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IMMUNOBIOLOGY
From the Department of Internal Medicine II, Chiba
University School of Medicine, Japan; Department of Hematopoietic
Factors, The Institute of Medical Science, University of Tokyo, Japan;
and Takara Shuzo, Biotechnology Research Laboratories, Shiga, Japan.
We have previously shown that CD4+ T cell-mediated
allergic inflammation is diminished in signal transducer and activator
of transcription (Stat)5a-deficient (Stat5a Newly activated CD4+ T helper cells
differentiate into at least 2 functionally distinct subsets as defined
by their pattern of cytokine production.1,2 T helper (Th)1
cells produce interferon Stat5a and Stat5b are cytosolic latent transcription factors that
are activated by a very wide range of cytokines, including immunologically important cytokines, IL-2 and IL-7.12,13
Although Stat5a and Stat5b are highly homologous, the different
phenotypes of Stat5a Recently, we have shown that antigen-induced IL-5 production and
subsequent eosinophil recruitment in the airways are severely decreased
in Stat5a Therefore, to determine whether Stat5a regulates Th1 and Th2 cell
differentiation, we studied T helper cell differentiation of
Stat5a Mice
T-cell purification
Cell culture Splenocytes (2 × 106 cells/mL) from wild-type mice, Stat5a / mice, or Stat5b/ mice
were stimulated with platebound anti-CD3 monoclonal antibody (mAb)
(5 µg/mL, clone 145-2C11, Pharmingen, San Diego, CA) in RPMI 1640 medium supplemented with 10% heat-inactivated fetal calf serum, 50 µM  -mercaptoethanol, 2 mM L-glutamine, and antibiotics in a
24-well microtiter plate at 37°C for 48 hours. Splenocytes (2 × 106 cells/mL) from DO10+ mice or
DO10+Stat5a/ mice were stimulated with
OVA323-339 peptide (0.1 µg/mL) at 37°C for 48 hours. Purified
splenic CD4+ T cells (1 × 106 cells/mL) from
DO10+ mice or DO10+Stat5a/ mice
were also stimulated with platebound anti-CD3 mAb plus anti-CD28 mAb
(5 µg/mL, clone 37.51, Pharmingen) at 37°C for 48 hours. Where
indicated, IL-12 (7.5 ng/mL, R&D Systems) was added to polarize toward
Th1 cells (Th1 condition), and IL-4 (15 ng/mL, R&D Systems),
anti-IL-12 mAb (15 µg/mL, clone C17.8, Pharmingen), and
anti-IFN- mAb (15 µg/mL, clone XMG1.2, Pharmingen) were added to
polarize toward Th2 cells (Th2 condition) as described
previously.23 Cells were then washed with
phosphate-buffered saline (PBS) and cultured for another 3 days in the
presence of indicated cytokines without antigen stimulation. IL-2 (20 U/mL, R&D Systems) was added in the second culture to support
the survival.
CFSE labeling of splenocytes Splenocytes from DO10+ mice or DO10+Stat5a/ mice were labeled with
5-(and-6)-carboxyfluorescein diacetate, succinimidyl ester (CFSE,
Molecular Probes, Eugene, OR) according to the manufacturer's instruction.
Retrovirus-mediated gene expression Retrovirus-mediated gene expression for splenic T cells was performed as described elsewhere.24 Bicistronic retrovirus vectors, pMX-Stat5a-IRES-GFP, pMX-Stat5b-IRES-GFP, and pMX-IRES-GFP (as a negative control), were constructed as previously described.25 High-titer retroviruses (average titer 1 × 107/mL) were produced with a transient retrovirus packaging cell line, Plat-E,26 and stored at 80°C
until use. A fraction of the retroviruses was tested for the ability of
infection on Ba/F3 cells, and the retroviruses that reached over 70%
of infection (as determined by green fluorescent protein
[GFP]+ population using fluorescence-activated cell
sorting [FACS]) were used in the following experiments. For infection
on T cells, after purified CD4+ T cells from
DO10+ mice or DO10+Stat5a/ mice
were stimulated with anti-CD3 mAb plus anti-CD28 mAb for 48 hours as
described above, cells were washed with PBS and incubated with 500 µL
of the retroviruses in the presence of IL-2 (20 U/mL) in a 24-well
microtiter plate that was coated with recombinant fibronectin fragment
CH-296, Retronectin (27 µg/mL, Takara, Ohtsu, Japan). After 4 hours
of infection, 500 µL fresh medium was added to the culture and cells
were allowed to grow for another 60 hours before being subjected to
intracellular cytokine analysis. Under these conditions, the efficiency
of infection to CD4+ T cells was 15% to 30% as assessed
by GFP+ cells by FACS.
