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Inducing and expanding regulatory T cell populations by foreign antigen

Abstract

Evidence suggests that regulatory T cells expressing the transcription factor Foxp3 develop extrathymically and intrathymically. Mechanisms of extrathymic induction require further scrutiny, especially as proliferation and/or phenotypic changes of preexisting suppressor cells must be distinguished from true de novo generation. Here we report the conversion of truly naive CD4+ T cells into suppressor cells expressing Foxp3 by targeting of peptide-agonist ligands to dendritic cells and by analysis of Foxp3 expression at the level of single cells. We show that conversion was achieved by minute antigen doses with suboptimal dendritic cell activation. The addition of transforming growth factor-β or the absence of interleukin 2 production, which reduces proliferation, enhanced the conversion rate. In addition, regulatory T cell populations induced in subimmunogenic conditions could subsequently be expanded by delivery of antigen in immunogenic conditions. The extrathymic generation and proliferation of regulatory T cells may contribute to self-tolerance as well as the poor immunogenicity of tumors and may be exploited clinically to prevent or reverse unwanted immunity.

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Figure 1: Conversion of naive CD4+CD25 T cells into CD4+CD25+ T cells.
Figure 2: Foxp3 expression and antigen-specific suppression by CD4+CD25+ T cells generated de novo.
Figure 3: Efficient induction of CD4+CD25+ regulatory T cells requires low doses of anti-DEC–HA and lack of costimulation.
Figure 4: Inverse relationship of cell division and CD25 expression.
Figure 5: Impaired TGF-βR signaling diminishes conversion of naive T cells into CD4+CD25+ regulatory T cells.
Figure 6: Costimulation of naive CD4+CD25 T cells in vitro by TCR and TGF-β receptor.
Figure 7: In vivo conversion of naive CD4+CD25 T cells costimulated in vitro with TCR and TGF-β.
Figure 8: Efficient conversion of Il2−/−CD4+CD25 naive T cells into regulatory T cells.

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References

  1. Asano, M., Toda, M., Sakaguchi, N. & Sakaguchi, S. Autoimmune disease as a consequence of developmental abnormality of a T cell subpopulation. J. Exp. Med. 184, 387–396 (1996).

    Article  CAS  PubMed  Google Scholar 

  2. Sakaguchi, S. et al. Immunologic tolerance maintained by CD25+ CD4+ regulatory T cells: their common role in controlling autoimmunity, tumor immunity, and transplantation tolerance. Immunol. Rev. 182, 18–32 (2001).

    Article  CAS  PubMed  Google Scholar 

  3. Seddon, B. & Mason, D. The third function of the thymus. Immunol. Today 21, 95–99 (2000).

    Article  CAS  PubMed  Google Scholar 

  4. 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  PubMed  Google Scholar 

  5. Walker, L.S., Chodos, A., Eggena, M., Dooms, H. & Abbas, A.K. Antigen-dependent proliferation of CD4+ CD25+ regulatory T cells in vivo. J. Exp. Med. 198, 249–258 (2003).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Apostolou, I., Sarukhan, A., Klein, L. & von Boehmer, H. Origin of regulatory T cells with known specificity for antigen. Nat. Immunol. 3, 756–763 (2002).

    Article  CAS  PubMed  Google Scholar 

  7. Tai, X., Cowan, M., Feigenbaum, L. & Singer, A. CD28 costimulation of developing thymocytes induces Foxp3 expression and regulatory T cell differentiation independently of interleukin 2. Nat. Immunol. 6, 152–162 (2005).

    Article  CAS  PubMed  Google Scholar 

  8. Apostolou, I. & von Boehmer, H. In vivo instruction of suppressor commitment in naive T cells. J. Exp. Med. 199, 1401–1408 (2004).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Chen, T.C., Waldmann, H. & Fairchild, P.J. Induction of dominant transplantation tolerance by an altered peptide ligand of the male antigen Dby. J. Clin. Invest. 113, 1754–1762 (2004).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Mahnke, K., Qian, Y., Knop, J. & Enk, A.H. Induction of CD4+CD25+ regulatory T cells by targeting of antigens to immature dendritic cells. Blood 101, 4862–4869 (2003).

    Article  CAS  PubMed  Google Scholar 

  11. Thorstenson, K.M. & Khoruts, A. Generation of anergic and potentially immunoregulatory CD25+CD4 T cells in vivo after induction of peripheral tolerance with intravenous or oral antigen. J. Immunol. 167, 188–195 (2001).

    Article  CAS  PubMed  Google Scholar 

  12. Liang, S. et al. Conversion of CD4+ CD25 cells into CD4+ CD25+ regulatory T cells in vivo requires B7 costimulation, but not the thymus. J. Exp. Med. 201, 127–137 (2005).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  14. Wan, Y.Y. & Flavell, R.A. Identifying Foxp3-expressing suppressor T cells with a bicistronic reporter. Proc. Natl. Acad. Sci. USA 102, 5126–5131 (2005).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Curotto de Lafaille, M.A., Lino, A.C., Kutchukhidze, N. & Lafaille, J.J. CD25 T cells generate CD25+Foxp3+ regulatory T cells by peripheral expansion. J. Immunol. 173, 7259–7268 (2004).

