,	NN	O	O
250	NN	O	O
,	NN	O	O
range	NN	O	O
:	NN	O	O
0	NN	O	O
to	NN	O	O
15	NN	O	O
,	NN	O	O
295	NN	O	O
)	NN	O	O
than	NN	O	O
the	NN	O	O
other	NN	O	O
leukemias	NN	O	O
.	NN	O	O

Of	NN	O	O
the	NN	O	O
15	NN	O	O
patients	NN	O	O
with	NN	O	O
CLL	NN	O	O
,	NN	O	O
six	NN	O	O
had	NN	O	O
received	NN	O	O
glucocorticoid	NN	O	O
treatment	NN	O	O
for	NN	O	O
3	NN	O	O
to	NN	O	O
5	NN	O	O
years	NN	O	O
.	NN	O	O

Their	NN	O	O
lymphocytes	NN	O	B-cell_type
had	NN	O	O
lower	NN	O	O
number	NN	O	O
of	NN	O	O
receptors	NN	O	O
(	NN	O	O
Median	NN	O	O
=	NN	O	O
2	NN	O	O
,	NN	O	O
000	NN	O	O
)	NN	O	O
than	NN	O	O
the	NN	O	O
other	NN	O	O
cases	NN	O	O
which	NN	O	O
were	NN	O	O
newly	NN	O	O
diagnosed	NN	O	O
(	NN	O	O
Median	NN	O	O
=	NN	O	O
4	NN	O	O
,	NN	O	O
500	NN	O	O
)	NN	O	O
.	NN	O	O

Four	NN	O	O
patients	NN	O	O
had	NN	O	O
ALL/AUL	NN	O	O
,	NN	O	O
three	NN	O	O
patients	NN	O	O
had	NN	O	O
blast	NN	O	O
crisis	NN	O	O
as	NN	O	O
terminal	NN	O	O
phase	NN	O	O
of	NN	O	O
CML	NN	O	O
,	NN	O	O
and	NN	O	O
seven	NN	O	O
had	NN	O	O
leukemic	NN	O	O
Non-Hodgkin	NN	O	O
lymphomas	NN	O	O
(	NN	O	O
Median	NN	O	O
=	NN	O	O
3	NN	O	O
,	NN	O	O
500	NN	O	O
sites/cell	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
24	NN	O	O
patients	NN	O	O
we	NN	O	O
have	NN	O	O
also	NN	O	O
studied	NN	O	O
the	NN	O	O
in	NN	O	O
vitro	NN	O	O
sensitivity	NN	O	O
of	NN	O	O
the	NN	O	O
leukemic	NN	O	B-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
dexamethasone	NN	O	O
.	NN	O	O

There	NN	O	O
was	NN	O	O
no	NN	O	O
marked	NN	O	O
correlation	NN	O	O
between	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
levels	NN	O	O
and	NN	O	O
in	NN	O	O
vitro	NN	O	O
sensitivity	NN	O	O
.	NN	O	O

An	NN	O	O
attempt	NN	O	O
to	NN	O	O
correlate	NN	O	O
receptor	NN	O	O
levels	NN	O	O
with	NN	O	O
clinical	NN	O	O
responsiveness	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
determination	NN	O	O
might	NN	O	O
be	NN	O	O
of	NN	O	O
value	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
lymphoid	NN	O	O
malignancies	NN	O	O
but	NN	O	O
probably	NN	O	O
not	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
other	NN	O	O
leukemias	NN	O	O
.	NN	O	O

-DOCSTART-	O

'Activation-labile	NN	O	B-protein
'	NN	O	I-protein
glucocorticoid-receptor	NN	O	I-protein
complexes	NN	O	I-protein
of	NN	O	O
a	NN	O	O
steroid-resistant	NN	O	O
variant	NN	O	O
of	NN	O	O
CEM-C7	NN	O	B-cell_line
human	NN	O	I-cell_line
lymphoid	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

For	NN	O	O
cytoplasmic	NN	O	B-protein
glucocorticoid-receptor	NN	O	I-protein
complexes	NN	O	I-protein
to	NN	O	O
enter	NN	O	O
and	NN	O	O
accumulate	NN	O	O
in	NN	O	O
the	NN	O	O
nucleus	NN	O	O
a	NN	O	O
temperature-dependent	NN	O	O
event	NN	O	O
,	NN	O	O
'activation	NN	O	O
'	NN	O	O
is	NN	O	O
required	NN	O	O
.	NN	O	O

Activation	NN	O	O
can	NN	O	O
be	NN	O	O
achieved	NN	O	O
in	NN	O	O
vitro	NN	O	O
by	NN	O	O
increased	NN	O	O
ionic	NN	O	O
strength	NN	O	O
,	NN	O	O
dilution	NN	O	O
or	NN	O	O
gel	NN	O	O
filtration	NN	O	O
and	NN	O	O
is	NN	O	O
manifested	NN	O	O
by	NN	O	O
an	NN	O	O
increased	NN	O	O
affinity	NN	O	O
of	NN	O	O
steroid-receptor	NN	O	B-protein
complex	NN	O	I-protein
for	NN	O	O
DNA	NN	O	B-DNA
and	NN	O	O
an	NN	O	O
altered	NN	O	O
elution	NN	O	O
profile	NN	O	O
from	NN	O	O
ion-exchange	NN	O	O
resins	NN	O	O
.	NN	O	O

Munck	NN	O	O
and	NN	O	O
Foley	NN	O	O
have	NN	O	O
shown	NN	O	O
that	NN	O	O
activated	NN	O	B-protein
complexes	NN	O	I-protein
isolated	NN	O	O
from	NN	O	O
thymocytes	NN	O	O
elute	NN	O	O
from	NN	O	O
DEAE-cellulose	NN	O	O
in	NN	O	O
a	NN	O	O
manner	NN	O	O
identical	NN	O	O
to	NN	O	O
complexes	NN	O	O
activated	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
here	NN	O	O
that	NN	O	O
DEAE-cellulose	NN	O	O
chromatography	NN	O	O
of	NN	O	O
steroid-receptor	NN	O	B-protein
complexes	NN	O	I-protein
from	NN	O	O
CEM-C7	NN	O	B-cell_line
,	NN	O	O
a	NN	O	O
cloned	NN	O	B-cell_line
human	NN	O	I-cell_line
leukaemic	NN	O	I-cell_line
T-cell	NN	O	I-cell_line
line	NN	O	I-cell_line
sensitive	NN	O	O
to	NN	O	O
the	NN	O	O
cytolytic	NN	O	O
action	NN	O	O
of	NN	O	O
glucocorticoids	NN	O	O
,	NN	O	O
and	NN	O	O
its	NN	O	O
steroid-resistant	NN	O	B-cell_line
subclone	NN	O	I-cell_line
4R4	NN	O	I-cell_line
demonstrated	NN	O	O
that	NN	O	O
steroid	NN	O	B-protein
receptors	NN	O	I-protein
of	NN	O	O
clone	NN	O	B-cell_line
4R4	NN	O	I-cell_line
can	NN	O	O
not	NN	O	O
form	NN	O	O
stable	NN	O	O
activated	NN	O	O
complexes	NN	O	O
.	NN	O	O

This	NN	O	O
defines	NN	O	O
a	NN	O	O
new	NN	O	O
defect	NN	O	O
in	NN	O	O
receptor	NN	O	O
action	NN	O	O
,	NN	O	O
activation	NN	O	O
lability	NN	O	O
(	NN	O	O
r+act1	NN	O	O
)	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
unlike	NN	O	O
either	NN	O	O
the	NN	O	O
r-	NN	O	B-cell_line
,	NN	O	I-cell_line
r+nt-	NN	O	I-cell_line
,	NN	O	I-cell_line
or	NN	O	I-cell_line
r+nti	NN	O	I-cell_line
phenotypes	NN	O	I-cell_line
previously	NN	O	O
described	NN	O	O
for	NN	O	O
mouse	NN	O	B-cell_line
lymphoid	NN	O	I-cell_line
variants	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

Granulocytes	NN	O	B-cell_type
in	NN	O	O
the	NN	O	O
endometrium	NN	O	O
of	NN	O	O
post-partum	NN	O	O
women	NN	O	O
.	NN	O	O

Endometrial	NN	O	O
samples	NN	O	O
of	NN	O	O
women	NN	O	O
at	NN	O	O
various	NN	O	O
stages	NN	O	O
of	NN	O	O
gonadal	NN	O	O
activity	NN	O	O
after	NN	O	O
parturition	NN	O	O
were	NN	O	O
examined	NN	O	O
for	NN	O	O
the	NN	O	O
presence	NN	O	O
and	NN	O	O
numbers	NN	O	O
of	NN	O	O
endometrial	NN	O	B-cell_type
granulocytes	NN	O	I-cell_type
.	NN	O	O

Although	NN	O	O
samples	NN	O	O
at	NN	O	O
all	NN	O	O
the	NN	O	O
stages	NN	O	O
contained	NN	O	O
significant	NN	O	O
numbers	NN	O	O
of	NN	O	O
the	NN	O	O
granulocytes	NN	O	B-cell_type
(	NN	O	O
i.e.	NN	O	O
greater	NN	O	O
than	NN	O	O
7/high-power	NN	O	O
field	NN	O	O
)	NN	O	O
,	NN	O	O
the	NN	O	O
100	NN	O	O
%	NN	O	O
values	NN	O	O
for	NN	O	O
late-proliferative	NN	O	O
and	NN	O	O
adaptation	NN	O	O
hyperplasia	NN	O	O
were	NN	O	O
significantly	NN	O	O
higher	NN	O	O
than	NN	O	O
the	NN	O	O
values	NN	O	O
for	NN	O	O
the	NN	O	O
resting	NN	O	O
(	NN	O	O
81.8	NN	O	O
%	NN	O	O
)	NN	O	O
,	NN	O	O
early	NN	O	O
(	NN	O	O
82.4	NN	O	O
%	NN	O	O
)	NN	O	O
and	NN	O	O
mid-	NN	O	O
(	NN	O	O
87.9	NN	O	O
%	NN	O	O
)	NN	O	O
proliferative	NN	O	O
and	NN	O	O
secretory	NN	O	O
(	NN	O	O
83.3	NN	O	O
%	NN	O	O
)	NN	O	O
phases	NN	O	O
.	NN	O	O

We	NN	O	O
suggest	NN	O	O
that	NN	O	O
this	NN	O	O
correlates	NN	O	O
with	NN	O	O
the	NN	O	O
suggestion	NN	O	O
that	NN	O	O
the	NN	O	O
granulocytes	NN	O	B-cell_type
constitute	NN	O	O
a	NN	O	O
receptor	NN	O	O
system	NN	O	O
for	NN	O	O
oestrogens	NN	O	O
.	NN	O	O

-DOCSTART-	O

Interaction	NN	O	O
of	NN	O	O
glucocorticoids	NN	O	O
with	NN	O	O
macrophages	NN	O	B-cell_type
.	NN	O	O

Identification	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
macrophages	NN	O	B-cell_type
.	NN	O	O

Glucocorticoid	NN	O	O
binding	NN	O	O
was	NN	O	O
measured	NN	O	O
in	NN	O	O
resident	NN	O	O
and	NN	O	O
thioglycollate-elicited	NN	O	B-cell_type
mouse	NN	O	I-cell_type
peritoneal	NN	O	I-cell_type
macrophages	NN	O	I-cell_type
,	NN	O	O
rabbit	NN	O	B-cell_type
alveolar	NN	O	I-cell_type
macrophages	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

Two	NN	O	O
assays	NN	O	O
of	NN	O	O
binding	NN	O	O
were	NN	O	O
used	NN	O	O
--	NN	O	O
an	NN	O	O
assay	NN	O	O
with	NN	O	O
intact	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
suspension	NN	O	O
or	NN	O	O
monolayers	NN	O	O
,	NN	O	O
and	NN	O	O
an	NN	O	O
assay	NN	O	O
of	NN	O	O
cytosol	NN	O	O
and	NN	O	O
nuclear	NN	O	O
forms	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
.	NN	O	O

The	NN	O	O
mononuclear	NN	O	B-cell_type
phagocytes	NN	O	I-cell_type
contained	NN	O	O
approximately	NN	O	O
equal	NN	O	O
to	NN	O	O
4	NN	O	O
--	NN	O	O
10	NN	O	O
X	NN	O	O
10	NN	O	O
(	NN	O	O
3	NN	O	O
)	NN	O	O
high	NN	O	B-protein
affinity	NN	O	I-protein
receptor	NN	O	I-protein
sites	NN	O	I-protein
per	NN	O	O
cell	NN	O	O
,	NN	O	O
with	NN	O	O
dissociation	NN	O	O
constants	NN	O	O
of	NN	O	O
approximately	NN	O	O
equal	NN	O	O
to	NN	O	O
2	NN	O	O
--	NN	O	O
8	NN	O	O
nM	NN	O	O
dexamethasone	NN	O	O
.	NN	O	O

The	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
saturable	NN	O	B-protein
sites	NN	O	I-protein
was	NN	O	O
specific	NN	O	O
for	NN	O	O
steroids	NN	O	O
with	NN	O	O
glucocorticoid	NN	O	O
or	NN	O	O
antiglucocorticoid	NN	O	O
activity	NN	O	O
.	NN	O	O

Cortisol	NN	O	O
,	NN	O	O
corticosterone	NN	O	O
,	NN	O	O
and	NN	O	O
progesterone	NN	O	O
competed	NN	O	O
with	NN	O	O
dexamethasone	NN	O	O
for	NN	O	O
binding	NN	O	O
,	NN	O	O
whereas	NN	O	O
estradiol	NN	O	O
,	NN	O	O
dihydrotestosterone	NN	O	O
,	NN	O	O
and	NN	O	O
11-epicortisol	NN	O	O
competed	NN	O	O
very	NN	O	O
little	NN	O	O
.	NN	O	O

Binding	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
to	NN	O	O
cytosol	NN	O	O
and	NN	O	O
nuclear	NN	O	O
forms	NN	O	O
of	NN	O	O
the	NN	O	O
receptor	NN	O	B-protein
complex	NN	O	I-protein
and	NN	O	O
temperature-sensitive	NN	O	O
translocation	NN	O	O
of	NN	O	O
cytosol	NN	O	O
forms	NN	O	O
to	NN	O	O
nuclear	NN	O	O
forms	NN	O	O
were	NN	O	O
shown	NN	O	O
.	NN	O	O

At	NN	O	O
37	NN	O	O
degrees	NN	O	O
C	NN	O	O
the	NN	O	O
predominant	NN	O	O
form	NN	O	O
of	NN	O	O
the	NN	O	O
hormone-receptor	NN	O	B-protein
complex	NN	O	I-protein
was	NN	O	O
nuclear	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
corticosteroids	NN	O	O
interact	NN	O	O
with	NN	O	O
macrophages	NN	O	B-cell_type
at	NN	O	O
physiological	NN	O	O
concentrations	NN	O	O
.	NN	O	O

-DOCSTART-	O

Nitric	NN	O	O
oxide	NN	O	O
signaling	NN	O	O
:	NN	O	O
a	NN	O	O
possible	NN	O	O
role	NN	O	O
for	NN	O	O
G	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
reported	NN	O	O
various	NN	O	O
inductive	NN	O	O
effects	NN	O	O
of	NN	O	O
nitric	NN	O	O
oxide	NN	O	O
on	NN	O	O
human	NN	O	B-cell_type
PBMC	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
describe	NN	O	O
a	NN	O	O
novel	NN	O	O
and	NN	O	O
potentially	NN	O	O
important	NN	O	O
mechanism	NN	O	O
of	NN	O	O
nitric	NN	O	O
oxide	NN	O	O
signaling-through	NN	O	O
direct	NN	O	O
activation	NN	O	O
of	NN	O	O
guanine	NN	O	B-protein
nucleotide-binding	NN	O	I-protein
proteins	NN	O	I-protein
(	NN	O	O
G	NN	O	B-protein
proteins	NN	O	I-protein
)	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
found	NN	O	O
that	NN	O	O
nitric	NN	O	O
oxide	NN	O	O
treatment	NN	O	O
of	NN	O	O
membranes	NN	O	O
isolated	NN	O	O
from	NN	O	O
fresh	NN	O	B-cell_type
human	NN	O	I-cell_type
PBMC	NN	O	I-cell_type
enhances	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
these	NN	O	O
membranes	NN	O	O
to	NN	O	O
hydrolyze	NN	O	O
[	NN	O	O
gamma-32P	NN	O	O
]	NN	O	O
GTP	NN	O	O
and	NN	O	O
bind	NN	O	O
[	NN	O	O
gamma-35S	NN	O	O
]	NN	O	O
GTP	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
treatment	NN	O	O
of	NN	O	O
whole	NN	O	O
cells	NN	O	O
with	NN	O	O
nitric	NN	O	O
oxide	NN	O	O
yielded	NN	O	O
membranes	NN	O	O
with	NN	O	O
enhanced	NN	O	O
GTPase	NN	O	O
activity	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
the	NN	O	O
GTPase	NN	O	O
activity	NN	O	O
of	NN	O	O
pure	NN	O	O
,	NN	O	O
recombinant	NN	O	O
Gs	NN	O	B-protein
alpha	NN	O	I-protein
,	NN	O	O
Gi	NN	O	B-protein
alpha	NN	O	I-protein
1	NN	O	I-protein
,	NN	O	O
and	NN	O	O
p21ras	NN	O	B-protein
was	NN	O	O
greatly	NN	O	O
enhanced	NN	O	O
by	NN	O	O
nitric	NN	O	O
oxide	NN	O	O
.	NN	O	O

In	NN	O	O
support	NN	O	O
of	NN	O	O
the	NN	O	O
existence	NN	O	O
of	NN	O	O
this	NN	O	O
pathway	NN	O	O
in	NN	O	O
whole	NN	O	O
cells	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
G	NN	O	O
protein	NN	O	O
inhibitor	NN	O	O
,	NN	O	O
GDP-beta-S	NN	O	O
,	NN	O	O
blocked	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
translocation	NN	O	O
induced	NN	O	O
by	NN	O	O
nitric	NN	O	O
oxide	NN	O	O
or	NN	O	O
LPS	NN	O	O
in	NN	O	O
permeabilized	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
nitric	NN	O	O
oxide	NN	O	O
greatly	NN	O	O
reduced	NN	O	O
the	NN	O	O
pertussis	NN	O	O
toxin-mediated	NN	O	O
ADP-ribosylation	NN	O	O
of	NN	O	O
45-	NN	O	B-protein
and	NN	O	I-protein
41-kDa	NN	O	I-protein
proteins	NN	O	I-protein
in	NN	O	O
membranes	NN	O	O
of	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

Because	NN	O	O
G	NN	O	B-protein
proteins	NN	O	I-protein
play	NN	O	O
a	NN	O	O
central	NN	O	O
role	NN	O	O
in	NN	O	O
many	NN	O	O
diverse	NN	O	O
signaling	NN	O	O
systems	NN	O	O
,	NN	O	O
activation	NN	O	O
by	NN	O	O
an	NN	O	O
endogenous	NN	O	O
and	NN	O	O
inducible	NN	O	O
oxidant	NN	O	O
may	NN	O	O
represent	NN	O	O
a	NN	O	O
novel	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
granulocyte-macrophage	NN	O	B-DNA
colony-stimulating	NN	O	I-DNA
factor	NN	O	I-DNA
promoter	NN	O	I-DNA
cis-acting	NN	O	I-DNA
element	NN	O	I-DNA
CLE0	NN	O	B-DNA
mediates	NN	O	O
induction	NN	O	O
signals	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
is	NN	O	O
recognized	NN	O	O
by	NN	O	O
factors	NN	O	O
related	NN	O	O
to	NN	O	O
AP1	NN	O	B-protein
and	NN	O	O
NFAT	NN	O	B-protein
.	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
gene	NN	O	I-DNA
in	NN	O	O
T	NN	O	O
cells	NN	O	O
is	NN	O	O
activated	NN	O	O
by	NN	O	O
the	NN	O	O
combination	NN	O	O
of	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
(	NN	O	O
phorbol	NN	O	O
myristate	NN	O	O
acetate	NN	O	O
)	NN	O	O
and	NN	O	O
calcium	NN	O	O
ionophore	NN	O	O
(	NN	O	O
A23187	NN	O	O
)	NN	O	O
,	NN	O	O
which	NN	O	O
mimic	NN	O	O
antigen	NN	O	O
stimulation	NN	O	O
through	NN	O	O
the	NN	O	O
T-cell	NN	O	B-protein
receptor	NN	O	I-protein
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
shown	NN	O	O
that	NN	O	O
a	NN	O	O
fragment	NN	O	O
containing	NN	O	O
bp	NN	O	B-DNA
-95	NN	O	I-DNA
to	NN	O	I-DNA
+27	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
mouse	NN	O	B-DNA
GM-CSF	NN	O	I-DNA
promoter	NN	O	I-DNA
can	NN	O	O
confer	NN	O	O
inducibility	NN	O	O
to	NN	O	O
reporter	NN	O	B-DNA
genes	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
Jurkat	NN	O	I-cell_line
T-cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

Here	NN	O	O
we	NN	O	O
use	NN	O	O
an	NN	O	O
in	NN	O	O
vitro	NN	O	O
transcription	NN	O	O
system	NN	O	O
to	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
a	NN	O	O
cis-acting	NN	O	B-DNA
element	NN	O	I-DNA
(	NN	O	O
positions	NN	O	B-DNA
-54	NN	O	I-DNA
to	NN	O	I-DNA
-40	NN	O	I-DNA
)	NN	O	O
,	NN	O	O
referred	NN	O	O
to	NN	O	O
as	NN	O	O
CLE0	NN	O	B-DNA
,	NN	O	O
is	NN	O	O
a	NN	O	O
target	NN	O	O
for	NN	O	O
the	NN	O	O
induction	NN	O	O
signals	NN	O	O
.	NN	O	O

We	NN	O	O
observed	NN	O	O
induction	NN	O	O
with	NN	O	O
templates	NN	O	O
containing	NN	O	O
intact	NN	O	O
CLE0	NN	O	B-DNA
but	NN	O	O
not	NN	O	O
with	NN	O	O
templates	NN	O	O
with	NN	O	O
deleted	NN	O	B-DNA
or	NN	O	I-DNA
mutated	NN	O	I-DNA
CLE0	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
also	NN	O	O
observed	NN	O	O
that	NN	O	O
two	NN	O	O
distinct	NN	O	O
signals	NN	O	O
were	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
stimulation	NN	O	O
through	NN	O	O
CLE0	NN	O	B-DNA
,	NN	O	O
since	NN	O	O
only	NN	O	O
extracts	NN	O	O
from	NN	O	O
cells	NN	O	O
treated	NN	O	O
with	NN	O	O
both	NN	O	O
phorbol	NN	O	O
myristate	NN	O	O
acetate	NN	O	O
and	NN	O	O
A23187	NN	O	O
supported	NN	O	O
optimal	NN	O	O
induction	NN	O	O
.	NN	O	O

Stimulation	NN	O	O
probably	NN	O	O
was	NN	O	O
mediated	NN	O	O
by	NN	O	O
CLE0-binding	NN	O	B-protein
proteins	NN	O	I-protein
because	NN	O	O
depletion	NN	O	O
of	NN	O	O
these	NN	O	O
proteins	NN	O	O
specifically	NN	O	O
reduced	NN	O	O
GM-CSF	NN	O	B-protein
transcription	NN	O	O
.	NN	O	O

One	NN	O	O
of	NN	O	O
the	NN	O	O
binding	NN	O	B-protein
factors	NN	O	I-protein
possessed	NN	O	O
biochemical	NN	O	O
and	NN	O	O
immunological	NN	O	O
features	NN	O	O
identical	NN	O	O
to	NN	O	O
those	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
AP1	NN	O	I-protein
.	NN	O	O

Another	NN	O	O
factor	NN	O	O
resembled	NN	O	O
the	NN	O	O
T-cell-specific	NN	O	B-protein
factor	NN	O	I-protein
NFAT	NN	O	I-protein
.	NN	O	O

The	NN	O	O
characteristics	NN	O	O
of	NN	O	O
these	NN	O	O
two	NN	O	O
factors	NN	O	O
are	NN	O	O
consistent	NN	O	O
with	NN	O	O
their	NN	O	O
involvement	NN	O	O
in	NN	O	O
GM-CSF	NN	O	O
induction	NN	O	O
.	NN	O	O

