| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
and IL-12 by STAT6-Dependent and -Independent Mechanisms1
The Biomedical Research Centre, University of British Columbia, Vancouver, BC, Canada
 |
Abstract |
|---|
 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
or IL-12 by macrophages
had both STAT6-dependent and -independent components, depending on the
stimuli. IL-4 failed to inhibit the release of TNF- or IL-12 from
STAT6 null macrophages stimulated with LPS alone. However, IL-4 still
induced significant inhibition of the production of TNF- and IL-12
from STAT6 null macrophages that were stimulated with the more
physiologically relevant combination of LPS and IFN-
. These data
show that STAT6 is required for the IL-4-mediated inhibition of the
production of TNF- and IL-12 stimulated by LPS alone, but that IL-4
also activates distinct, STAT6 independent mechanism(s) that inhibit
the IFN--mediated enhancement of IL-12 and TNF-
production. |
Introduction |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
and IL-12 may lead to new
ways to treat diseases like rheumatoid arthritis (1). The
antiinflammatory properties of IL-4 appear to be mediated at multiple
levels. IL-4 can directly suppress the production of proinflammatory
cytokines at the levels of transcription (2) or message stability (3, 4), as well as antagonize the proinflammatory effects of IFN- on
superoxide production by macrophages (5), expression of cell surface
Ags (6), and cytokine production (7, 8). The suppressive effects of
IL-4 on the production of IL-12 by accessory cells appears to be a
major mechanism that inhibits the generation of Th1 cells (9). However,
the intracellular signals through which IL-4 exerts these direct and
indirect effects are largely unknown.
Given its association with IL-4 effects, the STAT6 pathway is a good
candidate for the molecular mechanism that mediates the inhibitory
effects of IL-4 on macrophages. The analysis of mice that lack
functional STAT6 genes has shown that STAT6 plays an essential role in
many of the biological functions of IL-4, including the production of
Th2 cells, the switching of B-cells to the production of IgE, the
induction of Ag-dependent airway hyperresponsiveness, and in
IL-4-mediated up-regulation of cell-surface molecules, such as MHC
class II and CD23 (10, 11, 12, 13, 14). However, it is not clear whether the
suppressive effects of IL-4 and IL-13 on the production of TNF- and
IL-12 by macrophages are also dependent on STAT6.
During an infection, LPS stimulates the production of multiple
cytokines including TNF-, IL-12, and IL-18. This production of IL-12
and IL-18 in turn leads to the production of IFN- by NK cells (15).
The regulation of TNF- production is complex and occurs at the level
of transcription (16, 17) and message stability (4, 18) and is
suppressed by IL-4, IL-10, or IL-13 (2, 19, 20). As is the case for
TNF-, the production of IL-12 is greatly enhanced by IFN- (21)
and is suppressed by IL-4, IL-10, or IL-13 (22, 23).
To better understand the mechanistic basis of the antiinflammatory
effects of IL-4, we investigated whether activation of STAT6 was
required for the IL-4-mediated inhibition of the release of TNF- and
IL-12 from macrophages stimulated with LPS, or LPS in presence of
IFN-. Using STAT6 null macrophages, we found that STAT6 was
essential for the IL-4-mediated inhibition of the release of TNF-
and IL-12 from macrophages stimulated with LPS alone. However IL-4
activated another, STAT6-independent mechanism(s) that inhibited the
increased, and arguably more physiological important, production of
TNF- and IL-12 that occurred in macrophages that encountered LPS in
the presence of IFN-.
|
Materials and Methods |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
STAT6 null mice were obtained from Dr. Michael Grusby (Harvard
Medical School, Boston, MA) and wild-type BALB/c mice were obtained
from The Jackson Laboratory (Bar Harbor, ME). Age- and sex-matched
BALB/c or STAT6 null mice were injected with 2 ml of 2% (w/v)
thioglycollate broth (Difco, Detroit, MI) into the peritoneal cavity.
Five days later, peritoneal exudate cells were harvested by flushing
peritoneal cavities with PBS. Cells were plated in a 48-well plate
(Corning, Cambridge, MA) at an initial density of 2 x
106 cells/ml in a 500-µl volume of RPMI 1640 (Life
Technologies, Grand Island, NY), supplemented with 10% FCS (Intergen,
Purchase, NY) and 50 µM 2-ME. Sixteen hours later, the wells were
washed to remove all nonadherent cells and 200 µl of fresh medium was
added together with LPS and cytokines. This population of adherent
peritoneal exudate macrophages is referred to hereafter as
PEC.4 The femurs from the same mice were harvested and
flushed with 5 ml of PBS. Bone marrow cells were cultured in medium
that was supplemented with 20% L cell conditioned medium
(LCCM) as a convenient source of CSF-1.
