Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Instability of the transcription factor Foxp3 leads to the generation of pathogenic memory T cells in vivo

Nature Immunology volume 10, pages 1000–1007 (2009)Cite this article

Abstract

Regulatory T cells (Treg cells) are central to the maintenance of immune homeostasis. However, little is known about the stability of Treg cells in vivo. In this study, we demonstrate that a substantial percentage of cells had transient or unstable expression of the transcription factor Foxp3. These 'exFoxp3' T cells had an activated-memory T cell phenotype and produced inflammatory cytokines. Moreover, exFoxp3 cell numbers were higher in inflamed tissues in autoimmune conditions. Adoptive transfer of autoreactive exFoxp3 cells led to the rapid onset of diabetes. Finally, analysis of the T cell receptor repertoire suggested that exFoxp3 cells developed from both natural and adaptive Treg cells. Thus, the generation of potentially autoreactive effector T cells as a consequence of Foxp3 instability has important implications for understanding autoimmune disease pathogenesis.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Development of Foxp3+ T cells in Foxp3-GFP-Cre × R26-YFP–transgenic mice.
Figure 2: CD4+ YFP+ Foxp3− cells have a non–Treg cell surface phenotype.
Figure 3: CD4+ YFP+ Foxp3− cells have a non–Treg, memory cell surface phenotype and produce IFN-γ and IL-17.
Figure 4: The autoimmune microenvironment favors loss of Foxp3 expression.
Figure 5: Development of exFoxp3 cells from Treg cells after adoptive transfer.
Figure 6: Pathogenicity of exFoxp3 cells.
Figure 7: Development of exFoxp3 cells from both nTreg cells and aTreg cells.

Similar content being viewed by others

References

  1. Tang, Q. et al. Central role of defective interleukin-2 production in the triggering of islet autoimmune destruction. Immunity 28, 687–697 (2008).

    Article  CAS  Google Scholar 

  2. Bluestone, J.A., Tang, Q. & Sedwick, C.E. T regulatory cells in autoimmune diabetes: past challenges, future prospects. J. Clin. Immunol. 28, 677–684 (2008).

    Article  CAS  Google Scholar 

  3. Wan, Y.Y. & Flavell, R.A. Regulatory T-cell functions are subverted and converted owing to attenuated Foxp3 expression. Nature 445, 766–770 (2007).

    Article  CAS  Google Scholar 

  4. Hori, S., Nomura, T. & Sakaguchi, S. Control of regulatory T cell development by the transcription factor Foxp3. Science 299, 1057–1061 (2003).

    Article  CAS  Google Scholar 

  5. Bennett, C.L. & Ochs, H.D. IPEX is a unique X-linked syndrome characterized by immune dysfunction, polyendocrinopathy, enteropathy, and a variety of autoimmune phenomena. Curr. Opin. Pediatr. 13, 533–538 (2001).

    Article  CAS  Google Scholar 

  6. Bennett, C.L. et al. The immune dysregulation, polyendocrinopathy, enteropathy, X-linked syndrome (IPEX) is caused by mutations of FOXP3. Nat. Genet. 27, 20–21 (2001).

    Article  CAS  Google Scholar 

  7. Brunkow, M.E. 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).

    CAS  Google Scholar 

  8. Bluestone, J.A. & Abbas, A.K. Natural versus adaptive regulatory T cells. Nat. Rev. Immunol. 3, 253–257 (2003).

    Article  CAS  Google Scholar 

  9. Gavin, M.A. et al. Foxp3-dependent programme of regulatory T-cell differentiation. Nature 445, 771–775 (2007).

    Article  CAS  Google Scholar 

  10. Williams, L.M. & Rudensky, A.Y. Maintenance of the Foxp3-dependent developmental program in mature regulatory T cells requires continued expression of Foxp3. Nat. Immunol. 8, 277–284 (2007).

    Article  CAS  Google Scholar 

  11. Yang, X.O. et al. Molecular antagonism and plasticity of regulatory and inflammatory T cell programs. Immunity 29, 44–56 (2008).

    Article  CAS  Google Scholar 

  12. Zhou, X. et al. Selective miRNA disruption in T reg cells leads to uncontrolled autoimmunity. J. Exp. Med. 205, 1983–1991 (2008).

    Article  CAS  Google Scholar 

  13. Komatsu, N. et al. Heterogeneity of natural Foxp3+ T cells: a committed regulatory T-cell lineage and an uncommitted minor population retaining plasticity. Proc. Natl. Acad. Sci. USA 106, 1903–1908 (2009).

