| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
*
Deutsches Rheumaforschungszentrum, Berlin, Germany;
Millennium Pharmaceuticals, Cambridge, MA 021139; and
Universitätsklinikum Charité, Medizinische Klinik mit Schwerpunkt Rheumatologie/Klinische Immunologie, Berlin, Germany
 |
Abstract |
|---|
 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
enhanced
the expression of T1/ST2 on Th2 cells, and IL-6 was by far most
effective in this regard. However, the expression of T1/ST2 did not
depend on the presence of IL-6 and was also detected in IL-6-deficient
mice. Most important, cross-linking of T1/ST2 provided a costimulatory
signal for Th2 but not Th1 cells and directly induced proliferation and
type 2 cytokine production. Thus, T1/ST2 is not only a Th2 cell marker
but also plays an important role in the activation of Th2
cells. |
Introduction |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
, IL-2, and TNF-
and mediate delayed-type
hypersensitivity and protection against intracellular pathogens,
whereas Th2 cells produce mainly IL-4 and IL-5 and are implicated in
humoral and allergic immune responses (2, 3). Although
there are many well-documented Th1 and Th2 immune responses, it is now
widely accepted that Th cells can produce cytokine patterns different
from the canonical Th1/Th2 pattern and that the classical Th1 and Th2
subsets represent the end points of a continuous differentiation
process (2, 3, 4, 5). Th phenotype differentiation is influenced by a number of factors including different APC, MHC genes, non-MHC "background genes," costimulatory molecules, Ag dose or structure, and cytokines present during Th priming. The major factors for the induction of a Th1 or Th2 response seem to be IL-12 and IL-4, respectively (6, 7).
The transient nature of cytokine expression and the methodological limits of its detection for functional analysis of different Th cell subsets have stimulated an intensive search for further differences between Th1 and Th2 cells. The Th2-specific expression of two transcription factors, GATA3 and c-maf, and the Th1-specific expression of another transcription factor, T-bet (8), appear to represent, at least in part, the transcriptional basis for differential cytokine expression (9). In addition, some chemokine receptors such as CCR3, CCR4, and CCR8 have been reported to be predominantly expressed on Th2 cells, whereas others, such as CXCR 3 or CCR5, were found to be preferentially expressed on Th1 cells (10).
Recently, several groups, including ours, reported that T1/ST2, an orphan receptor with sequence homology to the IL-1R, is preferentially expressed on murine Th2 cells (11, 12, 13, 14) and also on mast cells (15). T1/ST2 is important for Th2 effector functions since treatment with Abs against T1/ST2 decreased Th2 effector functions in vivo (11, 12, 16). Here, we analyzed the kinetics and functional importance of T1/ST2 expression on Th2 cells in vitro. We found that the kinetics of type 2 cytokine expression in Th2 cells is faster than that of T1/ST2 expression. In vitro, T1/ST2 expression is enhanced by proinflammatory cytokines. Importantly, cross-linking of T1/ST2 induces proliferation and cytokine production in Th2 but not Th1 cells. Our results suggest that T1/ST2 expression is a late event during Th cell commitment to the Th2 phenotype and that signaling through T1/ST2 specifically enhances Th2 effector functions.
|
Materials and Methods |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
The OVA peptide OVA323–339 was
synthesized according to standard F-moc machine protocols with a
multiple peptide synthesizer (Abimed) and was kindly provided by A.
Kramer (Humboldt University, Berlin). Mouse recombinant IL-1 and
IL-1, TNF-, and IL-6 were purchased from TEBU (Frankfurt am Main,
Germany). Recombinant murine IL-13, anti-CD28 (37.51), and rat IgG1
isotype control mAb were purchased from PharMingen (Hamburg, Germany).
Recombinant IL-5 (X63IL5) at a specific activity of 158 U/µg and the
hamster anti-CD3 (145-2C11) mAb were obtained from institutional
facilities.
The recombinant baculovirus GP67T1S-Flag driving the synthesis of soluble murine T1/ST2 in insect cells was a gift from A. K. Werenskiold (Technische Universität München). Flag-tagged soluble T1/ST2 was purified from supernatants of GP67T1S-Flag baculovirus-infected Sf9 insect cells as described (17).