After 36–48 h, before the macrophages differentiated and became
adherent, the nonadherent cells were harvested and plated at an initial
density of 2 x 105 cells/ml in a 1-ml volume in
24-well plates (Nunc, Roskilde, Denmark). Day 5 ex vivo, when the
bottoms of the wells were covered with confluent layers of macrophages,
CSF-1 was removed by washing three times with medium, and 500 µl of
fresh medium was added together with LPS and cytokines. All cells were
cultured in humidified incubators at 37°C with 5% CO2.
This population of bone marrow-derived macrophages is hereafter
referred to as BMM
.
Stimulations
All stimulations were performed in triplicate as follows. LPS
(Escherichia coli strain 0111:B4; Difco) was used at 15
µg/ml; murine rIFN- (Genzyme, Cambridge, MA) was used at 100 U/ml;
murine rIL-4 (R&D Systems, Minneapolis, MN) was used at 20 ng/ml; and
murine rIL-10 (R&D Systems) was used at 10 ng/ml. The supernatants were
harvested 24 h later. Results with peritoneal macrophages are
representative of three independent experiments, and with bone
marrow-derived macrophages of five independent experiments.
Capture ELISAs
Maxisorp 96-well plates (Nunc) were coated overnight at 4°C
with capture Abs. Following washing, plates were blocked for 2 h
at room temperature with PBS and 3% BSA (blocking buffer). Standard
dilutions of cytokines were prepared (2000–31.25 pg/ml) and added to
wells in parallel with supernatants. Plates were incubated overnight at
4°C with shaking and were washed four times. Biotinylated detection
Ab was added, followed by incubation for 1 h at 4°C with
shaking. Plates were washed four times, and streptavidin-HRP (Genzyme)
was added for 15 min at 4°C with shaking. Plates were washed four
times, developed with tetramthylbenzadine (Sigma, St. Louis, MO), and
read at 370 nM. All washes were done with PBS and 0.05% Tween 20 using
a pressurized garden sprayer. Supernatants, standards, detection Abs,
and streptavidin-HRP were diluted in blocking buffer. Capture and
detection Abs were purchased from PharMingen (San Diego, CA):
anti-murine (m)IL-12 p40 (C15.6), anti-mIL-12 p40 biotin
(C17.8), anti-mTNF- (G281-2626), and anti-mTNF- biotin
(MP6-XT3). Standards of recombinant murine TNF- and recombinant
murine IL-12 were purchased from R&D Systems.
Biochemical analyses
Stimulations with cytokines were conducted as described (24).
Briefly, bone marrow-derived macrophages were derived from BALB/c or
STAT6 null mice by culture in medium supplemented with 20% LCCM on
10-cm dishes (Nunc) for 5–7 days. When the bottoms of the dishes were
almost completely covered with adherent macrophages, nonadherent cells
were removed by washing. Adherent macrophages were incubated overnight
in RPMI 1640 with 10% FCS with a reduced concentration of CSF-1 (2%
LCCM). Cells were washed three times with RPMI 1640 without FCS, and
incubated in 3 ml of RPMI without FCS for an additional hour at 37°C
in a humidified, gassed incubator. Cells were stimulated as indicated
with addition of synthetic mIL-4 (20 µg/ml) (Ian Clark-Lewis,
Biomedical Research Centre, Vancouver, BC), recombinant porcine insulin
(15 µg/ml) (Sigma), rmIFN- (1000 U/ml), or left unstimulated as a
control. The cells were lysed in lysis buffer (24). The amount of
protein in each lysate was normalized based on total protein content as
determined by the bicinchoninic acid protein assay (Pierce, Rockford,
IL). Insulin receptor substrate (IRS)-2, STAT1, or STAT6 were
immunoprecipitated from lysates by incubation with the appropriate Abs,
followed by adsorption to protein A-Sepharose (Pharmacia, Uppsala,
Sweden). Anti-IRS-2 Abs were purchased from Upstate Biotechnology (Lake
Placid, NY), and anti-STAT1 and -6 from Santa Cruz Biotechnology
(Santa Cruz, CA). The eluates were subjected to SDS-PAGE, and
immunoblotting. Membranes were blotted first with 4G10 (Upstate
Biotechnology) to determine the amount of tyrosine phosphorylation and
were subsequently stripped in stripping buffer (62.5 mM Tris
(pH 8.8), 2% SDS, and 100 mM 2-ME) for 1 h at 55°C and
reblotted with Abs to IRS-2, STAT1, or STAT6.