The	NN	O	O
presence	NN	O	O
of	NN	O	O
CLE0-like	NN	O	B-DNA
elements	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
promoters	NN	O	B-DNA
of	NN	O	O
interleukin-3	NN	O	B-DNA
(	NN	O	I-DNA
IL-3	NN	O	I-DNA
)	NN	O	I-DNA
,	NN	O	I-DNA
IL-4	NN	O	I-DNA
,	NN	O	I-DNA
IL-5	NN	O	I-DNA
,	NN	O	I-DNA
GM-CSF	NN	O	I-DNA
,	NN	O	I-DNA
and	NN	O	I-DNA
NFAT	NN	O	I-DNA
sites	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
suggests	NN	O	O
that	NN	O	O
the	NN	O	O
factors	NN	O	O
we	NN	O	O
detected	NN	O	O
,	NN	O	O
or	NN	O	O
related	NN	O	O
factors	NN	O	O
that	NN	O	O
recognize	NN	O	O
these	NN	O	O
sites	NN	O	O
,	NN	O	O
may	NN	O	O
account	NN	O	O
for	NN	O	O
the	NN	O	O
coordinate	NN	O	O
induction	NN	O	O
of	NN	O	O
these	NN	O	O
genes	NN	O	O
during	NN	O	O
T-cell	NN	O	B-cell_type
activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Identification	NN	O	O
and	NN	O	O
characterization	NN	O	O
of	NN	O	O
an	NN	O	O
Alu-containing	NN	O	B-DNA
,	NN	O	I-DNA
T-cell-specific	NN	O	I-DNA
enhancer	NN	O	I-DNA
located	NN	O	O
in	NN	O	O
the	NN	O	O
last	NN	O	O
intron	NN	O	B-DNA
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
CD8	NN	O	I-DNA
alpha	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
CD8	NN	O	I-DNA
alpha	NN	O	I-DNA
gene	NN	O	I-DNA
is	NN	O	O
restricted	NN	O	O
to	NN	O	O
cells	NN	O	O
of	NN	O	O
the	NN	O	O
lymphoid	NN	O	B-cell_type
lineage	NN	O	I-cell_type
and	NN	O	O
developmentally	NN	O	O
regulated	NN	O	O
during	NN	O	O
thymopoiesis	NN	O	O
.	NN	O	O

As	NN	O	O
an	NN	O	O
initial	NN	O	O
step	NN	O	O
towards	NN	O	O
understanding	NN	O	O
the	NN	O	O
molecular	NN	O	O
basis	NN	O	O
for	NN	O	O
tissue-specific	NN	O	O
expression	NN	O	O
of	NN	O	O
this	NN	O	O
gene	NN	O	O
,	NN	O	O
we	NN	O	O
surveyed	NN	O	O
the	NN	O	O
surrounding	NN	O	O
chromatin	NN	O	O
structure	NN	O	O
for	NN	O	O
potential	NN	O	O
cis-acting	NN	O	B-DNA
regulatory	NN	O	I-DNA
regions	NN	O	I-DNA
by	NN	O	O
DNase	NN	O	B-protein
I	NN	O	I-protein
hypersensitivity	NN	O	O
mapping	NN	O	O
and	NN	O	O
found	NN	O	O
four	NN	O	O
hypersensitive	NN	O	O
sites	NN	O	O
,	NN	O	O
three	NN	O	O
of	NN	O	O
which	NN	O	O
were	NN	O	O
T	NN	O	O
cell	NN	O	O
restricted	NN	O	O
.	NN	O	O

By	NN	O	O
using	NN	O	O
a	NN	O	O
reporter-based	NN	O	O
expression	NN	O	O
approach	NN	O	O
,	NN	O	O
a	NN	O	O
T-cell-specific	NN	O	B-DNA
enhancer	NN	O	I-DNA
was	NN	O	O
identified	NN	O	O
by	NN	O	O
its	NN	O	O
close	NN	O	O
association	NN	O	O
with	NN	O	O
a	NN	O	O
prominent	NN	O	O
T-cell-restricted	NN	O	B-DNA
hypersensitive	NN	O	I-DNA
sites	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
last	NN	O	O
intron	NN	O	O
of	NN	O	O
the	NN	O	O
CD8	NN	O	B-DNA
alpha	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

Deletion	NN	O	O
studies	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
minimal	NN	O	O
enhancer	NN	O	O
is	NN	O	O
adjacent	NN	O	O
to	NN	O	O
a	NN	O	O
negative	NN	O	O
regulatory	NN	O	O
element	NN	O	O
.	NN	O	O

DNA	NN	O	O
sequence	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
minimal	NN	O	B-DNA
enhancer	NN	O	I-DNA
revealed	NN	O	O
a	NN	O	O
striking	NN	O	O
cluster	NN	O	O
of	NN	O	O
consensus	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
for	NN	O	O
Ets-1	NN	O	B-protein
,	NN	O	I-protein
TCF-1	NN	O	I-protein
,	NN	O	I-protein
CRE	NN	O	I-protein
,	NN	O	I-protein
GATA-3	NN	O	I-protein
,	NN	O	I-protein
LyF-1	NN	O	I-protein
,	NN	O	I-protein
and	NN	O	I-protein
bHLH	NN	O	I-protein
proteins	NN	O	I-protein
which	NN	O	O
were	NN	O	O
verified	NN	O	O
by	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
the	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
end	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
enhancer	NN	O	B-DNA
was	NN	O	O
composed	NN	O	O
of	NN	O	O
an	NN	O	O
Alu	NN	O	O
repeat	NN	O	O
which	NN	O	O
contained	NN	O	O
the	NN	O	O
GATA-3	NN	O	B-DNA
,	NN	O	I-DNA
bHLH	NN	O	I-DNA
,	NN	O	I-DNA
and	NN	O	I-DNA
LyF-1	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
.	NN	O	O

Site-directed	NN	O	O
mutation	NN	O	O
of	NN	O	O
the	NN	O	O
Ets-1	NN	O	B-DNA
and	NN	O	I-DNA
GATA-3	NN	O	I-DNA
sites	NN	O	I-DNA
dramatically	NN	O	O
reduced	NN	O	O
enhancer	NN	O	O
activity	NN	O	O
.	NN	O	O

The	NN	O	O
functional	NN	O	O
importance	NN	O	O
of	NN	O	O
the	NN	O	O
other	NN	O	O
binding	NN	O	O
sites	NN	O	O
only	NN	O	O
became	NN	O	O
apparent	NN	O	O
when	NN	O	O
combinations	NN	O	O
of	NN	O	O
mutations	NN	O	O
were	NN	O	O
analyzed	NN	O	O
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
these	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
CD8	NN	O	I-DNA
alpha	NN	O	I-DNA
gene	NN	O	I-DNA
is	NN	O	O
regulated	NN	O	O
by	NN	O	O
the	NN	O	O
interaction	NN	O	O
of	NN	O	O
multiple	NN	O	O
T-cell	NN	O	B-protein
nuclear	NN	O	I-protein
proteins	NN	O	I-protein
with	NN	O	O
a	NN	O	O
transcriptional	NN	O	O
enhancer	NN	O	O
located	NN	O	O
in	NN	O	O
the	NN	O	O
last	NN	O	O
intron	NN	O	B-DNA
of	NN	O	O
the	NN	O	O
gene	NN	O	O
.	NN	O	O

Comparison	NN	O	O
of	NN	O	O
the	NN	O	O
CD8	NN	O	B-DNA
alpha	NN	O	I-DNA
enhancer	NN	O	I-DNA
with	NN	O	O
other	NN	O	O
recently	NN	O	O
identified	NN	O	O
T-cell-specific	NN	O	B-DNA
regulatory	NN	O	I-DNA
elements	NN	O	I-DNA
suggests	NN	O	O
that	NN	O	O
a	NN	O	O
common	NN	O	O
set	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
regulates	NN	O	O
several	NN	O	O
T-cell	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Molecular	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
IL-3	NN	O	I-DNA
gene	NN	O	I-DNA
:	NN	O	O
inducible	NN	O	O
T	NN	O	O
cell-restricted	NN	O	O
expression	NN	O	O
requires	NN	O	O
intact	NN	O	O
AP-1	NN	O	B-DNA
and	NN	O	I-DNA
Elf-1	NN	O	I-DNA
nuclear	NN	O	I-DNA
protein	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
.	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
a	NN	O	O
hematopoietic	NN	O	B-protein
stem-cell	NN	O	I-protein
growth	NN	O	I-protein
and	NN	O	I-protein
differentiation	NN	O	I-protein
factor	NN	O	I-protein
that	NN	O	O
is	NN	O	O
expressed	NN	O	O
solely	NN	O	O
in	NN	O	O
activated	NN	O	O
T	NN	O	B-cell_type
and	NN	O	I-cell_type
NK	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Studies	NN	O	O
to	NN	O	O
date	NN	O	O
have	NN	O	O
identified	NN	O	O
elements	NN	O	O
5	NN	O	O
'	NN	O	O
to	NN	O	O
the	NN	O	O
IL-3	NN	O	B-DNA
coding	NN	O	I-DNA
sequences	NN	O	I-DNA
that	NN	O	O
regulate	NN	O	O
its	NN	O	O
transcription	NN	O	O
,	NN	O	O
but	NN	O	O
the	NN	O	O
sequences	NN	O	O
that	NN	O	O
confer	NN	O	O
T	NN	O	O
cell-specific	NN	O	O
expression	NN	O	O
remain	NN	O	O
to	NN	O	O
be	NN	O	O
clearly	NN	O	O
defined	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
now	NN	O	O
identified	NN	O	O
DNA	NN	O	B-DNA
sequences	NN	O	I-DNA
that	NN	O	O
are	NN	O	O
required	NN	O	O
for	NN	O	O
T	NN	O	O
cell-restricted	NN	O	O
IL-3	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
.	NN	O	O

A	NN	O	O
series	NN	O	O
of	NN	O	O
transient	NN	O	O
transfections	NN	O	O
performed	NN	O	O
with	NN	O	O
human	NN	O	B-DNA
IL-3-chloramphenicol	NN	O	I-DNA
acetyltransferase	NN	O	I-DNA
(	NN	O	I-DNA
CAT	NN	O	I-DNA
)	NN	O	I-DNA
reporter	NN	O	I-DNA
plasmids	NN	O	I-DNA
in	NN	O	O
T	NN	O	B-cell_type
and	NN	O	I-cell_type
non-T	NN	O	I-cell_type
cells	NN	O	I-cell_type
revealed	NN	O	O
that	NN	O	O
a	NN	O	O
plasmid	NN	O	B-DNA
containing	NN	O	O
319	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
sequences	NN	O	I-DNA
was	NN	O	O
active	NN	O	O
exclusively	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Deletion	NN	O	O
analysis	NN	O	O
revealed	NN	O	O
that	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
specificity	NN	O	O
was	NN	O	O
conferred	NN	O	O
by	NN	O	O
a	NN	O	O
49-bp	NN	O	B-DNA
fragment	NN	O	I-DNA
(	NN	O	O
bp	NN	O	B-DNA
-319	NN	O	I-DNA
to	NN	O	I-DNA
-270	NN	O	I-DNA
)	NN	O	O
that	NN	O	O
included	NN	O	O
a	NN	O	O
potential	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
for	NN	O	O
AP-1	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
6	NN	O	O
bp	NN	O	O
upstream	NN	O	O
of	NN	O	O
a	NN	O	O
binding	NN	O	B-DNA
site	NN	O	I-DNA
for	NN	O	I-DNA
Elf-1	NN	O	I-DNA
,	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
Ets	NN	O	B-protein
family	NN	O	I-protein
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

DNaseI	NN	O	B-protein
footprint	NN	O	O
and	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
analyses	NN	O	O
performed	NN	O	O
with	NN	O	O
MLA-144	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
nuclear	NN	O	O
extracts	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
this	NN	O	O
49-bp	NN	O	B-DNA
region	NN	O	I-DNA
contains	NN	O	O
a	NN	O	O
nuclear	NN	O	B-DNA
protein	NN	O	I-DNA
binding	NN	O	I-DNA
region	NN	O	I-DNA
that	NN	O	O
includes	NN	O	O
consensus	NN	O	B-DNA
AP-1	NN	O	I-DNA
and	NN	O	I-DNA
Elf-1	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
extracts	NN	O	O
prepared	NN	O	O
from	NN	O	O
purified	NN	O	B-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
contained	NN	O	O
proteins	NN	O	O
that	NN	O	O
bound	NN	O	O
to	NN	O	O
synthetic	NN	O	O
oligonucleotides	NN	O	O
corresponding	NN	O	O
to	NN	O	O
the	NN	O	O
AP-1	NN	O	B-DNA
and	NN	O	I-DNA
Elf-1	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
vitro-transcribed	NN	O	O
and	NN	O	O
-translated	NN	O	O
Elf-1	NN	O	B-protein
protein	NN	O	I-protein
bound	NN	O	O
specifically	NN	O	O
to	NN	O	O
the	NN	O	O
Elf-1	NN	O	B-DNA
site	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
Elf-1	NN	O	O
antisera	NN	O	O
competed	NN	O	O
and	NN	O	O
super	NN	O	O
shifted	NN	O	O
nuclear	NN	O	O
protein	NN	O	O
complexes	NN	O	O
present	NN	O	O
in	NN	O	O
MLA-144	NN	O	B-cell_line
nuclear	NN	O	O
extracts	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
addition	NN	O	O
of	NN	O	O
anti-Jun	NN	O	O
family	NN	O	O
antiserum	NN	O	O
in	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
reactions	NN	O	O
completely	NN	O	O
blocked	NN	O	O
formation	NN	O	O
of	NN	O	O
the	NN	O	O
AP-1-related	NN	O	B-protein
complexes	NN	O	I-protein
.	NN	O	O

Transient	NN	O	O
transfection	NN	O	O
studies	NN	O	O
in	NN	O	O
MLA-144	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
revealed	NN	O	O
that	NN	O	O
constructs	NN	O	O
containing	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
AP-1	NN	O	B-DNA
site	NN	O	I-DNA
almost	NN	O	O
completely	NN	O	O
abolished	NN	O	O
CAT	NN	O	O
activity	NN	O	O
while	NN	O	O
mutation	NN	O	O
of	NN	O	O
the	NN	O	O
Elf-1	NN	O	B-DNA
site	NN	O	I-DNA
or	NN	O	O
the	NN	O	O
NF-IL-3	NN	O	B-DNA
site	NN	O	I-DNA
,	NN	O	O
a	NN	O	O
previously	NN	O	O
described	NN	O	O
nuclear	NN	O	B-DNA
protein	NN	O	I-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
(	NN	O	O
bp.	NN	O	B-DNA
-155	NN	O	I-DNA
to	NN	O	I-DNA
-148	NN	O	I-DNA
)	NN	O	O
in	NN	O	O
the	NN	O	O
IL-3	NN	O	B-DNA
promoter	NN	O	I-DNA
,	NN	O	O
reduced	NN	O	O
CAT	NN	O	O
activity	NN	O	O
to	NN	O	O
<	NN	O	O
25	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
activity	NN	O	O
given	NN	O	O
by	NN	O	O
wild-type	NN	O	O
constructs	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
IL-3	NN	O	I-DNA
gene	NN	O	I-DNA
requires	NN	O	O
the	NN	O	O
AP-1	NN	O	B-DNA
and	NN	O	I-DNA
Elf-1	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
;	NN	O	O
however	NN	O	O
,	NN	O	O
unlike	NN	O	O
other	NN	O	O
previously	NN	O	O
characterized	NN	O	O
cytokine	NN	O	B-DNA
genes	NN	O	I-DNA
such	NN	O	O
as	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
the	NN	O	O
AP-1	NN	O	B-protein
and	NN	O	I-protein
Elf-1	NN	O	I-protein
factors	NN	O	I-protein
can	NN	O	O
bind	NN	O	O
independently	NN	O	O
in	NN	O	O
the	NN	O	O
IL-3	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
400	NN	O	O
WORDS	NN	O	O
)	NN	O	O

-DOCSTART-	O

Combination	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
IL-4	NN	O	B-protein
reduces	NN	O	O
glucocorticoid	NN	O	O
receptor-binding	NN	O	O
affinity	NN	O	O
and	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
response	NN	O	O
to	NN	O	O
glucocorticoids	NN	O	O
.	NN	O	O

The	NN	O	O
mechanisms	NN	O	O
contributing	NN	O	O
to	NN	O	O
persistent	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
activation	NN	O	O
and	NN	O	O
poor	NN	O	O
response	NN	O	O
to	NN	O	O
glucocorticoids	NN	O	O
in	NN	O	O
chronic	NN	O	O
inflammatory	NN	O	O
illnesses	NN	O	O
such	NN	O	O
as	NN	O	O
steroid	NN	O	O
resistant	NN	O	O
(	NN	O	O
SR	NN	O	O
)	NN	O	O
asthma	NN	O	O
are	NN	O	O
poorly	NN	O	O
defined	NN	O	O
.	NN	O	O

We	NN	O	O
examined	NN	O	O
the	NN	O	O
possibility	NN	O	O
that	NN	O	O
certain	NN	O	O
cytokines	NN	O	O
,	NN	O	O
specifically	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
could	NN	O	O
affect	NN	O	O
T	NN	O	O
cell	NN	O	O
response	NN	O	O
to	NN	O	O
glucocorticoids	NN	O	O
.	NN	O	O

A	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
dexamethasone	NN	O	O
radioligand-binding	NN	O	O
assay	NN	O	O
was	NN	O	O
used	NN	O	O
to	NN	O	O
measure	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
(	NN	O	O
GR	NN	O	B-protein
)	NN	O	O
and	NN	O	O
dissociation	NN	O	O
constant	NN	O	O
(	NN	O	O
Kd	NN	O	O
)	NN	O	O
in	NN	O	O
PBMC	NN	O	B-cell_type
from	NN	O	O
normal	NN	O	O
donors	NN	O	O
and	NN	O	O
patients	NN	O	O
with	NN	O	O
SR	NN	O	O
asthma	NN	O	O
,	NN	O	O
cultured	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
these	NN	O	O
cytokines	NN	O	B-protein
.	NN	O	O

PBMC	NN	O	O
from	NN	O	O
normal	NN	O	O
donors	NN	O	O
incubated	NN	O	O
for	NN	O	O
48	NN	O	O
h	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
combination	NN	O	O
IL-2	NN	O	B-protein
+	NN	O	O
IL-4	NN	O	B-protein
had	NN	O	O
nuclear	NN	O	O
GR	NN	O	O
with	NN	O	O
significantly	NN	O	O
reduced	NN	O	O
binding	NN	O	O
affinity	NN	O	O
(	NN	O	O
GR	NN	O	O
Kd	NN	O	O
=	NN	O	O
36.1	NN	O	O
+/-	NN	O	O
1.63	NN	O	O
nM	NN	O	O
,	NN	O	O
mean	NN	O	O
+/-	NN	O	O
SEM	NN	O	O
;	NN	O	O
p	NN	O	O
=	NN	O	O
0.0001	NN	O	O
)	NN	O	O
as	NN	O	O
compared	NN	O	O
with	NN	O	O
PBMC	NN	O	B-cell_type
incubated	NN	O	O
with	NN	O	O
medium	NN	O	O
alone	NN	O	O
(	NN	O	O
GR	NN	O	O
Kd	NN	O	O
=	NN	O	O
6.74	NN	O	O
+/-	NN	O	O
0.46	NN	O	O
nM	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
cytosolic	NN	O	O
GR	NN	O	O
Kd	NN	O	O
remained	NN	O	O
unchanged	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
when	NN	O	O
PBMC	NN	O	B-cell_type
were	NN	O	O
incubated	NN	O	O
with	NN	O	O
IL-2	NN	O	B-protein
alone	NN	O	O
or	NN	O	O
IL-4	NN	O	B-protein
alone	NN	O	O
,	NN	O	O
no	NN	O	O
change	NN	O	O
in	NN	O	O
GR-binding	NN	O	O
affinity	NN	O	O
was	NN	O	O
observed	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
when	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
non-T	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
individually	NN	O	O
stimulated	NN	O	O
with	NN	O	O
combination	NN	O	O
IL-2	NN	O	O
+	NN	O	O
IL-4	NN	O	O
,	NN	O	O
a	NN	O	O
significant	NN	O	O
reduction	NN	O	O
in	NN	O	O
GR-binding	NN	O	O
affinity	NN	O	O
was	NN	O	O
observed	NN	O	O
only	NN	O	O
in	NN	O	O
the	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
population	NN	O	I-cell_type
(	NN	O	O
p	NN	O	O
=	NN	O	O
0.0001	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
IL-2	NN	O	O
+	NN	O	O
IL-4-induced	NN	O	O
alteration	NN	O	O
in	NN	O	O
PBMC	NN	O	O
GR	NN	O	O
Kd	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
GR	NN	O	O
number	NN	O	O
(	NN	O	O
8348	NN	O	O
+/-	NN	O	O
964	NN	O	O
vs	NN	O	O
1710	NN	O	O
+/-	NN	O	O
228	NN	O	O
sites/cell	NN	O	O
;	NN	O	O
p	NN	O	O
=	NN	O	O
0.0003	NN	O	O
)	NN	O	O
.	NN	O	O

More	NN	O	O
importantly	NN	O	O
,	NN	O	O
the	NN	O	O
alteration	NN	O	O
in	NN	O	O
PBMC	NN	O	O
GR-binding	NN	O	O
affinity	NN	O	O
with	NN	O	O
IL-2	NN	O	B-protein
+	NN	O	O
IL-4	NN	O	B-protein
was	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
functional	NN	O	O
change	NN	O	O
in	NN	O	O
T	NN	O	O
cell	NN	O	O
response	NN	O	O
to	NN	O	O
methylprednisolone	NN	O	O
MPN	NN	O	O
,	NN	O	O
i.e.	NN	O	O
,	NN	O	O
a	NN	O	O
reduced	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
of	NN	O	O
MPN	NN	O	O
on	NN	O	O
PMA/ionomycin-induced	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
proliferation	NN	O	O
.	NN	O	O

These	NN	O	O
effects	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
+	NN	O	O
IL-4	NN	O	B-protein
on	NN	O	O
PBMC	NN	O	B-cell_type
GR	NN	O	B-protein
affinity	NN	O	O
and	NN	O	O
response	NN	O	O
to	NN	O	O
MPN	NN	O	O
were	NN	O	O
blocked	NN	O	O
by	NN	O	O
co-incubation	NN	O	O
with	NN	O	O
IFN-gamma	NN	O	B-protein
.	NN	O	O

Freshly	NN	O	O
isolated	NN	O	O
PBMC	NN	O	B-cell_type
from	NN	O	O
four	NN	O	O
patients	NN	O	O
with	NN	O	O
SR	NN	O	O
asthma	NN	O	O
had	NN	O	O
a	NN	O	O
significantly	NN	O	O
reduced	NN	O	O
GR-binding	NN	O	O
affinity	NN	O	O
(	NN	O	O
Kd	NN	O	O
=	NN	O	O
40.0	NN	O	O
+/-	NN	O	O
2.68	NN	O	O
nM	NN	O	O
;	NN	O	O
p	NN	O	O
=	NN	O	O
0.0001	NN	O	O
)	NN	O	O
when	NN	O	O
compared	NN	O	O
with	NN	O	O
seven	NN	O	O
normal	NN	O	O
subjects	NN	O	O
(	NN	O	O
7.15	NN	O	O
+/-	NN	O	O
0.41	NN	O	O
nM	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
altered	NN	O	O
PBMC	NN	O	O
GR	NN	O	O
binding	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
SR	NN	O	O
asthma	NN	O	O
reversed	NN	O	O
to	NN	O	O
normal	NN	O	O
when	NN	O	O
incubated	NN	O	O
with	NN	O	O
medium	NN	O	O
alone	NN	O	O
,	NN	O	O
but	NN	O	O
was	NN	O	O
sustained	NN	O	O
with	NN	O	O
IL-2	NN	O	B-protein
+	NN	O	O
IL-4	NN	O	B-protein
.	NN	O	O

These	NN	O	O
observations	NN	O	O
suggest	NN	O	O
that	NN	O	O
with	NN	O	O
persistent	NN	O	O
inflammation	NN	O	O
certain	NN	O	O
cytokines	NN	O	B-protein
may	NN	O	O
contribute	NN	O	O
to	NN	O	O
an	NN	O	O
impaired	NN	O	O
response	NN	O	O
to	NN	O	O
glucocorticoids	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
IL-4	NN	O	B-protein
were	NN	O	O
blocked	NN	O	O
by	NN	O	O
IFN-gamma	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Characterization	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
CD4	NN	O	I-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
:	NN	O	O
transcription	NN	O	O
from	NN	O	O
the	NN	O	O
CD4	NN	O	B-DNA
gene	NN	O	I-DNA
core	NN	O	I-DNA
promoter	NN	O	I-DNA
is	NN	O	O
tissue-specific	NN	O	O
and	NN	O	O
is	NN	O	O
activated	NN	O	O
by	NN	O	O
Ets	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