Animals
Mice transgenic for the OVA-specific DO11.10 T cell Ag receptor (TCR) (18) were from D. Loh (Washington University, St. Louis, MO) and were maintained on the BALB/c background. IL-6-deficient C57BL/6 mice (19) and their heterozygous littermates were from T. Blankenstein (Max Delbrück Centrum, Berlin, Germany). Mice were kept under pathogen-free conditions in accordance with institutional and state guidelines.
Generation of Th1 and Th2 cells in vitro
CD4+CD62L+ spleen
cells (SC)5 from
DO11.10 TCR-transgenic mice were isolated by high-gradient magnetic
cell separation with MACS Multisort (Miltenyi Biotec, Bergisch
Gladbach, Germany) as described elsewhere (12). SC from
BALB/c depleted of T cells with CD4, CD8, and Thy1.2 microbeads
(Miltenyi Biotech) were used as APC. Cells were cultured in complete
RPMI 1640 containing 100 U/ml penicillin, 0,1 mg/ml streptomycin, 2 mM
glutamine (Biochrom, Berlin, Germany), 25 µM 2-ME, and 10% FCS
(Sigma, Deisenhofen, Germany). T cells were stimulated either with
OVA323–339 (0, 3 µM) and APC or with
immobilized anti-CD3 (1 µg) and anti-CD28 (5 µg/ml). For
Th1 phenotype development, recombinant murine IL-12 (gift from M.
Gately, Hoffman-LaRoche, Nutley, NJ) and neutralizing anti-IL-4 mAb
11B11 (20) were added, and for Th2 development recombinant
murine IL-4 (BioSource International, Camarillo, CA) and neutralizing
anti-murine IL-12 polyclonal Ab (M. Gately) or, when indicated,
recombinant murine IL-4 and neutralizing anti-IFN- mAb (RA-6A2 5
µg/ml) were added as described previously (12). In some
experiments, mouse recombinant IL-1 and IL-, TNF-, IL-5, IL-6,
and IL-13 at concentrations between 0.5 and 50 ng/ml were added as
indicated. Three days after Ag stimulation, recombinant human IL-2 (100
U/ml) was added. Cultures were maintained for 7 days. In some
experiments, several rounds of Ag stimulation and rest were performed
as indicated. In the experiments of IL-6-/-
mice and their IL-6+/- littermates,
CD4+ T cells were MACS isolated from peripheral
lymph nodes and Thy1.2-depleted, irradiated SC were used as APC.
Soluble anti-CD3 mAb (3 µg/ml) and anti-CD28 (2.5 µg/ml)
were added to the culture. For polarization of Th2 cells, recombinant
IL-4 (R&D Systems, Wiesbaden, Germany) and anti-IFN- mAb (5
µg/ml) and anti-IL-12 mAb (C17.15, 5 µg/ml) were added.
Abs and flow cytometry
For surface staining, the following rat anti-mouse mAbs were
used: CD4 (GK1.5), CD8 (53-6.7), CD62L (L-selectin; MEL-14), DO11.10
TCR (KJ.26.1) (21), and T1/ST2 (3E10) (12).
To prevent unspecific binding of the T1/ST2-specific mAb, all samples
were preincubated with blocking anti-FcR mAb 2.4G2/75 (100
µg/ml) and purified rat IgG (200 µg/ml; Dianova, Hamburg, Germany)
10 min before and during staining with digoxigenized 3E10 (1.5
µg/ml), FITC-conjugated 3E10 (3 µg/ml), or biotinylated 3E10 (3
µg/ml). Digoxigenized 3E10-labeled cells were detected by
anti-digoxigenin (DIG) Fab fragments (Boehringer Mannheim,
Mannheim, Germany) conjugated to PE or Cy5; biotinylated 3E10 was
detected with streptavidin-PE (PharMingen). Staining of 3E10 was
blocked by preincubating the cells with a 100-fold excess of
unconjugated 3E10. Samples were analyzed on a FACSCalibur (Becton
Dickinson, Mountain View, CA). Gates were set on viable cells according
to forward and sideward scatter and exclusion of propidium
iodide-binding particles (0.3 µg/ml).
