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CD28 costimulation of developing thymocytes induces Foxp3 expression and regulatory T cell differentiation independently of interleukin 2

Nature Immunology volume 6, pages 152–162 (2005)Cite this article

Abstract

Efficient generation of regulatory T cells (Treg cells) in the thymus requires CD28 costimulation, but it is not known why. Here, molecular mapping of CD28 costimulation showed that Treg cell generation requires a motif that binds the tyrosine kinase Lck, precisely the same motif that is required for CD28 costimulation of interleukin 2 production. Nevertheless, CD28 costimulation provides more than interleukin 2 to developing Treg cells, as CD28 costimulation of T cell receptor–signaled double-positive thymocytes induced expression of Foxp3, considered to be the Treg 'master gene', as well as GITR and CTLA-4, two proteins expressed on Treg cells. Thus, CD28 costimulation directly signals developing thymocytes to express Foxp3 and to initiate the Treg cell differentiation program.

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Figure 1: CD28 requirement for generation of Treg cells in the thymus.
Figure 2: Generation of CD28 transgenic mice.
Figure 3: Molecular mapping of the CD28 binding motifs required for CD28-dependent IL-2 production and for IL-2-dependent proliferation.
Figure 4: Generation of CD4+CD25+ T cells in B6 + Il2−/− → B6 mixed radiation bone marrow chimeras.
Figure 5: Generation of CD4+CD25+ T cells in mixed bone marrow chimeras.
Figure 6: Induction of Foxp3 expression in immature thymocytes.
Figure 7: CD28 cosimulation of TCR-signaled DP thymocytes initiates Treg cell differentiation.
Figure 8: Molecular mapping of CD28 costimulatory signals that induce Foxp3 and CTLA-4 expression in developing thymocytes.

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Acknowledgements

We thank A.S. Shaw for CD28 cDNA; S. Sharrow, L. Granger and T. Adam for flow cytometry; and E.M. Shevach, R.J. Hodes, A. Bhandoola and R. Bosselut for critically reading the manuscript.

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Authors and Affiliations

  1. Experimental Immunology Branch, National Cancer Institute, National Institutes of Health, Bethesda, 20892, Maryland, USA

    Xuguang Tai, Michelle Cowan & Alfred Singer

  2. SAIC Frederick, National Cancer Institute-Frederick Cancer Research and Development Center, Frederick, 21702, Maryland, USA

    Lionel Feigenbaum

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Correspondence to Alfred Singer.

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Supplementary information

Supplementary Fig. 1

Purified Bcl-2 Tg DP thymocytes were first signaled overnight with anti-TCR plus anti-CD28 mAb's in various combinations (expressed as μg/ml), and then transferred into overnight 'recovery' cultures with medium alone. (PDF 124 kb)

Supplementary Fig. 2

Purified Bcl-2 Tg+ DP thymocytes from indicated CD28 Tg mice were stimulated overnight with immobilized anti-TCR (2μg/ml) and anti-CD28 (25μg/ml) mAb's as indicated, and then transferred into overnight recovery cultures. (PDF 102 kb)

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Tai, X., Cowan, M., Feigenbaum, L. et al. CD28 costimulation of developing thymocytes induces Foxp3 expression and regulatory T cell differentiation independently of interleukin 2. Nat Immunol 6, 152–162 (2005). https://doi.org/10.1038/ni1160

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