We	NN	O	O
analyzed	NN	O	O
the	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
transcription	NN	O	I-DNA
control	NN	O	I-DNA
sequences	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
CD4	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
located	NN	O	O
the	NN	O	O
transcription	NN	O	B-DNA
initiation	NN	O	I-DNA
site	NN	O	I-DNA
and	NN	O	O
showed	NN	O	O
that	NN	O	O
the	NN	O	O
CD4	NN	O	B-DNA
core	NN	O	I-DNA
promoter	NN	O	I-DNA
(	NN	O	O
positions	NN	O	B-DNA
-40	NN	O	I-DNA
to	NN	O	I-DNA
+16	NN	O	I-DNA
)	NN	O	O
lacks	NN	O	O
a	NN	O	O
classical	NN	O	O
``	NN	O	O
TATA	NN	O	O
''	NN	O	O
or	NN	O	O
initiator	NN	O	B-DNA
positioning	NN	O	I-DNA
consensus	NN	O	I-DNA
sequence	NN	O	I-DNA
but	NN	O	O
directs	NN	O	O
precise	NN	O	O
and	NN	O	O
efficient	NN	O	O
transcription	NN	O	O
when	NN	O	O
coupled	NN	O	O
to	NN	O	O
the	NN	O	O
ubiquitously	NN	O	O
active	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
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
CD4	NN	O	B-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
correlated	NN	O	O
with	NN	O	O
CD4	NN	O	O
expression	NN	O	O
in	NN	O	O
various	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
the	NN	O	O
CD4	NN	O	B-DNA
core	NN	O	I-DNA
promoter	NN	O	I-DNA
also	NN	O	O
displayed	NN	O	O
a	NN	O	O
tissue-specific	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
.	NN	O	O

Within	NN	O	O
this	NN	O	O
fragment	NN	O	O
,	NN	O	O
three	NN	O	O
nucleic	NN	O	O
acid	NN	O	O
sequences	NN	O	O
are	NN	O	O
completely	NN	O	O
conserved	NN	O	O
in	NN	O	O
the	NN	O	O
murine	NN	O	B-DNA
CD4	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

One	NN	O	O
of	NN	O	O
these	NN	O	O
sequences	NN	O	O
contains	NN	O	O
a	NN	O	O
perfect	NN	O	B-DNA
ETS	NN	O	I-DNA
consensus	NN	O	I-DNA
sequence	NN	O	I-DNA
.	NN	O	O

Another	NN	O	O
ETS	NN	O	B-DNA
consensus	NN	O	I-DNA
sequence	NN	O	I-DNA
is	NN	O	O
located	NN	O	O
1060	NN	O	O
nt	NN	O	O
upstream	NN	O	O
.	NN	O	O

Electrophoretic-mobility-shift	NN	O	O
assays	NN	O	O
showed	NN	O	O
that	NN	O	O
the	NN	O	O
core	NN	O	O
promoter	NN	O	O
ETS	NN	O	O
motif	NN	O	O
binds	NN	O	O
an	NN	O	O
Ets-related	NN	O	B-protein
protein	NN	O	I-protein
specifically	NN	O	O
expressed	NN	O	O
at	NN	O	O
high	NN	O	O
levels	NN	O	O
in	NN	O	O
CD4+	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
in	NN	O	O
CD4-	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
overexpression	NN	O	O
of	NN	O	O
Ets-1	NN	O	B-protein
or	NN	O	O
Ets-2	NN	O	B-protein
efficiently	NN	O	O
and	NN	O	O
specifically	NN	O	O
activated	NN	O	O
transcription	NN	O	O
from	NN	O	O
the	NN	O	O
CD4	NN	O	B-DNA
promoter	NN	O	I-DNA
and	NN	O	O
core	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
Ets	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
play	NN	O	O
a	NN	O	O
central	NN	O	O
role	NN	O	O
in	NN	O	O
controlling	NN	O	O
CD4	NN	O	O
gene	NN	O	O
expression	NN	O	O
,	NN	O	O
by	NN	O	O
binding	NN	O	O
to	NN	O	O
both	NN	O	O
a	NN	O	O
classical	NN	O	B-DNA
remote	NN	O	I-DNA
site	NN	O	I-DNA
and	NN	O	O
an	NN	O	O
unusual	NN	O	O
proximal	NN	O	B-DNA
activator	NN	O	I-DNA
sequence	NN	O	I-DNA

-DOCSTART-	O

Glucocorticoid	NN	O	O
receptor	NN	O	O
activation	NN	O	O
and	NN	O	O
inactivation	NN	O	O
in	NN	O	O
c	NN	O	O
ultured	NN	O	B-cell_line
human	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
.	NN	O	O

Although	NN	O	O
glucocorticoids	NN	O	O
are	NN	O	O
not	NN	O	O
cytolytic	NN	O	O
for	NN	O	O
and	NN	O	O
do	NN	O	O
not	NN	O	O
inhibit	NN	O	O
the	NN	O	O
growth	NN	O	O
of	NN	O	O
the	NN	O	O
IM-9	NN	O	B-cell_line
line	NN	O	I-cell_line
of	NN	O	O
cultured	NN	O	B-cell_line
human	NN	O	I-cell_line
lymphoblasts	NN	O	I-cell_line
,	NN	O	O
these	NN	O	O
cells	NN	O	O
have	NN	O	O
a	NN	O	O
high	NN	O	O
steroid-binding	NN	O	O
capacity	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
used	NN	O	O
IM-9	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
order	NN	O	O
to	NN	O	O
examine	NN	O	O
whether	NN	O	O
unoccupied	NN	O	B-protein
glucocorticoid	NN	O	I-protein
receptors	NN	O	I-protein
are	NN	O	O
inactivated	NN	O	O
and	NN	O	O
activated	NN	O	O
in	NN	O	O
intact	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

when	NN	O	O
IM-9	NN	O	B-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
incubated	NN	O	O
in	NN	O	O
glucose-free	NN	O	O
medium	NN	O	O
in	NN	O	O
a	NN	O	O
nitrogen	NN	O	O
atmosphere	NN	O	O
,	NN	O	O
both	NN	O	O
their	NN	O	O
ability	NN	O	O
to	NN	O	O
bind	NN	O	O
triamcinolone	NN	O	O
acetonide	NN	O	O
and	NN	O	O
their	NN	O	O
ATP	NN	O	O
levels	NN	O	O
decline	NN	O	O
and	NN	O	O
,	NN	O	O
when	NN	O	O
glucose	NN	O	O
and	NN	O	O
oxygen	NN	O	O
are	NN	O	O
reintroduced	NN	O	O
,	NN	O	O
ATP	NN	O	O
levels	NN	O	O
and	NN	O	O
receptor	NN	O	O
activity	NN	O	O
return	NN	O	O
.	NN	O	O

The	NN	O	O
specific	NN	O	O
glucocorticoid-binding	NN	O	O
activity	NN	O	O
of	NN	O	O
cytosol	NN	O	O
prepared	NN	O	O
from	NN	O	O
cells	NN	O	O
exposed	NN	O	O
to	NN	O	O
various	NN	O	O
degrees	NN	O	O
of	NN	O	O
energy	NN	O	O
limitation	NN	O	O
is	NN	O	O
directly	NN	O	O
correlated	NN	O	O
with	NN	O	O
the	NN	O	O
ATP	NN	O	O
content	NN	O	O
.	NN	O	O

Receptor	NN	O	O
activation	NN	O	O
in	NN	O	O
intact	NN	O	B-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
rapid	NN	O	O
and	NN	O	O
independent	NN	O	O
of	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
.	NN	O	O

Cytosol	NN	O	O
prepared	NN	O	O
from	NN	O	O
inactivated	NN	O	O
cells	NN	O	O
can	NN	O	O
not	NN	O	O
be	NN	O	O
activated	NN	O	O
by	NN	O	O
addition	NN	O	O
of	NN	O	O
ATP	NN	O	O
.	NN	O	O

The	NN	O	O
inactivation	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
that	NN	O	O
occurs	NN	O	O
when	NN	O	O
cytosol	NN	O	O
from	NN	O	O
normal	NN	O	O
IM-9	NN	O	B-cell_line
cells	NN	O	I-cell_line
is	NN	O	O
incubated	NN	O	O
at	NN	O	O
25	NN	O	O
degrees	NN	O	O
C	NN	O	O
is	NN	O	O
inhibited	NN	O	O
by	NN	O	O
molybdate	NN	O	O
,	NN	O	O
vanadate	NN	O	O
,	NN	O	O
fluoride	NN	O	O
,	NN	O	O
ATP	NN	O	O
,	NN	O	O
and	NN	O	O
several	NN	O	O
other	NN	O	O
nucleotides	NN	O	O
.	NN	O	O

The	NN	O	O
experiments	NN	O	O
with	NN	O	O
intact	NN	O	B-cell_type
human	NN	O	I-cell_type
lymphoblasts	NN	O	I-cell_type
suggest	NN	O	O
that	NN	O	O
assays	NN	O	O
of	NN	O	O
specific	NN	O	O
glucocorticoid-binding	NN	O	O
capacity	NN	O	O
do	NN	O	O
not	NN	O	O
necessarily	NN	O	O
reflect	NN	O	O
the	NN	O	O
cellular	NN	O	O
content	NN	O	O
of	NN	O	O
receptor	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Glucocorticoids	NN	O	O
and	NN	O	O
lymphocytes	NN	O	B-cell_type
.	NN	O	O

II	NN	O	O
.	NN	O	O

Cell	NN	O	O
cycle-dependent	NN	O	O
changes	NN	O	O
in	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
content	NN	O	O
.	NN	O	O

To	NN	O	O
study	NN	O	O
variations	NN	O	O
in	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
levels	NN	O	O
during	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
separated	NN	O	O
mitogen-stimulated	NN	O	B-cell_line
human	NN	O	I-cell_line
peripheral	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
and	NN	O	O
rat	NN	O	B-cell_type
lymph	NN	O	I-cell_type
node	NN	O	I-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
unit	NN	O	O
gravity	NN	O	O
sedimentation	NN	O	O
and	NN	O	O
measured	NN	O	O
glucocorticoid	NN	O	O
binding	NN	O	O
in	NN	O	O
the	NN	O	O
resultant	NN	O	O
fractions	NN	O	O
.	NN	O	O

By	NN	O	O
morphologic	NN	O	O
criteria	NN	O	O
and	NN	O	O
thymidine	NN	O	O
incorporation	NN	O	O
,	NN	O	O
the	NN	O	O
fractions	NN	O	O
were	NN	O	O
separated	NN	O	O
into	NN	O	O
populations	NN	O	O
of	NN	O	O
G0	NN	O	B-cell_type
and	NN	O	I-cell_type
G1	NN	O	I-cell_type
phase	NN	O	I-cell_type
and	NN	O	I-cell_type
S	NN	O	I-cell_type
and	NN	O	I-cell_type
post-S	NN	O	I-cell_type
phase	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

A	NN	O	O
2-	NN	O	O
to	NN	O	O
3-fold	NN	O	O
increase	NN	O	O
in	NN	O	O
glucocorticoid	NN	O	O
receptor	NN	O	O
sites	NN	O	O
per	NN	O	O
cell	NN	O	O
,	NN	O	O
for	NN	O	O
cells	NN	O	O
in	NN	O	O
the	NN	O	O
S	NN	O	O
and	NN	O	O
post-S	NN	O	O
phase	NN	O	O
over	NN	O	O
those	NN	O	O
in	NN	O	O
G0	NN	O	O
and	NN	O	O
G1	NN	O	O
,	NN	O	O
was	NN	O	O
observed	NN	O	O
with	NN	O	O
both	NN	O	O
nonstimulated	NN	O	B-cell_type
rat	NN	O	I-cell_type
lymph	NN	O	I-cell_type
node	NN	O	I-cell_type
cell	NN	O	I-cell_type
suspensions	NN	O	I-cell_type
and	NN	O	O
concanavalin	NN	O	B-cell_line
A-stimulated	NN	O	I-cell_line
human	NN	O	I-cell_line
peripheral	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
.	NN	O	O

These	NN	O	O
observations	NN	O	O
together	NN	O	O
with	NN	O	O
those	NN	O	O
from	NN	O	O
other	NN	O	O
studies	NN	O	O
indicate	NN	O	O
that	NN	O	O
formation	NN	O	O
of	NN	O	O
new	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
near	NN	O	O
the	NN	O	O
S	NN	O	O
phase	NN	O	O
may	NN	O	O
be	NN	O	O
a	NN	O	O
general	NN	O	O
phenomenon	NN	O	O
in	NN	O	O
proliferating	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
propose	NN	O	O
that	NN	O	O
this	NN	O	O
increase	NN	O	O
in	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
during	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
may	NN	O	O
explain	NN	O	O
the	NN	O	O
increase	NN	O	O
in	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
mitogen-stimulated	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
leukocyte	NN	O	B-cell_type
migration	NN	O	O
inhibition	NN	O	O
response	NN	O	O
to	NN	O	O
certain	NN	O	O
breast	NN	O	B-protein
cancer-related	NN	O	I-protein
antigens	NN	O	I-protein
(	NN	O	O
MCF-7	NN	O	B-protein
and	NN	O	O
MuMTV	NN	O	B-protein
)	NN	O	O
:	NN	O	O
their	NN	O	O
potential	NN	O	O
as	NN	O	O
discriminants	NN	O	O
.	NN	O	O

Certain	NN	O	O
oncogenic	NN	O	O
viruses	NN	O	O
have	NN	O	O
been	NN	O	O
implicated	NN	O	O
in	NN	O	O
human	NN	O	O
breast	NN	O	O
cancer	NN	O	O
,	NN	O	O
including	NN	O	O
the	NN	O	O
murine	NN	O	O
mammary	NN	O	O
tumor	NN	O	O
virus	NN	O	O
(	NN	O	O
MuMTV	NN	O	O
)	NN	O	O
and	NN	O	O
the	NN	O	O
Mason-Pfizer	NN	O	O
monkey	NN	O	O
virus	NN	O	O
(	NN	O	O
MPMV	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
used	NN	O	O
the	NN	O	O
leukocyte	NN	O	O
migration	NN	O	O
inhibition	NN	O	O
(	NN	O	O
LMI	NN	O	O
)	NN	O	O
response	NN	O	O
to	NN	O	O
assay	NN	O	O
the	NN	O	O
response	NN	O	O
to	NN	O	O
several	NN	O	O
potential	NN	O	O
breast	NN	O	O
cancer-related	NN	O	O
antigens	NN	O	O
,	NN	O	O
including	NN	O	O
MuMTV	NN	O	B-protein
,	NN	O	O
MPMV	NN	O	B-protein
,	NN	O	O
and	NN	O	O
a	NN	O	O
breast	NN	O	B-cell_line
cancer	NN	O	I-cell_line
cultured	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
MCF-7	NN	O	B-cell_line
,	NN	O	O
in	NN	O	O
96	NN	O	O
breast	NN	O	O
cancer	NN	O	O
patients	NN	O	O
,	NN	O	O
in	NN	O	O
32	NN	O	O
women	NN	O	O
with	NN	O	O
benign	NN	O	O
breast	NN	O	O
disease	NN	O	O
,	NN	O	O
and	NN	O	O
in	NN	O	O
67	NN	O	O
normal	NN	O	O
women	NN	O	O
.	NN	O	O

The	NN	O	O
lowest	NN	O	O
tenth	NN	O	O
percentile	NN	O	O
of	NN	O	O
control	NN	O	O
(	NN	O	O
LMI	NN	O	O
)	NN	O	O
responses	NN	O	O
was	NN	O	O
used	NN	O	O
as	NN	O	O
the	NN	O	O
cutoff	NN	O	O
point	NN	O	O
to	NN	O	O
designate	NN	O	O
responders	NN	O	O
.	NN	O	O

Breast	NN	O	O
cancer	NN	O	O
patients	NN	O	O
showed	NN	O	O
significant	NN	O	O
responses	NN	O	O
to	NN	O	O
MuMTV	NN	O	B-protein
(	NN	O	O
49	NN	O	O
%	NN	O	O
and	NN	O	O
to	NN	O	O
MCF-7	NN	O	O
(	NN	O	O
50	NN	O	O
%	NN	O	O
)	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
to	NN	O	O
MPMV	NN	O	B-protein
(	NN	O	O
29	NN	O	O
%	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
a	NN	O	O
paired-antigen	NN	O	O
study	NN	O	O
using	NN	O	O
MuMTV	NN	O	B-protein
and	NN	O	O
MCF-7	NN	O	B-protein
,	NN	O	O
75	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
breast	NN	O	O
cancer	NN	O	O
patients	NN	O	O
responded	NN	O	O
,	NN	O	O
versus	NN	O	O
18	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
normal	NN	O	O
women	NN	O	O
(	NN	O	O
P	NN	O	O
less	NN	O	O
than	NN	O	O
0.0050	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
potential	NN	O	O
for	NN	O	O
this	NN	O	O
assay	NN	O	O
to	NN	O	O
distinguish	NN	O	O
``	NN	O	O
normal	NN	O	O
''	NN	O	O
from	NN	O	O
``	NN	O	O
breast	NN	O	O
cancer	NN	O	O
''	NN	O	O
was	NN	O	O
analyzed	NN	O	O
using	NN	O	O
a	NN	O	O
migration	NN	O	O
index	NN	O	O
derived	NN	O	O
from	NN	O	O
discriminant	NN	O	O
analysis	NN	O	O
.	NN	O	O

The	NN	O	O
ability	NN	O	O
of	NN	O	O
the	NN	O	O
assay	NN	O	O
to	NN	O	O
discriminate	NN	O	O
``	NN	O	O
normal	NN	O	O
''	NN	O	O
from	NN	O	O
``	NN	O	O
cancer	NN	O	O
''	NN	O	O
was	NN	O	O
significant	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
showed	NN	O	O
a	NN	O	O
sensitivity	NN	O	O
of	NN	O	O
detecting	NN	O	O
``	NN	O	O
cancer	NN	O	O
''	NN	O	O
of	NN	O	O
75	NN	O	O
%	NN	O	O
.	NN	O	O

The	NN	O	O
overall	NN	O	O
responses	NN	O	O
to	NN	O	O
MuMTV	NN	O	B-protein
and	NN	O	O
MCF-7	NN	O	B-protein
were	NN	O	O
analyzed	NN	O	O
with	NN	O	O
reference	NN	O	O
to	NN	O	O
certain	NN	O	O
prognostic	NN	O	O
factors	NN	O	O
,	NN	O	O
but	NN	O	O
showed	NN	O	O
no	NN	O	O
relation	NN	O	O
to	NN	O	O
age	NN	O	O
,	NN	O	O
menstrual	NN	O	O
status	NN	O	O
,	NN	O	O
estrogen	NN	O	O
receptor	NN	O	O
status	NN	O	O
,	NN	O	O
or	NN	O	O
stage	NN	O	O
of	NN	O	O
disease	NN	O	O
.	NN	O	O

The	NN	O	O
above	NN	O	O
reactions	NN	O	O
suggest	NN	O	O
that	NN	O	O
a	NN	O	O
large	NN	O	O
proportion	NN	O	O
of	NN	O	O
breast	NN	O	O
cancer	NN	O	O
patients	NN	O	O
exhibit	NN	O	O
presensitization	NN	O	O
to	NN	O	O
antigenfs	NN	O	O
found	NN	O	O
in	NN	O	O
MuMTV	NN	O	B-protein
and	NN	O	O
MCF-7	NN	O	B-protein
,	NN	O	O
which	NN	O	O
may	NN	O	O
be	NN	O	O
cross-reactive	NN	O	O
with	NN	O	O
antigens	NN	O	O
in	NN	O	O
the	NN	O	O
primary	NN	O	O
cancer	NN	O	O
.	NN	O	O

These	NN	O	O
responses	NN	O	O
appear	NN	O	O
to	NN	O	O
be	NN	O	O
independent	NN	O	O
of	NN	O	O
major	NN	O	O
prognostic	NN	O	O
variables	NN	O	O
.	NN	O	O

Further	NN	O	O
refinement	NN	O	O
of	NN	O	O
this	NN	O	O
assay	NN	O	O
may	NN	O	O
yield	NN	O	O
one	NN	O	O
which	NN	O	O
is	NN	O	O
more	NN	O	O
highly	NN	O	O
discriminating	NN	O	O
for	NN	O	O
breast	NN	O	O
cancer	NN	O	O
.	NN	O	O

-DOCSTART-	O

Chronic	NN	O	O
lymphatic	NN	O	O
leukaemia	NN	O	O
:	NN	O	O
cellular	NN	O	O
effects	NN	O	O
of	NN	O	O
glucocorticoids	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

Glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
levels	NN	O	O
and	NN	O	O
steroid	NN	O	O
induced	NN	O	O
inhibition	NN	O	O
of	NN	O	O
nucleic	NN	O	O
acid	NN	O	O
precursors	NN	O	O
have	NN	O	O
been	NN	O	O
examined	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
from	NN	O	O
27	NN	O	O
patients	NN	O	O
at	NN	O	O
different	NN	O	O
stages	NN	O	O
of	NN	O	O
chronic	NN	O	O
lymphatic	NN	O	O
leukaemia	NN	O	O
.	NN	O	O

No	NN	O	O
correlation	NN	O	O
can	NN	O	O
be	NN	O	O
found	NN	O	O
between	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
and	NN	O	O
the	NN	O	O
stage	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
.	NN	O	O

On	NN	O	O
the	NN	O	O
other	NN	O	O
hand	NN	O	O
,	NN	O	O
a	NN	O	O
significant	NN	O	O
difference	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
was	NN	O	O
found	NN	O	O
between	NN	O	O
stage	NN	O	O
O	NN	O	O
and	NN	O	O
stage	NN	O	O
III/IV	NN	O	O
patients	NN	O	O
,	NN	O	O
in	NN	O	O
terms	NN	O	O
of	NN	O	O
the	NN	O	O
in	NN	O	O
vitro	NN	O	O
effect	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
on	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
uridine	NN	O	O
incorporation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Pyrrolidine	NN	O	O
dithiocarbamate	NN	O	O
inhibits	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
mobilization	NN	O	O
and	NN	O	O
TNF	NN	O	O
production	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
human	NN	O	B-DNA
TNF	NN	O	I-DNA
promoter	NN	O	I-DNA
contains	NN	O	O
four	NN	O	O
potential	NN	O	O
nuclear	NN	O	B-DNA
factor-kappa	NN	O	I-DNA
B	NN	O	I-DNA
(	NN	O	I-DNA
NF-kappa	NN	O	I-DNA
B	NN	O	I-DNA
)	NN	O	I-DNA
-binding	NN	O	I-DNA
sites	NN	O	I-DNA
,	NN	O	O
with	NN	O	O
the	NN	O	O
strongest	NN	O	O
binding	NN	O	O
seen	NN	O	O
for	NN	O	O
the	NN	O	O
-605	NN	O	O
motif	NN	O	O
.	NN	O	O

Nuclear	NN	O	O
extracts	NN	O	O
from	NN	O	O
unstimulated	NN	O	B-cell_line
cells	NN	O	I-cell_line
of	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
monocytic	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
Mono	NN	O	B-cell_line
Mac	NN	O	I-cell_line
6	NN	O	I-cell_line
,	NN	O	O
contain	NN	O	O
one	NN	O	O
specific	NN	O	B-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
complex	NN	O	B-protein
II	NN	O	I-protein
)	NN	O	O
,	NN	O	O
consistent	NN	O	O
with	NN	O	O
a	NN	O	O
constitutive	NN	O	B-protein
p50	NN	O	I-protein
homodimer	NN	O	I-protein
.	NN	O	O

Stimulation	NN	O	O
of	NN	O	O
Mono	NN	O	B-cell_line
Mac	NN	O	I-cell_line
6	NN	O	I-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
LPS	NN	O	O
will	NN	O	O
increase	NN	O	O
complex	NN	O	B-protein
II	NN	O	I-protein
and	NN	O	O
will	NN	O	O
strongly	NN	O	O
induce	NN	O	O
a	NN	O	O
second	NN	O	O
specific	NN	O	O
complex	NN	O	O
(	NN	O	O
complex	NN	O	B-protein
I	NN	O	I-protein
)	NN	O	O
,	NN	O	O
which	NN	O	O
represents	NN	O	O
the	NN	O	O
p50/65	NN	O	B-protein
heterodimer	NN	O	I-protein
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
Mono	NN	O	B-cell_type
Mac	NN	O	I-cell_type
6	NN	O	I-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
pyrrolidine-dithiocarbamate	NN	O	O
(	NN	O	O
PDTC	NN	O	O
)	NN	O	O
at	NN	O	O
300	NN	O	O
microM	NN	O	O
will	NN	O	O
block	NN	O	O
the	NN	O	O
LPS-induced	NN	O	B-protein
complex	NN	O	I-protein
I	NN	O	I-protein
almost	NN	O	O
completely	NN	O	O
and	NN	O	O
will	NN	O	O
reduce	NN	O	O
complex	NN	O	B-protein
II	NN	O	I-protein
to	NN	O	O
the	NN	O	O
constitutive	NN	O	O
level	NN	O	O
.	NN	O	O