    Article  CAS  Google Scholar 

  14. Tsuji, M. et al. Preferential generation of follicular B helper T cells from Foxp3+ T cells in gut Peyer's patches. Science 323, 1488–1492 (2009).

    Article  CAS  Google Scholar 

  15. Srinivas, S. et al. Cre reporter strains produced by targeted insertion of EYFP and ECFP into the ROSA26 locus. BMC Dev. Biol. 1, 4 (2001).

    Article  CAS  Google Scholar 

  16. Fontenot, J.D. et al. Regulatory T cell lineage specification by the forkhead transcription factor foxp3. Immunity 22, 329–341 (2005).

    CAS  Google Scholar 

  17. Kishimoto, H. & Sprent, J. The thymus and negative selection. Immunol. Res. 21, 315–323 (2000).

    Article  CAS  Google Scholar 

  18. Jordan, M.S. et al. Thymic selection of CD4+CD25+ regulatory T cells induced by an agonist self-peptide. Nat. Immunol. 2, 301–306 (2001).

    Article  CAS  Google Scholar 

  19. Huehn, J., Polansky, J.K. & Hamann, A. Epigenetic control of FOXP3 expression: the key to a stable regulatory T-cell lineage? Nat. Rev. Immunol. 9, 83–89 (2009).

    Article  CAS  Google Scholar 

  20. Floess, S. et al. Epigenetic control of the foxp3 locus in regulatory T cells. PLoS Biol. 5, e38 (2007).

    Article  Google Scholar 

  21. Kim, H.P. & Leonard, W.J. CREB/ATF-dependent T cell receptor-induced FoxP3 gene expression: a role for DNA methylation. J. Exp. Med. 204, 1543–1551 (2007).

    Article  CAS  Google Scholar 

  22. Polansky, J.K. et al. DNA methylation controls Foxp3 gene expression. Eur. J. Immunol. 38, 1654–1663 (2008).

    Article  CAS  Google Scholar 

  23. Zhou, L. et al. TGF-β-induced Foxp3 inhibits TH17 cell differentiation by antagonizing RORγt function. Nature 453, 236–240 (2008).

    Article  CAS  Google Scholar 

  24. Katz, J.D., Wang, B., Haskins, K., Benoist, C. & Mathis, D. Following a diabetogenic T cell from genesis through pathogenesis. Cell 74, 1089–1100 (1993).

    Article  CAS  Google Scholar 

  25. Tang, Q. et al. In vitro-expanded antigen-specific regulatory T cells suppress autoimmune diabetes. J. Exp. Med. 199, 1455–1465 (2004).

    Article  CAS  Google Scholar 

  26. Wong, J., Mathis, D. & Benoist, C. TCR-based lineage tracing: no evidence for conversion of conventional into regulatory T cells in response to a natural self-antigen in pancreatic islets. J. Exp. Med. 204, 2039–2045 (2007).

    Article  CAS  Google Scholar 

  27. Tang, Q. & Bluestone, J.A. The Foxp3+ regulatory T cell: a jack of all trades, master of regulation. Nat. Immunol. 9, 239–244 (2008).

    Article  CAS  Google Scholar 

  28. Salomon, B. et al. B7/CD28 costimulation is essential for the homeostasis of the CD4+CD25+ immunoregulatory T cells that control autoimmune diabetes. Immunity 12, 431–440 (2000).

    Article  CAS  Google Scholar 

  29. Sakaguchi, S. et al. Foxp3+CD25+CD4+ natural regulatory T cells in dominant self-tolerance and autoimmune disease. Immunol. Rev. 212, 8–27 (2006).

    Article  CAS  Google Scholar 

  30. Hsieh, C.S., Zheng, Y., Liang, Y., Fontenot, J.D. & Rudensky, A.Y. An intersection between the self-reactive regulatory and nonregulatory T cell receptor repertoires. Nat. Immunol. 7, 401–410 (2006).

    Article  CAS  Google Scholar 

  31. Hsieh, C.S. & Rudensky, A.Y. The role of TCR specificity in naturally arising CD25+CD4+ regulatory T cell biology. Curr. Top. Microbiol. Immunol. 293, 25–42 (2005).

