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Intraclonal competition limits the fate determination of regulatory T cells in the thymus

Nature Immunology volume 10, pages 610–617 (2009)Cite this article

Abstract

Because the deletion of self-reactive T cells is incomplete, thymic development of natural Foxp3+CD4+ regulatory T cells (Treg cells) is required for preventing autoimmunity. However, the function of T cell antigen receptor (TCR) specificity in thymic Treg cell development remains controversial. To address this issue, we generated a transgenic line expressing a naturally occurring Treg cell–derived TCR. Unexpectedly, we found that efficient thymic Treg cell development occurred only when the antigen-specific Treg cell precursors were present at low clonal frequency (<1%) in a normal thymus. Using retroviral vectors and bone marrow chimeras, we observed similar activity with two other Treg cell–derived TCRs. Our data demonstrate that thymic Treg cell development is a 'TCR-instructive' process involving a niche that can be saturable at much lower clonal frequencies than is the niche for positive selection.

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Figure 1: Characterization of TCR-transgenic lines by flow cytometry.
Figure 2: Inverse relationship between TCR-transgenic cell frequency and thymic Treg cell development.
Figure 3: The generation of Foxp3+ G113 Treg cells is saturable.
Figure 4: Peripheral versus thymic G113 Treg cell development.
Figure 5: No evidence of proliferation or negative selection during TCR-dependent G113 Treg cell development.
Figure 6: Varying efficiencies of TCR-dependent Treg cell development.

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References

  1. Sakaguchi, S., Yamaguchi, T., Nomura, T. & Ono, M. Regulatory T cells and immune tolerance. Cell 133, 775–787 (2008).

    Article  CAS  Google Scholar 

  2. Lahl, K. et al. Selective depletion of Foxp3+ regulatory T cells induces a scurfy-like disease. J. Exp. Med. 204, 57–63 (2007).

    Article  CAS  Google Scholar 

  3. Kim, J.M., Rasmussen, J.P. & Rudensky, A.Y. Regulatory T cells prevent catastrophic autoimmunity throughout the lifespan of mice. Nat. Immunol. 8, 191–197 (2007).

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  5. Curotto de Lafaille, M.A. et al. Adaptive Foxp3+ regulatory T cell-dependent and -independent control of allergic inflammation. Immunity 29, 114–126 (2008).

    Article  CAS  Google Scholar 

  6. Knoechel, B., Lohr, J., Kahn, E., Bluestone, J.A. & Abbas, A.K. Sequential development of interleukin 2–dependent effector and regulatory T cells in response to endogenous systemic antigen. J. Exp. Med. 202, 1375–1386 (2005).

    Article  CAS  Google Scholar 

  7. Sakaguchi, S. & Sakaguchi, N. Thymus and autoimmunity: capacity of the normal thymus to produce pathogenic self-reactive T cells and conditions required for their induction of autoimmune disease. J. Exp. Med. 172, 537–545 (1990).

    Article  CAS  Google Scholar 

  8. Powrie, F., Leach, M.W., Mauze, S., Caddle, L.B. & Coffman, R.L. Phenotypically distinct subsets of CD4+ T cells induce or protect from chronic intestinal inflammation. Int. Immunol. 5, 1461–1471 (1993).

    Article  CAS  Google Scholar 

  9. 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  Google Scholar 

  10. Wong, J. et al. Adaptation of TCR repertoires to self-peptides in regulatory and nonregulatory CD4+ T cells. J. Immunol. 178, 7032–7041 (2007).

    Article  CAS  Google Scholar 

  11. Hsieh, C.-S. et al. Recognition of the peripheral self by naturally arising CD25+ CD4+ T cell receptors. Immunity 21, 267–277 (2004).

    Article  CAS  Google Scholar 

  12. 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 

  13. Pacholczyk, R., Ignatowicz, H., Kraj, P. & Ignatowicz, L. Origin and T cell receptor diversity of Foxp3+CD4+CD25+ T cells. Immunity 25, 249–259 (2006).

    Article  CAS  Google Scholar 

  14. 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 

  15. 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  Google Scholar 

  16. van Santen, H.-M., Benoist, C. & Mathis, D. Number of T reg cells that differentiate does not increase upon encounter of agonist ligand on thymic epithelial cells. J. Exp. Med. 200, 1221–1230 (2004).