Binding	NN	O	O
activity	NN	O	O
of	NN	O	O
other	NN	O	O
nuclear	NN	O	B-protein
factors	NN	O	I-protein
that	NN	O	O
recognize	NN	O	O
the	NN	O	O
SP-1	NN	O	O
and	NN	O	O
c/EBP	NN	O	O
motifs	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
TNF	NN	O	I-DNA
promoter	NN	O	I-DNA
is	NN	O	O
not	NN	O	O
affected	NN	O	O
by	NN	O	O
such	NN	O	O
treatment	NN	O	O
.	NN	O	O

Northern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
demonstrates	NN	O	O
that	NN	O	O
PDTC	NN	O	O
treatment	NN	O	O
will	NN	O	O
strongly	NN	O	O
reduce	NN	O	O
LPS-induced	NN	O	B-protein
TNF	NN	O	I-protein
transcripts	NN	O	I-protein
.	NN	O	O

Secreted	NN	O	O
TNF	NN	O	B-protein
protein	NN	O	I-protein
as	NN	O	O
detected	NN	O	O
in	NN	O	O
the	NN	O	O
Wehi	NN	O	O
164S/ActD	NN	O	O
bioassay	NN	O	O
and	NN	O	O
in	NN	O	O
a	NN	O	O
sandwich	NN	O	O
immunoassay	NN	O	O
was	NN	O	O
similarly	NN	O	O
reduced	NN	O	O
by	NN	O	O
PDTC	NN	O	O
.	NN	O	O

Kinetic	NN	O	O
analyses	NN	O	O
show	NN	O	O
that	NN	O	O
after	NN	O	O
LPS	NN	O	O
stimulation	NN	O	O
,	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
will	NN	O	O
peak	NN	O	O
at	NN	O	O
1	NN	O	O
h	NN	O	O
,	NN	O	O
TNF	NN	O	O
transcript	NN	O	O
prevalence	NN	O	O
at	NN	O	O
2	NN	O	O
h	NN	O	O
,	NN	O	O
and	NN	O	O
TNF	NN	O	B-protein
protein	NN	O	I-protein
at	NN	O	O
4	NN	O	O
h	NN	O	O
.	NN	O	O

PDTC	NN	O	O
did	NN	O	O
not	NN	O	O
shift	NN	O	O
this	NN	O	O
response	NN	O	O
to	NN	O	O
LPS	NN	O	O
to	NN	O	O
a	NN	O	O
later	NN	O	O
time	NN	O	O
,	NN	O	O
but	NN	O	O
suppressed	NN	O	O
NF-kappa	NN	O	O
B	NN	O	O
mobilization	NN	O	O
,	NN	O	O
TNF	NN	O	B-protein
transcripts	NN	O	I-protein
,	NN	O	O
and	NN	O	O
TNF	NN	O	B-protein
protein	NN	O	I-protein
over	NN	O	O
the	NN	O	O
entire	NN	O	O
8-h	NN	O	O
observation	NN	O	O
period	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
freshly	NN	O	O
isolated	NN	O	O
,	NN	O	O
LPS-stimulated	NN	O	B-cell_type
blood	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
showed	NN	O	O
a	NN	O	O
similar	NN	O	O
blockade	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
in	NN	O	O
these	NN	O	O
primary	NN	O	O
cells	NN	O	O
,	NN	O	O
induction	NN	O	O
of	NN	O	O
TNF	NN	O	B-protein
transcripts	NN	O	I-protein
,	NN	O	O
as	NN	O	O
determined	NN	O	O
by	NN	O	O
Northern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
and	NN	O	O
by	NN	O	O
quantitative	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
,	NN	O	O
was	NN	O	O
prevented	NN	O	O
by	NN	O	O
PDTC	NN	O	O
as	NN	O	O
was	NN	O	O
TNF	NN	O	B-protein
protein	NN	O	I-protein
production	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
show	NN	O	O
that	NN	O	O
dithiocarbamates	NN	O	O
can	NN	O	O
profoundly	NN	O	O
affect	NN	O	O
cytokine	NN	O	O
expression	NN	O	O
and	NN	O	O
suggest	NN	O	O
that	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
is	NN	O	O
involved	NN	O	O
in	NN	O	O
LPS-induced	NN	O	O
TNF	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Comparing	NN	O	O
regions	NN	O	O
of	NN	O	O
the	NN	O	O
Epstein-Barr	NN	O	B-protein
virus	NN	O	I-protein
ZEBRA	NN	O	I-protein
protein	NN	O	I-protein
which	NN	O	O
function	NN	O	O
as	NN	O	O
transcriptional	NN	O	B-DNA
activating	NN	O	I-DNA
sequences	NN	O	I-DNA
in	NN	O	O
Saccharomyces	NN	O	O
cerevisiae	NN	O	O
and	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
ZEBRA	NN	O	B-protein
protein	NN	O	I-protein
activates	NN	O	O
expression	NN	O	O
of	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
early-lytic-cycle	NN	O	B-DNA
genes	NN	O	I-DNA
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

Here	NN	O	O
it	NN	O	O
is	NN	O	O
shown	NN	O	O
that	NN	O	O
ZEBRA	NN	O	B-protein
also	NN	O	O
behaves	NN	O	O
as	NN	O	O
a	NN	O	O
sequence-specific	NN	O	B-protein
transcriptional	NN	O	I-protein
activator	NN	O	I-protein
in	NN	O	O
Saccharomyces	NN	O	O
cerevisiae	NN	O	O
.	NN	O	O

Deletional	NN	O	O
mutagenesis	NN	O	O
defined	NN	O	O
three	NN	O	O
regions	NN	O	O
of	NN	O	O
ZEBRA	NN	O	B-protein
that	NN	O	O
participate	NN	O	O
in	NN	O	O
activation	NN	O	O
in	NN	O	O
S.	NN	O	O
cerevisiae	NN	O	O
.	NN	O	O

These	NN	O	O
regions	NN	O	O
are	NN	O	O
designated	NN	O	O
YI	NN	O	B-protein
(	NN	O	O
amino	NN	O	B-protein
acids	NN	O	I-protein
[	NN	O	I-protein
aa	NN	O	I-protein
]	NN	O	I-protein
1	NN	O	I-protein
to	NN	O	I-protein
25	NN	O	I-protein
)	NN	O	O
,	NN	O	O
YII	NN	O	B-protein
(	NN	O	O
aa	NN	O	B-protein
51	NN	O	I-protein
to	NN	O	I-protein
102	NN	O	I-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
YIII	NN	O	B-protein
(	NN	O	O
aa	NN	O	B-protein
228	NN	O	I-protein
to	NN	O	I-protein
245	NN	O	I-protein
)	NN	O	O
.	NN	O	O

Two	NN	O	O
of	NN	O	O
the	NN	O	O
three	NN	O	O
regions	NN	O	O
of	NN	O	O
the	NN	O	O
native	NN	O	O
ZEBRA	NN	O	B-protein
protein	NN	O	I-protein
act	NN	O	O
together	NN	O	O
to	NN	O	O
mediate	NN	O	O
activation	NN	O	O
when	NN	O	O
assayed	NN	O	O
on	NN	O	O
ZEBRA	NN	O	B-protein
binding	NN	O	I-protein
sites	NN	O	I-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
when	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
GAL	NN	O	B-protein
4	NN	O	O
and	NN	O	O
assayed	NN	O	O
on	NN	O	O
GAL4	NN	O	B-protein
binding	NN	O	I-protein
sites	NN	O	I-protein
,	NN	O	O
regions	NN	O	B-protein
YII	NN	O	I-protein
and	NN	O	I-protein
YIII	NN	O	I-protein
were	NN	O	O
each	NN	O	O
sufficient	NN	O	O
to	NN	O	O
confer	NN	O	O
activation	NN	O	O
in	NN	O	O
S.	NN	O	O
cerevisiae	NN	O	O
.	NN	O	O

Regions	NN	O	O
of	NN	O	O
ZEBRA	NN	O	B-protein
which	NN	O	O
affected	NN	O	O
activation	NN	O	O
in	NN	O	O
S.	NN	O	O
cerevisiae	NN	O	O
were	NN	O	O
also	NN	O	O
required	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
amino-terminal	NN	O	B-protein
region	NN	O	I-protein
of	NN	O	O
ZEBRA	NN	O	B-protein
(	NN	O	O
aa	NN	O	B-protein
1	NN	O	I-protein
to	NN	O	I-protein
98	NN	O	I-protein
)	NN	O	O
was	NN	O	O
required	NN	O	O
for	NN	O	O
activation	NN	O	O
both	NN	O	O
in	NN	O	O
S.	NN	O	O
cerevisiae	NN	O	O
and	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
;	NN	O	O
deletion	NN	O	O
of	NN	O	O
the	NN	O	O
carboxy-terminal	NN	O	B-protein
18	NN	O	I-protein
aa	NN	O	I-protein
also	NN	O	O
significantly	NN	O	O
reduced	NN	O	O
activation	NN	O	O
in	NN	O	O
both	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
behavior	NN	O	O
of	NN	O	O
ZEBRA	NN	O	B-protein
in	NN	O	O
human	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
S.	NN	O	O
cerevisiae	NN	O	O
suggests	NN	O	O
that	NN	O	O
the	NN	O	O
protein	NN	O	O
contains	NN	O	O
universal	NN	O	B-protein
activation	NN	O	I-protein
motifs	NN	O	I-protein
that	NN	O	O
interact	NN	O	O
with	NN	O	O
conserved	NN	O	O
components	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	O
machinery	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
certain	NN	O	O
deletion	NN	O	B-protein
mutants	NN	O	I-protein
of	NN	O	O
ZEBRA	NN	O	B-protein
containing	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
N-terminal	NN	O	B-protein
region	NN	O	I-protein
exhibited	NN	O	O
discordant	NN	O	O
behaviors	NN	O	O
in	NN	O	O
S.	NN	O	O
cerevisiae	NN	O	O
and	NN	O	O
in	NN	O	O
B	NN	O	O
cells	NN	O	O
.	NN	O	O

For	NN	O	O
example	NN	O	O
,	NN	O	O
deletion	NN	O	O
of	NN	O	O
ZEBRA	NN	O	B-protein
aa	NN	O	I-protein
26	NN	O	I-protein
to	NN	O	I-protein
51	NN	O	I-protein
impaired	NN	O	O
activation	NN	O	O
to	NN	O	O
a	NN	O	O
great	NN	O	O
extent	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
but	NN	O	O
had	NN	O	O
little	NN	O	O
or	NN	O	O
no	NN	O	O
effect	NN	O	O
in	NN	O	O
S.	NN	O	O
cerevisiae	NN	O	O
.	NN	O	O

The	NN	O	O
discordant	NN	O	O
mutants	NN	O	O
may	NN	O	O
reflect	NN	O	O
interactions	NN	O	O
with	NN	O	O
a	NN	O	O
variable	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
a	NN	O	O
conserved	NN	O	O
component	NN	O	O
or	NN	O	O
unique	NN	O	O
interactions	NN	O	O
with	NN	O	O
specialized	NN	O	O
components	NN	O	O
of	NN	O	O
the	NN	O	O
basal	NN	O	O
transcription	NN	O	O
apparatus	NN	O	O
in	NN	O	O
different	NN	O	O
cells	NN	O	O
.	NN	O	O

-DOCSTART-	O

Functional	NN	O	O
interaction	NN	O	O
of	NN	O	O
the	NN	O	O
v-Rel	NN	O	B-protein
and	NN	O	I-protein
c-Rel	NN	O	I-protein
oncoproteins	NN	O	I-protein
with	NN	O	O
the	NN	O	O
TATA-binding	NN	O	B-protein
protein	NN	O	I-protein
and	NN	O	O
association	NN	O	O
with	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
IIB	NN	O	I-protein
.	NN	O	O

Rel	NN	O	B-protein
family	NN	O	I-protein
proteins	NN	O	I-protein
regulate	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
genes	NN	O	O
linked	NN	O	O
to	NN	O	O
kappa	NN	O	O
B-binding	NN	O	O
motifs	NN	O	O
.	NN	O	O

Little	NN	O	O
is	NN	O	O
known	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
of	NN	O	O
the	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
they	NN	O	O
enhance	NN	O	O
transcription	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
investigated	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
the	NN	O	O
v-Rel	NN	O	B-protein
and	NN	O	I-protein
c-Rel	NN	O	I-protein
oncoproteins	NN	O	I-protein
to	NN	O	O
interact	NN	O	O
with	NN	O	O
components	NN	O	O
of	NN	O	O
the	NN	O	O
basal	NN	O	O
transcription	NN	O	O
machinery	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
both	NN	O	O
the	NN	O	O
acidic	NN	O	O
transcription	NN	O	O
activation	NN	O	O
domain	NN	O	O
mapping	NN	O	O
to	NN	O	O
the	NN	O	O
unique	NN	O	O
C	NN	O	B-protein
terminus	NN	O	I-protein
of	NN	O	O
chicken	NN	O	B-protein
c-Rel	NN	O	I-protein
and	NN	O	O
the	NN	O	O
F9	NN	O	B-protein
cell-specific	NN	O	I-protein
activation	NN	O	I-protein
region	NN	O	I-protein
common	NN	O	O
to	NN	O	O
both	NN	O	O
v-Rel	NN	O	B-protein
and	NN	O	O
c-Rel	NN	O	B-protein
interact	NN	O	O
with	NN	O	O
the	NN	O	O
TATA-binding	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	O
TBP	NN	O	B-protein
)	NN	O	O
and	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
IIB	NN	O	I-protein
(	NN	O	O
TFIIB	NN	O	B-protein
)	NN	O	O
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

We	NN	O	O
also	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
TPB	NN	O	B-protein
interaction	NN	O	O
with	NN	O	O
Rel	NN	O	B-protein
activation	NN	O	I-protein
regions	NN	O	I-protein
leads	NN	O	O
to	NN	O	O
synergistic	NN	O	O
activation	NN	O	O
of	NN	O	O
transcription	NN	O	O
of	NN	O	O
a	NN	O	O
kappa	NN	O	B-DNA
B-linked	NN	O	I-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

Combined	NN	O	O
with	NN	O	O
the	NN	O	O
observation	NN	O	O
that	NN	O	O
the	NN	O	O
mouse	NN	O	B-protein
c-Rel	NN	O	I-protein
and	NN	O	I-protein
human	NN	O	I-protein
RelA	NN	O	I-protein
proteins	NN	O	I-protein
also	NN	O	O
interact	NN	O	O
with	NN	O	O
TBP	NN	O	B-protein
and	NN	O	O
TFIIB	NN	O	B-protein
in	NN	O	O
vitro	NN	O	O
,	NN	O	O
these	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
association	NN	O	O
with	NN	O	O
basal	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
is	NN	O	O
important	NN	O	O
for	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
activities	NN	O	O
of	NN	O	O
Rel	NN	O	B-protein
family	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

p21ras	NN	O	B-protein
and	NN	O	O
calcineurin	NN	O	O
synergize	NN	O	O
to	NN	O	O
regulate	NN	O	O
the	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

In	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
triggering	NN	O	O
of	NN	O	O
the	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
TCR	NN	O	B-protein
)	NN	O	O
induces	NN	O	O
several	NN	O	O
signaling	NN	O	O
cascades	NN	O	O
which	NN	O	O
ultimately	NN	O	O
synergize	NN	O	O
to	NN	O	O
induce	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
the	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-cell_line
)	NN	O	O
,	NN	O	O
a	NN	O	O
DNA	NN	O	B-protein
binding	NN	O	I-protein
complex	NN	O	I-protein
critical	NN	O	O
to	NN	O	O
the	NN	O	O
inducibility	NN	O	O
and	NN	O	O
T	NN	O	O
cell	NN	O	O
specificity	NN	O	O
of	NN	O	O
the	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
growth	NN	O	I-protein
factor	NN	O	I-protein
interleukin	NN	O	B-protein
2	NN	O	I-protein
.	NN	O	O

One	NN	O	O
immediate	NN	O	O
consequence	NN	O	O
of	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
via	NN	O	O
the	NN	O	O
TCR	NN	O	B-protein
is	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
cytosolic	NN	O	O
calcium	NN	O	O
.	NN	O	O

Calcium	NN	O	O
signals	NN	O	O
are	NN	O	O
important	NN	O	O
for	NN	O	O
NFAT	NN	O	B-cell_line
induction	NN	O	O
,	NN	O	O
and	NN	O	O
recent	NN	O	O
studies	NN	O	O
have	NN	O	O
identified	NN	O	O
calcineurin	NN	O	B-protein
,	NN	O	O
a	NN	O	O
calcium-calmodulin	NN	O	B-protein
dependent	NN	O	I-protein
serine-threonine	NN	O	I-protein
phosphatase	NN	O	I-protein
,	NN	O	O
as	NN	O	O
a	NN	O	O
prominent	NN	O	O
component	NN	O	O
of	NN	O	O
the	NN	O	O
calcium	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

A	NN	O	O
second	NN	O	O
important	NN	O	O
molecule	NN	O	O
in	NN	O	O
TCR	NN	O	O
signal	NN	O	O
transduction	NN	O	O
is	NN	O	O
the	NN	O	O
guanine	NN	O	B-protein
nucleotide	NN	O	I-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
p21ras	NN	O	B-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
coupled	NN	O	O
to	NN	O	O
the	NN	O	O
TCR	NN	O	B-protein
by	NN	O	O
a	NN	O	O
protein	NN	O	B-protein
tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
dependent	NN	O	O
mechanism	NN	O	O
.	NN	O	O

The	NN	O	O
experiments	NN	O	O
presented	NN	O	O
here	NN	O	O
show	NN	O	O
that	NN	O	O
expression	NN	O	O
by	NN	O	O
transfection	NN	O	O
of	NN	O	O
mutationally	NN	O	O
activated	NN	O	O
calcineurin	NN	O	B-protein
or	NN	O	O
activated	NN	O	O
p21ras	NN	O	B-protein
alone	NN	O	O
is	NN	O	O
insufficient	NN	O	O
for	NN	O	O
NFAT	NN	O	B-cell_type
transactivation	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
coexpression	NN	O	O
of	NN	O	O
the	NN	O	O
activated	NN	O	O
calcineurin	NN	O	O
with	NN	O	O
activated	NN	O	O
p21ras	NN	O	O
could	NN	O	O
mimic	NN	O	O
TCR	NN	O	O
signals	NN	O	O
in	NN	O	O
NFAT	NN	O	B-cell_type
induction	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
identify	NN	O	O
calcineurin	NN	O	B-protein
and	NN	O	O
p21ras	NN	O	B-protein
as	NN	O	O
cooperative	NN	O	O
partners	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Expression	NN	O	O
of	NN	O	O
the	NN	O	O
chicken	NN	O	B-protein
GATA	NN	O	I-protein
factor	NN	O	I-protein
family	NN	O	I-protein
during	NN	O	O
early	NN	O	O
erythroid	NN	O	O
development	NN	O	O
and	NN	O	O
differentiation	NN	O	O
.	NN	O	O

The	NN	O	O
DNA	NN	O	O
motif	NN	O	O
WGATAR	NN	O	O
has	NN	O	O
been	NN	O	O
identified	NN	O	O
within	NN	O	O
transcriptional	NN	O	B-DNA
regulatory	NN	O	I-DNA
domains	NN	O	I-DNA
of	NN	O	O
globin	NN	O	B-DNA
and	NN	O	O
other	NN	O	O
erythroid-specific	NN	O	B-DNA
genes	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
activator	NN	O	B-protein
proteins	NN	O	I-protein
that	NN	O	O
bind	NN	O	O
to	NN	O	O
this	NN	O	O
regulatory	NN	O	B-DNA
element	NN	O	I-DNA
,	NN	O	O
the	NN	O	O
GATA	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
belong	NN	O	O
to	NN	O	O
a	NN	O	O
multi-gene	NN	O	B-protein
family	NN	O	I-protein
that	NN	O	O
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
chicken	NN	O	B-cell_type
erythroid	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
,	NN	O	O
as	NN	O	O
in	NN	O	O
chickens	NN	O	O
,	NN	O	O
multiple	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
GATA	NN	O	B-protein
factor	NN	O	I-protein
family	NN	O	I-protein
are	NN	O	O
expressed	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
and	NN	O	I-cell_type
murine	NN	O	I-cell_type
erythroid	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

During	NN	O	O
the	NN	O	O
early	NN	O	O
stages	NN	O	O
of	NN	O	O
chicken	NN	O	O
embryogenesis	NN	O	O
(	NN	O	O
well	NN	O	O
before	NN	O	O
blood	NN	O	O
island	NN	O	O
formation	NN	O	O
)	NN	O	O
,	NN	O	O
each	NN	O	O
of	NN	O	O
the	NN	O	O
GATA	NN	O	B-protein
family	NN	O	I-protein
members	NN	O	I-protein
is	NN	O	O
transcribed	NN	O	O
with	NN	O	O
a	NN	O	O
unique	NN	O	O
temporal	NN	O	O
and	NN	O	O
spatial	NN	O	O
pattern	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
primitive	NN	O	B-cell_type
erythroid	NN	O	I-cell_type
lineage	NN	O	I-cell_type
,	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
embryonic	NN	O	B-DNA
epsilon-globin	NN	O	I-DNA
gene	NN	O	I-DNA
parallels	NN	O	O
GATA-1	NN	O	O
expression	NN	O	O
while	NN	O	O
the	NN	O	O
switch	NN	O	O
to	NN	O	O
beta-globin	NN	O	O
transcription	NN	O	O
in	NN	O	O
definitive	NN	O	B-cell_type
erythroid	NN	O	I-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
directly	NN	O	O
preceded	NN	O	O
by	NN	O	O
a	NN	O	O
pronounced	NN	O	O
increase	NN	O	O
in	NN	O	O
GATA-3	NN	O	B-protein
accumulation	NN	O	O
.	NN	O	O

The	NN	O	O
timing	NN	O	O
and	NN	O	O
pattern	NN	O	O
of	NN	O	O
expression	NN	O	O
of	NN	O	O
these	NN	O	O
different	NN	O	O
mRNAs	NN	O	B-RNA
during	NN	O	O
avian	NN	O	O
erythroid	NN	O	O
development	NN	O	O
and	NN	O	O
differentiation	NN	O	O
suggests	NN	O	O
that	NN	O	O
temporally	NN	O	O
regulated	NN	O	O
changes	NN	O	O
in	NN	O	O
GATA	NN	O	B-protein
factor	NN	O	I-protein
expression	NN	O	O
are	NN	O	O
required	NN	O	O
for	NN	O	O
vertebrate	NN	O	O
hematopoiesis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Molecular	NN	O	O
regulation	NN	O	O
of	NN	O	O
human	NN	O	B-protein
interleukin	NN	O	I-protein
2	NN	O	I-protein
and	NN	O	O
T-cell	NN	O	B-cell_type
function	NN	O	O
by	NN	O	O
interleukin	NN	O	B-protein
4	NN	O	I-protein
.	NN	O	O

Distinct	NN	O	O
functional	NN	O	O
T-cell	NN	O	B-cell_type
subsets	NN	O	I-cell_type
,	NN	O	O
differing	NN	O	O
in	NN	O	O
the	NN	O	O
patterns	NN	O	O
of	NN	O	O
lymphokines	NN	O	B-protein
produced	NN	O	O
,	NN	O	O
regulate	NN	O	O
cell-mediated	NN	O	O
and	NN	O	O
humoral	NN	O	O
immune	NN	O	O
responses	NN	O	O
.	NN	O	O

The	NN	O	O
two	NN	O	O
major	NN	O	O
types	NN	O	O
and	NN	O	O
their	NN	O	O
principal	NN	O	O
products	NN	O	O
,	NN	O	O
interleukin	NN	O	B-protein
4	NN	O	I-protein
and	NN	O	O
interferon	NN	O	B-protein
gamma	NN	O	I-protein
(	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
IFN-gamma	NN	O	B-protein
)	NN	O	O
,	NN	O	O
are	NN	O	O
reciprocally	NN	O	O
negatively	NN	O	O
interactive	NN	O	O
.	NN	O	O