Flow cytometric analysis Cells were stained and analyzed on a FACScaliber (Becton Dickinson, San Jose, CA) using CellQuest software. The following antibodies were purchased: anti-CD4 fluorescein isothiocynate (FITC), Allophycocyanin (APC), peridinin chlorophyll protein (PerCP) (RM4-5, Pharmingen), anti-CD25 FITC (7D4, Pharmingen), anti-CD25 phycoerythrin (PE) (PC61, Pharmingen), anti-c PE (4G2, Pharmingen), anti-CDw124 (IL-4R ) (1688-01, Genzyme, Cambridge, MA), KJI-26 FITC,22 and antirat IgG2a FITC (RG7/1.30, Pharmingen).
Prior to staining, Fc receptors were blocked with anti-CD16/32 antibody (2.4G2, Pharmingen). Negative controls consisted of isotype-matched, directly conjugated, nonspecific antibodies (Pharmingen).
Intracellular cytokine analysis Cultured cells were washed with PBS and restimulated with platebound anti-CD3 mAb at 37°C for 6 hours in the presence of IL-2 (10 U/mL), with monensin (2 µM) (Sigma, St Louis, MO) added for the final 4 hours to prevent cytokine release. Cells were harvested, washed with PBS, and stained with anti-CD4 PerCP for 30 minutes at 4°C. Cells were washed with PBS, fixed with IC Fix (Biosource International, Camarillo, CA), permeabilized with IC Perm (Biosource International), and stained with anti-IL-4 PE (BVD4-1D11, Pharmingen) and anti-IFN- APC (XMG1.2, Pharmingen) for 30 minutes at 4°C. After
washing, cytokine profile (IL-4 vs IFN-) on CD4+ cells
or CD4+GFP+ cells (in the case of retrovirus
experiments) was analyzed on a FACScaliber using CellQuest software. In
preliminary experiments, we found that these procedures did not
significantly decrease the frequency of GFP+ cells within
CD4+ T cells.
Western blotting Cells were washed with PBS and lysed in lysis buffer (1% Nonidet P-40, 20 mM Tris-HCl [pH 8.0], 50 mM NaCl, 2 mM dithiothreitol, 4 mM EGTA, 10 mM NaF, 1 mM Na3VO4, 5 µg/mL aprotinin, 5 µg/mL leupeptin, 2 µg/mL pepstatin, 0.5 mM phenylmethylsulfonyl fluoride, and 10% glycerol) on ice for 30 minutes, and cell lysates were prepared by centrifugation. A total of 15 µg of cell lysates was separated on 6% or 10% sodium dodecyl sulfate polyacrylamide gels and transferred to Immobilon-P membranes (Millipore, Bedford, MA). After blocking with PBS containing 0.15% Tween 20 and 3% bovine serum albumin for 1 hour at room temperature, the membranes were incubated with antisera to mouse Stat5 (Transduction Laboratories, Lexington, KY), mouse Stat6 (M-200) (Santa Cruz Biotechnology, Santa Cruz, CA), phospho-Stat5 (New England Biolabs, Beverly, MA), or phospho-Stat6 (New England Biolabs) for 1 hour at room temperature. After washing 3 times with PBS containing 0.15% Tween 20, the membranes were incubated with antirabbit IgG or antimouse IgG antibodies conjugated with horseradish peroxidase (Amersham Pharmacia Biotech, Little Chalfont, United Kingdom) in PBS containing 0.15% Tween 20 and 3% bovine serum albumin for 1 hour at room temperature. The membranes were then washed with PBS containing 0.15% Tween 20 and developed with an enhanced chemiluminescent substrate (Roche Diagnostics, Mannheim, Germany).Depletion of CD4+CD25+ T cells from splenocytes Splenocytes from DO10+ mice or DO10+Stat5a/ mice were incubated with
biotinylated anti-CD25 antibody (clone PC61, Immunotech, Marseilles,
France) at 4°C for 20 minutes. After washing with PBS, cells were
incubated with streptavidin-magnetic microbeads (Miltenyi Biotec,
Sunnyvale, CA) at 4°C for 20 minutes, washed 3 times with PBS, and
passed through the magnetic-activated cell separation BS column
(Miltenyi Biotec) according to the manufacturer's instructions. As a
control, cells were incubated with isotype-matched biotinylated
antibody (Pharmingen), followed by the incubation with
streptavidin-magnetic microbeads, and passed through the BS column. The
efficiency of depletion was evaluated by FACS with anti-CD4 APC and
anti-CD25 FITC (clone 7D4, Pharmingen), which recognizes the different
epitope of murine CD25 from that of PC61.27 More than 99%
of CD4+CD25+ T cells were removed from
splenocytes by this procedure, whereas other cell types, including
CD4+CD25 T cells, CD8+ cells,
B220+ cells, Mac-1+ cells, and NK cells, were
not eliminated (data not shown).