    Article  CAS  Google Scholar 

  17. Pennington, D.J. et al. Early events in the thymus affect the balance of effector and regulatory T cells. Nature 444, 1073–1077 (2006).

    Article  CAS  Google Scholar 

  18. Pacholczyk, R. et al. Nonself-antigens are the cognate specificities of Foxp3+ regulatory T cells. Immunity 27, 493–504 (2007).

    Article  CAS  Google Scholar 

  19. Lio, C.W. & Hsieh, C.S. A two-step process for thymic regulatory T cell development. Immunity 28, 100–111 (2008).

    Article  CAS  Google Scholar 

  20. Lathrop, S.K., Santacruz, N.A., Pham, D., Luo, J. & Hsieh, C.S. Antigen-specific peripheral shaping of the natural regulatory T cell population. J. Exp. Med. 205, 3105–3117 (2008).

    Article  CAS  Google Scholar 

  21. Malek, T.R., Yu, A., Vincek, V., Scibelli, P. & Kong, L. CD4 regulatory T cells prevent lethal autoimmunity in IL-2Rβ-deficient mice. Implications for the nonredundant function of IL-2. Immunity 17, 167–178 (2002).

    Article  CAS  Google Scholar 

  22. Burchill, M.A., Yang, J., Vogtenhuber, C., Blazar, B.R. & Farrar, M.A. IL-2 receptor β-dependent STAT5 activation is required for the development of Foxp3+ regulatory T cells. J. Immunol. 178, 280–290 (2007).

    Article  CAS  Google Scholar 

  23. Cabarrocas, J. et al. Foxp3+ CD25+ regulatory T cells specific for a neo-self-antigen develop at the double-positive thymic stage. Proc. Natl. Acad. Sci. USA 103, 8453–8458 (2006).

    Article  CAS  Google Scholar 

  24. Wong, P., Goldrath, A.W. & Rudensky, A.Y. Competition for specific intrathymic ligands limits positive selection in a TCR transgenic model of CD4+ T cell development. J. Immunol. 164, 6252–6259 (2000).

    Article  CAS  Google Scholar 

  25. Canelles, M., Park, M.L., Schwartz, O.M. & Fowlkes, B.J. The influence of the thymic environment on the CD4-versus-CD8 T lineage decision. Nat. Immunol. 4, 756–764 (2003).

    Article  CAS  Google Scholar 

  26. Huesmann, M., Scott, B., Kisielow, P. & von Boehmer, H. Kinetics and efficacy of positive selection in the thymus of normal and T cell receptor transgenic mice. Cell 66, 533–540 (1991).

    Article  CAS  Google Scholar 

  27. Liu, V.C. et al. Tumor evasion of the immune system by converting CD4+CD25− T cells into CD4+CD25+ T regulatory cells: role of tumor-derived TGF-beta. J. Immunol. 178, 2883–2892 (2007).

    Article  CAS  Google Scholar 

  28. Hill, J.A. et al. Foxp3 transcription-factor-dependent and -independent regulation of the regulatory T cell transcriptional signature. Immunity 27, 786–800 (2007).

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  30. Lin, W. et al. Regulatory T cell development in the absence of functional Foxp3. Nat. Immunol. 8, 359–368 (2007).

    Article  CAS  Google Scholar 

  31. Feuerer, M. et al. Enhanced thymic selection of FoxP3+ regulatory T cells in the NOD mouse model of autoimmune diabetes. Proc. Natl. Acad. Sci. USA 104, 18181–18186 (2007).

    Article  CAS  Google Scholar 

  32. 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  Google Scholar 

  33. Troy, A.E. & Shen, H. Cutting edge: homeostatic proliferation of peripheral T lymphocytes is regulated by clonal competition. J. Immunol. 170, 672–676 (2003).

    Article  CAS  Google Scholar 

  34. Moses, C.T., Thorstenson, K.M., Jameson, S.C. & Khoruts, A. Competition for self ligands restrains homeostatic proliferation of naive CD4 T cells. Proc. Natl. Acad. Sci. USA 100, 1185–1190 (2003).

    Article  CAS  Google Scholar 

  35. Hataye, J., Moon, J.J., Khoruts, A., Reilly, C. & Jenkins, M.K. Naive and memory CD4+ T cell survival controlled by clonal abundance. Science 312, 114–116 (2006).

