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T-bet is a STAT1-induced regulator of IL-12R expression in naïve CD4+ T cells

Abstract

T helper type 1 (TH1) cell development involves interferon-γ (IFN-γ) signaling through signal transducer and activator of transcription 1 (STAT1) and interleukin-12 (IL-12) signaling through STAT4 activation. We examined here T-bet regulation and evaluated the actions of T-bet in STAT1- and STAT4-dependent TH1 development processes. We found that T-bet expression during T cell activation was strongly dependent on IFN-γ signaling and STAT1 activation, but was independent of STAT4. Ectopic T-bet expression strongly increased IFN-γ production in TH2 cells activated by PMA-ionomycin, but weakly increased IFN-γ production in TH2 cells stimulated by IL-12–IL-18 or OVA peptide–antigen-presenting cell stimulation. In contrast, IL-12–IL-18–induced IFN-γ production remained STAT4-dependent despite ectopic T-bet expression. Ectopic T-bet expression selectively induced expression of IL-12Rβ2, but not IL-18Rα, in wild-type and STAT1−/− TH2 cells, but did not extinguish expression of GATA-3 and TH2 cytokines. Finally, ectopic T-bet did not directly induce expression of endogenous T- bet independently of IFN-γ or STAT1. Thus, T-bet is induced by IFN-γ and STAT1 signaling during T cell activation. In addition, T-bet mediates STAT1-dependent processes of TH1 development, including the induction of IL-12Rβ2.

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Figure 1: PMA-ionomycin enhances STAT4-independent IFN-γ production.
Figure 2: Expression of the gene encoding T-bet is STAT1-dependent.
Figure 3: The effect of T-bet on cytokine production in STAT4−/− and STAT1−/− T cells.
Figure 4: T-bet expression is compatible with TH2 cytokine production.
Figure 5: T-bet induces IL-12Rβ2 expression by a STAT1-independent mechanism.

References

  1. Glimcher, L.H. & Murphy, K.M. Lineage commitment in the immune system: the T helper lymphocyte grows up. Genes Dev. 14, 1693–1711 (2000).

    CAS  PubMed  Google Scholar 

  2. Murphy, K.M. et al. Signaling and transcription in T helper development. Annu. Rev. Immunol. 18, 451–494 (2000).

    Article  CAS  Google Scholar 

  3. Szabo, S.J., Dighe, A.S., Gubler, U. & Murphy, K.M. Regulation of the interleukin (IL)-12Rβ2 subunit expression in developing T helper 1 (TH1) and TH2 cells. J. Exp. Med. 185, 817–824 (1997).

    Article  CAS  Google Scholar 

  4. Presky, D.H. et al. A functional interleukin 12 receptor complex is composed of two β-type cytokine receptor subunits. Proc. Natl. Acad. Sci. USA 93, 14002–14007 (1996).

    Article  CAS  Google Scholar 

  5. Carter, L.L. & Murphy, K.M. Lineage-specific requirement for signal transducer and activator of transcription (Stat)4 in interferon γ production from CD4+versus CD8+ T cells. J. Exp. Med. 189, 1355–1360 (1999).

    Article  CAS  Google Scholar 

  6. Kaplan, M.H., Wurster, A.L., Smiley, S.T. & Grusby, M.J. STAT6-dependent and -independent pathways for IL-4 production. J. Immunol. 163, 6536–6540 (1999).

    CAS  PubMed  Google Scholar 

  7. Lawless, V.A. et al. STAT4 regulates multiple components of IFN-γ-inducing signaling pathways. J. Immunol. 165, 6803–6808 (2000).

    Article  CAS  Google Scholar 

  8. Hsieh, C.S., Macatonia, S.E., O'Garra, A. & Murphy, K.M. Pathogen-induced TH1 phenotype development in CD4+ αβ-TCR transgenic T cells is macrophage dependent. Int. Immunol. 5, 371–382 (1993).

    Article  CAS  Google Scholar 

  9. Thierfelder, W.E. et al. Requirement for STAT4 in interleukin-12-mediated responses of natural killer and T cells. Nature 382, 171–174 (1996).

    Article  CAS  Google Scholar 

  10. Kaplan, M.H., Sun, Y.L., Hoey, T. & Grusby, M.J. Impaired IL-12 responses and enhanced development of TH2 cells in STAT4-deficient mice. Nature 382, 174–177 (1996).

    Article  CAS  Google Scholar 

  11. Szabo, S.J. et al. A novel transcription factor, T-bet, directs TH1 lineage commitment. Cell 100, 655–669 (2000).

    Article  CAS  Google Scholar 

  12. Rengarajan, J., Szabo, S.J. & Glimcher, L.H. Transcriptional regulation of TH1/TH2 polarization. Immunol. Today 21, 479–483 (2000).