Data analysis Data are summarized as mean ± SD. The statistical analysis of the results was performed by the unpaired t test. P values below .05 were considered significant.
Development of Th2 cells from splenocytes is decreased in
Stat5a / mice,17 suggesting
that Th2 cell-mediated immune responses are diminished in
Stat5a/ mice. Therefore, to determine whether Stat5a
plays a regulatory role in T helper cell differentiation, we first
examined T helper cell differentiation in Stat5a/ mice
at single-cell levels by intracellular cytokine analysis. When
stimulated with anti-CD3 mAb, CD4+ T cells that produced
IL-4 but not IFN- (Th2 cells) were significantly decreased in
Stat5a/ mice as compared with those in wild-type mice
(wild-type mice 6.0% ± 1.1% vs Stat5a/ mice
0.2% ± 0.1%, mean ± SD, n = 5 mice in each group,
P < .001) (Figure 1). In
contrast, CD4+ T cells that produced IFN- but not IL-4
(Th1 cells) were significantly increased in Stat5a/
mice (wild-type mice 25.8% ± 3.7% vs Stat5a/ mice
42.0% ± 4.9%, P < .001) (Figure 1), suggesting that
T helper cell differentiation is biased toward Th1 cells in
Stat5a/ mice. In contrast, both Th1 cells and Th2 cells
were decreased in Stat5b/ mice as compared with those
in wild-type mice (Figure 1).
To address the role of Stat5a in T helper cell differentiation in
more detail, Stat5a
We further examined the effect of Th1 or Th2 polarization on T helper
cell differentiation in DO10+Stat5a Development of Th2 cells from purified CD4+ T cells is
impaired in Stat5a / mice. While Th2 cells were
readily observed when CD4+ T cells were stimulated with
antigenic peptide in the presence of antigen-presenting cells (Figure
2A), almost no Th2 cells were observed when purified CD4+ T
cells were stimulated with anti-CD3 mAb plus anti-CD28 mAb (Figure
3A). As a result, no significant
difference was observed in the frequency of Th1 cells and Th2 cells in
DO10+ mice and DO10+Stat5a/
mice in nonpolarizing Th0 condition (n = 4 mice, each) (Figure 3A,B).
In addition, similar frequency of Th1 cells was observed in
DO10+ mice and DO10+Stat5a/
mice in Th1-polarizing condition (Figure 3C and 3D). Interestingly, in
contrast to Th1 cell differentiation, Th2 cell differentiation was
significantly decreased in DO10+Stat5a/
mice even when purified CD4+ T cells were stimulated with
anti-CD3 mAb plus anti-CD28 mAb in Th2-polarizing condition
(DO10+ mice 30.0% ± 3.8% vs
DO10+Stat5a/ mice 13.7% ± 2.3%,
n = 4, P < .001) (Figure 3E,F). These results indicate
that the intrinsic expression of Stat5a in CD4+ T cells is
required for the IL-4-dependent Th2 cell differentiation.