Analysis of cytokine production by flow cytometry and ELISA
Analysis of intracellular cytokines and surface markers were
performed as described elsewhere (12). Cells
(106/ml) were stimulated with PMA (5 ng/ml) and
ionomycin (0.5 µg/ml; Sigma) for 5 h. At 2 h, brefeldin A
(Sigma) was added at 5 µg/ml. Before fixation, cells were stained
with biotinylated mAb against DO11.10 TCR (KJ26.1) and digoxigenized
mAb 3E10 against T1/ST2. Cells from IL-6-/- or
IL-6+/- mice were stained before fixation with
anti-CD4 FITC. Stained cells were fixed with 2% formaldehyde.
DO11.10 TCR and T1/ST2 were detected on the surface of fixed cells by
streptavidin coupled to PerCP (PharMingen) and anti-DIG Fab
fragments (Boehringer Mannheim) conjugated to Cy5, respectively. Cells
were permeabilized and incubated with two of the following mAbs
(PharMingen): anti-IL-4-PE (1D11; 3 µg/ml), anti-IL-4-FITC
(1D11; 5 µg/ml), anti-IL-4 DIG (11B11, 1.3 µg/ml),
anti-IL-5-PE (TRFK5; 3 µg/ml), anti-IL-6-PE (MP5-20F3, 2
µg/ml; PharMingen), anti-IL-10-FITC (5 µg/ml), or
anti-IFN--FITC (XMG1.2, 5 µg/ml), followed where indicated by
incubation with Cy5-conjugated anti-DIG Fab fragments. FITC- or
PE-labeled isotype control mAbs (PharMingen) were used at 5 and 3
µg/ml, respectively. Samples were analyzed by four-color flow
cytometry. Concentrations of IL-4, IL-5, and IL-10 in culture
supernatants were determined by ELISA as described previously
(12). IFN- was determined with a commercially available
kit according to the manufacturer’s instructions (Genzyme, Cambridge,
MA). Lower detection limits for each ELISA were as follows: IL-4 and
IFN-, 40 pg/ml; IL-5, 10 U/ml; and IL-10, 0.3 ng/ml.
Measurement of T cell proliferation and cytokine expression
mAbs were immobilized to 96-well round-bottom cell culture plates (Costar, Bodenheim, Germany) in a final volume of 0.15 ml PBS for 2 h at 37°C. Ten to 12 days after the last Ag stimulation, 1 x 105 T cells/well were cultured in complete medium at 37°C in 5% CO2. Supernatants were collected at 48 h for analysis by sandwich ELISA. Proliferation was measured by 16-h incorporation of 1 µCi [3H]thymidine on day 3. Results were calculated from triplicate cultures and expressed as the mean ± SEM.
Statistical analysis of intracellular cytokine coexpression and surface T1/ST2
The observed value for cytokine-coexpressing cells in percent
was compared with the expected value calculated for random coincidence
of two independent variables. Correlations of cytokine coexpression in
the total DO11.10 TCR+
CD4+, T1/ST2+, and
T1/ST2- DO11.10 TCR+
CD4+ T cells were calculated using the test for
correlation coefficients (22) as described elsewhere
(13). Coefficients of 
-0.1 or 
0.1 were considered
to be significant in this analysis.
|
Results |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
To determine the kinetics of T1/ST2 expression on Th2 cells naive
(CD62Lhigh) CD4+ spleen
cells from DO11.10 TCR-transgenic mice were subjected to several rounds
of in vitro Ag stimulation and rest. T cells were stimulated with OVA
peptide (OVA323–339) and APC in the presence of
IL-4 plus anti-IL-12. Seven days after each Ag-stimulation, T1/ST2
expression on DO11.10-TCR+ cells and
intracellular cytokine expression upon restimulation with PMA/ionomycin
was analyzed by flow cytometry. As shown in Fig. 1
A, the number of
T1/ST2+ cells increased during continual Ag
stimulation under Th2-polarizing conditions. In contrast, neither
Th1-polarized T cells (IL-12 plus anti-IL-4) nor T cells primed
with peptide alone expressed T1/ST2 (Ref. 16 and data not
shown). Following priming under Th2-polarizing conditions, the
expression of T1/ST2 was not dependent on further addition of exogenous
IL-4 during the subsequent rounds of restimulation and rest as
restimulation with peptide alone resulted in the same kinetics of
T1/ST2 expression. Even when endogenous IL-4 was neutralized with mAbs
during restimulation, a delayed yet steady increase of T1/ST2
expression was observed (data not shown).
