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The transcription factor musculin promotes the unidirectional development of peripheral Treg cells by suppressing the TH2 transcriptional program

Nature Immunology volume 18, pages 344353 (2017) | Download Citation


Although master transcription factors (TFs) are key to the development of specific T cell subsets, whether additional transcriptional regulators are induced by the same stimuli that dominantly repress the development of other, non-specific T cell lineages has not been fully elucidated. Through the use of regulatory T cells (Treg cells) induced by transforming growth factor-β (TGF-β), we identified the TF musculin (MSC) as being critical for the development of induced Treg cells (iTreg cells) by repression of the T helper type 2 (TH2) transcriptional program. Loss of MSC reduced expression of the Treg cell master TF Foxp3 and induced TH2 differentiation even under iTreg-cell-differentiation conditions. MSC interrupted binding of the TF GATA-3 to the locus encoding TH2-cell-related cytokines and diminished intrachromosomal interactions within that locus. MSC-deficient (Msc−/−) iTreg cells were unable to suppress TH2 responses, and Msc−/− mice spontaneously developed gut and lung inflammation with age. MSC therefore enforced Foxp3 expression and promoted the unidirectional induction of iTreg cells by repressing the TH2 developmental program.

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Referenced accessions

Gene Expression Omnibus


  1. 1.

    & Effector T cell plasticity: flexibility in the face of changing circumstances. Nat. Immunol. 11, 674–680 (2010).

  2. 2.

    , & Regulatory T cells: mechanisms of differentiation and function. Annu. Rev. Immunol. 30, 531–564 (2012).

  3. 3.

    , , & FOXP3+ regulatory T cells in the human immune system. Nat. Rev. Immunol. 10, 490–500 (2010).

  4. 4.

    et al. Disruption of a new forkhead/winged-helix protein, scurfin, results in the fatal lymphoproliferative disorder of the scurfy mouse. Nat. Genet. 27, 68–73 (2001).

  5. 5.

    & Foxp3 in control of the regulatory T cell lineage. Nat. Immunol. 8, 457–462 (2007).

  6. 6.

    & Natural versus adaptive regulatory T cells. Nat. Rev. Immunol. 3, 253–257 (2003).

  7. 7.

    , & Foxp3 programs the development and function of CD4+CD25+ regulatory T cells. Nat. Immunol. 4, 330–336 (2003).

  8. 8.

    , , & TGF-beta1 maintains suppressor function and Foxp3 expression in CD4+CD25+ regulatory T cells. J. Exp. Med. 201, 1061–1067 (2005).

  9. 9.

    , , & Cutting Edge: IL-2 is essential for TGF-β-mediated induction of Foxp3+ T regulatory cells. J. Immunol. 178, 4022–4026 (2007).

  10. 10.

    et al. Transforming growth factor-β is a potent immunosuppressive agent that inhibits IL-1-dependent lymphocyte proliferation. J. Immunol. 140, 3026–3032 (1988).

  11. 11.

    et al. Regulatory effects of transforming growth factor-β on IL-2- and IL-4-dependent T cell-cycle progression. J. Immunol. 144, 1767–1776 (1990).

  12. 12.

    et al. Intrinsic inhibition of transcription factor E2A by HLH proteins ABF-1 and Id2 mediates reprogramming of neoplastic B cells in Hodgkin lymphoma. Nat. Immunol. 7, 207–215 (2006).

  13. 13.

    et al. Control of facial muscle development by MyoR and capsulin. Science 298, 2378–2381 (2002).

  14. 14.

    , , , & Myor/ABF-1 mRNA expression marks follicular helper T cells but is dispensable for Tfh cell differentiation and function in vivo. PLoS One 8, e84415 (2013).

  15. 15.

    et al. The cytokines interleukin 27 and interferon-γ promote distinct Treg cell populations required to limit infection-induced pathology. Immunity 37, 511–523 (2012).

  16. 16.

    et al. A validated regulatory network for Th17 cell specification. Cell 151, 289–303 (2012).

  17. 17.

    et al. Compensatory dendritic cell development mediated by BATF-IRF interactions. Nature 490, 502–507 (2012).