    Article  CAS  Google Scholar 

  36. Tang, Q. et al. Visualizing regulatory T cell control of autoimmune responses in nonobese diabetic mice. Nat. Immunol. 7, 83–92 (2006).

    Article  CAS  Google Scholar 

  37. Garcia, Z. et al. Competition for antigen determines the stability of T cell-dendritic cell interactions during clonal expansion. Proc. Natl. Acad. Sci. USA 104, 4553–4558 (2007).

    Article  CAS  Google Scholar 

  38. Romagnoli, P., Hudrisier, D. & van Meerwijk, J.P. Molecular signature of recent thymic selection events on effector and regulatory CD4+ T lymphocytes. J. Immunol. 175, 5751–5758 (2005).

    Article  CAS  Google Scholar 

  39. You, S., Slehoffer, G., Barriot, S., Bach, J.F. & Chatenoud, L. Unique role of CD4+CD62L+ regulatory T cells in the control of autoimmune diabetes in T cell receptor transgenic mice. Proc. Natl. Acad. Sci. USA 101 (Suppl 2), 14580–14585 (2004).

    Article  CAS  Google Scholar 

  40. Olivares-Villagómez, D., Wang, Y. & Lafaille, J.J. Regulatory CD4+ T cells expressing endogenous T cell receptor chains protect myelin basic protein-specific transgenic mice from spontaneous autoimmune encephalomyelitis. J. Exp. Med. 188, 1883–1894 (1998).

    Article  Google Scholar 

  41. Fontenot, J.D. et al. Regulatory T cell lineage specification by the forkhead transcription factor Foxp3. Immunity 22, 329–341 (2005).

    Article  CAS  Google Scholar 

  42. Barnden, M.J., Allison, J., Heath, W.R. & Carbone, F.R. Defective TCR expression in transgenic mice constructed using cDNA-based α- and β-chain genes under the control of heterologous regulatory elements. Immunol. Cell Biol. 76, 34–40 (1998).

    Article  CAS  Google Scholar 

  43. Kurts, C. et al. Constitutive class I-restricted exogenous presentation of self antigens in vivo. J. Exp. Med. 184, 923–930 (1996).

    Article  CAS  Google Scholar 

  44. Zhumabekov, T., Corbella, P., Tolaini, M. & Kioussis, D. Improved version of a human CD2 minigene based vector for T cell-specific expression in transgenic mice. J. Immunol. Methods 185, 133–140 (1995).

    Article  CAS  Google Scholar 

  45. Gough, P.J. & Raines, E.W. Gene therapy of apolipoprotein E-deficient mice using a novel macrophage-specific retroviral vector. Blood 101, 485–491 (2003).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank N. Santacruz and J. Hunn for technical assistance, and K. Murphy, W. Yokoyama, P. Allen, W. Swat, J. Scott-Brown and J. Fontenot for discussions and critical review of the manuscript. Supported by the Arthritis Foundation, Burroughs Wellcome Fund and the US National Institutes of Health (C.-S.H.).

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Author notes

  1. Yuqiong Liang & Alexander Y Rudensky

    Present address: Present addresses: Center for Hepatitis Research, University of Texas Medical Branch, Galveston, Texas, USA (Y.L.) and Department of Immunology, Memorial Sloan-Kettering Cancer Center, New York, New York, USA (A.Y.R.).,

Authors and Affiliations

  1. Department of Medicine, Division of Rheumatology, Washington University School of Medicine, St. Louis, Missouri, USA

    Jhoanne L Bautista, Chan-Wang J Lio, Stephanie K Lathrop & Chyi-Song Hsieh

  2. Howard Hughes Medical Institute and Department of Immunology, University of Washington, Seattle, Washington, USA

    Katherine Forbush, Yuqiong Liang & Alexander Y Rudensky

  3. Division of Biostatistics, Washington University School of Medicine, St. Louis, Missouri, USA

    Jingqin Luo

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Correspondence to Chyi-Song Hsieh.

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Bautista, J., Lio, CW., Lathrop, S. et al. Intraclonal competition limits the fate determination of regulatory T cells in the thymus. Nat Immunol 10, 610–617 (2009). https://doi.org/10.1038/ni.1739

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