Dichotomous response to transforming growth factor β after T cell receptor activation by naive CD4+ T cells from DBA/1 mice: Enhanced retinoic acid receptor-related orphan nuclear receptor γt expression yet reduced FoxP3 expression

Yoshiaki Morita, Doaa M. Ismail, Keith B. Elkon, Cong-Qiu Chu

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Objective To investigate the molecular mechanism for biased interleukin-17 (IL-17) production by DBA/1 CD4+ T cells upon T cell receptor (TCR) and transforming growth factor β (TGFβ) stimulation. Methods Purified naive CD4+ T cells were stimulated with anti-CD3/CD28 under Th1, Th2, Th17, and induced T regulatory (iTreg) cell conditions. Cytokine production was assayed by intracellular staining and enzyme-linked immunosorbent assay. Expression of transcription factors was determined by reverse transcription-polymerase chain reaction, flow cytometry, and immunoblotting techniques. Results Naive CD4+ T cells from DBA/1 mice produced more IL-17 under Th17 cell polarizing conditions as compared with those from C57BL/6 or BALB/c mice. Further investigation revealed no difference among the strains in terms of CD4+ T cell survival, upstream TCR signaling, or CD69 expression or in the phosphorylation of STAT-3 and expression of suppressor of cytokine signaling 3 that positively or negatively regulate IL-17 cell production. However, DBA/1 CD4+ T cells expressed increased levels of retinoic acid-related orphan receptor γt (RORγt). Furthermore, under iTreg cell polarizing conditions, DBA/1 CD4+ T cells showed a strikingly reduced level of FoxP3 expression. When interferon-γ and IL-4 were blocked, FoxP3 expression increased but remained lower in DBA/1 CD4+ T cells following exposure to TGFβ as compared with C57BL/6 CD4+ T cells. Moreover, DBA/1 CD4+ T cells showed reduced phosphorylation of Smad2 and Smad3 under both Th17 and iTreg cell polarizing conditions. Conclusion These results indicate that naive CD4+ T cells from DBA/1 mice have a dichotomous response to TGFβ: enhanced RORγt, yet reduced FoxP3, up-regulation. This observation may help to elucidate the branch point of TGFβ signaling that leads to skewed Th17, but reduced iTreg, cell differentiation.

Original languageEnglish (US)
Pages (from-to)118-126
Number of pages9
JournalArthritis and Rheumatism
Volume63
Issue number1
DOIs
StatePublished - Jan 2011

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Orphan Nuclear Receptors
Inbred DBA Mouse
Retinoic Acid Receptors
Transforming Growth Factors
T-Cell Antigen Receptor
T-Lymphocytes
Interleukin-17
Regulatory T-Lymphocytes
Phosphorylation
Cytokines
Th17 Cells
Tretinoin
Immunoblotting
Interleukin-4
Interferons
Reverse Transcription
Interleukin-2
Cell Differentiation
Cell Survival
Flow Cytometry

ASJC Scopus subject areas

  • Immunology
  • Immunology and Allergy
  • Rheumatology
  • Pharmacology (medical)

Cite this

@article{75a0036b82e84dc78de63c7bd1cae83e,
title = "Dichotomous response to transforming growth factor β after T cell receptor activation by naive CD4+ T cells from DBA/1 mice: Enhanced retinoic acid receptor-related orphan nuclear receptor γt expression yet reduced FoxP3 expression",
abstract = "Objective To investigate the molecular mechanism for biased interleukin-17 (IL-17) production by DBA/1 CD4+ T cells upon T cell receptor (TCR) and transforming growth factor β (TGFβ) stimulation. Methods Purified naive CD4+ T cells were stimulated with anti-CD3/CD28 under Th1, Th2, Th17, and induced T regulatory (iTreg) cell conditions. Cytokine production was assayed by intracellular staining and enzyme-linked immunosorbent assay. Expression of transcription factors was determined by reverse transcription-polymerase chain reaction, flow cytometry, and immunoblotting techniques. Results Naive CD4+ T cells from DBA/1 mice produced more IL-17 under Th17 cell polarizing conditions as compared with those from C57BL/6 or BALB/c mice. Further investigation revealed no difference among the strains in terms of CD4+ T cell survival, upstream TCR signaling, or CD69 expression or in the phosphorylation of STAT-3 and expression of suppressor of cytokine signaling 3 that positively or negatively regulate IL-17 cell production. However, DBA/1 CD4+ T cells expressed increased levels of retinoic acid-related orphan receptor γt (RORγt). Furthermore, under iTreg cell polarizing conditions, DBA/1 CD4+ T cells showed a strikingly reduced level of FoxP3 expression. When interferon-γ and IL-4 were blocked, FoxP3 expression increased but remained lower in DBA/1 CD4+ T cells following exposure to TGFβ as compared with C57BL/6 CD4+ T cells. Moreover, DBA/1 CD4+ T cells showed reduced phosphorylation of Smad2 and Smad3 under both Th17 and iTreg cell polarizing conditions. Conclusion These results indicate that naive CD4+ T cells from DBA/1 mice have a dichotomous response to TGFβ: enhanced RORγt, yet reduced FoxP3, up-regulation. This observation may help to elucidate the branch point of TGFβ signaling that leads to skewed Th17, but reduced iTreg, cell differentiation.",
author = "Yoshiaki Morita and Ismail, {Doaa M.} and Elkon, {Keith B.} and Cong-Qiu Chu",
year = "2011",
month = "1",
doi = "10.1002/art.27759",
language = "English (US)",
volume = "63",
pages = "118--126",
journal = "Arthritis and Rheumatology",
issn = "2326-5191",
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T1 - Dichotomous response to transforming growth factor β after T cell receptor activation by naive CD4+ T cells from DBA/1 mice