To	NN	O	O
analyze	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanism	NN	O	O
of	NN	O	O
IL-4-mediated	NN	O	O
suppression	NN	O	O
of	NN	O	O
cell-mediated	NN	O	O
immunity	NN	O	O
we	NN	O	O
studied	NN	O	O
its	NN	O	O
effects	NN	O	O
on	NN	O	O
expression	NN	O	O
of	NN	O	O
interleukin	NN	O	B-protein
2	NN	O	I-protein
(	NN	O	O
IL-2	NN	O	B-protein
)	NN	O	O
and	NN	O	O
IFN-gamma	NN	O	B-protein
.	NN	O	O

IL-4	NN	O	B-protein
pretreatment	NN	O	O
of	NN	O	O
Jurkat	NN	O	B-cell_type
cells	NN	O	I-cell_type
prior	NN	O	O
to	NN	O	O
stimulation	NN	O	O
resulted	NN	O	O
in	NN	O	O
a	NN	O	O
decrease	NN	O	O
in	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
IL2	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

IL-4	NN	O	B-protein
suppressed	NN	O	O
IL-2	NN	O	O
and	NN	O	O
IFN-gamma	NN	O	O
mRNA	NN	O	O
levels	NN	O	O
in	NN	O	O
primary	NN	O	B-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
addition	NN	O	O
of	NN	O	O
anti-CD28	NN	O	B-protein
antibodies	NN	O	I-protein
relieved	NN	O	O
this	NN	O	O
suppression	NN	O	O
.	NN	O	O

Using	NN	O	O
enhancer-reporter	NN	O	B-DNA
constructs	NN	O	I-DNA
,	NN	O	O
IL-4	NN	O	B-protein
specifically	NN	O	O
down-regulated	NN	O	O
the	NN	O	O
NFIL-2B	NN	O	B-DNA
element	NN	O	I-DNA
.	NN	O	O

Electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
using	NN	O	O
a	NN	O	O
DNA	NN	O	O
oligomer	NN	O	O
containing	NN	O	O
the	NN	O	O
NFIL-2B	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
indicated	NN	O	O
that	NN	O	O
IL-4	NN	O	B-protein
inhibited	NN	O	O
the	NN	O	O
NFIL-2B	NN	O	B-protein
complex	NN	O	I-protein
and	NN	O	O
that	NN	O	O
the	NN	O	O
NFIL-2B	NN	O	B-protein
DNA	NN	O	I-protein
binding	NN	O	I-protein
factor	NN	O	I-protein
is	NN	O	O
distinct	NN	O	O
from	NN	O	O
AP-	NN	O	B-protein
1	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
IL-4	NN	O	B-protein
may	NN	O	O
regulate	NN	O	O
development	NN	O	O
and	NN	O	O
function	NN	O	O
of	NN	O	O
T-cell	NN	O	B-cell_type
subsets	NN	O	I-cell_type
involved	NN	O	O
in	NN	O	O
cell-mediated	NN	O	O
immunity	NN	O	O
in	NN	O	O
part	NN	O	O
by	NN	O	O
inhibiting	NN	O	B-protein
factors	NN	O	I-protein
required	NN	O	O
for	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
IL2	NN	O	B-DNA
gene	NN	O	I-DNA

-DOCSTART-	O

Immunochemical	NN	O	O
differences	NN	O	O
between	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
from	NN	O	O
corticoid-sensitive	NN	O	B-cell_type
and	NN	O	I-cell_type
-resistant	NN	O	I-cell_type
malignant	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
have	NN	O	O
explored	NN	O	O
the	NN	O	O
possibility	NN	O	O
of	NN	O	O
using	NN	O	O
antibodies	NN	O	B-protein
against	NN	O	O
purified	NN	O	O
rat	NN	O	O
liver	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
to	NN	O	O
study	NN	O	O
the	NN	O	O
immunochemical	NN	O	O
properties	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
from	NN	O	O
murine	NN	O	B-cell_type
and	NN	O	I-cell_type
human	NN	O	I-cell_type
malignant	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

For	NN	O	O
this	NN	O	O
purpose	NN	O	O
,	NN	O	O
purified	NN	O	B-protein
immune	NN	O	I-protein
immunoglobulin	NN	O	I-protein
G	NN	O	I-protein
was	NN	O	O
covalently	NN	O	O
linked	NN	O	O
to	NN	O	O
Sepharose	NN	O	B-protein
CL-4B	NN	O	I-protein
.	NN	O	O

We	NN	O	O
then	NN	O	O
examined	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
the	NN	O	O
affinity	NN	O	O
gel	NN	O	O
to	NN	O	O
recognize	NN	O	O
cytosolic	NN	O	B-protein
[	NN	O	I-protein
3H	NN	O	I-protein
]	NN	O	I-protein
triamcinolone	NN	O	I-protein
acetonide-receptor	NN	O	I-protein
complexes	NN	O	I-protein
from	NN	O	O
the	NN	O	O
corticoid-sensitive	NN	O	B-cell_line
(	NN	O	I-cell_line
CS	NN	O	I-cell_line
)	NN	O	I-cell_line
and	NN	O	I-cell_line
-resistant	NN	O	I-cell_line
strains	NN	O	I-cell_line
of	NN	O	I-cell_line
mouse	NN	O	I-cell_line
lymphoma	NN	O	I-cell_line
P1798	NN	O	I-cell_line
,	NN	O	O
from	NN	O	O
CS	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
of	NN	O	O
patients	NN	O	O
with	NN	O	O
chronic	NN	O	O
lymphatic	NN	O	O
leukemia	NN	O	O
,	NN	O	O
and	NN	O	O
from	NN	O	O
a	NN	O	O
CS	NN	O	B-cell_line
clone	NN	O	I-cell_line
of	NN	O	O
human	NN	O	B-cell_type
leukemic	NN	O	I-cell_type
lymphoblasts	NN	O	I-cell_type
in	NN	O	O
tissue	NN	O	O
culture	NN	O	O
(	NN	O	O
CH6	NN	O	B-cell_line
)	NN	O	O
.	NN	O	O

Mouse	NN	O	O
thymus	NN	O	O
was	NN	O	O
used	NN	O	O
as	NN	O	O
a	NN	O	O
source	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
from	NN	O	O
normal	NN	O	B-cell_type
CS	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Whereas	NN	O	O
the	NN	O	O
immunoaffinity	NN	O	O
column	NN	O	O
retained	NN	O	O
70	NN	O	O
to	NN	O	O
84	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
58-	NN	O	O
to	NN	O	O
62-A	NN	O	O
(	NN	O	O
Stokes	NN	O	O
radius	NN	O	O
)	NN	O	O
[	NN	O	B-protein
3H	NN	O	I-protein
]	NN	O	I-protein
triamcinolone	NN	O	I-protein
acetonide-receptor	NN	O	I-protein
complexes	NN	O	I-protein
characteristic	NN	O	O
of	NN	O	O
the	NN	O	O
CS	NN	O	B-cell_line
mouse	NN	O	I-cell_line
and	NN	O	I-cell_line
human	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
,	NN	O	O
it	NN	O	O
failed	NN	O	O
to	NN	O	O
recognize	NN	O	O
the	NN	O	O
27-	NN	O	O
to	NN	O	O
28-A	NN	O	O
(	NN	O	O
Stokes	NN	O	O
radius	NN	O	O
)	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
present	NN	O	O
in	NN	O	O
corticoid-resistant	NN	O	O
mouse	NN	O	B-cell_line
lymphoma	NN	O	I-cell_line
P1798	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
under	NN	O	O
appropriate	NN	O	O
experimental	NN	O	O
conditions	NN	O	O
,	NN	O	O
it	NN	O	O
was	NN	O	O
possible	NN	O	O
to	NN	O	O
demonstrate	NN	O	O
cross-reactivity	NN	O	O
between	NN	O	O
the	NN	O	O
antiserum	NN	O	O
against	NN	O	O
rat	NN	O	B-protein
liver	NN	O	I-protein
glucocorticoid	NN	O	I-protein
receptor	NN	O	I-protein
and	NN	O	O
the	NN	O	O
58-	NN	O	O
to	NN	O	O
62-A	NN	O	O
(	NN	O	O
Stokes	NN	O	O
radius	NN	O	O
)	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
from	NN	O	O
species	NN	O	O
as	NN	O	O
diverse	NN	O	O
as	NN	O	O
mouse	NN	O	O
and	NN	O	O
humans	NN	O	O
.	NN	O	O

-DOCSTART-	O

Heterogeneity	NN	O	O
of	NN	O	O
the	NN	O	O
in	NN	O	O
vitro	NN	O	O
responses	NN	O	O
to	NN	O	O
glucocorticoids	NN	O	O
in	NN	O	O
acute	NN	O	O
leukemia	NN	O	O
.	NN	O	O

In	NN	O	O
leukocyte	NN	O	B-cell_type
population	NN	O	I-cell_type
freshly	NN	O	O
isolated	NN	O	O
from	NN	O	O
the	NN	O	O
blood	NN	O	O
of	NN	O	O
26	NN	O	O
patients	NN	O	O
with	NN	O	O
acute	NN	O	O
leukemia	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
measured	NN	O	O
several	NN	O	O
parameters	NN	O	O
including	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
,	NN	O	O
nucleoside	NN	O	O
incorporation	NN	O	O
,	NN	O	O
percentage	NN	O	O
of	NN	O	O
cells	NN	O	O
in	NN	O	O
S	NN	O	O
phase	NN	O	O
,	NN	O	O
and	NN	O	O
steroid-induced	NN	O	O
cell	NN	O	O
lysis	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
in	NN	O	O
some	NN	O	O
cases	NN	O	O
,	NN	O	O
the	NN	O	O
short-term	NN	O	O
response	NN	O	O
to	NN	O	O
steroid	NN	O	O
therapy	NN	O	O
was	NN	O	O
determined	NN	O	O
.	NN	O	O

Although	NN	O	O
,	NN	O	O
in	NN	O	O
all	NN	O	O
the	NN	O	O
patients	NN	O	O
studied	NN	O	O
,	NN	O	O
leukocytes	NN	O	B-cell_type
were	NN	O	O
found	NN	O	O
to	NN	O	O
contain	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
,	NN	O	O
we	NN	O	O
failed	NN	O	O
to	NN	O	O
demonstrate	NN	O	O
any	NN	O	O
correlation	NN	O	O
between	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
binding	NN	O	O
sites	NN	O	O
and	NN	O	O
the	NN	O	O
in	NN	O	O
vitro	NN	O	O
or	NN	O	O
in	NN	O	O
vivo	NN	O	O
response	NN	O	O
to	NN	O	O
glucocorticoids	NN	O	O
.	NN	O	O

This	NN	O	O
absence	NN	O	O
of	NN	O	O
correlation	NN	O	O
could	NN	O	O
be	NN	O	O
in	NN	O	O
part	NN	O	O
explained	NN	O	O
by	NN	O	O
the	NN	O	O
marked	NN	O	O
heterogeneity	NN	O	O
of	NN	O	O
the	NN	O	O
steroid	NN	O	O
response	NN	O	O
demonstrated	NN	O	O
in	NN	O	O
leukocyte	NN	O	O
subpopulations	NN	O	O
.	NN	O	O

It	NN	O	O
appears	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
that	NN	O	O
the	NN	O	O
degree	NN	O	O
of	NN	O	O
steroid	NN	O	O
action	NN	O	O
in	NN	O	O
vitro	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
extent	NN	O	O
of	NN	O	O
spontaneous	NN	O	O
and	NN	O	O
dexamethasone-induced	NN	O	O
cell	NN	O	O
death	NN	O	O
may	NN	O	O
be	NN	O	O
related	NN	O	O
to	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
cells	NN	O	O
in	NN	O	O
the	NN	O	O
S	NN	O	O
phase	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
.	NN	O	O

-DOCSTART-	O

Glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
cytosol	NN	O	O
and	NN	O	O
nuclear	NN	O	O
extract	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
.	NN	O	O

Cortisol	NN	O	O
binding	NN	O	O
by	NN	O	O
cytosol	NN	O	O
and	NN	O	O
0.4	NN	O	O
M	NN	O	O
KCl	NN	O	O
extract	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	O
fraction	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
were	NN	O	O
studied	NN	O	O
by	NN	O	O
gel	NN	O	O
chromatography	NN	O	O
and	NN	O	O
ion	NN	O	O
exchange	NN	O	O
filtration	NN	O	O
on	NN	O	O
DEAE	NN	O	O
cellulose	NN	O	O
.	NN	O	O

The	NN	O	O
cytoplasmic	NN	O	B-protein
cortisol	NN	O	I-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
has	NN	O	O
a	NN	O	O
molecular	NN	O	O
weight	NN	O	O
95	NN	O	O
000	NN	O	O
and	NN	O	O
the	NN	O	O
soluble	NN	O	B-protein
nuclear	NN	O	I-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
50	NN	O	O
000	NN	O	O
.	NN	O	O

The	NN	O	O
absence	NN	O	O
of	NN	O	O
the	NN	O	O
uptake	NN	O	O
of	NN	O	O
radioactive	NN	O	O
cortisol	NN	O	O
by	NN	O	O
isolated	NN	O	O
nuclei	NN	O	O
and	NN	O	O
the	NN	O	O
apparent	NN	O	O
requirement	NN	O	O
of	NN	O	O
the	NN	O	O
cytosol	NN	O	O
for	NN	O	O
glucocorticoid	NN	O	O
specific	NN	O	O
binding	NN	O	O
in	NN	O	O
nuclear	NN	O	O
receptor	NN	O	O
sites	NN	O	O
was	NN	O	O
observed	NN	O	O
.	NN	O	O

The	NN	O	O
association	NN	O	O
constant	NN	O	O
characterising	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
cortisol	NN	O	O
to	NN	O	O
cytosol	NN	O	O
was	NN	O	O
KA	NN	O	O
=	NN	O	O
3.5	NN	O	O
.	NN	O	O
10	NN	O	O
(	NN	O	O
9	NN	O	O
)	NN	O	O
l/mol	NN	O	O
.	NN	O	O

-DOCSTART-	O

Corticosteroid-induced	NN	O	O
lymphopenia	NN	O	O
,	NN	O	O
immunosuppression	NN	O	O
,	NN	O	O
and	NN	O	O
body	NN	O	O
defense	NN	O	O
.	NN	O	O

The	NN	O	O
apparent	NN	O	O
paradox	NN	O	O
of	NN	O	O
heightened	NN	O	O
adrenal	NN	O	O
corticosteroid	NN	O	O
levels	NN	O	O
associated	NN	O	O
with	NN	O	O
reduction	NN	O	O
in	NN	O	O
the	NN	O	O
competence	NN	O	O
of	NN	O	O
the	NN	O	O
body	NN	O	O
's	NN	O	O
defensive	NN	O	O
apparatus	NN	O	O
to	NN	O	O
cope	NN	O	O
with	NN	O	O
exposure	NN	O	O
to	NN	O	O
new	NN	O	O
microbial	NN	O	B-protein
antigens	NN	O	I-protein
is	NN	O	O
considered	NN	O	O
.	NN	O	O

The	NN	O	O
question	NN	O	O
is	NN	O	O
asked	NN	O	O
how	NN	O	O
this	NN	O	O
lowered	NN	O	O
defensive	NN	O	O
capability	NN	O	O
,	NN	O	O
which	NN	O	O
occurs	NN	O	O
in	NN	O	O
the	NN	O	O
face	NN	O	O
of	NN	O	O
a	NN	O	O
threat	NN	O	O
to	NN	O	O
body	NN	O	O
integrity	NN	O	O
,	NN	O	O
is	NN	O	O
consistent	NN	O	O
with	NN	O	O
Cannon	NN	O	O
's	NN	O	O
principals	NN	O	O
of	NN	O	O
the	NN	O	O
``	NN	O	O
wisdom	NN	O	O
of	NN	O	O
the	NN	O	O
body.	NN	O	O
''	NN	O	O

The	NN	O	O
suggestion	NN	O	O
is	NN	O	O
offered	NN	O	O
that	NN	O	O
the	NN	O	O
immunologic	NN	O	O
response	NN	O	O
to	NN	O	O
self-antigens	NN	O	B-protein
exposed	NN	O	O
by	NN	O	O
disease	NN	O	O
or	NN	O	O
trauma	NN	O	O
may	NN	O	O
be	NN	O	O
suppressed	NN	O	O
by	NN	O	O
corticosteroid	NN	O	O
to	NN	O	O
offset	NN	O	O
the	NN	O	O
likelihood	NN	O	O
of	NN	O	O
autoimmune	NN	O	O
attack	NN	O	O
.	NN	O	O

-DOCSTART-	O

Protein	NN	O	B-protein
tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
activation	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
lipopolysaccharide	NN	O	O
induction	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
in	NN	O	O
human	NN	O	B-cell_type
blood	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

Bacterial	NN	O	O
LPS	NN	O	O
induce	NN	O	O
production	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
such	NN	O	O
as	NN	O	O
IL-1	NN	O	B-protein
,	NN	O	O
IL-6	NN	O	B-protein
,	NN	O	O
and	NN	O	O
TNF	NN	O	B-protein
in	NN	O	O
mononuclear	NN	O	B-cell_type
phagocytes	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
this	NN	O	O
represents	NN	O	O
a	NN	O	O
central	NN	O	O
component	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
septic	NN	O	O
shock	NN	O	O
syndrome	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
by	NN	O	O
which	NN	O	O
LPS	NN	O	O
activates	NN	O	O
these	NN	O	O
cells	NN	O	O
to	NN	O	O
express	NN	O	O
cytokines	NN	O	B-protein
are	NN	O	O
not	NN	O	O
completely	NN	O	O
characterized	NN	O	O
.	NN	O	O

The	NN	O	O
present	NN	O	O
study	NN	O	O
addressed	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
different	NN	O	O
protein	NN	O	B-protein
kinases	NN	O	I-protein
in	NN	O	O
the	NN	O	O
LPS	NN	O	O
induction	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
.	NN	O	O

It	NN	O	O
is	NN	O	O
shown	NN	O	O
that	NN	O	O
LPS	NN	O	O
induced	NN	O	O
a	NN	O	O
12-	NN	O	O
to	NN	O	O
16-fold	NN	O	O
increase	NN	O	O
in	NN	O	O
IL-1	NN	O	O
beta	NN	O	O
,	NN	O	O
IL-6	NN	O	O
,	NN	O	O
and	NN	O	O
TNF-alpha	NN	O	O
mRNA	NN	O	O
levels	NN	O	O
,	NN	O	O
and	NN	O	O
this	NN	O	O
was	NN	O	O
completely	NN	O	O
or	NN	O	O
more	NN	O	O
than	NN	O	O
80	NN	O	O
%	NN	O	O
blocked	NN	O	O
by	NN	O	O
the	NN	O	O
protein	NN	O	O
tyrosine	NN	O	O
kinase	NN	O	O
specific	NN	O	O
inhibitors	NN	O	O
herbimycin	NN	O	O
A	NN	O	O
and	NN	O	O
genistein	NN	O	O
at	NN	O	O
the	NN	O	O
concentrations	NN	O	O
of	NN	O	O
1.7	NN	O	O
and	NN	O	O
37	NN	O	O
microM	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

Protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
inhibition	NN	O	O
by	NN	O	O
staurosporine	NN	O	O
reduced	NN	O	O
LPS	NN	O	O
induction	NN	O	O
of	NN	O	O
TNF-alpha	NN	O	B-protein
,	NN	O	O
whereas	NN	O	O
it	NN	O	O
had	NN	O	O
no	NN	O	O
effects	NN	O	O
on	NN	O	O
IL-6	NN	O	B-protein
and	NN	O	O
IL-1	NN	O	O
beta	NN	O	O
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
A	NN	O	I-protein
by	NN	O	O
H89	NN	O	O
reduced	NN	O	O
IL-6	NN	O	B-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
but	NN	O	O
did	NN	O	O
not	NN	O	O
detectably	NN	O	O
change	NN	O	O
IL-1	NN	O	O
beta	NN	O	O
or	NN	O	O
TNF-alpha	NN	O	O
mRNA	NN	O	O
levels	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
LPS	NN	O	O
did	NN	O	O
not	NN	O	O
increase	NN	O	O
leukemia	NN	O	B-RNA
inhibitory	NN	O	I-RNA
factor	NN	O	I-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
which	NN	O	O
was	NN	O	O
constitutively	NN	O	O
expressed	NN	O	O
and	NN	O	O
not	NN	O	O
significantly	NN	O	O
reduced	NN	O	O
by	NN	O	O
these	NN	O	O
inhibitors	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
to	NN	O	O
cytokine	NN	O	B-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
,	NN	O	O
LPS-induced	NN	O	O
IL-6	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
and	NN	O	O
IL-6	NN	O	O
bioactivity	NN	O	O
were	NN	O	O
also	NN	O	O
reduced	NN	O	O
to	NN	O	O
baseline	NN	O	O
levels	NN	O	O
by	NN	O	O
the	NN	O	O
PTK	NN	O	O
inhibitors	NN	O	O
herbimycin	NN	O	O
A	NN	O	O
and	NN	O	O
genistein	NN	O	O
.	NN	O	O

Both	NN	O	O
PTK	NN	O	O
inhibitors	NN	O	O
also	NN	O	O
reduced	NN	O	O
the	NN	O	O
LPS	NN	O	O
activation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
a	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
involved	NN	O	O
in	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
cytokine	NN	O	B-RNA
genes	NN	O	I-RNA
such	NN	O	O
as	NN	O	O
IL-6	NN	O	B-DNA
and	NN	O	O
TNF-alpha	NN	O	B-DNA
.	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
was	NN	O	O
also	NN	O	O
reduced	NN	O	O
by	NN	O	O
H89	NN	O	O
,	NN	O	O
whereas	NN	O	O
staurosporine	NN	O	O
had	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
this	NN	O	O
response	NN	O	O
.	NN	O	O

In	NN	O	O
summary	NN	O	O
,	NN	O	O
these	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
and	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
A	NN	O	I-protein
appear	NN	O	O
to	NN	O	O
have	NN	O	O
selective	NN	O	O
effects	NN	O	O
in	NN	O	O
the	NN	O	O
LPS	NN	O	O
induction	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
,	NN	O	O
whereas	NN	O	O
PTK	NN	O	B-protein
is	NN	O	O
required	NN	O	O
for	NN	O	O
LPS	NN	O	O
induction	NN	O	O
of	NN	O	O
a	NN	O	O
broad	NN	O	O
spectrum	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

-DOCSTART-	O

In	NN	O	O
vivo	NN	O	O
control	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
by	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
.	NN	O	O

The	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
is	NN	O	O
stored	NN	O	O
in	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
in	NN	O	O
complexes	NN	O	O
with	NN	O	O
the	NN	O	O
inhibitor	NN	O	B-protein
protein	NN	O	I-protein
I	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
.	NN	O	O

It	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
in	NN	O	O
vitro	NN	O	O
that	NN	O	O
dissociation	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
from	NN	O	O
these	NN	O	O
complexes	NN	O	O
results	NN	O	O
in	NN	O	O
active	NN	O	B-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
-induced	NN	O	O
activation	NN	O	O
of	NN	O	O
B	NN	O	B-cell_type
or	NN	O	I-cell_type
pre-B	NN	O	I-cell_type
cells	NN	O	I-cell_type
results	NN	O	O
in	NN	O	O
loss	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
from	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
complexes	NN	O	I-protein
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

Many	NN	O	O
liberated	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
dimers	NN	O	I-protein
reached	NN	O	O
the	NN	O	O
nucleus	NN	O	O
,	NN	O	O
where	NN	O	O
increased	NN	O	O
c-rel	NN	O	B-protein
,	NN	O	O
p65	NN	O	O
and	NN	O	O
p50	NN	O	O
were	NN	O	O
detected	NN	O	O
by	NN	O	O
immunoblotting	NN	O	O
and	NN	O	O
by	NN	O	O
DNA	NN	O	O
binding	NN	O	O
assays	NN	O	O
.	NN	O	O

Some	NN	O	O
liberated	NN	O	B-protein
dimers	NN	O	I-protein
were	NN	O	O
retained	NN	O	O
in	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
through	NN	O	O
binding	NN	O	O
to	NN	O	O
newly	NN	O	O
synthesized	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
,	NN	O	O
a	NN	O	O
finding	NN	O	O
which	NN	O	O
strongly	NN	O	O
suggests	NN	O	O
(	NN	O	O
i	NN	O	O
)	NN	O	O
that	NN	O	O
the	NN	O	O
LPS-induced	NN	O	O
signal	NN	O	O
causes	NN	O	O
dissociation	NN	O	O
of	NN	O	O
complexes	NN	O	O
rather	NN	O	O
than	NN	O	O
preventing	NN	O	O
their	NN	O	O
association	NN	O	O
and	NN	O	O
(	NN	O	O
ii	NN	O	O
)	NN	O	O
that	NN	O	O
dissociation	NN	O	O
results	NN	O	O
from	NN	O	O
modification	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
and	NN	O	O
not	NN	O	O
of	NN	O	O
c-rel	NN	O	B-protein
or	NN	O	O
p65	NN	O	B-protein
.	NN	O	O