|
Results |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
production
We compared the ability of IL-4 to inhibit the production of
TNF- from macrophages derived from the bone marrow of wild-type
BALB/c or STAT6 null mice (12). When bone marrow cells were cultured
for 5 days in medium supplemented with CSF-1, wells became
confluent with adherent morphologically differentiatedmacrophages.
Wells were washed free of nonadherent cells and CSF-1, and the
remaining adherent macrophages were stimulated by addition of LPS, with
or without IL-4. Supernatants were collected 24 h later and
analyzed by capture ELISA for the presence of TNF-. Consistent with
previous reports (2, 19, 25), the presence of IL-4 resulted in a
50–65% reduction in the amount of TNF- produced by wild-type
macrophages (Fig. 1
A). In
contrast, the amount of TNF- produced by STAT6 null macrophages was
not reduced in the presence of IL-4.
|
by macrophages harvested from the peritoneal cavity (Fig. 1
B). In three independent experiments, the IL-4-mediated
reduction in the amount of TNF- produced by wild-type cells ranged
from 10 to 20%. In parallel cultures of STAT6 null peritoneal
macrophages stimulated with LPS, IL-4 exhibited no inhibitory effect
(Fig. 1B).
A STAT6-independent component of IL-4-mediated inhibition of
TNF- production in the presence of IFN-
IFN- enhances the ability of macrophages to produce
proinflammatory cytokines, such as TNF- and IL-12, in response to
LPS (18, 21). Moreover, in vivo bacterial infections are normally
associated with the production of IFN- from sources such as NK cells
(26) and, except at the earliest times, macrophages will normally
encounter LPS in the presence of IFN-. Therefore, we examined the
ability of IL-4 to inhibit the LPS-induced production of TNF- by
macrophages in the presence of IFN-. Addition of IFN- resulted in
a modest (2- to 5-fold) increase in the amount of TNF- produced in
response to LPS in both wild-type and STAT6 null macrophages (Fig. 1C). The presence of IL-4 resulted in a 72% reduction in
the amount of TNF- released by wild-type bone marrow-derived
macrophages in response to LPS (Fig. 1C). However, in
contrast to results seen with STAT6 null bone marrow macrophages
stimulated with LPS alone (Fig. 1A), in the presence of
IFN-, the STAT6 null macrophages were no longer refractory to the
inhibitory effects of IL-4, which induced a significant reduction
(43%) in TNF- production. Addition of IL-10, another potent
inhibitor of the production of proinflammatory cytokines, including
TNF- (27), completely suppressed the production of TNF- in both
wild-type and STAT6 null cells (Fig. 1C). Although, in the
experiment presented in Fig. 1C, STAT6 null macrophages
produced slightly less TNF- than did their wild-type counterparts,
in five independent experiments there were no consistent differences
between cells derived from wild-type or STAT6 null mice in the absolute
amounts of TNF- produced with or without IFN-.
We repeated these experiments with peritoneal macrophages, and as was
the case with cells stimulated in the absence of IFN-, IL-4 had
little or no effect on TNF- production. Once again IL-4 induced only
a marginal (10–20%) reduction in TNF- production by wild-type
peritoneal macrophages and had no significant effect on TNF-
production by STAT6 null peritoneal macrophages (Fig. 1D).
Addition of IL-10, however, resulted in the complete inhibition of the
production of TNF- from both wild-type and STAT6 null
peritoneal-exudate macrophages (Fig. 1D).