T2 - Enhanced retinoic acid receptor-related orphan nuclear receptor γt expression yet reduced FoxP3 expression

AU - Morita, Yoshiaki

AU - Ismail, Doaa M.

AU - Elkon, Keith B.

AU - Chu, Cong-Qiu

PY - 2011/1

Y1 - 2011/1

N2 - Objective To investigate the molecular mechanism for biased interleukin-17 (IL-17) production by DBA/1 CD4+ T cells upon T cell receptor (TCR) and transforming growth factor β (TGFβ) stimulation. Methods Purified naive CD4+ T cells were stimulated with anti-CD3/CD28 under Th1, Th2, Th17, and induced T regulatory (iTreg) cell conditions. Cytokine production was assayed by intracellular staining and enzyme-linked immunosorbent assay. Expression of transcription factors was determined by reverse transcription-polymerase chain reaction, flow cytometry, and immunoblotting techniques. Results Naive CD4+ T cells from DBA/1 mice produced more IL-17 under Th17 cell polarizing conditions as compared with those from C57BL/6 or BALB/c mice. Further investigation revealed no difference among the strains in terms of CD4+ T cell survival, upstream TCR signaling, or CD69 expression or in the phosphorylation of STAT-3 and expression of suppressor of cytokine signaling 3 that positively or negatively regulate IL-17 cell production. However, DBA/1 CD4+ T cells expressed increased levels of retinoic acid-related orphan receptor γt (RORγt). Furthermore, under iTreg cell polarizing conditions, DBA/1 CD4+ T cells showed a strikingly reduced level of FoxP3 expression. When interferon-γ and IL-4 were blocked, FoxP3 expression increased but remained lower in DBA/1 CD4+ T cells following exposure to TGFβ as compared with C57BL/6 CD4+ T cells. Moreover, DBA/1 CD4+ T cells showed reduced phosphorylation of Smad2 and Smad3 under both Th17 and iTreg cell polarizing conditions. Conclusion These results indicate that naive CD4+ T cells from DBA/1 mice have a dichotomous response to TGFβ: enhanced RORγt, yet reduced FoxP3, up-regulation. This observation may help to elucidate the branch point of TGFβ signaling that leads to skewed Th17, but reduced iTreg, cell differentiation.

AB - Objective To investigate the molecular mechanism for biased interleukin-17 (IL-17) production by DBA/1 CD4+ T cells upon T cell receptor (TCR) and transforming growth factor β (TGFβ) stimulation. Methods Purified naive CD4+ T cells were stimulated with anti-CD3/CD28 under Th1, Th2, Th17, and induced T regulatory (iTreg) cell conditions. Cytokine production was assayed by intracellular staining and enzyme-linked immunosorbent assay. Expression of transcription factors was determined by reverse transcription-polymerase chain reaction, flow cytometry, and immunoblotting techniques. Results Naive CD4+ T cells from DBA/1 mice produced more IL-17 under Th17 cell polarizing conditions as compared with those from C57BL/6 or BALB/c mice. Further investigation revealed no difference among the strains in terms of CD4+ T cell survival, upstream TCR signaling, or CD69 expression or in the phosphorylation of STAT-3 and expression of suppressor of cytokine signaling 3 that positively or negatively regulate IL-17 cell production. However, DBA/1 CD4+ T cells expressed increased levels of retinoic acid-related orphan receptor γt (RORγt). Furthermore, under iTreg cell polarizing conditions, DBA/1 CD4+ T cells showed a strikingly reduced level of FoxP3 expression. When interferon-γ and IL-4 were blocked, FoxP3 expression increased but remained lower in DBA/1 CD4+ T cells following exposure to TGFβ as compared with C57BL/6 CD4+ T cells. Moreover, DBA/1 CD4+ T cells showed reduced phosphorylation of Smad2 and Smad3 under both Th17 and iTreg cell polarizing conditions. Conclusion These results indicate that naive CD4+ T cells from DBA/1 mice have a dichotomous response to TGFβ: enhanced RORγt, yet reduced FoxP3, up-regulation. This observation may help to elucidate the branch point of TGFβ signaling that leads to skewed Th17, but reduced iTreg, cell differentiation.

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