|
and Table I). Thus, 1
wk after priming most IL-4 producers were T1/ST2 negative. After the
second round of Ag stimulation and rest IL-4 was produced by 51.9
± 8.7% of T cells, whereas 25.2 ± 5.4% of the T cells
expressed T1/ST2 (Table I). Over the course of repeated antigenic
stimulation, the percentage of T1/ST2+ Th cells
gradually approached the percentage of IL-4 producers. After four to
five rounds of in vitro Ag stimulation and rest, the percentages of
OVA-TCR+ cells expressing T1/ST2 or IL-4 were
approximately equal (79.4 ± 3.4 IL-4 producers and 69.3 ±
5.1 T1/ST2+ cells after 4 wk; 76.6 ± 3.7
IL-4 producers and 81.6 ± 5.9 T1/ST2+ cells
after 5 wk). The frequency of IL-10-producing cells (4.9 ± 1.4) 1
wk after priming was higher than that of T1/ST2+
cells (Table I). However, already after the second Ag stimulation, the
percentages of OVA-TCR+ cells expressing T1/ST2
or IL-10 were approximately equal. IL-5 expression was low throughout
the experiment and ranged from 0.1 to 7.9% positive cells. The
percentage of cytokine-producing cells was always higher in the
T1/ST2+ population than in the
T1/ST2- population (Fig. 2), although the percentage of
T1/ST2+ cells was initially low (Table I and
Fig. 2).
|
|
Naive DO11.10 TCR+
CD4+ T cells were primed under Th2-polarizing
conditions for several rounds. Seven days after each Ag stimulation, an
aliquot of the cells was restimulated with PMA and ionomycin and
stained for T1/ST2 and intracellular cytokines. To determine whether
coexpression of type 2 cytokines was stochastic or coordinate, the
observed frequencies of cytokine-coexpressing cells were compared with
those calculated for expected values (i.e., random coincidence). The
observed values for cytokine expression and the correlation
coefficients for cytokine coexpression in
T1/ST2- and T1/ST2+
DO11.10 TCR+ T cells are shown in Fig. 2. After
two rounds of Ag stimulation, the production of IL-4 was strongly
associated with IL-10 production in both the
T1/ST2+ and T1/ST2-
fractions (Fig. 2A: 0.32 in
T1/ST2+ cells and 0.25 in
T1/ST2- cells). Similar values were found after
the fourth Ag stimulation (Fig. 2B). In contrast, although
IL-5 producers were enriched in the T1/ST2+
fraction (3.2% in T1/ST2+ cells and 0.46% in
T1/ST2- cells after 2 wk and 6.3% in
T1/ST2+ cells and 3.1% in
T1/ST2- cells after 4 wk), coexpression of IL-5
with either IL-4 or IL-10 was random in both
T1/ST2+ and T1/ST2- cells
(Fig. 2). The coexpression of type 1 and type 2 cytokines was not
determined since there were very few if any IFN-- or IL-2-producing
Th cells in these in vitro-generated Th2 lines.
IL-6, IL-1, TNF-, and IL-5 increase T1/ST2 expression
We next asked whether cytokines produced by either APC or T cells
influence T1/ST2 expression. Naive (CD62Lhigh)
CD4+ splenic T cells from
DO11.10-TCR+ mice were stimulated with peptide
and T cell-depleted BALB/c SC under Th2-polarizing conditions in the
presence or absence of recombinant TNF-, IL-1, IL-5, IL-6, or
IL-13. In preliminary experiments, the optimal concentrations for each
cytokine were determined. After 7 days, cells were counted and stained
for T1/ST2. Expression of T1/ST2 was enhanced most effectively by IL-6
and to a lesser extent by IL-5, IL-1, and TNF-. The number of
T1/ST2+ cells was increased 
6-fold (6.1
± 0.8, mean ± SEM, data from at least three independent
experiments) by IL-6; 3-fold (3 ± 0.4) by IL-5; and 2-fold
(2.1 ± 0.3) by IL-1 or TNF- (Fig. 3). Studies were also performed with
IL-1 and virtually identical results were obtained as with IL-1
(data not shown). The effect of IL-6 on T1/ST2 expression was
detectable even at a concentration as low as 500 pg/ml. IL-13 had no
effect on T1/ST2 expression at any of the tested concentrations
(0.5–50 ng/ml, data not shown). Combinations of different cytokines
had additive but not synergistic effects on T1/ST2 expression (Fig. 3).