Impaired T helper cell differentiation in
Stat5a /CD4+ T cells might be
involved in the increased Th1 cell differentiation in
Stat5a/ mice. Therefore, we examined cell-cycle
progression of activated CD4+ T cells in
DO10+Stat5a/ mice using CFSE labeling
methods.30 As shown in Figure
4, the frequency of cell division was
indistinguishable between DO10+CD4+ T cells and
DO10+Stat5a/CD4+ T cells
(Figure 4A vs 4B). Moreover, the frequency of IL-4-producing cells was
decreased but that of IFN--producing cells was increased in
DO10+Stat5a/ mice as compared with those in
DO10+ mice regardless of the rounds of cell-cycle
progression (Figure 4C vs 4D for IL-4, Figure 4E vs 4F for IFN-).
Thus, these results indicate that the impaired T helper cell
differentiation in Stat5a/CD4+ T cells is
independent of cell-cycle progression.
IL-4 normally phosphorylates Stat6 in the absence of Stat5a Because the development of Th2 cells was decreased in DO10+Stat5a/CD4+ T cells even
in the presence of IL-4 (Figures 2 and 3), it was possible that IL-4
signaling was impaired in Stat5a/CD4+ T
cells. Therefore, we analyzed the expression of IL-4 receptor components (IL-4R and c) and IL-4-induced Stat6 phosphorylation in DO10+Stat5a/ mice. As shown in Figure
5, no significant difference was observed in the expression of IL-4R on CD4+ T cells between
DO10+ mice and DO10+Stat5a/
mice (Figure 5A for unstimulated CD4+ T cells and Figure 5B
for activated CD4+ T cells). The expression of c was
also normal in CD4+ T cells from
DO10+Stat5a/ mice (Figure 5A,B). Moreover,
Stat6 was equally expressed and phosphorylated upon IL-4 stimulation in
CD4+ T cells from DO10+ mice and
DO10+Stat5a/ mice (Figure 5C).
IL-4 does not phosphorylate Stat5 even in the absence of Stat6 Recently, it was reported that IL-4 activated Stat5 in a pro-B cell line and that the activation of Stat5 was involved in IL-4-induced cell proliferation.31 Thus, it was possible that Stat5a was activated directly by IL-4 and enhanced the IL-4-induced Th2 cell expansion. To determine whether IL-4 activates Stat5 in CD4+ T cells, we analyzed the phosphorylation of Stat5 in IL-4-stimulated CD4+ T cells. Inconsistent with the previous report,31 no Stat5 phosphorylation was detected by IL-4 stimulation in wild-type CD4+ T cells (Figure 6). To exclude the possibility that Stat5 activation via IL-4R (IL-4 receptor) was blocked by the presence of Stat6, we analyzed IL-4-induced phosphorylation of Stat5 in Stat6/CD4+ T cells. As shown in Figure
6, IL-4 did not phosphorylate Stat5 even in the absence of Stat6. In
contrast, IL-2 efficiently phosphorylated Stat5 in both wild-type
CD4+ T cells and Stat6/CD4+ T
cells (Figure 6). As expected, both Stat5a and Stat5b were equally
expressed in wild-type CD4+ T cells and
Stat6/CD4+ T cells (Figure 6). Therefore,
it is unlikely that the impaired Th2 cell differentiation results from
the direct involvement of Stat5a in IL-4 signaling.
Retrovirus-mediated expression of Stat5a restores Th2 cell
differentiation in Stat5a / mice,
we employed a bicistronic retrovirus-mediated gene expression system in
which infected cells were identified by coexpressed GFP.32
Purified CD4+ T cells from
DO10+Stat5a/ mice were stimulated with
anti-CD3 mAb plus anti-CD28 mAb and infected with either Stat5a or
control retrovirus in either Th0-, Th1-, or Th2-polarizing condition.