No	NN	O	O
effect	NN	O	O
of	NN	O	O
LPS	NN	O	O
treatment	NN	O	O
was	NN	O	O
detected	NN	O	O
on	NN	O	O
p105	NN	O	B-protein
or	NN	O	O
p100	NN	O	B-protein
,	NN	O	O
which	NN	O	O
also	NN	O	O
retain	NN	O	O
rel	NN	O	B-protein
family	NN	O	I-protein
members	NN	O	I-protein
in	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
.	NN	O	O

Quite	NN	O	O
unexpectedly	NN	O	O
,	NN	O	O
we	NN	O	O
also	NN	O	O
found	NN	O	O
that	NN	O	O
in	NN	O	O
unstimulated	NN	O	B-cell_type
cells	NN	O	I-cell_type
there	NN	O	O
is	NN	O	O
a	NN	O	O
constant	NN	O	O
ongoing	NN	O	O
process	NN	O	O
of	NN	O	O
degradation	NN	O	O
and	NN	O	O
replacement	NN	O	O
of	NN	O	O
complexed	NN	O	B-protein
I	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
.	NN	O	O

We	NN	O	O
propose	NN	O	O
that	NN	O	O
this	NN	O	O
turnover	NN	O	O
results	NN	O	O
in	NN	O	O
the	NN	O	O
low	NN	O	O
level	NN	O	O
of	NN	O	O
active	NN	O	B-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
presumably	NN	O	O
necessary	NN	O	O
even	NN	O	O
in	NN	O	O
the	NN	O	O
unstimulated	NN	O	O
cell	NN	O	O
,	NN	O	O
and	NN	O	O
that	NN	O	O
the	NN	O	O
high	NN	O	O
rate	NN	O	O
of	NN	O	O
synthesis	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
provides	NN	O	O
the	NN	O	O
ability	NN	O	O
to	NN	O	O
turn	NN	O	O
off	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activity	NN	O	O
rapidly	NN	O	O
as	NN	O	O
soon	NN	O	O
as	NN	O	O
the	NN	O	O
activating	NN	O	O
signal	NN	O	O
ceases	NN	O	O
.	NN	O	O

-DOCSTART-	O

Identification	NN	O	O
of	NN	O	O
a	NN	O	O
killer	NN	O	B-DNA
cell-specific	NN	O	I-DNA
regulatory	NN	O	I-DNA
element	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
mouse	NN	O	B-DNA
perforin	NN	O	I-DNA
gene	NN	O	I-DNA
:	NN	O	O
an	NN	O	O
Ets-binding	NN	O	O
site-homologous	NN	O	O
motif	NN	O	O
that	NN	O	O
interacts	NN	O	O
with	NN	O	O
Ets-related	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

The	NN	O	O
gene	NN	O	O
encoding	NN	O	O
the	NN	O	O
cytolytic	NN	O	B-protein
protein	NN	O	I-protein
perforin	NN	O	B-protein
is	NN	O	O
selectively	NN	O	O
expressed	NN	O	O
by	NN	O	O
activated	NN	O	B-cell_type
killer	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

To	NN	O	O
understand	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
underlying	NN	O	O
the	NN	O	O
cell-type-specific	NN	O	O
expression	NN	O	O
of	NN	O	O
this	NN	O	O
gene	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
characterized	NN	O	O
the	NN	O	O
regulatory	NN	O	O
functions	NN	O	O
and	NN	O	O
the	NN	O	O
DNA-protein	NN	O	O
interactions	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
mouse	NN	O	B-DNA
perforin	NN	O	I-DNA
gene	NN	O	I-DNA
(	NN	O	O
Pfp	NN	O	B-DNA
)	NN	O	O
.	NN	O	O

A	NN	O	O
region	NN	O	O
extending	NN	O	O
from	NN	O	O
residues	NN	O	B-DNA
+62	NN	O	I-DNA
through	NN	O	I-DNA
-141	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
possesses	NN	O	O
the	NN	O	O
essential	NN	O	O
promoter	NN	O	O
activity	NN	O	O
,	NN	O	O
and	NN	O	O
regions	NN	O	O
further	NN	O	O
upstream	NN	O	O
,	NN	O	O
which	NN	O	O
are	NN	O	O
able	NN	O	O
to	NN	O	O
either	NN	O	O
enhance	NN	O	O
or	NN	O	O
suppress	NN	O	O
gene	NN	O	O
expression	NN	O	O
,	NN	O	O
were	NN	O	O
identified	NN	O	O
.	NN	O	O

The	NN	O	O
region	NN	O	O
between	NN	O	O
residues	NN	O	B-DNA
-411	NN	O	I-DNA
and	NN	O	I-DNA
-566	NN	O	I-DNA
was	NN	O	O
chosen	NN	O	O
for	NN	O	O
further	NN	O	O
characterization	NN	O	O
,	NN	O	O
since	NN	O	O
it	NN	O	O
contains	NN	O	O
an	NN	O	O
enhancer-like	NN	O	O
activity	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
identified	NN	O	O
a	NN	O	O
32-mer	NN	O	O
sequence	NN	O	O
(	NN	O	O
residues	NN	O	B-DNA
-491	NN	O	I-DNA
to	NN	O	I-DNA
-522	NN	O	I-DNA
)	NN	O	O
which	NN	O	O
appeared	NN	O	O
to	NN	O	O
be	NN	O	O
capable	NN	O	O
of	NN	O	O
enhancing	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
a	NN	O	O
killer	NN	O	O
cell-specific	NN	O	O
manner	NN	O	O
.	NN	O	O

Within	NN	O	O
this	NN	O	O
segment	NN	O	O
,	NN	O	O
a	NN	O	O
9-mer	NN	O	O
motif	NN	O	O
(	NN	O	O
5'-ACAGGAAGT-3	NN	O	O
'	NN	O	O
,	NN	O	O
residues	NN	O	B-DNA
-505	NN	O	I-DNA
to	NN	O	I-DNA
-497	NN	O	I-DNA
;	NN	O	O
designated	NN	O	O
NF-P	NN	O	O
motif	NN	O	O
)	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
highly	NN	O	O
homologous	NN	O	O
to	NN	O	O
the	NN	O	O
Ets	NN	O	B-DNA
proto-oncoprotein-binding	NN	O	I-DNA
site	NN	O	I-DNA
,	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
interact	NN	O	O
with	NN	O	O
two	NN	O	O
proteins	NN	O	O
,	NN	O	O
NF-P1	NN	O	B-protein
and	NN	O	O
NF-P2	NN	O	B-protein
.	NN	O	O

NF-P2	NN	O	B-protein
appears	NN	O	O
to	NN	O	O
be	NN	O	O
induced	NN	O	O
by	NN	O	O
reagents	NN	O	O
known	NN	O	O
to	NN	O	O
up-regulate	NN	O	O
the	NN	O	O
perforin	NN	O	O
message	NN	O	O
level	NN	O	O
and	NN	O	O
is	NN	O	O
present	NN	O	O
exclusively	NN	O	O
in	NN	O	O
killer	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
and	NN	O	O
UV	NN	O	O
cross-linking	NN	O	O
experiments	NN	O	O
revealed	NN	O	O
that	NN	O	O
NF-P1	NN	O	B-protein
and	NN	O	O
NF-P2	NN	O	B-protein
may	NN	O	O
possess	NN	O	O
common	NN	O	O
DNA-binding	NN	O	B-protein
subunits	NN	O	I-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
larger	NN	O	O
native	NN	O	O
molecular	NN	O	O
mass	NN	O	O
of	NN	O	O
NF-P1	NN	O	B-protein
suggests	NN	O	O
that	NN	O	O
NF-P1	NN	O	B-protein
contains	NN	O	O
an	NN	O	O
additional	NN	O	O
non-DNA-binding	NN	O	B-protein
subunit	NN	O	I-protein
(	NN	O	O
s	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
view	NN	O	O
of	NN	O	O
the	NN	O	O
homology	NN	O	O
between	NN	O	O
the	NN	O	O
NF-P	NN	O	B-protein
motif	NN	O	I-protein
and	NN	O	O
other	NN	O	O
Ets	NN	O	B-protein
proto-oncoprotein-binding	NN	O	I-protein
sites	NN	O	I-protein
,	NN	O	O
it	NN	O	O
is	NN	O	O
postulated	NN	O	O
that	NN	O	O
NF-P1	NN	O	B-protein
and	NN	O	O
NF-P2	NN	O	B-protein
belong	NN	O	O
to	NN	O	O
the	NN	O	O
Ets	NN	O	B-protein
protein	NN	O	I-protein
family	NN	O	I-protein
.	NN	O	O

Results	NN	O	O
obtained	NN	O	O
from	NN	O	O
the	NN	O	O
binding	NN	O	O
competition	NN	O	O
assay	NN	O	O
,	NN	O	O
nevertheless	NN	O	O
,	NN	O	O
suggest	NN	O	O
that	NN	O	O
NF-P1	NN	O	B-protein
and	NN	O	O
NF-P2	NN	O	B-protein
are	NN	O	O
related	NN	O	O
to	NN	O	O
but	NN	O	O
distinct	NN	O	O
from	NN	O	O
Ets	NN	O	B-protein
proteins	NN	O	I-protein
,	NN	O	O
e.g.	NN	O	O
,	NN	O	O
Ets-1	NN	O	B-protein
,	NN	O	O
Ets-2	NN	O	B-protein
,	NN	O	O
and	NN	O	O
NF-AT/Elf-1	NN	O	B-protein
,	NN	O	O
known	NN	O	O
to	NN	O	O
be	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

-DOCSTART-	O

The	NN	O	O
role	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-DNA
B1	NN	O	I-DNA
(	NN	O	I-DNA
p50/p105	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
activation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
blood	NN	O	I-cell_type
T-lymphocytes	NN	O	I-cell_type
via	NN	O	O
CD2	NN	O	B-protein
and	NN	O	I-protein
CD28	NN	O	I-protein
adhesion	NN	O	I-protein
molecules	NN	O	I-protein
.	NN	O	O

Stimulation	NN	O	O
of	NN	O	O
primary	NN	O	B-cell_type
human	NN	O	I-cell_type
T-lymphocytes	NN	O	I-cell_type
via	NN	O	O
CD2	NN	O	B-protein
and	NN	O	I-protein
CD28	NN	O	I-protein
adhesion	NN	O	I-protein
molecules	NN	O	I-protein
induces	NN	O	O
a	NN	O	O
long-lasting	NN	O	O
proliferation	NN	O	O
(	NN	O	O
>	NN	O	O
3	NN	O	O
weeks	NN	O	O
)	NN	O	O
.	NN	O	O

This	NN	O	O
potent	NN	O	O
activation	NN	O	O
does	NN	O	O
not	NN	O	O
require	NN	O	O
accessory	NN	O	O
cells	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
monocytes	NN	O	B-cell_type
,	NN	O	O
but	NN	O	O
depends	NN	O	O
on	NN	O	O
persistent	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
secretion	NN	O	O
and	NN	O	O
receptivity	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
high	NN	O	O
and	NN	O	O
prolonged	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
inducible	NN	O	O
CD25/IL-2	NN	O	B-DNA
receptor	NN	O	I-DNA
alpha	NN	O	I-DNA
(	NN	O	I-DNA
IL-2R	NN	O	I-DNA
alpha	NN	O	I-DNA
)	NN	O	I-DNA
chain	NN	O	I-DNA
gene	NN	O	I-DNA
.	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
participates	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
both	NN	O	O
IL-2	NN	O	B-DNA
and	NN	O	I-DNA
IL-2R	NN	O	I-DNA
alpha	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
multiple	NN	O	B-DNA
cellular	NN	O	I-DNA
genes	NN	O	I-DNA
involved	NN	O	O
in	NN	O	O
T-cell	NN	O	O
proliferation	NN	O	O
.	NN	O	O

To	NN	O	O
evaluate	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
T-lymphocytes	NN	O	I-cell_type
,	NN	O	O
we	NN	O	O
previously	NN	O	O
analyzed	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B-related	NN	O	I-protein
complexes	NN	O	I-protein
in	NN	O	O
response	NN	O	O
to	NN	O	O
CD2+CD28	NN	O	O
costimulation	NN	O	O
.	NN	O	O

We	NN	O	O
demonstrated	NN	O	O
a	NN	O	O
long-term	NN	O	O
induction	NN	O	O
of	NN	O	O
p50/p65	NN	O	B-protein
heterodimer	NN	O	I-protein
,	NN	O	O
a	NN	O	O
putative	NN	O	B-protein
p65/c-Rel	NN	O	I-protein
heterodimer	NN	O	I-protein
,	NN	O	O
and	NN	O	O
a	NN	O	O
constitutive	NN	O	O
nuclear	NN	O	O
expression	NN	O	O
of	NN	O	O
KBF1/p50	NN	O	B-protein
homodimers	NN	O	I-protein
.	NN	O	I-protein

As	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
p50	NN	O	B-protein
remains	NN	O	O
unclear	NN	O	O
,	NN	O	O
we	NN	O	O
focused	NN	O	O
our	NN	O	O
present	NN	O	O
study	NN	O	O
on	NN	O	O
NF-kappa	NN	O	B-protein
B1	NN	O	I-protein
(	NN	O	O
p50/p105	NN	O	B-protein
)	NN	O	O
gene	NN	O	O
regulation	NN	O	O
.	NN	O	O

Using	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
and	NN	O	O
Western	NN	O	O
and	NN	O	O
Northern	NN	O	O
blot	NN	O	O
analyses	NN	O	O
,	NN	O	O
we	NN	O	O
studied	NN	O	O
NF-kappa	NN	O	B-DNA
B1	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
during	NN	O	O
T-cell	NN	O	O
stimulation	NN	O	O
via	NN	O	O
CD2+CD28	NN	O	B-protein
.	NN	O	O

We	NN	O	O
observed	NN	O	O
a	NN	O	O
transient	NN	O	O
4-	NN	O	O
to	NN	O	O
5-fold	NN	O	O
increase	NN	O	O
of	NN	O	O
NF-kappa	NN	O	O
B1	NN	O	O
gene	NN	O	O
expression	NN	O	O
at	NN	O	O
both	NN	O	O
the	NN	O	O
mRNA	NN	O	O
and	NN	O	O
protein	NN	O	O
levels	NN	O	O
,	NN	O	O
lasting	NN	O	O
for	NN	O	O
at	NN	O	O
least	NN	O	O
24	NN	O	O
h	NN	O	O
.	NN	O	O

p50	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
apparently	NN	O	O
stays	NN	O	O
highly	NN	O	O
controlled	NN	O	O
when	NN	O	O
p105	NN	O	O
expression	NN	O	O
is	NN	O	O
enhanced	NN	O	O
by	NN	O	O
a	NN	O	O
physiological	NN	O	O
stimulus	NN	O	O
of	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
T-cells	NN	O	I-cell_type
.	NN	O	O

Partial	NN	O	O
inhibition	NN	O	O
of	NN	O	O
p50	NN	O	O
and	NN	O	O
p105	NN	O	O
expression	NN	O	O
by	NN	O	O
NF-kappa	NN	O	B-protein
B1	NN	O	I-protein
antisense	NN	O	O
oligonucleotides	NN	O	O
significantly	NN	O	O
reduced	NN	O	O
T-cell	NN	O	O
proliferation	NN	O	O
and	NN	O	O
CD25/IL-2R	NN	O	B-protein
alpha	NN	O	I-protein
cell	NN	O	O
surface	NN	O	O
expression	NN	O	O
.	NN	O	O

(	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

Vitamin	NN	O	B-protein
D	NN	O	I-protein
receptor	NN	O	I-protein
quantitation	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
health	NN	O	O
and	NN	O	O
disease	NN	O	O
.	NN	O	O

Vitamin	NN	O	B-protein
D	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
VDR	NN	O	O
)	NN	O	O
concentration	NN	O	O
was	NN	O	O
quantitated	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
from	NN	O	O
patients	NN	O	O
with	NN	O	O
absorptive	NN	O	O
hypercalciuria	NN	O	O
(	NN	O	O
AH	NN	O	O
)	NN	O	O
and	NN	O	O
patients	NN	O	O
with	NN	O	O
high	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
due	NN	O	O
to	NN	O	O
acquired	NN	O	O
or	NN	O	O
transient	NN	O	O
disease	NN	O	O
states	NN	O	O
and	NN	O	O
the	NN	O	O
results	NN	O	O
compared	NN	O	O
to	NN	O	O
those	NN	O	O
in	NN	O	O
normal	NN	O	O
subjects	NN	O	O
.	NN	O	O

VDR	NN	O	O
concentration	NN	O	O
in	NN	O	O
resting	NN	O	B-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
not	NN	O	O
different	NN	O	O
between	NN	O	O
the	NN	O	O
three	NN	O	O
groups	NN	O	O
and	NN	O	O
represented	NN	O	O
constitutive	NN	O	O
receptor	NN	O	O
expression	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

Following	NN	O	O
activation	NN	O	O
with	NN	O	O
phytohemagglutinin	NN	O	B-protein
,	NN	O	O
patients	NN	O	B-cell_type
with	NN	O	O
hypercalcitriolemia	NN	O	O
demonstrated	NN	O	O
significantly	NN	O	O
greater	NN	O	O
VDR	NN	O	O
concentrations	NN	O	O
.	NN	O	O

Patients	NN	O	O
with	NN	O	O
AH	NN	O	O
demonstrated	NN	O	O
a	NN	O	O
normal	NN	O	O
value	NN	O	O
for	NN	O	O
the	NN	O	O
group	NN	O	O
,	NN	O	O
but	NN	O	O
6	NN	O	O
patients	NN	O	O
had	NN	O	O
significantly	NN	O	O
greater	NN	O	O
concentrations	NN	O	O
of	NN	O	O
VDR	NN	O	B-protein
despite	NN	O	O
normal	NN	O	O
plasma	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
four	NN	O	O
of	NN	O	O
the	NN	O	O
patients	NN	O	O
.	NN	O	O

Proliferation	NN	O	O
,	NN	O	O
as	NN	O	O
assessed	NN	O	O
from	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
thymidine	NN	O	O
incorporation	NN	O	O
was	NN	O	O
inversely	NN	O	O
correlated	NN	O	O
with	NN	O	O
serum	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
and	NN	O	O
was	NN	O	O
significant	NN	O	O
(	NN	O	O
R	NN	O	O
=	NN	O	O
-0.299	NN	O	O
,	NN	O	O
p	NN	O	O
=	NN	O	O
0.048	NN	O	O
)	NN	O	O
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
the	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
PBMC	NN	O	B-cell_type
provide	NN	O	O
a	NN	O	O
useful	NN	O	O
system	NN	O	O
for	NN	O	O
studying	NN	O	O
VDR	NN	O	B-protein
status	NN	O	O
in	NN	O	O
transient	NN	O	O
or	NN	O	O
acquired	NN	O	O
states	NN	O	O
of	NN	O	O
hypercalcitriolemia	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
the	NN	O	O
studies	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
absorptive	NN	O	O
hypercalciuria	NN	O	O
disclosed	NN	O	O
it	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
heterogeneous	NN	O	O
disorder	NN	O	O
,	NN	O	O
characterized	NN	O	O
by	NN	O	O
both	NN	O	O
vitamin	NN	O	O
D-dependent	NN	O	O
and	NN	O	O
D-independent	NN	O	O
forms	NN	O	O
of	NN	O	O
receptor	NN	O	O
up-regulation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Reactive	NN	O	O
oxygen	NN	O	O
intermediates	NN	O	O
activate	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
a	NN	O	O
tyrosine	NN	O	O
kinase-dependent	NN	O	O
mechanism	NN	O	O
and	NN	O	O
in	NN	O	O
combination	NN	O	O
with	NN	O	O
vanadate	NN	O	O
activate	NN	O	O
the	NN	O	O
p56lck	NN	O	B-protein
and	NN	O	I-protein
p59fyn	NN	O	I-protein
tyrosine	NN	O	I-protein
kinases	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
observed	NN	O	O
that	NN	O	O
ionizing	NN	O	O
radiation	NN	O	O
induces	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
B-lymphocyte	NN	O	I-cell_type
precursors	NN	O	I-cell_type
by	NN	O	O
stimulation	NN	O	O
of	NN	O	O
unidentified	NN	O	O
tyrosine	NN	O	B-protein
kinases	NN	O	I-protein
and	NN	O	O
this	NN	O	O
phosphorylation	NN	O	O
is	NN	O	O
substantially	NN	O	O
augmented	NN	O	O
by	NN	O	O
vanadate	NN	O	O
.	NN	O	O

Ionizing	NN	O	O
radiation	NN	O	O
generates	NN	O	O
reactive	NN	O	O
oxygen	NN	O	O
intermediates	NN	O	O
(	NN	O	O
ROI	NN	O	O
)	NN	O	O
.	NN	O	O

Because	NN	O	O
H2O2	NN	O	O
is	NN	O	O
a	NN	O	O
potent	NN	O	O
ROI	NN	O	O
generator	NN	O	O
that	NN	O	O
readily	NN	O	O
crosses	NN	O	O
the	NN	O	O
plasma	NN	O	O
membrane	NN	O	O
,	NN	O	O
we	NN	O	O
used	NN	O	O
H2O2	NN	O	O
to	NN	O	O
examine	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
ROI	NN	O	O
on	NN	O	O
signal	NN	O	O
transduction	NN	O	O
.	NN	O	O

We	NN	O	O
now	NN	O	O
provide	NN	O	O
evidence	NN	O	O
that	NN	O	O
the	NN	O	O
tyrosine	NN	O	O
kinase	NN	O	O
inhibitor	NN	O	O
herbimycin	NN	O	O
A	NN	O	O
and	NN	O	O
the	NN	O	O
free	NN	O	O
radical	NN	O	O
scavenger	NN	O	O
N-acetyl-cysteine	NN	O	O
inhibit	NN	O	O
both	NN	O	O
radiation-induced	NN	O	O
and	NN	O	O
H2O2-induced	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
indicating	NN	O	O
that	NN	O	O
activation	NN	O	O
triggered	NN	O	O
by	NN	O	O
ROI	NN	O	O
is	NN	O	O
dependent	NN	O	O
on	NN	O	O
tyrosine	NN	O	O
kinase	NN	O	O
activity	NN	O	O
.	NN	O	O

H2O2	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
stimulate	NN	O	O
Ins-1	NN	O	B-protein
,	NN	O	I-protein
4	NN	O	I-protein
,	NN	O	I-protein
5-P3	NN	O	I-protein
production	NN	O	O
in	NN	O	O
a	NN	O	O
tyrosine	NN	O	O
kinase-dependent	NN	O	O
manner	NN	O	O
and	NN	O	O
to	NN	O	O
induce	NN	O	O
calcium	NN	O	O
signals	NN	O	O
that	NN	O	O
were	NN	O	O
greatly	NN	O	O
augmented	NN	O	O
by	NN	O	O
vanadate	NN	O	O
.	NN	O	O

The	NN	O	O
synergistic	NN	O	O
induction	NN	O	O
of	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
by	NN	O	O
H2O2	NN	O	O
plus	NN	O	O
vanadate	NN	O	O
included	NN	O	O
physiologically	NN	O	O
relevant	NN	O	O
proteins	NN	O	O
such	NN	O	O
as	NN	O	O
PLC	NN	O	B-protein
gamma	NN	O	I-protein
1	NN	O	I-protein
.	NN	O	O

Although	NN	O	O
treatment	NN	O	O
of	NN	O	O
cells	NN	O	O
with	NN	O	O
H2O2	NN	O	O
alone	NN	O	O
did	NN	O	O
not	NN	O	O
affect	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
src	NN	O	B-protein
family	NN	O	I-protein
kinases	NN	O	I-protein
,	NN	O	O
treatment	NN	O	O
with	NN	O	O
H2O2	NN	O	O
plus	NN	O	O
vanadate	NN	O	O
led	NN	O	O
to	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
p56lck	NN	O	B-protein
and	NN	O	I-protein
p59fyn	NN	O	I-protein
tyrosine	NN	O	I-protein
kinases	NN	O	I-protein
.	NN	O	O

The	NN	O	O
combined	NN	O	O
inhibition	NN	O	O
of	NN	O	O
phosphatases	NN	O	B-protein
and	NN	O	O
activation	NN	O	O
of	NN	O	O
kinases	NN	O	B-protein
provides	NN	O	O
a	NN	O	O
potent	NN	O	O
mechanism	NN	O	O
for	NN	O	O
the	NN	O	O
synergistic	NN	O	O
effects	NN	O	O
of	NN	O	O
H2O2	NN	O	O
plus	NN	O	O
vanadate	NN	O	O
.	NN	O	O

