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Foxp3 programs the development and function of CD4+CD25+ regulatory T cells

Nature Immunology volume 4, pages 330–336 (2003)Cite this article

Abstract

CD4+CD25+ regulatory T cells are essential for the active suppression of autoimmunity. Here we report that the forkhead transcription factor Foxp3 is specifically expressed in CD4+CD25+ regulatory T cells and is required for their development. The lethal autoimmune syndrome observed in Foxp3-mutant scurfy mice and Foxp3-null mice results from a CD4+CD25+ regulatory T cell deficiency and not from a cell-intrinsic defect of CD4+CD25− T cells. CD4+CD25+ regulatory T cells rescue disease development and preferentially expand when transferred into neonatal Foxp3-deficient mice. Furthermore, ectopic expression of Foxp3 confers suppressor function on peripheral CD4+CD25− T cells. Thus, Foxp3 is a critical regulator of CD4+CD25+ regulatory T cell development and function.

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Figure 1: Foxp3 is specifically expressed in CD4+CD25+ regulatory T cells.
Figure 2: Generation and analysis of Foxp3 conditional mutant mice.
Figure 3: Foxp3 is required for development of CD4+CD25+ regulatory T cells and does not affect CD4+CD25− T cell proliferation.
Figure 4: Neonatal transfer of CD4+CD25+ regulatory T cells rescues the lymphoproliferative syndrome in Foxp3-deficient mice.
Figure 5: Expression of Foxp3 confers suppressor phenotype and function upon peripheral CD4+CD25− T cells.

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Acknowledgements

We thank F. Ramsdell and R. Khattri for sharing data before publication, discussions and Foxp3 cosmid DNA, cDNA, real-time PCR primers, recombinant Foxp3 protein and rabbit polyclonal antiserum; C. Beers and K. Honey for technical assistance; C. Hsieh for discussions; P. DeRoos, K. Forebush and E. Nergou for technical support; C. Plata and N. Li for expert mouse colony management; C. Ware for ES cell electroporation and blastocyst injection; W. Pear for the MSCV MigR1 retroviral construct; B. Sopher for CMV-Cre TG mice; and P. Soriano for the KO targeting vector. Supported by training grants from the National Institutes of Health, the Cancer Research Institute (J.D.F.) and the Howard Hughes Medical Institute, and a grant from the National Institutes of Health.

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  1. Department of Immunology, Howard Hughes Medical Institute, University of Washington, Box 357370, Seattle, 98195, WA, USA

    Jason D. Fontenot, Marc A. Gavin & Alexander Y. Rudensky

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  1. Jason D. Fontenot
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  2. Marc A. Gavin
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Correspondence to Alexander Y. Rudensky.

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Supplementary information

Supplementary Fig. 1.

Cotransfer of CD4+CD25+ regulatory T cells suppresses development of Foxp3-deficient CD4+ T cell-mediated autoimmunity in RAG-/- Mice. 1×106 purified CD4+ T cells from 15-20 day old Foxp3sf mice were injected i.v. into 8 week old RAG1-/- mice alone or in combination with 5×105 wild-type CD4+CD25+ T cells or CD4+CD25+ T cells. Host mice were monitored for the development of colitis and wasting disease by weight loss. Representative of 2 experiments with at least 3 mice per group. (PDF 17 kb)

Supplementary Fig. 2.

Increased Foxp3 expression in anergic cells generated in vivo upon engagement of TCR by high affinity self peptide-MHC complexes. CD4+ T cells from TCR transgenic mice (TEa) co-expressing cognate ligand (pEα:Ii) are anergic and upregulate Foxp3 and IL-10 mRNA. Anergic CD4+ T cells were generated by crossing TEa TCR transgenic mice with mice expressing the TEa TCR cognate peptide ligand (pEα:Ii). (a) Flow cytometric analysis of thymocytes demonstrated substantial negative selection and transgenic TCR downregulation in these mice. However, a small number of mature CD4+ thymocytes expressing the transgenic TCR (Vβ6+Vα2+) were readily detectable. (b) Proliferative responses of sorted CD4+ T cells from TEa or Tea-pEα mice. TCR transgene positive Vβ6+Vα2+ or Vβ6+Vα2– CD4+ T cells were stimulated in vitro with B6 or pEα-expressing splenocytes. (c) Quantitative real time PCR for Foxp3 and IL-10 mRNA expression was using cDNA from sorted T cell populations. (PDF 30 kb)

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Fontenot, J., Gavin, M. & Rudensky, A. Foxp3 programs the development and function of CD4+CD25+ regulatory T cells. Nat Immunol 4, 330–336 (2003). https://doi.org/10.1038/ni904

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