As a control, CD4+ T cells from DO10+ mice were
stimulated with anti-CD3 mAb plus anti-CD28 mAb and infected with
control retrovirus. Three days after infection, cells were analyzed for
intracellular cytokines (IL-4 vs IFN-) on GFP-expressing
CD4+ T cells (GFP+CD4+ T cells)
(Figure 7). As shown in the upper panels
of Figure 7, the expression of Stat5a itself did not induce Th2 cell
differentiation in
DO10+Stat5a/CD4+ T cells in Th0
condition (Figure 7). However, the retrovirus-mediated expression of
Stat5a rescued Th2 cell differentiation in
DO10+Stat5a/CD4+ T cells at the
similar levels to that in DO10+CD4+ T cells in
Th2-polarizing condition (n = 4 experiments) (Figure 7, lower
panels). In contrast to infected GFP+CD4+ T
cells, uninfected GFPCD4+ T cells were not
affected for their T helper cell differentiation by the presence of
GFP+CD4+ T cells (data not shown). These
results suggest that the intrinsic expression of Stat5a is required for
the appropriate Th2 cell differentiation of CD4+ T cells
and that soluble factor(s) from Stat5a-expressing
GFP+CD4+ T cells is not sufficient for the Th2
cell differentiation of uninfected GFPCD4+ T
cells. On the other hand, the retrovirus-mediated expression of Stat5a
did not affect the Th2 cell differentiation of CD4+ T cells
in DO10+ mice (data not shown), suggesting that the
endogenous levels of Stat5 protein are sufficient for Th2 cell
differentiation. Moreover, the enforced expression of Stat5a did not
affect the Th1 cell differentiation in
DO10+Stat5a/CD4+ T cells in
Th1-polarizing condition (Figure 7).
Retrovirus-mediated expression of Stat5b also restores Th2 cell
differentiation in Stat5a / mice. Interestingly, the
retrovirus-mediated expression of Stat5b rescued Th2 cell
differentiation in
DO10+Stat5a/CD4+ T cells as
efficiently as Stat5a-expressing retroviruses (Figure 8). These results suggest that the role
of Stat5a in Th2 cell differentiation is shared with that of Stat5b and
that the expression levels of Stat5 proteins may be important for the
regulation of Th2 cell differentiation.
CD4+CD25+ immunoregulatory T cells
are decreased in Stat5a /
mice. Interestingly, the number of CD4+CD25+ T
cells was significantly decreased in
DO10+Stat5a/ mice as compared with
that in DO10+ mice (DO10+ mice 5.4% ± 0.8%
vs DO10+Stat5a/ mice 1.7% ± 0.3% in
CD4+ T cells, n = 5 mice in each group,
P < .001) (Figure 9).
Depletion of CD4+CD25+ T cells prevents Th2 cell differentiation We have recently shown that Th2 cell-mediated eosinophil recruitment into the airways is decreased by the depletion of CD4+CD25+ immunoregulatory T cells (A. Suto, unpublished data, 2000), suggesting that CD4+CD25+ immunoregulatory T cells modulate T helper cell differentiation toward Th2-type. To determine whether CD4+CD25+ T cells regulate the differentiation of Th1 and Th2 cells in the presence or absence of Stat5a, we further examined the effect of CD4+CD25+ T-cell depletion on antigen-induced Th1 and Th2 cell differentiation from splenocytes in DO10+ mice and DO10+Stat5a/ mice. As shown in the left
panels of Figure 10, the depletion of
CD4+CD25+ T cells from DO10+
splenocytes significantly decreased the development of Th2 cells (41.0% vs 8.1%) but increased the development of Th1 cells (10.5% vs
44.2%). However, the depletion of CD4+CD25+ T
cells from DO10+Stat5a/ splenocytes had no
effect on the differentiation of Th1 cells and Th2 cells (Figure 10,
right panels). Taken together, these results suggest that the decreased
CD4+CD25+ T cells may account in part for the
decreased Th2 cell differentiation of CD4+ T cells in
DO10+Stat5a/ mice.