STAT6 is required for IL-4-mediated inhibition of LPS-stimulated IL-12 production
Next, we evaluated the involvement of STAT6 in IL-4-mediated
suppression of IL-12 production. We stimulated wild-type or STAT6 null
bone marrow-derived macrophages with LPS in the presence or absence of
IL-4 for 24 h and determined the amount of IL-12 (p40) in the
supernatant by capture ELISA. In wild-type cells, IL-4 induced a marked
(99%) inhibition of the LPS-stimulated production of IL-12 (Fig. 2A). In keeping with our
observations that IL-4 failed to inhibit the production of TNF-
by bone marrow-derived macrophages from STAT6 null mice (Fig. 1A), IL-4 also failed to inhibit the production of IL-12
by bone marrow-derived macrophages from STAT6 null mice (Fig. 2A). We repeated these experiments using peritoneal-exudate
macrophages. In striking contrast to the marginal effects of IL-4 on
the production of TNF- by wild-type peritoneal macrophages (Fig. 1B), IL-4 induced a marked inhibition (94%) of
LPS-stimulated production of IL-12 p40. This marked
inhibitory effect of IL-4 on the production of IL-12 by peritoneal
macrophages was abrogated in macrophages lacking STAT6 (Fig. 2B).
|
Next, we examined the effects of IL-4 on IL-12 production in
macrophages that had been stimulated with LPS in the presence of
IFN-. The presence of IFN- in cultures of bone marrow-derived
macrophages resulted in a much greater (5- to 10-fold) increase in the
amount of IL-12 p40 secreted than it did in the amount of TNF-
produced (2- to 5-fold). There was no consistent difference in the
absolute amount of IL-12 produced by wild-type or STAT6 null
macrophages that had been stimulated with or without IFN-. In
wild-type bone marrow-derived macrophages, IL-4 induced a marked
reduction (76%) in the amount of IL-12 p40 released in response to LPS
and IFN- (Fig. 2C). Moreover, the increased production of
IL-12 p40 by STAT6 null bone marrow-derived macrophages stimulated in
the presence of IFN- was significantly inhibited (57%) by IL-4
(Fig. 2C). Virtually all of the increase in IL-12 p40
production stimulated by IFN- was eliminated in the presence of
IL-4.
Very similar results were obtained with macrophages from peritoneal
exudates. IL-4 markedly inhibited the production of IL-12 p40 in
response by both wild-type cells (78%) and STAT6 null cells (56%)
(Fig. 2D). Addition of IL-10, which has previously been
shown to inhibit the production of IL-12 (23), completely suppressed
the release of IL-12 p40 by both cell types.
IL-4 induced signaling in STAT6 null cells
In that the promoter for the IL-4R gene contains a
STAT6-consensus binding site (28), we wanted to determine whether the
defect in IL-4-mediated inhibition of cytokine production in STAT6 null
cells reflected simply a lack of expression of IL-4R. Therefore, we
examined the ability of IL-4 to induce tyrosine phosphorylation of the
IRS-2 in bone marrow-derived macrophages. Cells were starved of CSF-1
and left unstimulated as a control, or stimulated with insulin or IL-4,
two factors that we have previously shown stimulate phosphorylation of
IRS-2 in this cell-type (24). Insulin signaling should be independent
of any effects on expression of the IL-4R that result from the lack
of STAT6. As shown in Fig. 3, wild-type
and STAT6 null bone marrow-derived macrophages responded equivalently
to insulin and IL-4 in terms of stimulation of phosphorylation of
IRS-2. These results are consistent with those of Kaplan et al. (29) in
lymphocytes. Thus, the defect in IL-4 signaling in STAT6 null
macrophages does not reflect a deficiency in the basal levels of
IL-4R on STAT6 null macrophages.
|
to
stimulate tyrosine phosphorylation of STAT1
One of the potential mechanisms by which IL-4 could antagonize the
production of proinflammatory cytokines by macrophages stimulated with
LPS and IFN- is by affecting the activation of STAT1 by IFN-.
Ohmori and Hamilton (30) observed that pretreatment of a murine
macrophage cell line with IL-4 did not inhibit the ability of IFN-
to stimulate phosphorylation of STAT1; nor did IFN- affect
IL-4-stimulated phosphorylation of STAT6. However, Dickensheets and
Donnelly (31) observed a marked reduction in the IL-4-induced tyrosine
phosphorylation, nuclear translocation, and DNA-binding activity of
STAT6 in monocytes that had been pretreated with IFN-. We wished to
evaluate the effects of pretreatment with IL-4 or IFN- on
phosphorylation of STAT1 or STAT6, respectively, in our model system
bone marrow-derived macrophages. Pretreatment for 1 h with IFN-
failed to effect the IL-4-stimulated tyrosine phosphorylation of STAT6
(Fig. 4). Similarly, pretreatment with
IL-4 for 1 h did not affect the ability of IFN- to stimulate
tyrosine phosphorylation of STAT1.