Induction	NN	O	O
of	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
by	NN	O	O
ROI	NN	O	O
may	NN	O	O
thus	NN	O	O
lead	NN	O	O
to	NN	O	O
many	NN	O	O
of	NN	O	O
the	NN	O	O
pleiotropic	NN	O	O
effects	NN	O	O
of	NN	O	O
ROI	NN	O	O
in	NN	O	O
lymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
including	NN	O	O
downstream	NN	O	O
activation	NN	O	O
of	NN	O	O
PLC	NN	O	B-protein
gamma	NN	O	I-protein
1	NN	O	I-protein
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein

-DOCSTART-	O

Influence	NN	O	O
of	NN	O	O
sex	NN	O	B-protein
hormone	NN	O	I-protein
binding	NN	O	I-protein
globulin	NN	O	I-protein
and	NN	O	O
serum	NN	O	B-protein
albumin	NN	O	I-protein
on	NN	O	O
the	NN	O	O
conversion	NN	O	O
of	NN	O	O
androstenedione	NN	O	O
to	NN	O	O
testosterone	NN	O	O
by	NN	O	O
human	NN	O	B-cell_type
erythrocytes	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
influence	NN	O	O
of	NN	O	O
human	NN	O	B-protein
serum	NN	O	I-protein
albumin	NN	O	I-protein
and	NN	O	O
sex	NN	O	B-protein
hormone	NN	O	I-protein
binding	NN	O	I-protein
globulin	NN	O	I-protein
(	NN	O	O
SHBG	NN	O	B-protein
)	NN	O	O
on	NN	O	O
the	NN	O	O
enzymic	NN	O	O
conversion	NN	O	O
of	NN	O	O
androstenedione	NN	O	O
to	NN	O	O
testosterone	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
erythrocytes	NN	O	I-cell_type
was	NN	O	O
investigated	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

Total	NN	O	O
plasma	NN	O	O
and	NN	O	O
albumin	NN	O	B-protein
delayed	NN	O	O
the	NN	O	O
conversion	NN	O	O
rate	NN	O	O
of	NN	O	O
androstenedione	NN	O	O
,	NN	O	O
while	NN	O	O
SHBG	NN	O	B-protein
increased	NN	O	O
it	NN	O	O
markedly	NN	O	O
.	NN	O	O

The	NN	O	O
effect	NN	O	O
of	NN	O	O
SHBG	NN	O	B-protein
was	NN	O	O
largely	NN	O	O
abolished	NN	O	O
by	NN	O	O
heating	NN	O	O
to	NN	O	O
60	NN	O	O
degrees	NN	O	O
C	NN	O	O
for	NN	O	O
1	NN	O	O
h	NN	O	O
and	NN	O	O
by	NN	O	O
saturating	NN	O	O
its	NN	O	O
binding	NN	O	O
sites	NN	O	O
by	NN	O	O
DHT	NN	O	O
.	NN	O	O

The	NN	O	O
effect	NN	O	O
of	NN	O	O
both	NN	O	O
proteins	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
their	NN	O	O
concentration	NN	O	O
.	NN	O	O

It	NN	O	O
appears	NN	O	O
that	NN	O	O
the	NN	O	O
binding	NN	O	O
sites	NN	O	O
of	NN	O	O
albumin	NN	O	B-protein
provide	NN	O	O
a	NN	O	O
mechanism	NN	O	O
for	NN	O	O
retarding	NN	O	O
androstenedione	NN	O	O
uptake	NN	O	O
by	NN	O	O
the	NN	O	O
erythrocytes	NN	O	B-cell_type
and	NN	O	O
that	NN	O	O
the	NN	O	O
high	NN	O	O
binding	NN	O	O
affinity	NN	O	O
of	NN	O	O
SHBG	NN	O	B-protein
for	NN	O	O
testosterone	NN	O	O
facilitates	NN	O	O
the	NN	O	O
diffusion	NN	O	O
of	NN	O	O
this	NN	O	O
steroid	NN	O	O
out	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	O
and	NN	O	O
thus	NN	O	O
,	NN	O	O
displaces	NN	O	O
the	NN	O	O
chemical	NN	O	O
equilibrium	NN	O	O
within	NN	O	O
the	NN	O	O
cell	NN	O	O
.	NN	O	O

-DOCSTART-	O

Glucocorticoids	NN	O	O
and	NN	O	O
lymphocytes	NN	O	B-cell_type
.	NN	O	O

I	NN	O	O
.	NN	O	O

Increased	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
levels	NN	O	O
in	NN	O	O
antigen-stimulated	NN	O	B-cell_line
lymphocytes	NN	O	I-cell_line
.	NN	O	O

Recently	NN	O	O
a	NN	O	O
2-	NN	O	O
to	NN	O	O
3-fold	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
has	NN	O	O
been	NN	O	O
noted	NN	O	O
after	NN	O	O
in	NN	O	O
vitro	NN	O	O
mitogen	NN	O	O
stimulation	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
extend	NN	O	O
these	NN	O	O
observations	NN	O	O
to	NN	O	O
in	NN	O	O
vivo	NN	O	O
immunization	NN	O	O
.	NN	O	O

After	NN	O	O
unilateral	NN	O	O
immunization	NN	O	O
of	NN	O	O
adrenalectomized	NN	O	O
male	NN	O	O
rats	NN	O	O
,	NN	O	O
a	NN	O	O
50	NN	O	O
%	NN	O	O
increase	NN	O	O
in	NN	O	O
glucocorticoid	NN	O	O
receptor	NN	O	O
sites	NN	O	O
per	NN	O	O
cell	NN	O	O
,	NN	O	O
determined	NN	O	O
by	NN	O	O
binding	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
,	NN	O	O
was	NN	O	O
observed	NN	O	O
in	NN	O	O
cell	NN	O	O
suspensions	NN	O	O
of	NN	O	O
homolateral	NN	O	O
lymph	NN	O	O
nodes	NN	O	O
over	NN	O	O
those	NN	O	O
from	NN	O	O
the	NN	O	O
contralateral	NN	O	O
nonimmunized	NN	O	O
side	NN	O	O
of	NN	O	O
the	NN	O	O
same	NN	O	O
animal	NN	O	O
.	NN	O	O

The	NN	O	O
association	NN	O	O
constant	NN	O	O
for	NN	O	O
dexamethasone	NN	O	O
was	NN	O	O
similar	NN	O	O
in	NN	O	O
both	NN	O	O
groups	NN	O	O
,	NN	O	O
as	NN	O	O
was	NN	O	O
the	NN	O	O
stereospecificity	NN	O	O
for	NN	O	O
various	NN	O	O
steroids	NN	O	O
,	NN	O	O
the	NN	O	O
time	NN	O	O
course	NN	O	O
of	NN	O	O
cytoplasmic	NN	O	O
and	NN	O	O
nuclear	NN	O	O
association	NN	O	O
,	NN	O	O
and	NN	O	O
cytoplasmic-to-nuclear	NN	O	O
translocation	NN	O	O
.	NN	O	O

Despite	NN	O	O
a	NN	O	O
50	NN	O	O
%	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	O
receptor	NN	O	O
sites	NN	O	O
per	NN	O	O
cell	NN	O	O
,	NN	O	O
the	NN	O	O
cells	NN	O	O
from	NN	O	O
the	NN	O	O
homolateral	NN	O	O
and	NN	O	O
controlateral	NN	O	O
lymph	NN	O	O
nodes	NN	O	O
were	NN	O	O
equally	NN	O	O
sensitive	NN	O	O
to	NN	O	O
the	NN	O	O
inhibitory	NN	O	O
effects	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
,	NN	O	O
as	NN	O	O
determined	NN	O	O
by	NN	O	O
measurements	NN	O	O
of	NN	O	O
the	NN	O	O
incorporation	NN	O	O
of	NN	O	O
radiolabeled	NN	O	O
precursors	NN	O	O
of	NN	O	O
protein	NN	O	O
,	NN	O	O
RNA	NN	O	O
,	NN	O	O
and	NN	O	O
DNA	NN	O	O
,	NN	O	O
or	NN	O	O
measurements	NN	O	O
of	NN	O	O
in	NN	O	O
vitro	NN	O	O
cell	NN	O	O
survival	NN	O	O
.	NN	O	O

-DOCSTART-	O

Glucocorticoid	NN	O	O
receptors	NN	O	O
and	NN	O	O
actions	NN	O	O
in	NN	O	O
rat	NN	O	B-cell_type
thymocytes	NN	O	I-cell_type
and	NN	O	O
immunologically	NN	O	O
stimulated	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

After	NN	O	O
reviewing	NN	O	O
briefly	NN	O	O
our	NN	O	O
earlier	NN	O	O
studies	NN	O	O
on	NN	O	O
glucocorticoid	NN	O	O
receptors	NN	O	O
and	NN	O	O
mechanisms	NN	O	O
in	NN	O	O
thymus	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
we	NN	O	O
have	NN	O	O
outlined	NN	O	O
results	NN	O	O
from	NN	O	O
the	NN	O	O
following	NN	O	O
two	NN	O	O
areas	NN	O	O
of	NN	O	O
current	NN	O	O
interest	NN	O	O
in	NN	O	O
our	NN	O	O
laboratories	NN	O	O
:	NN	O	O
the	NN	O	O
``	NN	O	O
life-cycle	NN	O	O
''	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	O
receptors	NN	O	O
and	NN	O	O
complexes	NN	O	O
in	NN	O	O
thymus	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	O
receptors	NN	O	O
and	NN	O	O
sensitivity	NN	O	O
in	NN	O	O
immunologically	NN	O	B-cell_type
stimulated	NN	O	I-cell_type
human	NN	O	I-cell_type
peripheral	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Several	NN	O	O
of	NN	O	O
our	NN	O	O
results	NN	O	O
on	NN	O	O
energetics	NN	O	O
and	NN	O	O
kinetics	NN	O	O
of	NN	O	O
hormone	NN	O	O
binding	NN	O	O
to	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
rat	NN	O	B-cell_type
thymus	NN	O	I-cell_type
cells	NN	O	I-cell_type
seem	NN	O	O
to	NN	O	O
require	NN	O	O
extension	NN	O	O
of	NN	O	O
the	NN	O	O
simplest	NN	O	O
model	NN	O	O
of	NN	O	O
hormone-receptor	NN	O	O
transformations	NN	O	O
in	NN	O	O
intact	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

ATP-depletion	NN	O	O
experiments	NN	O	O
suggest	NN	O	O
the	NN	O	O
existence	NN	O	O
of	NN	O	O
a	NN	O	O
nonbinding	NN	O	O
form	NN	O	O
of	NN	O	O
the	NN	O	O
receptor	NN	O	O
;	NN	O	O
``	NN	O	O
chase	NN	O	O
''	NN	O	O
experiments	NN	O	O
suggest	NN	O	O
reaction	NN	O	O
of	NN	O	O
hormone	NN	O	O
directly	NN	O	O
with	NN	O	O
nuclear-bound	NN	O	O
receptor	NN	O	O
;	NN	O	O
experiments	NN	O	O
on	NN	O	O
depletion	NN	O	O
and	NN	O	O
replenishment	NN	O	O
of	NN	O	O
cytoplasmic	NN	O	B-protein
receptor	NN	O	I-protein
using	NN	O	O
cortisol	NN	O	O
and	NN	O	O
dexamethasone	NN	O	O
suggest	NN	O	O
the	NN	O	O
existence	NN	O	O
of	NN	O	O
at	NN	O	O
least	NN	O	O
two	NN	O	O
subpopulations	NN	O	O
of	NN	O	O
nuclear-bound	NN	O	B-protein
hormone-receptor	NN	O	I-protein
complex	NN	O	I-protein
.	NN	O	O

We	NN	O	O
have	NN	O	O
found	NN	O	O
that	NN	O	O
mitogen	NN	O	O
or	NN	O	O
immunologic	NN	O	O
stimulation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
in	NN	O	O
culture	NN	O	O
leads	NN	O	O
within	NN	O	O
24	NN	O	O
h	NN	O	O
or	NN	O	O
so	NN	O	O
to	NN	O	O
a	NN	O	O
striking	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
sites	NN	O	O
per	NN	O	O
cell	NN	O	O
.	NN	O	O

We	NN	O	O
believe	NN	O	O
this	NN	O	O
increase	NN	O	O
may	NN	O	O
be	NN	O	O
due	NN	O	O
to	NN	O	O
partial	NN	O	O
synchronization	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	O
population	NN	O	O
in	NN	O	O
a	NN	O	O
phase	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
in	NN	O	O
which	NN	O	O
receptor	NN	O	O
content	NN	O	O
is	NN	O	O
high	NN	O	O
.	NN	O	O

Contrary	NN	O	O
to	NN	O	O
the	NN	O	O
widely	NN	O	O
held	NN	O	O
view	NN	O	O
that	NN	O	O
mitogen-stimulated	NN	O	B-cell_type
cells	NN	O	I-cell_type
become	NN	O	O
insensitive	NN	O	O
to	NN	O	O
glucocorticoids	NN	O	O
,	NN	O	O
our	NN	O	O
experiments	NN	O	O
show	NN	O	O
that	NN	O	O
with	NN	O	O
respect	NN	O	O
to	NN	O	O
inhibition	NN	O	O
of	NN	O	O
thymidine	NN	O	O
and	NN	O	O
uridine	NN	O	O
incorporation	NN	O	O
and	NN	O	O
glucose	NN	O	O
uptake	NN	O	O
,	NN	O	O
the	NN	O	O
cells	NN	O	O
are	NN	O	O
highly	NN	O	O
sensitive	NN	O	O
to	NN	O	O
dexamethasone	NN	O	O
at	NN	O	O
24	NN	O	O
,	NN	O	O
48	NN	O	O
,	NN	O	O
and	NN	O	O
72	NN	O	O
h	NN	O	O
after	NN	O	O
stimulation	NN	O	O
with	NN	O	O
concanavalin	NN	O	B-protein
A	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Nuclear	NN	O	O
glucocorticoid	NN	O	O
binding	NN	O	O
in	NN	O	O
chronic	NN	O	B-cell_type
lymphatic	NN	O	I-cell_type
leukemia	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

A	NN	O	O
reliable	NN	O	O
procedure	NN	O	O
is	NN	O	O
described	NN	O	O
for	NN	O	O
isolating	NN	O	O
3H-triamcinolone	NN	O	B-protein
acetonide	NN	O	I-protein
(	NN	O	I-protein
3H-TA	NN	O	I-protein
)	NN	O	I-protein
receptor	NN	O	I-protein
complexes	NN	O	I-protein
from	NN	O	O
purified	NN	O	O
chronic	NN	O	O
lymphatic	NN	O	B-cell_type
leukemia	NN	O	I-cell_type
(	NN	O	I-cell_type
CLL	NN	O	I-cell_type
)	NN	O	I-cell_type
lymphocyte	NN	O	I-cell_type
nuclei	NN	O	O
,	NN	O	O
based	NN	O	O
on	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
carbobenzoxy-L-phenylalanine	NN	O	O
(	NN	O	O
CBZ-L-phe	NN	O	O
)	NN	O	O
to	NN	O	O
prevent	NN	O	O
breakdown	NN	O	O
of	NN	O	O
hormone-receptor	NN	O	B-protein
complexes	NN	O	I-protein
during	NN	O	O
extraction	NN	O	O
of	NN	O	O
nuclei	NN	O	O
with	NN	O	O
0.6M	NN	O	O
KCl	NN	O	O
.	NN	O	O

Using	NN	O	O
this	NN	O	O
method	NN	O	O
,	NN	O	O
specific	NN	O	O
nuclear	NN	O	O
glucocorticoid	NN	O	O
binding	NN	O	O
was	NN	O	O
demonstrated	NN	O	O
in	NN	O	O
14/14	NN	O	O
patients	NN	O	O
with	NN	O	O
untreated	NN	O	O
CLL	NN	O	O
.	NN	O	O

No	NN	O	O
correlation	NN	O	O
was	NN	O	O
found	NN	O	O
between	NN	O	O
levels	NN	O	O
of	NN	O	O
nuclear-associated	NN	O	O
3H-TA	NN	O	O
and	NN	O	O
peripheral	NN	O	O
white	NN	O	O
blood	NN	O	O
cell	NN	O	O
count	NN	O	O
or	NN	O	O
rosetting	NN	O	O
ability	NN	O	O
of	NN	O	O
circulating	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Multiple	NN	O	O
closely-linked	NN	O	O
NFAT/octamer	NN	O	B-DNA
and	NN	O	O
HMG	NN	O	B-DNA
I	NN	O	I-DNA
(	NN	O	I-DNA
Y	NN	O	I-DNA
)	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
are	NN	O	O
part	NN	O	O
of	NN	O	O
the	NN	O	O
interleukin-4	NN	O	O
promoter	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
here	NN	O	O
that	NN	O	O
the	NN	O	O
immediate	NN	O	B-DNA
upstream	NN	O	I-DNA
region	NN	O	I-DNA
(	NN	O	O
from	NN	O	O
position	NN	O	O
-12	NN	O	O
to	NN	O	O
-270	NN	O	O
)	NN	O	O
of	NN	O	O
the	NN	O	O
murine	NN	O	B-DNA
interleukin	NN	O	I-DNA
4	NN	O	I-DNA
(	NN	O	I-DNA
Il-4	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
harbors	NN	O	O
a	NN	O	O
strong	NN	O	O
cell-type	NN	O	O
specific	NN	O	O
transcriptional	NN	O	B-DNA
enhancer	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
T	NN	O	B-cell_type
lymphoma	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
Il-4	NN	O	B-DNA
promoter/enhancer	NN	O	I-DNA
is	NN	O	O
stimulated	NN	O	O
by	NN	O	O
phorbol	NN	O	O
esters	NN	O	O
,	NN	O	O
Ca++	NN	O	O
ionophores	NN	O	O
and	NN	O	O
agonists	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
A	NN	O	I-protein
and	NN	O	O
inhibited	NN	O	O
by	NN	O	O
low	NN	O	O
doses	NN	O	O
of	NN	O	O
the	NN	O	O
immunosuppressant	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
.	NN	O	O

The	NN	O	O
Il-4	NN	O	B-DNA
promoter/enhancer	NN	O	I-DNA
is	NN	O	O
transcriptionally	NN	O	O
inactive	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
lymphoma	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
HeLa	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

DNase	NN	O	O
I	NN	O	O
footprint	NN	O	O
protection	NN	O	O
experiments	NN	O	O
revealed	NN	O	O
six	NN	O	O
sites	NN	O	O
of	NN	O	O
the	NN	O	O
Il-4	NN	O	B-DNA
promoter/enhancer	NN	O	I-DNA
to	NN	O	O
be	NN	O	O
bound	NN	O	O
by	NN	O	O
nuclear	NN	O	B-protein
proteins	NN	O	I-protein
from	NN	O	O
lymphoid	NN	O	B-cell_type
and	NN	O	I-cell_type
myeloid	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Among	NN	O	O
them	NN	O	O
are	NN	O	O
four	NN	O	O
purine	NN	O	O
boxes	NN	O	O
which	NN	O	O
have	NN	O	O
been	NN	O	O
described	NN	O	O
to	NN	O	O
be	NN	O	O
important	NN	O	O
sequence	NN	O	O
motifs	NN	O	O
of	NN	O	O
the	NN	O	O
Il-2	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

They	NN	O	O
contain	NN	O	O
the	NN	O	O
motif	NN	O	O
GGAAA	NN	O	O
and	NN	O	O
are	NN	O	O
recognized	NN	O	O
by	NN	O	O
the	NN	O	O
inducible	NN	O	B-protein
and	NN	O	I-protein
cyclosporin	NN	O	I-protein
A-sensitive	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
NFAT-1	NN	O	I-protein
.	NN	O	O

Three	NN	O	O
of	NN	O	O
the	NN	O	O
Il-4	NN	O	B-DNA
NFAT-1	NN	O	I-DNA
sites	NN	O	I-DNA
are	NN	O	O
closely	NN	O	O
linked	NN	O	O
to	NN	O	O
weak	NN	O	O
binding	NN	O	O
sites	NN	O	O
of	NN	O	O
Octamer	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

Several	NN	O	O
purine	NN	O	O
boxes	NN	O	O
and	NN	O	O
an	NN	O	O
AT-rich	NN	O	B-DNA
protein-binding	NN	O	I-DNA
site	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
Il-4	NN	O	O
promoter	NN	O	O
are	NN	O	O
also	NN	O	O
recognized	NN	O	O
by	NN	O	O
the	NN	O	O
high	NN	O	B-protein
mobility	NN	O	I-protein
group	NN	O	I-protein
protein	NN	O	I-protein
HMG	NN	O	B-protein
I	NN	O	I-protein
(	NN	O	I-protein
Y	NN	O	I-protein
)	NN	O	O
.	NN	O	O

Whereas	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
NFAT-1	NN	O	B-protein
and	NN	O	O
Octamer	NN	O	B-protein
factors	NN	O	I-protein
enhance	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
Il-4	NN	O	B-DNA
promoter	NN	O	I-DNA
,	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
HMG	NN	O	O
I	NN	O	O
(	NN	O	O
Y	NN	O	O
)	NN	O	O
suppresses	NN	O	O
its	NN	O	O
activity	NN	O	O
and	NN	O	O
,	NN	O	O
therefore	NN	O	O
,	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
suppression	NN	O	O
of	NN	O	O
Il-4	NN	O	B-protein
transcription	NN	O	O
in	NN	O	O
resting	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Reversibility	NN	O	O
of	NN	O	O
the	NN	O	O
differentiated	NN	O	O
state	NN	O	O
in	NN	O	O
somatic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
de	NN	O	O
novo	NN	O	O
gene	NN	O	O
activation	NN	O	O
in	NN	O	O
multinucleated	NN	O	B-cell_type
heterokaryons	NN	O	I-cell_type
has	NN	O	O
shown	NN	O	O
that	NN	O	O
the	NN	O	O
differentiated	NN	O	O
state	NN	O	O
,	NN	O	O
although	NN	O	O
stable	NN	O	O
,	NN	O	O
is	NN	O	O
not	NN	O	O
irreversible	NN	O	O
,	NN	O	O
and	NN	O	O
can	NN	O	O
be	NN	O	O
reprogrammed	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
appropriate	NN	O	O
combinations	NN	O	O
of	NN	O	O
trans-acting	NN	O	B-protein
regulatory	NN	O	I-protein
molecules	NN	O	I-protein
.	NN	O	O

These	NN	O	O
properties	NN	O	O
have	NN	O	O
been	NN	O	O
exploited	NN	O	O
to	NN	O	O
design	NN	O	O
strategies	NN	O	O
for	NN	O	O
identifying	NN	O	O
novel	NN	O	O
regulators	NN	O	O
of	NN	O	O
cellular	NN	O	O
differentiation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Phosphatidylcholine	NN	O	O
hydrolysis	NN	O	O
activates	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
increases	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
replication	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
and	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
tested	NN	O	O
whether	NN	O	O
breakdown	NN	O	O
of	NN	O	O
phosphatidylcholine	NN	O	O
(	NN	O	O
PC	NN	O	O
)	NN	O	O
initiated	NN	O	O
by	NN	O	O
exogenous	NN	O	O
addition	NN	O	O
of	NN	O	O
a	NN	O	O
PC-specific	NN	O	B-protein
phospholipase	NN	O	I-protein
C	NN	O	I-protein
(	NN	O	O
PC-PLC	NN	O	B-protein
)	NN	O	O
from	NN	O	O
Bacillus	NN	O	O
cereus	NN	O	O
or	NN	O	O
by	NN	O	O
endogenous	NN	O	O
overexpression	NN	O	O
of	NN	O	O
PC-PLC	NN	O	B-protein
induces	NN	O	O
functional	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
increases	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
activity	NN	O	O
.	NN	O	O

PC-PLC-activated	NN	O	O
hydrolysis	NN	O	O
of	NN	O	O
PC	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
induce	NN	O	O
bona	NN	O	O
fide	NN	O	O
p50/p65	NN	O	B-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
binding	NN	O	O
activity	NN	O	O
in	NN	O	O
three	NN	O	O
different	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
of	NN	O	O
human	NN	O	O
or	NN	O	O
murine	NN	O	O
origin	NN	O	O
.	NN	O	O