    Article  CAS  Google Scholar 

  13. Rao, A. & Avni, O. Molecular aspects of T-cell differentiation. Brit. Med. Bull. 56, 969–984 (2000).

    Article  CAS  Google Scholar 

  14. Mullen, A.C. et al. Role of T-bet in commitment of TH1 cells before IL-12-dependent selection. Science 292, 1907–1910 (2001).

    Article  CAS  Google Scholar 

  15. Lighvani, A.A. et al. T-bet is rapidly induced by interferon-γ in lymphoid and myeloid cells. Proc. Natl. Acad. Sci. USA 98, 15137–15142 (2001).

    Article  CAS  Google Scholar 

  16. Robinson, D. et al. IGIF does not drive TH1 development but synergizes with IL-12 for interferon-γ production and activates IRAK and NFκB. Immunity 7, 571–581 (1997).

    Article  CAS  Google Scholar 

  17. Yang, J., Murphy, T.L., Ouyang, W. & Murphy, K.M. Induction of interferon-γ production in TH1 CD4+ T cells: evidence for two distinct pathways for promoter activation. Eur. J. Immunol. 29, 548–555 (1999).

    Article  CAS  Google Scholar 

  18. Yang, J., Zhu, H., Murphy, T.L., Ouyang, W. & Murphy, K.M., IL-18-stimulated GADD45β required in cytokine-induced, but not TCR-induced, IFN-γ production. Nature Immunol. 2, 157–164 (2001).

    Article  CAS  Google Scholar 

  19. Wenner, C.A., Guler, M.L., Macatonia, S.E., O'Garra, A. & Murphy, K.M. Roles of IFN-γ and IFN-α in IL-12-induced T helper cell-1 development. J. Immunol. 156, 1442–1447 (1996).

    CAS  PubMed  Google Scholar 

  20. Jacobson, N.G., Szabo, S.J., Guler, M.L., Gorham, J.D. & Murphy, K.M. Regulation of interleukin-12 signal transduction during T helper phenotype development. Res. Immunol. 146, 446–456 (1995).

    Article  CAS  Google Scholar 

  21. Wu, C. et al. IL-12 receptor β2 (IL-12Rβ2)-deficient mice are defective in IL-12-mediated signaling despite the presence of high affinity IL-12 binding sites. J. Immunol. 165, 6221–6228 (2000).

    Article  CAS  Google Scholar 

  22. Magram, J. et al. IL-12-deficient mice are defective in IFN-γ production and type 1 cytokine responses. Immunity 4, 471–481 (1996).

    Article  CAS  Google Scholar 

  23. Hsieh, C.S. et al. Development of TH1 CD4+ T cells through IL-12 produced by Listeria-induced macrophages. Science 260, 547–549 (1993).

    Article  CAS  Google Scholar 

  24. Manetti, R. et al. Interleukin 12 induces stable priming for interferon γ (IFN-γ) production during differentiation of human T helper (Th) cells and transient IFN-γ production in established TH2 cell clones. J. Exp. Med. 179, 1273–1283 (1994).

    Article  CAS  Google Scholar 

  25. Kaplan, M.H., Wurster, A.L. & Grusby, M.J. A signal transducer and activator of transcription (Stat)4-independent pathway for the development of T helper type 1 cells. J. Exp. Med. 188, 1191–1196 (1998).

    Article  CAS  Google Scholar 

  26. Bird, J.J. et al. Helper T cell differentiation is controlled by the cell cycle. Immunity 9, 229–237 (1998).

    Article  CAS  Google Scholar 

  27. Xu, D. et al. Selective expression and functions of interleukin 18 receptor on T helper (Th) type 1 but not TH2 cells. J. Exp. Med. 188, 1485–1492 (1998).

    Article  CAS  Google Scholar 

  28. Smeltz, R.B., Chen, J., Hu-Li, J. & Shevach, E.M. Regulation of interleukin (IL)-18 receptor α chain expression on CD4+ T cells during T helper (Th)1/TH2 differentiation. Critical downregulatory role of IL-4. J. Exp. Med. 194, 143–153 (2001).

    Article  CAS  Google Scholar 

  29. Yoshimoto, T. et al. IL-12 up-regulates IL-18 receptor expression on T cells, TH1 cells, and B cells: synergism with IL-18 for IFN-γ production. J. Immunol. 161, 3400–3407 (1998).

    CAS  PubMed  Google Scholar 

  30. Ahn, H.J. et al. A mechanism underlying synergy between IL-12 and IFN-γ-inducing factor in enhanced production of IFN-γ. J. Immunol. 159, 2125–2131 (1997).

    CAS  PubMed  Google Scholar 

  31. Ouyang, W. et al. STAT6-independent GATA-3 autoactivation directs IL-4-independent TH2 development and commitment. Immunity 12, 27–37 (2000).