|
|
Discussion |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
and IL-12 by murine macrophages from peritoneal exudates or generated
in vitro from bone marrow. When bone marrow-derived macrophages were
stimulated with LPS alone, there was an absolute requirement for the
presence of STAT6 for IL-4 to inhibit the production of either TNF-
or IL-12 (Figs. 1A and 2A). In contrast, when
IFN- was present, STAT6 was not required for IL-4 to significantly
inhibit the production of TNF- or IL-12 (Fig. 1C and
2C). On average, the IL-4-mediated inhibition of the
production of TNF- or IL-12 in bone marrow macrophages stimulated in
the presence of IFN- was slightly less (43–57%) in STAT6 null
cells than in wild-type cells (72–76%), indicating that a minor part
of the inhibitory effect of IL-4 depended on STAT6, even in
IFN--treated cells.
Macrophages from peritoneal cavity exudates similarly exhibited
STAT6-dependent and -independent mechanisms of IL-4-mediated inhibition
of IL-12 production. In the absence of IFN-, IL-4 almost completely
suppressed the production of IL-12 by wild-type cells (94%), but had
no effect on the production of IL-12 by STAT6 null peritoneal cells
(Fig. 2B). The presence of IFN- resulted in a large
increase in the LPS-stimulated production of IL-12 (10x), that was
suppressed by IL-4 in both wild-type (78%) and STAT6 null cells (56%)
(Fig. 2D).
In contrast to our observations in bone marrow-derived macrophages, in
peritoneal macrophages, the effects of IL-4 on TNF- production did
not parallel those on production of IL-12. Thus, IL-4 had little or no
inhibitory effect on the amount of TNF- produced by wild-type or
STAT6 null cells in response to LPS, whether or not IFN- was present
(Figs. 1B and 1D), despite the significant
inhibitory effects of IL-4 on the production of IL-12 in the same
cultures (Figs. 2B and 2D). This failure of IL-4
to significantly inhibit TNF- production in macrophages from
peritoneal exudates is consistent with the observations of Oswald et
al. (32), and suggests that different mechanisms regulate TNF-
production in bone marrow-derived and peritoneal macrophages.
Collectively, these data indicate that IL-4 inhibits the production of
IL-12 and TNF- through distinct mechanisms.
In the absence of IFN-, IL-4-mediated inhibition of TNF- and
IL-12 was absolutely dependent on STAT6 (Figs. 1A,
2A, and 2B). STAT6 could potentially influence
the production of proinflammatory cytokines by competing with NF-
B
(33, 34) DNA-binding activity or inducing expression of proteins that
down-regulate cytokine levels (7, 35, 36, 37).
We did not see defects in the acute biochemical responses to IL-4 in
STAT6 null macrophages, as judged by phosphorylation of IRS-2 (Fig. 3)
in keeping with other evidence that the constitutive level of IL-4R
expression is not dependent on expression of STAT6 (12). However,
expression of the IL-4R-chain is increased following stimulation by
IL-4 through a STAT6-dependent mechanism (28, 38). Thus, it is
conceivable that the IL-4-mediated inhibition of production of TNF-
and IL-12 by LPS-stimulated macrophages requires this up-regulation of
IL-4R expression to generate the necessary strength or duration of
this signal.
IL-4 induces increased production of IL-10 (39), raising the possibility that some antiinflammatory effects of IL-4 are secondary to the production of IL-10. However, in our hands, IL-4 induced only a modest enhancement of IL-10 production, and this was equivalent in wild-type and STAT6 null macrophages (data not shown).
The presence of IFN- resulted in the expected large increase in the
LPS-stimulated production of IL-12 (5–10x) and a modest increase
(2–5x) in the production of TNF-, which, in both cases, was
inhibited by IL-4 (Figs. 1C, 2C, and
2D). Our results show that 70–100% of this IFN--induced
increase in the LPS-stimulated production of TNF- or IL-12 was
inhibited by IL-4 via a mechanism that did not require STAT6 (Figs. 1C, 2C, and 2D). In vivo, bacterial
products directly induce the production of IL-18 and IL-12, resulting
in the early production of IFN- from NK cells (15, 26). Thus, the
STAT6-independent mechanism of IL-4-mediated inhibition of TNF- and
IL-12 production that we observed when IFN- was added to our
cultures is likely to be physiologically significant. Our data suggest
that the inhibitory effects of IL-4 on the production of
proinflammatory cytokines during infections, where IFN- is present,
will be largely independent of STAT6.