The cytokine-induced increase in T1/ST2 expression was not due to
enhanced T cell proliferation or survival, since similar numbers of
viable T cells were recovered from cytokine-treated and control
cultures.
|
, the frequency of
CD4+ T1/ST2+ cells in
IL-6-/- mice was similar compared with their
heterozygous littermates (0.9 and 1.3%, respectively, Fig. 4).
Moreover, when lymph node cells from IL-6-/-
mice or IL-6+/- heterozygous littermates were
polyclonally stimulated in vitro, the kinetics of both IL-4 production
and surface expression of T1/ST2 were identical (Fig. 5).
|
|
. Under those conditions, none of the investigated cytokines
induced Th2 phenotype development or T1/ST2 expression (Fig. 3 and data
not shown).
To test directly the importance of APC for T1/ST2 induction, naive
splenic CD4+ T cells from
DO11.10-TCR+ mice were stimulated with
immobilized anti-CD3 plus anti-CD28 mAb in the presence of IL-4
and anti-IFN- mAb for three rounds of stimulation and rest. The
differentiation of T cells under these conditions resulted in similar
kinetics of T1/ST2 expression as observed with Ag stimulation in the
presence of APC (Fig. 6). Thus, T1/ST2
expression is neither dependent on the presence of APC-derived
cytokines, nor on cell-cell contact between T cells and APC.
|
In a murine model of Th2-dependent allergic airway inflammation,
administration of either anti-T1/ST2 mAb or T1/ST2-Ig fusion
protein inhibited the secretion of Th2 cytokines and infiltration of
eosinophils into the airways (12, 16). We therefore
hypothesized that T1/ST2 could act as a costimulatory molecule for Th2
responses. To test whether T1/ST2 influences T cell proliferation and
cytokine production, we examined the effects of T1/ST2 cross-linking on
APC-independent and Ag-specific T cell activation in vitro. In
vitro-polarized DO11.10 TCR+ Th2 cells were used
for proliferation and cytokine assays. Cross-linking of T1/ST2 enhanced
proliferation (3- to 6-fold) as well as IL-4 and IL-5 production (4- to
10-fold) of Th2 cells that were stimulated with suboptimal
concentrations of anti-CD3 mAb (<100 ng/ml, Fig. 7A) alone or in combination
with anti-CD28 mAb (data not shown).
|
100 ng/ml), T1/ST2
cross-linking did not further increase the proliferation or type 2
cytokine production induced by anti-CD3 (Fig. 7A) or
anti-CD3/anti-CD28 (data not shown). In contrast, immobilized
anti-T1/ST2 mAb alone or in combination with anti-CD3 had no
effect on proliferation and cytokine production by Th1 cells (Fig. 7B).
When in vitro-polarized DO11.10 TCR+ Th2 cells
were restimulated with APC and OVA, cross-linking of T1/ST2 enhanced
IL-4 production of Th2 cells that were stimulated with suboptimal
concentrations of OVA. Furthermore, cross-linking T1/ST2 strongly
induced Th2 cell proliferation that was not further enhanced by the
addition of OVA (Fig. 7C).
Immobilized but not soluble anti-T1/ST2 mAb 3E10 alone induced
proliferation and IL-4 and IL-5 production in a dose-dependent manner
(Fig. 8). Both proliferation and cytokine
production were significantly inhibited by a 4-fold molar excess of
soluble recombinant T1/ST2. Proliferation was not inhibited in the
presence of neutralizing anti-IL-4 mAb or anti-IL-2 mAb (data
not shown).
|
|
Discussion |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
We therefore assessed the potential of several APC- or T cell-derived
cytokines to increase T1/ST2 expression during Th2 effector
differentiation. The strongest effect (up to 8-fold increase of T1/ST2
expression) was seen with IL-6. Other cytokines, e.g., IL-1, TNF-,
and IL-5, had reproducible but smaller effects on T1/ST2 expression.