    Article  CAS  Google Scholar 

  32. Ranganath, S. & Murphy, K.M. Structure and specificity of GATA proteins in TH2 development. Mol. Cell Biol. 21, 2716–2725 (2001).

    Article  CAS  Google Scholar 

  33. Zhou, M. et al. Friend of GATA-1 represses GATA-3-dependent activity in CD4+ T cells. J. Exp. Med. 194, 1461–1471 (2001).

    Article  CAS  Google Scholar 

  34. Lee, H.J. et al. GATA-3 induces T helper cell type 2 (TH2) cytokine expression and chromatin remodeling in committed TH1 cells. J. Exp. Med. 192, 105–115 (2000).

    Article  CAS  Google Scholar 

  35. Ouyang, W. et al. The Ets transcription factor ERM is TH1-specific and induced by IL-12 through a STAT4-dependent pathway. Proc. Natl. Acad. Sci. USA 96, 3888–3893 (1999).

    Article  CAS  Google Scholar 

  36. Szabo, S.J. et al. Distinct effects of T-bet in TH1 lineage commitment and IFN-γ production in CD4 and CD8 T cells. Science 295, 338–342 (2002).

    Article  CAS  Google Scholar 

  37. Ouyang, W. et al. Inhibition of TH1 development mediated by GATA-3 through an IL-4-independent mechanism. Immunity 9, 745–755 (1998).

    Article  CAS  Google Scholar 

  38. Nishikomori, R., Ehrhardt, R.O. & Strober, W. T helper type 2 cell differentiation occurs in the presence of interleukin 12 receptor β2 chain expression and signaling. J. Exp. Med. 191, 847–858 (2000).

    Article  CAS  Google Scholar 

  39. Zhang, W.X. & Yang, S.Y. Cloning and characterization of a new member of the T-box gene family. Genomics 70, 41–48 (2000).

    Article  CAS  Google Scholar 

  40. Ranganath, S. et al. GATA-3-dependent enhancer activity in IL-4 gene regulation. J. Immunol. 161, 3822–3826 (1998).

    CAS  PubMed  Google Scholar 

  41. Farrar, J.D. et al. Selective loss of type I interferon-induced STAT4 activation caused by a minisatellite insertion in mouse STAT2. Nature Immunol. 1, 65–69 (2000).

    Article  CAS  Google Scholar 

  42. Assenmacher, M., Schmitz, J. & Radbruch, A. Flow cytometric determination of cytokines in activated murine T helper lymphocytes: expression of interleukin-10 in interferon-γ and in interleukin-4-expressing cells. Eur. J. Immunol. 24, 1097–1101 (1994).

    Article  CAS  Google Scholar 

  43. Wittwer, C.T., Herrmann, M.G., Moss, A.A. & Rasmussen, R.P. Continuous fluorescence monitoring of rapid cycle DNA amplification. Biotechniques 22, 130–131 (1900).

    Article  Google Scholar 

  44. Wittwer, C.T. et al. The LightCycler: a microvolume multisample fluorimeter with rapid temperature control. Biotechniques 22, 176–181 (1997).

    Article  CAS  Google Scholar 

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Acknowledgements

We thank K. Fredrick for animal husbandry. Supported by grants from the NIH.

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Correspondence to Kenneth M. Murphy.

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The authors declare no competing financial interests.

Supplementary information

Web Fig. 1.

STAT1 and STAT4 differentially regulate IL-12–induced TH1 development: assessment by ELISA. CD4+ DO11.10 lymph node T cells from wild-type, STAT1-/- or STAT4-/- mice were activated with 0.3 µM OVA peptide in the presence or absence cytokines or anti-cytokine antibodies as indicated for 7 days, collected and restimulated with OVA peptide–APCs without cytokine manipulations for 48 h. Supernatants were collected at 48 h and IL-4 and IFN-γ were measured by ELISA. (GIF 8 kb)

Web Fig. 2.

STAT1 and STAT4 differentially regulate IL-12–induced TH1 development: assessment by RNA blotting. Cells in Web Fig. 1 were lysed on day 7 and total RNA was prepared and analyzed by RNA blot for IL-12Rβ2 and GAPDH expression as described3. (JPG 25 kb)

Web Fig. 3.

Conditional requirement for STAT1 in IL-12Rβ2 expression during TH1 development. Mel-14hi CD4+ cells were sorted from wild-type or STAT1-/- DO11.10 lymph nodes and spleens and activated in the presence of the indicated cytokines and neutralizing anti-cytokine antibodies as described in the Methods. On days 1, 2, 3, 4 and 7 post-activation, cells were collected and stained with isotype control (dashed lines) or anti–IL-12Rβ2 (solid lines). (GIF 28 kb)

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Afkarian, M., Sedy, J., Yang, J. et al. T-bet is a STAT1-induced regulator of IL-12R expression in naïve CD4+ T cells. Nat Immunol 3, 549–557 (2002). https://doi.org/10.1038/ni794

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