In this study, we first show that Stat5a modulates T helper cell
differentiation of CD4+ T cells toward Th2 cells. We found
that Th2 cell differentiation of CD4+ T cells was
significantly decreased in DO10+Stat5a Second, we also show that Stat5a is involved in the development of
CD4+CD25+ immunoregulatory T cells, as
indicated by a significant decrease in the number of
CD4+CD25+ T cells in
DO10+Stat5a Because Th2 cell differentiation was impaired in CD4+ T
cells from Stat5a The precise mechanisms by which Stat5a promotes Th2 cell
differentiation are unclear. Stat5 has been shown to up-regulate a
number of molecules, including cytokine-inducible SH2 proteins (CIS family, also referred to as the SOCS or SSI
family).37 Accumulating evidence suggests that some CIS
family proteins might be involved in the cross-regulation of cytokine
network and may regulate Th1 cell and Th2 cell
differentiation.38,39 CIS1, a prototype of CIS family
proteins, is induced by Stat5 and inhibits Stat5 activation by blocking
the interaction between Stat5 and cytokine receptors.37
Thus, CIS1 seems to function in a classical negative feedback of Stat5
signaling. Recently, the CIS1 transgenic mice were reported and the
phenotype of these mice remarkably resembled that found in
Stat5a Stat5a is known to potently regulate the expression of CD25 by directly
binding to the 5' regulatory region of the CD25 gene.33,34 Our findings that the development of CD4+CD25+
T cells was impaired in DO10+Stat5a We demonstrate that CD4+CD25+ T cells play an
important regulatory role in Th2 cell differentiation (Figure 10). We
have also recently found that CD4+CD25+ T cells
enhance Th2 cell-mediated allergic inflammation in vivo (A. Suto,
unpublished data 2000). Regarding Th2 cell differentiation induced by the presence of CD4+CD25+ T cells,
cytokines produced by CD4+CD25+ T cells may not
significantly affect the differentiation of
CD4+CD25 We show that both Th1 cells and Th2 cells are decreased in
Stat5b In conclusion, our results indicate that the expression of Stat5a is required both for the appropriate differentiation of Th2 cells upon antigen stimulation and for the development of CD4+CD25+ immunoregulatory T cells that modulate T helper cell differentiation toward Th2 cells. Thus, specific inactivation of Stat5a could potentially represent a basis for the treatment of Th2-biased, allergic diseases such as asthma.
We thank Drs L. Hennighausen and X. Liu for
Stat5a
Submitted August 23, 2000; accepted December 12, 2000.
Supported in part by grants from the Ministry of Education, Science and Culture of Japan, the Japan Allergy Foundation, and a Chiba University research grant.
The publication costs of this article were defrayed in part by page charge payment. Therefore, and solely to indicate this fact, this article is hereby marked "advertisement" in accordance with 18 U.S.C. section 1734.
Reprints: Hiroshi Nakajima, Dept of Internal Medicine II, Chiba University School of Medicine, 1-8-1 Inohana, Chiba City, Chiba 260-8670, Japan; e-mail: nakajimh{at}intmed02.m.chiba-u.ac.jp.
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  S. Furuta, S.-i. Kagami, T. Tamachi, K. Ikeda, M. Fujiwara, A. Suto, K. Hirose, N. Watanabe, Y. Saito, I. Iwamoto, et al. Overlapping and Distinct Roles of STAT4 and T-bet in the Regulation of T Cell Differentiation and Allergic Airway Inflammation J. Immunol., May 15, 2008; 180(10): 6656 - 6662. [Abstract] [Full Text] [PDF]
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M. Omori and S. Ziegler Induction of IL-4 Expression in CD4+ T Cells by Thymic Stromal Lymphopoietin J. Immunol., February 1, 2007; 178(3): 1396 - 1404. [Abstract] [Full Text] [PDF]
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A. Suto, H. Nakajima, N. Tokumasa, H. Takatori, S.-i. Kagami, K. Suzuki, and I. Iwamoto Murine Plasmacytoid Dendritic Cells Produce IFN-{gamma} upon IL-4 Stimulation J. Immunol., November 1, 2005; 175(9): 5681 - 5689. [Abstract] [Full Text] [PDF]
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H. Takatori, H. Nakajima, S.-i. Kagami, K. Hirose, A. Suto, K. Suzuki, M. Kubo, A. Yoshimura, Y. Saito, and I. Iwamoto Stat5a Inhibits IL-12-Induced Th1 Cell Differentiation through the Induction of Suppressor of Cytokine Signaling 3 Expression J. Immunol., April 1, 2005; 174(7): 4105 - 4112. [Abstract] [Full Text] [PDF]