Nevertheless, the induction of T1/ST2 on Th2 cells is independent of
IL-6 since the frequency of T1/ST2+
CD4+ cells is similar in
IL-6-/- mice and their heterozygous littermates
(Fig. 4). Our data also show that APC are not necessary for T1/ST2
induction in vitro since virtually the same T1/ST2 expression kinetics
were observed in T cells stimulated either with peptide/APC or with
anti-CD3/anti-CD28 under Th2-polarizing conditions.
What then, could be the significance of the enhanced T1/ST2 expression
by APC-derived cytokines such as IL-1, TNF-, and IL-6? We have shown
recently that T1/ST2-Ig fusion protein inhibited Th2 phenotype
development in vitro. In contrast, inhibition of T1/ST2 failed to
modify IFN- production when cells were primed in the presence of
IL-12 (16). These results suggest, that inhibition of
T1/ST2 signaling results in skewing of the immune response from a Th2
to a Th1 phenotype. It is, therefore, tempting to speculate that in
vivo depending on the pathogen-induced polarizing cytokine environment,
the up-regulation of T1/ST2 expression on T cells by cytokines such as
IL-6 favors the differentiation of naive Th cells into Th2 cells. It is
clear, however, that Th2 development can occur in vivo even in the
absence of T1/ST2 (23, 24, 25).
The mechanisms by which IL-6 and other cytokines increase T1/ST2
expression remain elusive. IL-1, TNF-, and in particular IL-6 can
function as growth and/or survival factors for Th2 cells in vitro and
in vivo (26, 27). Therefore, the increased frequency of
T1/ST2+ cells observed in the presence of
exogenous IL-1, TNF-, and IL-6 could be due to enhanced T cell
proliferation. However, we did not observe an increased proliferation
in the presence of these cytokines, suggesting a direct effect on
T1/ST2 gene expression. In vitro, but not in vivo, Th2 differentiation
strictly depends on the presence of IL-4 (28). It was
therefore not surprising that none of the cytokines investigated here
induced T1/ST2 expression or enhanced IL-4 expression in naive Th cells
primed in the absence of exogenous IL-4. In vivo, however, IL-4 is
dispensable for the induction of T1/ST2 expression, which is
unperturbed in IL-4 knockout mice (12, 14).
The coexpression of different type 2 cytokines in vitro differs
somewhat from the coexpression patterns during a Th2-dominated immune
response in vivo. In pulmonary granulomas induced by schistosome eggs,
the production of IL-4 was positively correlated with IL-10 production
in T1/ST2+ but not in
T1/ST2- CD4+ cells
(13). This positive association was even stronger in
T1/ST2+ cells which did not produce IL-5
concomitantly (our unpublished observations). In vitro, however, we
found a strong positive correlation between IL-4 and IL-10 production
in both T1/ST2+ and
T1/ST2- subsets at any time point of in vitro
culture, although consistently higher correlation values were found
in those cells stained most brightly for T1/ST2 (5% brightest cells,
Fig. 2). In contrast, we found random coexpression of IL-4 and IL-5 as
well as of IL-5 and IL-10, again in both T1/ST2+
and T1/ST2- cells. Thus, under Th2-polarizing
conditions in vitro, the cytokine coexpression patterns of
T1/ST2+ or T1/ST2-
CD4+ cells do not differ significantly.