The mechanism of this STAT6-independent suppression of IFN--enhanced
production of TNF- and IL-12 is unclear. Stimulation with IL-4 leads
to a modest increase in tyrosine phosphorylation of STAT3 in both
wild-type or STAT6-null bone marrow-derived macrophages (data not
shown). Analysis of STAT3 null macrophages will be necessary to
determine whether the STAT6-independent inhibition of the production of
proinflammatory cytokines by IL-4 is mediated by STAT3.
Although the greater part of IL-4-mediated inhibition of the production
of TNF- and IL-12 that was stimulated by the combination of LPS and
IFN- was not dependent on STAT6, we consistently observed that the
IL-4-mediated inhibition of IFN--enhanced production of TNF- or
IL-12 was 
20% less in STAT6 null cells than in wild-type cells
(compare 72% vs 43%, Fig. 1C; 76% vs 57%, Fig. 2C; and 78% vs 56%, Fig. 2D). Acute treatment
with IL-4 did not inhibit the tyrosine phosphorylation of STAT1 induced
by IFN- in bone marrow-derived macrophages (Fig. 4). However, IL-4
might induce the STAT6-dependent production of proteins, such as
members of the suppressor of cytokine synthesis family (40) or the
protein inhibitors of cytokine signaling family (41), and, thereby,
inhibit the levels of STAT1 activity over time. STAT6 may also compete
with STAT1 for binding to STAT1 sites (30), as STAT6 can bind to the
TTC(N3)GAA sequence recognized by STAT1 (42).
In conclusion, we have shown that IL-4 can inhibit the production of
TNF- and IL-12 through STAT6-dependent and -independent mechanisms.
The STAT6-independent mechanisms only operated on the enhanced
production of TNF- and IL-12 induced by IFN-. However, LPS
stimulates the production of IFN-, and, thus, this STAT6-independent
pathway is likely to be important for the inhibitory effects of IL-4 on
the production of proinflammatory cytokines in vivo. Future
characterization of this new pathway will be an important step toward
understanding the mechanisms through which IL-4 suppresses the
production of cytokines that promote the generation of Th1 cells and
inflammation.
|
Acknowledgments |
|---|
|
Footnotes |
|---|
2 Current address: Telethon Institute for Gene Therapy, San Raffaele Research Institute, Milan, Italy.
3 Address correspondence and reprint requests to Dr. John W. Schrader, The Biomedical Research Centre, University of British Columbia, 2222 Health Sciences Mall, Vancouver, BC, V6T 1Z3 Canada. E-mail address:
4 Abbreviations used in this paper: PEC, adherent peritoneal exudate cells; LCCM, L cell conditioned medium; m, murine; IRS, insulin receptor substrate-2. BMM, bone marrow-derived macrophages.
Received for publication October 2, 1998. Accepted for publication February 16, 1999.
|
References |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
, interleukin 1, and prostaglandin E2. Proc. Natl. Acad. Sci. USA 86:3803.B (NF B) activation in human monocytes: IL-10 and IL-4 suppress cytokine synthesis by different mechanisms. J. Biol. Chem. 270:9558. mRNA translational activation in lipopolysaccharide-induced mouse macrophages. J. Biol. Chem. 272:14394.R membrane expression and FcR-mediated cytotoxic activity of human monocytes. J. Immunol. 144:3046.[Abstract]
and/or IL- 2 in murine peritoneal macrophages. J. Immunol. 148:1725.[Abstract]
B and IB: an inducible regulatory system in endothelial activation. J. Exp. Med. 179:503. transcription in macrophages: involvement of four B-like motifs and of constitutive and inducible forms of NF-B. Mol. Cell. Biol. 10:1498. priming of monocytes enhances LPS-induced TNF production by augmenting both transcription and MRNA stability. Cytokine 7:427.[Medline]
or IL-10. J. Immunol. 151:6370.[Abstract]
of lipopolysaccharide-inducible p35 and p40 genes. Blood 86:646. production. J. Exp. Med. 181:537., and IL-6 by human monocytes. Blood 76:1392.). Curr. Opin. Immunol. 9:17.[Medline]
-stimulated STAT1-dependent transcription in mouse macrophages. J. Immunol. 159:5474.[Abstract]