  18. 18.

    et al. Neuropilin 1 is expressed on thymus-derived natural regulatory T cells, but not mucosa-generated induced Foxp3+ T reg cells. J. Exp. Med. 209, 1723–1742 (2012).

  19. 19.

    et al. Neuropilin-1 distinguishes natural and inducible regulatory T cells among regulatory T cell subsets in vivo. J. Exp. Med. 209, 1713–1722 (2012).

  20. 20.

    et al. Smad2 and Smad3 are redundantly essential for the TGF-β-mediated regulation of regulatory T plasticity and Th1 development. J. Immunol. 185, 842–855 (2010).

  21. 21.

    et al. Positive and negative transcriptional regulation of the Foxp3 gene is mediated by access and binding of the Smad3 protein to enhancer I. Immunity 33, 313–325 (2010).

  22. 22.

    & CD103+ GALT DCs promote Foxp3+ regulatory T cells. Mucosal Immunol. 1 (Suppl. 1), S34–S38 (2008).

  23. 23.

    et al. A functionally specialized population of mucosal CD103+ DCs induces Foxp3+ regulatory T cells via a TGF-β and retinoic acid-dependent mechanism. J. Exp. Med. 204, 1757–1764 (2007).

  24. 24.

    , , & MyoR: a muscle-restricted basic helix-loop-helix transcription factor that antagonizes the actions of MyoD. Proc. Natl. Acad. Sci. USA 96, 552–557 (1999).

  25. 25.

    et al. IL-4 inhibits TGF-β-induced Foxp3+ T cells and, together with TGF-β, generates IL-9+IL-10+Foxp3 effector T cells. Nat. Immunol. 9, 1347–1355 (2008).

  26. 26.

    , , , & TGF-β induces the expression of the adaptor Ndfip1 to silence IL-4 production during iTreg cell differentiation. Nat. Immunol. 13, 77–85 (2011).

  27. 27.

    et al. Histone methyltransferases direct different degrees of methylation to define distinct chromatin domains. Mol. Cell 12, 1591–1598 (2003).

  28. 28.

    et al. Neutralization of IL-4 and IFN-γ facilitates inducing TGF-β-induced CD4+Foxp3+ regulatory cells. Int. J. Biomed. Sci. 4, 52–57 (2008).

  29. 29.

    & The transcription factor GATA-3 is necessary and sufficient for Th2 cytokine gene expression in CD4 T cells. Cell 89, 587–596 (1997).

  30. 30.

    et al. Characterization of ABF-1, a novel basic helix-loop-helix transcription factor expressed in activated B lymphocytes. Mol. Cell. Biol. 18, 3130–3139 (1998).

  31. 31.

    , , , & The cell type-specific expression of the murine IL-13 gene is regulated by GATA-3. J. Immunol. 167, 4414–4420 (2001).

  32. 32.

    et al. The enhancer HS2 critically regulates GATA-3-mediated Il4 transcription in TH2 cells. Nat. Immunol. 12, 77–85 (2011).

  33. 33.

    & Long-range intrachromosomal interactions in the T helper type 2 cytokine locus. Nat. Immunol. 5, 1017–1027 (2004).

  34. 34.

    , , & The long-range interaction landscape of gene promoters. Nature 489, 109–113 (2012).

  35. 35.

    et al. Critical role for the transcription regulator CCCTC-binding factor in the control of Th2 cytokine expression. J. Immunol. 182, 999–1010 (2009).

  36. 36.

    et al. Treg cells expressing the coinhibitory molecule TIGIT selectively inhibit proinflammatory Th1 and Th17 cell responses. Immunity 40, 569–581 (2014).

  37. 37.

    et al. Conversion of peripheral CD4+CD25 naive T cells to CD4+CD25+ regulatory T cells by TGF-β induction of transcription factor Foxp3. J. Exp. Med. 198, 1875–1886 (2003).

  38. 38.

    et al. Genome-wide analyses of transcription factor GATA-3-mediated gene regulation in distinct T cell types. Immunity 35, 299–311 (2011).

  39. 39.

    et al. Adoptive transfer of induced-Treg cells effectively attenuates murine airway allergic inflammation. PLoS One 7, e40314 (2012).

  40. 40.

    Regulatory T cells and infection: a dangerous necessity. Nat. Rev. Immunol. 7, 875–888 (2007).

  41. 41.

    , , & The polarization of immune cells in the tumour environment by TGFβ. Nat. Rev. Immunol. 10, 554–567 (2010).

  42. 42.

    et al. Role of conserved non-coding DNA elements in the Foxp3 gene in regulatory T-cell fate. Nature 463, 808–812 (2010).

  43. 43.

    et al. Extrathymically generated regulatory T cells control mucosal TH2 inflammation. Nature 482, 395–399 (2012).

  44. 44.

    et al. Control of the differentiation of regulatory T cells and TH17 cells by the DNA-binding inhibitor Id3. Nat. Immunol. 12, 86–95 (2011).

  45. 45.

    An overview of the basic helix-loop-helix proteins. Genome Biol. 5, 226 (2004).

  46. 46.

    & Master regulators or lineage-specifying? Changing views on CD4+ T cell transcription factors. Nat. Rev. Immunol. 12, 799–804 (2012).

  47. 47.

    et al. STAT6 activation confers upon T helper cells resistance to suppression by regulatory T cells. J. Immunol. 183, 155–163 (2009).

  48. 48.

    et al. GATA-3 controls Foxp3 regulatory T cell fate during inflammation in mice. J. Clin. Invest. 121, 4503–4515 (2011).

  49. 49.

    et al. Targeted disruption of Smad3 reveals an essential role in transforming growth factor β-mediated signal transduction. Mol. Cell. Biol. 19, 2495–2504 (1999).

  50. 50.

    et al. Tim-3 inhibits T helper type 1-mediated auto- and alloimmune responses and promotes immunological tolerance. Nat. Immunol. 4, 1093–1101 (2003).

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We thank D. Kozoriz for cell sorting; T. Chatila for advice; M. Collins for advice and editing the manuscript; E.N. Olson (University of Texas Southwestern Medical Center at Dallas) for Msc−/− mice; L. Maggio-Price (University of Washington) for Smad3−/− mice; and A. Yoshimura (Keio University School of Medicine) for the plasmid pCMV5-Smad3. Supported by the National Multiple Sclerosis Society (Career Transition Award TA 3059-A-2 to C.W.) and the US National Institutes of Health (K99 NIH Pathway to Independence Award KAI110649A to C.W.; K01DK090105 to S.X.; and P01AI073748, P01AI056299, P01AI039671, R01NS045937 and R01 NS030843 to V.K.).

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Author notes

    • Chuan Wu
    • , Valerie Dardalhon
    •  & Rafael F Franca

    Present addresses: Experimental Immunology Branch, National Cancer Institute, US National Institutes of Health, Bethesda, Maryland, USA (C.W.), GMM-UMR5535-CNRS, Montpellier, France (V.D.), and Department of Pharmacology, University of São Paulo, São Paulo, Ribeirao Preto, Brazil (R.F.F.).

    • Chuan Wu
    •  & Zuojia Chen

    These authors contributed equally to this work.


  1. Evergrande Center for Immunologic Diseases, Harvard Medical School and Brigham and Women's Hospital, Boston, Massachusetts, USA.

    • Chuan Wu
    • , Zuojia Chen
    • , Valerie Dardalhon
    • , Sheng Xiao
    • , Theresa Thalhamer
    • , Asaf Madi
    • , Rafael F Franca
    • , Timothy Han
    •  & Vijay Kuchroo
  2. Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, USA.

    • Mengyang Liao
  3. Center for Immunity and Immunotherapies, Seattle Children's Research Institute, Seattle, Washington, USA.

    • Mohammed Oukka


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C.W. designed and performed experiments and wrote the manuscript; Z.C. performed experiments and wrote the manuscript; V.D., S.X., T.T., M.L., A.M., R.F.F., T.H. and M.O. performed experiments; A.M. analyzed the data; and V.K. supervised the study and edited the manuscript.

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The authors declare no competing financial interests.

Corresponding authors

Correspondence to Chuan Wu or Vijay Kuchroo.

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