Using anti-T1/ST2 mAb as a surrogate ligand, we have investigated the functional consequences of T1/ST2 receptor triggering in vitro. Plate-bound, but not soluble anti-T1/ST2 mAb 3E10 increased both proliferation and IL-4 and IL-5 production of Th2 cells stimulated with suboptimal concentrations of either anti-CD3 alone or CD3 in combination with anti-CD28, or with APC and antigenic peptide. These results are in accordance with our recent findings in a murine model of bronchial hyperreactivity where administration of either T1/ST2-Ig fusion protein or anti-T1/ST2 mAb before Ag challenge attenuated eosinophilic airway inflammation and reduced type 2 cytokine production (12, 16). It should be noted that administration of the 3E10 mAb in vivo did not result in the depletion of Ag-specific Th2 cells, suggesting that the reduction of Th2 effector function by anti-T1/ST2-treatment as well as by T1/ST2-Ig fusion protein was due to inhibition of T1/ST2 signaling by preventing its putative ligand from binding to the T1/ST2 receptor. This may explain the apparently different effects of the anti-T1/ST2 mAb observed in vivo and in vitro.
In addition to its costimulatory effects, plate-bound mAb 3E10 induced
proliferation and cytokine production of Th2 cells in the absence of
any further stimuli. Similar findings were reported by Cocks et al.
(29), showing that ligation of the signaling lymphocytic
activation molecule in the absence of TCR triggering induced
proliferation and cytokine production in T cell lines and clones. The
mechanisms by which T1/ST2 triggers proliferation and cytokine
production of Th2 cells are currently unknown. Using a chimeric IL-1R
type I expressing the intracellular domain of T1/ST2, Mitcham et al.
(30) observed IL-1-like signaling including NF-
B
translocation into the cell upon binding of IL-1, demonstrating not
only structural but also functional homology of T1/ST2 with the IL-1RI
molecule. Whether T1/ST2 like several other IL-1R family members
utilizes a signaling pathway using MyD88, IRAK, TRAF6, or c-Jun
N-terminal kinase (31) remains to be assessed.
In summary, we have shown that the production of type 2 cytokines precedes the expression of T1/ST2 in Th2 cells polarized in vitro. Cytokines such as IL-6 can up-regulate T1/ST2 expression. Most important, cross-linking of T1/ST2 provides a costimulatory signal for Th2 but not Th1 cells and directly induces proliferation and type 2 cytokine production. Thus, T1/ST2 is not only a Th2 cell marker but also plays an important role in the activation of Th2 cells.
|
Acknowledgments |
|---|
|
Footnotes |
|---|
2 C.M. and K.B. contributed equally to this work.
3 Current address: Universitätsklinikum Charité, Institut für Medizinische Immunologie, 10098 Berlin, Germany.
4 Address correspondence and reprint requests to Dr. Thomas Kamradt, Deutsches Rheumaforschungszentrum, Schumannstrasse 21/22, 10117 Berlin, Germany.
5 Abbreviations used in this paper: SC, spleen cell; DIG, digoxigenin.
Received for publication December 20, 2000. Accepted for publication January 6, 2000.
|
References |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
This article has been cited by other articles:
|
|
 |
|
  K. Hagenow, K. A. Gelderman, M. Hultqvist, P. Merky, J. Backlund, O. Frey, T. Kamradt, and R. Holmdahl Ncf1-Associated Reduced Oxidative Burst Promotes IL-33R+ T Cell-Mediated Adjuvant-Free Arthritis in Mice J. Immunol., July 15, 2009; 183(2): 874 - 881. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Bulek, S. Swaidani, J. Qin, Y. Lu, M. F. Gulen, T. Herjan, B. Min, R. A. Kastelein, M. Aronica, M. Kosz-Vnenchak, et al. The Essential Role of Single Ig IL-1 Receptor-Related Molecule/Toll IL-1R8 in Regulation of Th2 Immune Response J. Immunol., March 1, 2009; 182(5): 2601 - 2609. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. Pecaric-Petkovic, S. A. Didichenko, S. Kaempfer, N. Spiegl, and C. A. Dahinden Human basophils and eosinophils are the direct target leukocytes of the novel IL-1 family member IL-33 Blood, February 12, 2009; 113(7): 1526 - 1534. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. D. Smithgall, M. R. Comeau, B.-R. Park Yoon, D. Kaufman, R. Armitage, and D. E. Smith IL-33 amplifies both Th1- and Th2-type responses through its activity on human basophils, allergen-reactive Th2 cells, iNKT and NK Cells Int. Immunol., August 1, 2008; 20(8): 1019 - 1030. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. H. Ho, T. Ohno, K. Oboki, N. Kajiwara, H. Suto, M. Iikura, Y. Okayama, S. Akira, H. Saito, S. J. Galli, et al. IL-33 induces IL-13 production by mouse mast cells independently of IgE-Fc{epsilon}RI signals J. Leukoc. Biol., December 1, 2007; 82(6): 1481 - 1490. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Amatucci, T. Novobrantseva, K. Gilbride, M. Brickelmaier, P. Hochman, and A. Ibraghimov Recombinant ST2 boosts hepatic Th2 response in vivo J. Leukoc. Biol., July 1, 2007; 82(1): 124 - 132. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Beauregard, C. Stevens, E. Mayhew, and J. Y. Niederkorn Cutting Edge: Atopy Promotes Th2 Responses to Alloantigens and Increases the Incidence and Tempo of Corneal Allograft Rejection J. Immunol., June 1, 2005; 174(11): 6577 - 6581. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
W. J. Simmons, M. Koneru, M. Mohindru, R. Thomas, S. Cutro, P. Singh, R. H. DeKruyff, G. Inghirami, A. J. Coyle, B. S. Kim, et al. Tim-3+ T-bet+ Tumor-Specific Th1 Cells Colocalize with and Inhibit Development and Growth of Murine Neoplasms J. Immunol., February 1, 2005; 174(3): 1405 - 1415. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. G. Agadjanyan, A. Ghochikyan, I. Petrushina, V. Vasilevko, N. Movsesyan, M. Mkrtichyan, T. Saing, and D. H. Cribbs Prototype Alzheimer's Disease Vaccine Using the Immunodominant B Cell Epitope from {beta}-Amyloid and Promiscuous T Cell Epitope Pan HLA DR-Binding Peptide J. Immunol., February 1, 2005; 174(3): 1580 - 1586. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Svendsen, C. B. Andersen, N. Willcox, A. J. Coyle, R. Holmdahl, T. Kamradt, and L. Fugger Tracking of Proinflammatory Collagen-Specific T Cells in Early and Late Collagen-Induced Arthritis in Humanized Mice J. Immunol., December 1, 2004; 173(11): 7037 - 7045. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. Schubert, B. Maier, L. Morawietz, V. Krenn, and T. Kamradt Immunization with Glucose-6-Phosphate Isomerase Induces T Cell-Dependent Peripheral Polyarthritis in Genetically Unaltered Mice J. Immunol., April 1, 2004; 172(7): 4503 - 4509. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. K. Brint, K. A. Fitzgerald, P. Smith, A. J. Coyle, J.-C. Gutierrez-Ramos, P. G. Fallon, and L. A. J. O'Neill Characterization of Signaling Pathways Activated by the Interleukin 1 (IL-1) Receptor Homologue T1/ST2. A ROLE FOR JUN N-TERMINAL KINASE IN IL-4 INDUCTION J. Biol. Chem., December 13, 2002; 277(51): 49205 - 49211. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. Kropf, S. Herath, R. Tewari, N. Syed, R. Klemenz, and I. Muller Identification of Two Distinct Subpopulations of Leishmania major-Specific T Helper 2 Cells Infect. Immun., October 1, 2002; 70(10): 5512 - 5520. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. A. Ritz, M. J. Cundall, B. U. Gajewska, D. Alvarez, J.-C. Gutierrez-Ramos, A. J. Coyle, A. N. J. McKenzie, M. R. Stampfli, and M. Jordana Granulocyte Macrophage Colony-Stimulating Factor-Driven Respiratory Mucosal Sensitization Induces Th2 Differentiation and Function Independently of Interleukin-4 Am. J. Respir. Cell Mol. Biol., October 1, 2002; 27(4): 428 - 435. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. C. Nawijn, G. M. Dingjan, R. Ferreira, B. N. Lambrecht, A. Karis, F. Grosveld, H. Savelkoul, and R. W. Hendriks Enforced Expression of GATA-3 in Transgenic Mice Inhibits Th1 Differentiation and Induces the Formation of a T1/ST2-Expressing Th2-Committed T Cell Compartment In Vivo J. Immunol., July 15, 2001; 167(2): 724 - 732. [Abstract] [Full Text] [PDF]
|
|
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