and IL-10 inhibit induction of IL-1 receptor type I and type II gene expression by IL-4 and IL-13 in human monocytes. J. Immunol. 159:6226.[Abstract]
B activity in monocytes and fibroblasts. J. Immunol. 151:5603.[Abstract]
-stimulated E- selectin gene transcription is mediated by STAT6 antagonism of NF-B. J. Biol. Chem. 272:10212. gene expression by human T lymphocytes and monocytes. Pathobiology 60:100.[Medline]
promoter region. Cell. Immunol. 161:125.[Medline]
This article has been cited by other articles:
|
|
 |
|
  K. Yamaji-Kegan, Q. Su, D. J. Angelini, and R. A. Johns IL-4 Is Proangiogenic in the Lung under Hypoxic Conditions J. Immunol., May 1, 2009; 182(9): 5469 - 5476. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 N. Thawani, M. Tam, and M. M. Stevenson STAT6-mediated suppression of erythropoiesis in an experimental model of malarial anemia Haematologica, February 1, 2009; 94(2): 195 - 204. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. Wang, N. van Panhuys, J. Hu-Li, S. Kim, G. Le Gros, and B. Min Blimp-1 Induced by IL-4 Plays a Critical Role in Suppressing IL-2 Production in Activated CD4 T Cells J. Immunol., October 15, 2008; 181(8): 5249 - 5256. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Iwamoto, S.-i. Iwai, K. Tsujiyama, C. Kurahashi, K. Takeshita, M. Naoe, A. Masunaga, Y. Ogawa, K. Oguchi, and A. Miyazaki TNF-{alpha} Drives Human CD14+ Monocytes to Differentiate into CD70+ Dendritic Cells Evoking Th1 and Th17 Responses J. Immunol., August 1, 2007; 179(3): 1449 - 1457. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Marcais, C.-A. Coupet, T. Walzer, M. Tomkowiak, R. Ghittoni, and J. Marvel Cell-Autonomous CCL5 Transcription by Memory CD8 T Cells Is Regulated by IL-4 J. Immunol., October 1, 2006; 177(7): 4451 - 4457. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Saito, M. Matsuura, and Y. Hirai Regulation of Lipopolysaccharide-Induced Interleukin-12 Production by Activation of Repressor Element GA-12 through Hyperactivation of the ERK Pathway. Clin. Vaccine Immunol., August 1, 2006; 13(8): 876 - 883. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T.A. Silva, V.S. Lara, J.S. Silva, S.H.P. Oliveira, W.T. Butler, and F.Q. Cunha Macrophages and Mast Cells Control the Neutrophil Migration Induced by Dentin Proteins Journal of Dental Research, January 1, 2005; 84(1): 79 - 83. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. C. Morrison, C. B. Wilson, M. Ray, and P. H. Correll Macrophage-Stimulating Protein, the Ligand for the Stem Cell-Derived Tyrosine Kinase/RON Receptor Tyrosine Kinase, Inhibits IL-12 Production by Primary Peritoneal Macrophages Stimulated with IFN-{gamma} and Lipopolysaccharide J. Immunol., February 1, 2004; 172(3): 1825 - 1832. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. L. Oleszak, J. R. Chang, H. Friedman, C. D. Katsetos, and C. D. Platsoucas Theiler's Virus Infection: a Model for Multiple Sclerosis Clin. Microbiol. Rev., January 1, 2004; 17(1): 174 - 207. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. Suzuki, H. Nakajima, K. Ikeda, Y. Maezawa, A. Suto, H. Takatori, Y. Saito, and I. Iwamoto IL-4-Stat6 Signaling Induces Tristetraprolin Expression and Inhibits TNF-{alpha} Production in Mast Cells J. Exp. Med., December 1, 2003; 198(11): 1717 - 1727. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Smith, F. Santoro, G. Di Lullo, L. Dagna, A. Verani, and P. Lusso Selective suppression of IL-12 production by human herpesvirus 6 Blood, October 15, 2003; 102(8): 2877 - 2884. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. Huaux, T. Liu, B. McGarry, M. Ullenbruch, and S. H. Phan Dual Roles of IL-4 in Lung Injury and Fibrosis J. Immunol., February 15, 2003; 170(4): 2083 - 2092. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Finnegan, M. J. Grusby, C. D. Kaplan, S. K. O'Neill, H. Eibel, T. Koreny, M. Czipri, K. Mikecz, and J. Zhang IL-4 and IL-12 Regulate Proteoglycan-Induced Arthritis Through Stat-Dependent Mechanisms J. Immunol., September 15, 2002; 169(6): 3345 - 3352. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Rodriguez-Sosa, J. R. David, R. Bojalil, A. R. Satoskar, and L. I. Terrazas Cutting Edge: Susceptibility to the Larval Stage of the Helminth Parasite Taenia crassiceps Is Mediated by Th2 Response Induced Via STAT6 Signaling J. Immunol., April 1, 2002; 168(7): 3135 - 3139. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Major, J. E. Fletcher, and T. A. Hamilton IL-4 Pretreatment Selectively Enhances Cytokine and Chemokine Production in Lipopolysaccharide-Stimulated Mouse Peritoneal Macrophages J. Immunol., March 1, 2002; 168(5): 2456 - 2463. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Luft, M. Jefford, P. Luetjens, H. Hochrein, K.-A. Masterman, C. Maliszewski, K. Shortman, J. Cebon, and E. Maraskovsky IL-1{beta} Enhances CD40 Ligand-Mediated Cytokine Secretion by Human Dendritic Cells (DC): A Mechanism for T Cell-Independent DC Activation J. Immunol., January 15, 2002; 168(2): 713 - 722. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Malaviya and F. M. Uckun Role of STAT6 in IgE Receptor/Fc{varepsilon}RI-Mediated Late Phase Allergic Responses of Mast Cells J. Immunol., January 1, 2002; 168(1): 421 - 426. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. Qadir, A. Metwali, A. M. Blum, J. Li, D. E. Elliott, and J. V. Weinstock TGF-beta and IL-10 regulation of IFN-gamma produced in Th2-type schistosome granulomas requires IL-12 Am J Physiol Gastrointest Liver Physiol, October 1, 2001; 281(4): G940 - G946. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Becker, S. Wirtz, X. Ma, M. Blessing, P. R. Galle, and M. F. Neurath Regulation of IL-12 p40 Promoter Activity in Primary Human Monocytes: Roles of NF-{kappa}B, CCAAT/Enhancer-Binding Protein {beta}, and PU.1 and Identification of a Novel Repressor Element (GA-12) That Responds to IL-4 and Prostaglandin E2 J. Immunol., September 1, 2001; 167(5): 2608 - 2618. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. D. Ahlers, I. M. Belyakov, S. Matsui, and J. A. Berzofsky Mechanisms of cytokine synergy essential for vaccine protection against viral challenge Int. Immunol., July 1, 2001; 13(7): 897 - 908. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. K. Levings, R. Sangregorio, F. Galbiati, S. Squadrone, R. de Waal Malefyt, and M.-G. Roncarolo IFN-{{alpha}} and IL-10 Induce the Differentiation of Human Type 1 T Regulatory Cells J. Immunol., May 1, 2001; 166(9): 5530 - 5539. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
V. T. Nguyen and E. N. Benveniste IL-4-Activated STAT-6 Inhibits IFN-{gamma}-Induced CD40 Gene Expression in Macrophages/Microglia J. Immunol., December 1, 2000; 165(11): 6235 - 6243. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Kato, H. Yoshidome, M. J. Edwards, and A. B. Lentsch Regulation of Liver Inflammatory Injury by Signal Transducer and Activator of Transcription-6 Am. J. Pathol., July 1, 2000; 157(1): 297 - 302. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Yoshidome, A. Kato, M. Miyazaki, M. J. Edwards, and A. B. Lentsch IL-13 Activates STAT6 and Inhibits Liver Injury Induced by Ischemia/Reperfusion Am. J. Pathol., October 1, 1999; 155(4): 1059 - 1064. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Mirmonsef, C. P. Shelburne, C. Fitzhugh Yeatman II, H. J. Chong, and J. J. Ryan Inhibition of Kit Expression by IL-4 and IL-10 in Murine Mast Cells: Role of STAT6 and Phosphatidylinositol 3'-Kinase J. Immunol., September 1, 1999; 163(5): 2530 - 2539. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. L. Orchansky, R. Kwan, F. Lee, and J. W. Schrader Characterization of the Cytoplasmic Domain of Interleukin-13 Receptor-alpha J. Biol. Chem., July 23, 1999; 274(30): 20818 - 20825. [Abstract] [Full Text] [PDF]
|
|
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
) or were harvested from
the peritoneal cavities after stimulation with thioglycollate (PEC).
A and B, Macrophages were cultured in
triplicate with LPS (15 µg/ml) in the presence or absence of IL-4.
C and D, Macrophages were cultured in
triplicate with LPS (15 µg/ml) and IFN-
0.05) IL-4-mediated decrease in the amount of
TNF-