No	NN	O	O
significant	NN	O	O
changes	NN	O	O
in	NN	O	O
the	NN	O	O
turnover	NN	O	O
of	NN	O	O
other	NN	O	O
cellular	NN	O	O
phospholipids	NN	O	O
were	NN	O	O
detected	NN	O	O
in	NN	O	O
PC-PLC-treated	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	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
PC-PLC	NN	O	B-protein
did	NN	O	O
not	NN	O	O
depend	NN	O	O
on	NN	O	O
de	NN	O	O
novo	NN	O	O
synthesis	NN	O	O
of	NN	O	O
proteins	NN	O	O
or	NN	O	O
autocrine	NN	O	O
secretion	NN	O	O
of	NN	O	O
either	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
or	NN	O	O
interleukin	NN	O	B-protein
1	NN	O	I-protein
.	NN	O	O

In	NN	O	O
human	NN	O	B-cell_line
monocytic	NN	O	I-cell_line
and	NN	O	I-cell_line
lymphoblastoid	NN	O	I-cell_line
T-cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	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
PC-PLC	NN	O	B-protein
resulted	NN	O	O
in	NN	O	O
clear	NN	O	O
induction	NN	O	O
of	NN	O	O
luciferase	NN	O	B-protein
expression	NN	O	O
vectors	NN	O	O
placed	NN	O	O
under	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
synthetic	NN	O	B-DNA
kappa	NN	O	I-DNA
B	NN	O	I-DNA
enhancers	NN	O	I-DNA
or	NN	O	O
wild	NN	O	O
type	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
kappa	NN	O	O
B-mutated	NN	O	O
,	NN	O	O
HIV	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
constructs	NN	O	I-DNA
.	NN	O	O

HIV	NN	O	O
replication	NN	O	O
was	NN	O	O
increased	NN	O	O
by	NN	O	O
PC-PLC	NN	O	B-protein
in	NN	O	O
chronically	NN	O	O
infected	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
T	NN	O	O
lymphocytes	NN	O	B-cell_type
.	NN	O	O

NF-kappa	NN	O	O
B	NN	O	O
activation	NN	O	O
promoted	NN	O	O
by	NN	O	O
addition	NN	O	O
of	NN	O	O
exogenous	NN	O	B-protein
PC-PLC	NN	O	I-protein
correlated	NN	O	O
with	NN	O	O
an	NN	O	O
intense	NN	O	O
production	NN	O	O
of	NN	O	O
diacylglycerol	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
addition	NN	O	O
of	NN	O	O
a	NN	O	O
phosphatidylinositol-specific	NN	O	B-protein
PLC	NN	O	I-protein
from	NN	O	O
B.cereus	NN	O	O
also	NN	O	O
induced	NN	O	O
diacylglycerol	NN	O	O
but	NN	O	O
did	NN	O	O
not	NN	O	O
activate	NN	O	O
kappa	NN	O	B-protein
B	NN	O	I-protein
enhancer-directed	NN	O	I-protein
vectors	NN	O	I-protein
.	NN	O	O

PC-PLC-induced	NN	O	O
NF-kappa	NN	O	O
B	NN	O	O
activation	NN	O	O
could	NN	O	O
not	NN	O	O
be	NN	O	O
blocked	NN	O	O
by	NN	O	O
a	NN	O	O
specific	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
phorbol	NN	O	B-protein
ester-inducible	NN	O	I-protein
protein	NN	O	I-protein
kinases	NN	O	I-protein
C	NN	O	I-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
a	NN	O	O
cellular	NN	O	O
transduction	NN	O	O
pathway	NN	O	O
,	NN	O	O
dependent	NN	O	O
on	NN	O	O
specific	NN	O	O
PC	NN	O	O
breakdown	NN	O	O
,	NN	O	O
is	NN	O	O
functional	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
may	NN	O	O
be	NN	O	O
used	NN	O	O
by	NN	O	O
various	NN	O	O
transmembrane	NN	O	O
receptors	NN	O	O
to	NN	O	O
activate	NN	O	O
HIV	NN	O	O
transcription	NN	O	O
through	NN	O	O
NF-kappa	NN	O	O
B-dependent	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
HIV	NN	O	B-DNA
enhancer	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Novel	NN	O	O
mechanism	NN	O	O
for	NN	O	O
inhibition	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
glucocorticoids	NN	O	O
.	NN	O	O

Glucocorticoids	NN	O	O
inhibit	NN	O	O
signal	NN	O	O
transduction	NN	O	O
through	NN	O	O
IL-2	NN	O	B-protein
receptor	NN	O	I-protein
.	NN	O	O

Interaction	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
with	NN	O	O
its	NN	O	O
high	NN	O	B-protein
affinity	NN	O	I-protein
membrane	NN	O	I-protein
receptor	NN	O	I-protein
complex	NN	O	I-protein
(	NN	O	O
IL-2R	NN	O	B-protein
)	NN	O	O
present	NN	O	O
on	NN	O	O
activated	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
induces	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
and	NN	O	O
mediates	NN	O	O
effector	NN	O	O
functions	NN	O	O
.	NN	O	O

Glucocorticoids	NN	O	O
inhibit	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
by	NN	O	O
inhibiting	NN	O	O
TCR-mediated	NN	O	O
signal	NN	O	O
transduction	NN	O	O
.	NN	O	O

We	NN	O	O
asked	NN	O	O
whether	NN	O	O
they	NN	O	O
also	NN	O	O
inhibit	NN	O	O
the	NN	O	O
action	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
by	NN	O	O
inhibiting	NN	O	O
signal	NN	O	O
transduction	NN	O	O
through	NN	O	O
IL-2R	NN	O	B-protein
.	NN	O	O

Human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
stimulated	NN	O	O
with	NN	O	O
PMA	NN	O	O
for	NN	O	O
48	NN	O	O
h	NN	O	O
(	NN	O	O
PMA	NN	O	B-cell_line
blasts	NN	O	I-cell_line
)	NN	O	O
,	NN	O	O
were	NN	O	O
incubated	NN	O	O
with	NN	O	O
IL-2	NN	O	B-protein
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
incremental	NN	O	O
dosages	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
(	NN	O	O
Dex	NN	O	O
;	NN	O	O
10	NN	O	O
(	NN	O	O
-5	NN	O	O
)	NN	O	O
-10	NN	O	O
(	NN	O	O
-9	NN	O	O
)	NN	O	O
M	NN	O	O
)	NN	O	O
.	NN	O	O

Dex	NN	O	O
inhibited	NN	O	O
the	NN	O	O
IL-2-dependent	NN	O	O
proliferation	NN	O	O
of	NN	O	O
PMA	NN	O	B-cell_line
blasts	NN	O	I-cell_line
in	NN	O	O
a	NN	O	O
dose-dependent	NN	O	O
fashion	NN	O	O
(	NN	O	O
IC50	NN	O	O
,	NN	O	O
5	NN	O	O
x	NN	O	O
10	NN	O	O
(	NN	O	O
-8	NN	O	O
)	NN	O	O
M	NN	O	O
)	NN	O	O
.	NN	O	O

Cell	NN	O	O
surface	NN	O	O
expression	NN	O	O
of	NN	O	O
IL-2R	NN	O	B-protein
alpha-	NN	O	I-protein
and	NN	O	I-protein
beta-chains	NN	O	I-protein
as	NN	O	O
determined	NN	O	O
by	NN	O	O
immunofluorescence	NN	O	O
analysis	NN	O	O
was	NN	O	O
not	NN	O	O
affected	NN	O	O
by	NN	O	O
Dex	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
Scatchard	NN	O	O
plot	NN	O	O
analysis	NN	O	O
of	NN	O	O
125I-labeled	NN	O	B-protein
IL-2	NN	O	I-protein
showed	NN	O	O
that	NN	O	O
Dex	NN	O	O
did	NN	O	O
not	NN	O	O
affect	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
thus	NN	O	O
suggesting	NN	O	O
that	NN	O	O
inhibition	NN	O	O
is	NN	O	O
due	NN	O	O
to	NN	O	O
a	NN	O	O
postreceptor	NN	O	O
effect	NN	O	O
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
T	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
by	NN	O	O
Dex	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
decreased	NN	O	O
IL-2-dependent	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
several	NN	O	O
intracellular	NN	O	B-protein
proteins	NN	O	I-protein
and	NN	O	O
decreased	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
the	NN	O	O
retinoblastoma	NN	O	B-protein
gene	NN	O	I-protein
product	NN	O	I-protein
Rb	NN	O	I-protein
,	NN	O	O
a	NN	O	O
protein	NN	O	O
essential	NN	O	O
for	NN	O	O
controlling	NN	O	O
the	NN	O	O
progression	NN	O	O
of	NN	O	O
cells	NN	O	O
through	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
.	NN	O	O

IL-2-dependent	NN	O	O
IL-2R	NN	O	O
alpha	NN	O	O
expression	NN	O	O
in	NN	O	O
PMA	NN	O	B-cell_line
blasts	NN	O	I-cell_line
and	NN	O	O
NF-kB	NN	O	B-protein
induction	NN	O	O
in	NN	O	O
resting	NN	O	B-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
also	NN	O	O
inhibited	NN	O	O
by	NN	O	O
Dex	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
glucocorticoids	NN	O	O
inhibit	NN	O	O
preactivated	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
by	NN	O	O
down-regulating	NN	O	O
signal	NN	O	O
transduction	NN	O	O
through	NN	O	O
IL-2R	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Chronic	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
infection	NN	O	O
stimulates	NN	O	O
distinct	NN	O	O
NF-kappa	NN	O	O
B/rel	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activities	NN	O	O
in	NN	O	O
myelomonoblastic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
relationship	NN	O	O
between	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
(	NN	O	O
HIV-1	NN	O	O
)	NN	O	O
infection	NN	O	O
and	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
NF-kappa	NN	O	O
B	NN	O	O
binding	NN	O	O
activity	NN	O	O
was	NN	O	O
examined	NN	O	O
in	NN	O	O
a	NN	O	O
myeloid	NN	O	B-cell_line
cell	NN	O	I-cell_line
model	NN	O	I-cell_line
of	NN	O	O
HIV-1	NN	O	O
infection	NN	O	O
derived	NN	O	O
from	NN	O	O
the	NN	O	O
PLB-985	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

Chronic	NN	O	O
infection	NN	O	O
of	NN	O	O
PLB-985	NN	O	B-cell_line
cells	NN	O	I-cell_line
led	NN	O	O
to	NN	O	O
increased	NN	O	O
monocyte-specific	NN	O	O
surface	NN	O	O
marker	NN	O	O
expression	NN	O	O
,	NN	O	O
increased	NN	O	O
c-fms	NN	O	O
gene	NN	O	O
transcription	NN	O	O
,	NN	O	O
and	NN	O	O
morphological	NN	O	O
alterations	NN	O	O
consistent	NN	O	O
with	NN	O	O
differentiation	NN	O	O
along	NN	O	O
the	NN	O	O
monocytic	NN	O	O
pathway	NN	O	O
.	NN	O	O

PLB-IIIB	NN	O	B-cell_line
cells	NN	O	I-cell_line
displayed	NN	O	O
a	NN	O	O
constitutive	NN	O	O
NF-kappa	NN	O	O
B-like	NN	O	O
binding	NN	O	O
activity	NN	O	O
that	NN	O	O
was	NN	O	O
distinct	NN	O	O
from	NN	O	O
that	NN	O	O
induced	NN	O	O
by	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
or	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
treatment	NN	O	O
of	NN	O	O
the	NN	O	O
parental	NN	O	B-cell_line
PLB-985	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

This	NN	O	O
unique	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
consisted	NN	O	O
of	NN	O	O
proteins	NN	O	O
of	NN	O	O
70	NN	O	B-protein
,	NN	O	I-protein
90	NN	O	I-protein
,	NN	O	I-protein
and	NN	O	I-protein
100	NN	O	I-protein
kDa	NN	O	I-protein
with	NN	O	O
a	NN	O	O
high	NN	O	O
degree	NN	O	O
of	NN	O	O
binding	NN	O	O
specificity	NN	O	O
for	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
site	NN	O	I-protein
within	NN	O	O
the	NN	O	O
PRDII	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
beta	NN	O	B-protein
interferon	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
characterize	NN	O	O
the	NN	O	O
nature	NN	O	O
of	NN	O	O
these	NN	O	O
proteins	NN	O	O
and	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
binding	NN	O	O
of	NN	O	O
these	NN	O	O
proteins	NN	O	O
is	NN	O	O
also	NN	O	O
induced	NN	O	O
following	NN	O	O
Sendai	NN	O	O
paramyxovirus	NN	O	O
infection	NN	O	O
.	NN	O	O

The	NN	O	O
70-kDa	NN	O	B-protein
protein	NN	O	I-protein
corresponds	NN	O	O
to	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
RelA	NN	O	I-protein
(	NN	O	I-protein
p65	NN	O	I-protein
)	NN	O	I-protein
subunit	NN	O	I-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
activated	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
an	NN	O	O
acute	NN	O	O
paramyxovirus	NN	O	O
infection	NN	O	O
or	NN	O	O
a	NN	O	O
chronic	NN	O	O
HIV-1	NN	O	O
infection	NN	O	O
.	NN	O	O

Virus	NN	O	O
infection	NN	O	O
does	NN	O	O
not	NN	O	O
appear	NN	O	O
to	NN	O	O
alter	NN	O	O
the	NN	O	O
amount	NN	O	O
of	NN	O	O
RelA	NN	O	B-protein
(	NN	O	O
p65	NN	O	B-protein
)	NN	O	O
or	NN	O	O
NFKB1	NN	O	B-protein
(	NN	O	O
p50	NN	O	B-protein
)	NN	O	O
but	NN	O	O
rather	NN	O	O
affects	NN	O	O
the	NN	O	O
capacity	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
sequester	NN	O	O
RelA	NN	O	B-protein
(	NN	O	I-protein
p65	NN	O	I-protein
)	NN	O	I-protein
,	NN	O	O
therefore	NN	O	O
leading	NN	O	O
to	NN	O	O
constitutive	NN	O	O
levels	NN	O	O
of	NN	O	O
RelA	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
and	NN	O	O
to	NN	O	O
increased	NN	O	O
levels	NN	O	O
of	NN	O	O
NF-kappa	NN	O	O
B-dependent	NN	O	O
gene	NN	O	O
activity	NN	O	O
.	NN	O	O

The	NN	O	O
virally	NN	O	O
induced	NN	O	O
90-	NN	O	B-protein
to	NN	O	I-protein
100-kDa	NN	O	I-protein
proteins	NN	O	I-protein
have	NN	O	O
a	NN	O	O
distinct	NN	O	O
binding	NN	O	O
specificity	NN	O	O
for	NN	O	O
the	NN	O	O
PRDII	NN	O	B-DNA
domain	NN	O	I-DNA
and	NN	O	O
an	NN	O	O
AT-rich	NN	O	B-DNA
sequence	NN	O	I-DNA
but	NN	O	O
do	NN	O	O
not	NN	O	O
cross-react	NN	O	O
with	NN	O	O
NF-kappa	NN	O	O
B	NN	O	O
subunit-specific	NN	O	O
antisera	NN	O	O
directed	NN	O	O
against	NN	O	O
NFKB1	NN	O	B-protein
(	NN	O	O
p105	NN	O	B-protein
or	NN	O	O
p50	NN	O	B-protein
)	NN	O	O
,	NN	O	O
NFKB2	NN	O	B-protein
(	NN	O	O
p100	NN	O	B-protein
or	NN	O	O
p52	NN	O	B-protein
)	NN	O	O
,	NN	O	O
RelA	NN	O	B-protein
(	NN	O	O
p65	NN	O	B-protein
)	NN	O	O
,	NN	O	O
or	NN	O	O
c-rel	NN	O	B-protein
.	NN	O	O

DNA	NN	O	O
binding	NN	O	O
of	NN	O	O
the	NN	O	O
90-	NN	O	B-protein
to	NN	O	I-protein
100-kDa	NN	O	I-protein
proteins	NN	O	I-protein
was	NN	O	O
not	NN	O	O
inhibited	NN	O	O
by	NN	O	O
recombinant	NN	O	B-protein
I	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha/MAD-3	NN	O	I-protein
and	NN	O	O
was	NN	O	O
resistant	NN	O	O
to	NN	O	O
tryptic	NN	O	O
digestion	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
these	NN	O	O
proteins	NN	O	O
may	NN	O	O
not	NN	O	O
be	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
related	NN	O	I-protein
.	NN	O	O

Transient	NN	O	O
cotransfection	NN	O	O
experiments	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
RelA	NN	O	O
and	NN	O	O
NFKB1	NN	O	O
expression	NN	O	O
maximally	NN	O	O
stimulated	NN	O	O
HIV-1	NN	O	B-DNA
LTR-	NN	O	I-DNA
and	NN	O	I-DNA
NF-kappa	NN	O	I-DNA
B-dependent	NN	O	I-DNA
reporter	NN	O	I-DNA
genes	NN	O	I-DNA
;	NN	O	O
differences	NN	O	O
in	NN	O	O
NF-kappa	NN	O	O
B-like	NN	O	O
binding	NN	O	O
activity	NN	O	O
were	NN	O	O
also	NN	O	O
reflected	NN	O	O
in	NN	O	O
higher	NN	O	O
constitutive	NN	O	O
levels	NN	O	O
of	NN	O	O
NF-kappa	NN	O	O
B-regulated	NN	O	O
gene	NN	O	O
expression	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
.	NN	O	O

-DOCSTART-	O

Presence	NN	O	O
of	NN	O	O
estrogen-binding	NN	O	O
sites	NN	O	O
on	NN	O	O
macrophage-like	NN	O	B-cell_type
synoviocytes	NN	O	I-cell_type
and	NN	O	O
CD8+	NN	O	B-cell_line
,	NN	O	I-cell_line
CD29+	NN	O	I-cell_line
,	NN	O	I-cell_line
CD45RO+	NN	O	I-cell_line
T	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
in	NN	O	O
normal	NN	O	O
and	NN	O	O
rheumatoid	NN	O	O
synovium	NN	O	O
.	NN	O	O

OBJECTIVE	NN	O	O
.	NN	O	O

To	NN	O	O
study	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
estrogen-binding	NN	O	O
sites	NN	O	O
(	NN	O	O
EBS	NN	O	O
)	NN	O	O
in	NN	O	O
the	NN	O	O
synovial	NN	O	O
tissues	NN	O	O
of	NN	O	O
male	NN	O	O
and	NN	O	O
female	NN	O	O
patients	NN	O	O
with	NN	O	O
rheumatoid	NN	O	O
arthritis	NN	O	O
(	NN	O	O
RA	NN	O	O
)	NN	O	O
and	NN	O	O
in	NN	O	O
age-	NN	O	O
and	NN	O	O
sex-matched	NN	O	O
healthy	NN	O	O
controls	NN	O	O
.	NN	O	O

METHODS	NN	O	O
.	NN	O	O

Both	NN	O	O
type	NN	O	O
1	NN	O	O
(	NN	O	O
high	NN	O	O
affinity	NN	O	O
,	NN	O	O
low	NN	O	O
binding	NN	O	O
capacity	NN	O	O
)	NN	O	O
and	NN	O	O
type	NN	O	O
2	NN	O	O
(	NN	O	O
reduced	NN	O	O
affinity	NN	O	O
,	NN	O	O
higher	NN	O	O
binding	NN	O	O
capacity	NN	O	O
)	NN	O	O
EBS	NN	O	O
were	NN	O	O
investigated	NN	O	O
in	NN	O	O
both	NN	O	O
soluble	NN	O	O
and	NN	O	O
nuclear	NN	O	O
fractions	NN	O	O
of	NN	O	O
homogenized	NN	O	O
synovial	NN	O	O
tissue	NN	O	O
samples	NN	O	O
by	NN	O	O
a	NN	O	O
dextran-coated	NN	O	O
charcoal	NN	O	O
method	NN	O	O
.	NN	O	O

To	NN	O	O
determine	NN	O	O
what	NN	O	O
type	NN	O	O
of	NN	O	O
synovial	NN	O	B-cell_type
cell	NN	O	I-cell_type
was	NN	O	O
positive	NN	O	O
for	NN	O	O
EBS	NN	O	O
,	NN	O	O
cryosections	NN	O	O
of	NN	O	O
synovial	NN	O	O
tissues	NN	O	O
were	NN	O	O
immunostained	NN	O	O
with	NN	O	O
a	NN	O	O
specific	NN	O	O
monoclonal	NN	O	B-protein
anti-estrogen	NN	O	I-protein
receptor	NN	O	I-protein
antibody	NN	O	I-protein
(	NN	O	O
anti-ER	NN	O	B-protein
MAb	NN	O	I-protein
)	NN	O	O
using	NN	O	O
both	NN	O	O
immunofluorescence	NN	O	O
and	NN	O	O
immunoperoxidase	NN	O	O
techniques	NN	O	O
.	NN	O	O

Double	NN	O	O
immunostaining	NN	O	O
with	NN	O	O
the	NN	O	O
anti-ER	NN	O	B-protein
MAb	NN	O	I-protein
and	NN	O	O
with	NN	O	O
specific	NN	O	B-protein
MAb	NN	O	I-protein
to	NN	O	O
detect	NN	O	O
different	NN	O	O
macrophage	NN	O	B-protein
antigens	NN	O	I-protein
(	NN	O	O
Ber-MAC3	NN	O	B-protein
,	NN	O	O
MAC387	NN	O	B-protein
,	NN	O	O
CD68	NN	O	B-protein
)	NN	O	O
and	NN	O	O
CD8+	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
subsets	NN	O	I-cell_line
(	NN	O	O
CD29+	NN	O	B-cell_line
,	NN	O	O
CD45RO+	NN	O	B-cell_line
and	NN	O	O
CD29-	NN	O	B-cell_line
,	NN	O	O
CD45RO-	NN	O	B-cell_line
)	NN	O	O
was	NN	O	O
performed	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
.	NN	O	O

Higher	NN	O	O
affinity	NN	O	O
EBS	NN	O	O
were	NN	O	O
found	NN	O	O
mostly	NN	O	O
in	NN	O	O
nuclear	NN	O	O
cell	NN	O	O
fractions	NN	O	O
of	NN	O	O
either	NN	O	O
RA	NN	O	O
or	NN	O	O
control	NN	O	O
synovial	NN	O	O
tissues	NN	O	O
(	NN	O	O
28	NN	O	O
of	NN	O	O
the	NN	O	O
33	NN	O	O
)	NN	O	O
.	NN	O	O

These	NN	O	O
EBS	NN	O	O
were	NN	O	O
present	NN	O	O
to	NN	O	O
a	NN	O	O
lesser	NN	O	O
extent	NN	O	O
in	NN	O	O
soluble	NN	O	O
cell	NN	O	O
fractions	NN	O	O
(	NN	O	O
11	NN	O	O
of	NN	O	O
the	NN	O	O
33	NN	O	O
)	NN	O	O
.	NN	O	O

Immunostaining	NN	O	O
showed	NN	O	O
the	NN	O	O
estrogen	NN	O	B-cell_type
receptor-positive	NN	O	I-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
be	NN	O	O
the	NN	O	O
macrophage-like	NN	O	B-cell_type
synoviocytes	NN	O	I-cell_type
and	NN	O	O
the	NN	O	O
CD8+	NN	O	B-cell_line
,	NN	O	O
CD29+	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
both	NN	O	O
in	NN	O	O
RA	NN	O	O
and	NN	O	O
in	NN	O	O
control	NN	O	O
synovial	NN	O	O
tissues	NN	O	O
.	NN	O	O

Higher	NN	O	O
nuclear	NN	O	O
content	NN	O	O
of	NN	O	O
EBS	NN	O	O
was	NN	O	O
consistent	NN	O	O
with	NN	O	O
more	NN	O	O
intense	NN	O	O
nuclear	NN	O	O
staining	NN	O	O
of	NN	O	O
synoviocytes	NN	O	B-cell_type
and	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

CONCLUSION	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
conceivable	NN	O	O
that	NN	O	O
the	NN	O	O
immunomodulatory	NN	O	O
activity	NN	O	O
exerted	NN	O	O
by	NN	O	O
estrogens	NN	O	O
is	NN	O	O
at	NN	O	O
least	NN	O	O
partly	NN	O	O
mediated	NN	O	O
through	NN	O	O
their	NN	O	O
interaction	NN	O	O
with	NN	O	O
EBS	NN	O	O
that	NN	O	O
are	NN	O	O
present	NN	O	O
on	NN	O	O
macrophage-like	NN	O	B-cell_type
synoviocytes	NN	O	I-cell_type
,	NN	O	O
functioning	NN	O	O
as	NN	O	O
antigen-processing	NN	O	B-cell_type
and	NN	O	I-cell_type
antigen-presenting	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
on	NN	O	O
antigen-experienced	NN	O	B-cell_line
(	NN	O	I-cell_line
memory	NN	O	I-cell_line
)	NN	O	I-cell_line
CD8+	NN	O	I-cell_line
T	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
(	NN	O	O
CD29+	NN	O	B-cell_line
,	NN	O	O
CD45RO+	NN	O	B-cell_line