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H. Takatori, H. Nakajima, K. Hirose, S.-i. Kagami, T. Tamachi, A. Suto, K. Suzuki, Y. Saito, and I. Iwamoto Indispensable Role of Stat5a in Stat6-Independent Th2 Cell Differentiation and Allergic Airway Inflammation J. Immunol., March 15, 2005; 174(6): 3734 - 3740. [Abstract] [Full Text] [PDF]
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M. Koyanagi, K.'i. Imanishi, Y. Arimura, H. Kato, J. Yagi, and T. Uchiyama Immunologic immaturity, but high IL-4 productivity, of murine neonatal thymic CD4 single-positive T cells in the last stage of maturation Int. Immunol., February 1, 2004; 16(2): 315 - 326. [Abstract] [Full Text] [PDF]
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J. W. Snow, N. Abraham, M. C. Ma, B. G. Herndier, A. W. Pastuszak, and M. A. Goldsmith Loss of Tolerance and Autoimmunity Affecting Multiple Organs in STAT5A/5B-Deficient Mice J. Immunol., November 15, 2003; 171(10): 5042 - 5050. [Abstract] [Full Text] [PDF]
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P. Anderson, A. Sundstedt, L. Li, E. J. O'Neill, S. Li, D. C. Wraith, and P. Wang Differential activation of signal transducer and activator of transcription (STAT)3 and STAT5 and induction of suppressors of cytokine signalling in Th1 and Th2 cells Int. Immunol., November 1, 2003; 15(11): 1309 - 1317. [Abstract] [Full Text] [PDF]
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S. Thomas, R. Kumar, A. Preda-Pais, S. Casares, and T.-D. Brumeanu A Model for Antigen-Specific T-Cell Anergy: Displacement of CD4-p56lck Signalosome from the Lipid Rafts by a Soluble, Dimeric Peptide-MHC Class II Chimera J. Immunol., June 15, 2003; 170(12): 5981 - 5992. [Abstract] [Full Text] [PDF]
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A. Kunisato, S. Chiba, E. Nakagami-Yamaguchi, K. Kumano, T. Saito, S. Masuda, T. Yamaguchi, M. Osawa, R. Kageyama, H. Nakauchi, et al. HES-1 preserves purified hematopoietic stem cells ex vivo and accumulates side population cells in vivo Blood, March 1, 2003; 101(5): 1777 - 1783. [Abstract] [Full Text] [PDF]
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Y. Seto, H. Nakajima, A. Suto, K. Shimoda, Y. Saito, K. I. Nakayama, and I. Iwamoto Enhanced Th2 Cell-Mediated Allergic Inflammation in Tyk2-Deficient Mice J. Immunol., January 15, 2003; 170(2): 1077 - 1083. [Abstract] [Full Text] [PDF]
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D. Xu, H. Liu, M. Komai-Koma, C. Campbell, C. McSharry, J. Alexander, and F. Y. Liew CD4+CD25+ Regulatory T Cells Suppress Differentiation and Functions of Th1 and Th2 Cells, Leishmania major Infection, and Colitis in Mice J. Immunol., January 1, 2003; 170(1): 394 - 399. [Abstract] [Full Text] [PDF]
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A. Suto, H. Nakajima, K. Hirose, K. Suzuki, S.-i. Kagami, Y. Seto, A. Hoshimoto, Y. Saito, D. C. Foster, and I. Iwamoto Interleukin 21 prevents antigen-induced IgE production by inhibiting germ line Cepsilon transcription of IL-4-stimulated B cells Blood, December 15, 2002; 100(13): 4565 - 4573. [Abstract] [Full Text] [PDF]
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E. S. Hwang, I. A. White, and I-C. Ho An IL-4-independent and CD25-mediated function of c-maf in promoting the production of Th2 cytokines PNAS, October 1, 2002; 99(20): 13026 - 13030. [Abstract] [Full Text] [PDF]
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 G. Kunstle, J. Laine, G. Pierron, S.-i. Kagami, H. Nakajima, F. Hoh, C. Roumestand, M.-H. Stern, and M. Noguchi Identification of Akt Association and Oligomerization Domains of the Akt Kinase Coactivator TCL1 Mol. Cell. Biol., March 1, 2002; 22(5): 1513 - 1525. [Abstract] [Full Text] [PDF]
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A. Suto, H. Nakajima, K. Ikeda, S. Kubo, T. Nakayama, M. Taniguchi, Y. Saito, and I. Iwamoto CD4+CD25+ T-cell development is regulated by at least 2 distinct mechanisms Blood, January 15, 2002; 99(2): 555 - 560. [Abstract] [Full Text] [PDF]
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Abstract
Introduction
T cells and the impaired IL-2R