    Article  CAS  PubMed  Google Scholar 

  16. Hawiger, D., Masilamani, R.F., Bettelli, E., Kuchroo, V.K. & Nussenzweig, M.C. Immunological unresponsiveness characterized by increased expression of CD5 on peripheral T cells induced by dendritic cells in vivo. Immunity 20, 695–705 (2004).

    Article  CAS  PubMed  Google Scholar 

  17. Hawiger, D. et al. Dendritic cells induce peripheral T cell unresponsiveness under steady state conditions in vivo. J. Exp. Med. 194, 769–779 (2001).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Klein, L., Khazaie, K. & von Boehmer, H. In vivo dynamics of antigen-specific regulatory T cells not predicted from behavior in vitro. Proc. Natl. Acad. Sci. USA 100, 8886–8891 (2003).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Itoh, M. et al. Thymus and autoimmunity: production of CD25+CD4+ naturally anergic and suppressive T cells as a key function of the thymus in maintaining immunologic self-tolerance. J. Immunol. 162, 5317–5326 (1999).

    CAS  PubMed  Google Scholar 

  20. Zheng, S.G., Gray, J.D., Ohtsuka, K., Yamagiwa, S. & Horwitz, D.A. Generation ex vivo of TGF-β-producing regulatory T cells from CD4+CD25 precursors. J. Immunol. 169, 4183–4189 (2002).

    Article  CAS  PubMed  Google Scholar 

  21. Chen, W. 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).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Fantini, M.C. et al. Cutting edge: TGF-β induces a regulatory phenotype in CD4+CD25 T cells through Foxp3 induction and down-regulation of Smad7. J. Immunol. 172, 5149–5153 (2004).

    Article  CAS  PubMed  Google Scholar 

  23. Huber, S. et al. Cutting edge: TGF-β signaling is required for the in vivo expansion and immunosuppressive capacity of regulatory CD4+CD25+ T cells. J. Immunol. 173, 6526–6531 (2004).

    Article  CAS  PubMed  Google Scholar 

  24. Marie, J.C., Letterio, J.J., Gavin, M. & Rudensky, A.Y. TGF-β1 maintains suppressor function and Foxp3 expression in CD4+CD25+ regulatory T cells. J. Exp. Med. 201, 1061–1067 (2005).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Steinman, R.M., Hawiger, D. & Nussenzweig, M.C. Tolerogenic dendritic cells. Annu. Rev. Immunol. 21, 685–711 (2003).

    Article  CAS  PubMed  Google Scholar 

  26. Inaba, K. et al. Tissue distribution of the DEC-205 protein that is detected by the monoclonal antibody NLDC-145. I. Expression on dendritic cells and other subsets of mouse leukocytes. Cell. Immunol. 163, 148–156 (1995).

    Article  CAS  PubMed  Google Scholar 

  27. Peng, Y., Laouar, Y., Li, M.O., Green, E.A. & Flavell, R.A. TGF-β regulates in vivo expansion of Foxp3-expressing CD4+CD25+ regulatory T cells responsible for protection against diabetes. Proc. Natl. Acad. Sci. USA 101, 4572–4577 (2004).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Gorelik, L. & Flavell, R.A. Abrogation of TGF-β signaling in T cells leads to spontaneous T cell differentiation and autoimmune disease. Immunity 12, 171–181 (2000).

    Article  CAS  PubMed  Google Scholar 

  29. Shi, Y. & Massague, J. Mechanisms of TGF-β signaling from cell membrane to the nucleus. Cell 113, 685–700 (2003).

    Article  CAS  PubMed  Google Scholar 

  30. Schorle, H., Holtschke, T., Hunig, T., Schimpl, A. & Horak, I. Development and function of T cells in mice rendered interleukin-2 deficient by gene targeting. Nature 352, 621–624 (1991).

    Article  CAS  PubMed  Google Scholar 

  31. Rolink, A., Melchers, F. & Andersson, J. The SCID but not the RAG-2 gene product is required for Sμ-Sε heavy chain class switching. Immunity 5, 319–330 (1996).

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

We thank A. Krueger and P. Ruchala (von Boehmer lab) and all members of the Flow Cytometry Facility at the Dana-Farber Cancer Institute for technical help in cell sorting; A. Rolink (University of Basel, Basel, Switzerland) for providing the FGK 45 hybridoma; J. Polansky for antibody purification; and T. Hunig (University of Wurzburg, Wurzburg, Germany) for the gift of IL-2-deficient mice. Supported by the German Research Foundation (KR2316/1-1to K.K.) and the National Institutes of Health (R37 AI53102).

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Correspondence to Harald von Boehmer.

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Kretschmer, K., Apostolou, I., Hawiger, D. et al. Inducing and expanding regulatory T cell populations by foreign antigen. Nat Immunol 6, 1219–1227 (2005). https://doi.org/10.1038/ni1265

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