    CAS  PubMed  Google Scholar 

  32. Liu, W. et al. CD127 expression inversely correlates with FoxP3 and suppressive function of human CD4+ T reg cells. J. Exp. Med. 203, 1701–1711 (2006).

    Article  CAS  Google Scholar 

  33. Lu, L.F. et al. Foxp3-dependent microRNA155 confers competitive fitness to regulatory T cells by targeting SOCS1 protein. Immunity 30, 80–91 (2009).

    Article  CAS  Google Scholar 

  34. Lochner, M. et al. In vivo equilibrium of proinflammatory IL-17+ and regulatory IL-10+Foxp3+RORγt+ T cells. J. Exp. Med. 205, 1381–1393 (2008).

    Article  CAS  Google Scholar 

  35. Lal, G. et al. Epigenetic regulation of Foxp3 expression in regulatory T cells by DNA methylation. J. Immunol. 182, 259–273 (2009).

    Article  CAS  Google Scholar 

  36. Zheng, Y. et al. Regulatory T-cell suppressor program co-opts transcription factor IRF4 to control TH2 responses. Nature 458, 351–356 (2009).

    Article  CAS  Google Scholar 

  37. Koch, M.A. et al. The transcription factor T-bet controls regulatory T cell homeostasis and function during type 1 inflammation. Nat. Immunol. 10, 595–602 (2009).

    Article  CAS  Google Scholar 

  38. Cobb, B.S. et al. A role for Dicer in immune regulation. J. Exp. Med. 203, 2519–2527 (2006).

    Article  CAS  Google Scholar 

  39. Rose, N.R. The adjuvant effect in infection and autoimmunity. Clin. Rev. Allergy Immunol. 34, 279–282 (2008).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank C.J. McArthur, M. Lee, N. Grewal, S. Jiang, J. Beilke and S. Zhu for technical assistance; N. Martinier and D. Fuentes for animal husbandry; and A. Abbas, Q. Tang, M. Anderson, R. Locksley and members of the Bluestone laboratory for discussions and reading the manuscript. Supported by the US National Institutes of Health (P01 AI35297 and U19 AI056388 to J.A.B., and P30 DK63720 for core support), the American Diabetes Association (X.Z.), the Swiss National Science Foundation (PBBSB-118644 to L.T.J.), the Roche Research Foundation (L.T.J.), the Novartis Foundation (formerly Ciba-Geigy Jubilee Foundation; L.T.J.) and the US National Institute of General Medical Sciences (1 R25 GM56847 to C.P.).

Author information

Author notes

  1. Marc Martínez-Llordella

    Present address: Present address: Liver Transplant Unit, Hospital Clínic Barcelona–August Pi Sunyer Institute for Biomedical Investigations, Barcelona, Spain.,

  2. Xuyu Zhou, Samantha L Bailey-Bucktrout and Lukas T Jeker: These authors contributed equally to this work.

Authors and Affiliations

  1. Diabetes Center and the Department of Medicine, University of California, San Francisco, California, USA

    Xuyu Zhou, Samantha L Bailey-Bucktrout, Lukas T Jeker, Cristina Penaranda, Meredith Ashby, Wendy Rosenthal & Jeffrey A Bluestone

  2. Barbara Davis Center for Childhood Diabetes, University of Colorado Denver, Aurora, Colorado, USA

    Maki Nakayama

Authors
  1. Xuyu Zhou
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Samantha L Bailey-Bucktrout
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Lukas T Jeker
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Cristina Penaranda
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Marc Martínez-Llordella
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Meredith Ashby
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Maki Nakayama
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Wendy Rosenthal
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Jeffrey A Bluestone
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

X.Z. and J.A.B. initiated the study; X.Z., S.L.B.-B. and L.T.J. designed and did experiments, analyzed data and wrote the manuscript; C.P. and M.M.-L. designed and did experiments; M.A. did bioinformatic analysis of TCR sequences with help from X.Z., S.L.B.-B. and M.N.; M.N. and W.R. did experiments; and J.A.B. designed experiments, supervised the work, analyzed data and wrote the manuscript.

Corresponding author

Correspondence to Jeffrey A Bluestone.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–3 and Supplementary Tables 1–2 (PDF 1925 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Zhou, X., Bailey-Bucktrout, S., Jeker, L. et al. Instability of the transcription factor Foxp3 leads to the generation of pathogenic memory T cells in vivo. Nat Immunol 10, 1000–1007 (2009). https://doi.org/10.1038/ni.1774

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/ni.1774

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing