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Regulatory T cell development in the absence of functional Foxp3

Nature Immunology volume 8, pages 359–368 (2007)Cite this article

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Abstract

Although the development of regulatory T cells (Treg cells) in the thymus is defined by expression of the lineage marker Foxp3, the precise function of Foxp3 in Treg cell lineage commitment is unknown. Here we examined Treg cell development and function in mice with a Foxp3 allele that directs expression of a nonfunctional fusion protein of Foxp3 and enhanced green fluorescent protein (Foxp3ΔEGFP). Thymocyte development in Foxp3ΔEGFP male mice and Foxp3ΔEGFP/+ female mice recapitulated that of wild-type mice. Although mature EGFP+ CD4+ T cells from Foxp3ΔEGFP mice lacked suppressor function, they maintained the characteristic Treg cell 'genetic signature' and failed to develop from EGFP− CD4+ T cells when transferred into lymphopenic hosts, indicative of their common ontogeny with Treg cells. Our results indicate that Treg cell effector function but not lineage commitment requires the expression of functional Foxp3 protein.

NOTE: In the version of this article initially published online, the ‘Foxp2EGFP’ and ‘Foxp2ΔEGFP’ labels in Figure 7 are incorrect. The correct labels should be ‘Foxp3EGFP’ and ‘Foxp3ΔEGFP’, respectively. Also, Supplementary Tables 1-3 truncated early. The errors have been corrected for all versions of the article.

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Figure 1: Foxp3ΔEGFP mice develop a fatal lymphoproliferative disease similar to that of Foxp3-deficient mice.
Figure 2: Characterization of peripheral T cells in Foxp3ΔEGFP male mice.
Figure 3: Thymocyte development in Foxp3ΔEGFP mice.
Figure 4: Population expansion of EGFP+ CD4+ T cells from Foxp3ΔEGFP male mice.
Figure 5: Peripheral depletion of EGFP+ CD4+ T cells in Foxp3ΔEGFP/+ female mice.
Figure 6: EGFP− CD4+ T cells from Foxp3ΔEGFP mice do not convert to EGFP+ CD4+ T cells after adoptive transfer into lymphopenic hosts.
Figure 7: EGFP+ CD4+ T cells from Foxp3ΔEGFP and Foxp3EGFP male mice share a common 'genetic signature'.

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  • 18 February 2007

    NOTE: In the version of this article initially published online, the ‘Foxp2EGFP’ and ‘Foxp2ΔEGFP’ labels in Figure 7 are incorrect. The correct labels should be ‘Foxp3EGFP’ and ‘Foxp3ΔEGFP’, respectively. Also, Supplementary Tables 1-3 truncated early. The errors have been corrected for all versions of the article.

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Acknowledgements

We thank J. Booth, J. Ziegelbauer and B. Edwards for animal care; I. Williams-McClain for cell sorting; and W. Grossman, J. Verbsky and J.M. Routes for critical reading of the manuscript. Supported by the National Institutes of Health (2R01 AI065617 to T.A.C. and R01 AI47154 to C.B.W.), the Nickolett and the D.B. and Marjorie Reinhart Family Foundations (C.B.W.) and the American Heart Association (0525142Y to W.L.).

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Authors and Affiliations

  1. Division of Immunology, Department of Pediatrics, Allergy and Rheumatology, The David Geffen School of Medicine at the University of California at Los Angeles, Los Angeles, 90095, California, USA

    Wen Lin, Nga Truong & Talal A Chatila

  2. Division of Rheumatology, Department of Pediatrics, Medical College of Wisconsin, Milwaukee, 53226, Wisconsin, USA

    Dipica Haribhai, Lance M Relland & Calvin B Williams

  3. Department of Human Genetics, The David Geffen School of Medicine at the University of California at Los Angeles, Los Angeles, 90095, California, USA

    Marc R Carlson

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Correspondence to Calvin B Williams or Talal A Chatila.

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Supplementary information

Supplementary Fig. 1

Generation of Foxp3ΔEGFP mice. (PDF 251 kb)

Supplementary Table 1

Overlap among gene expression profiles of Foxp3ΔEGFP/+ and Foxp3EGFP/+ T cells. (PDF 113 kb)

Supplementary Table 2

Overlap among gene expression profiles of Foxp3ΔEGFP/+ and Foxp3EGFP/+ T cells. (PDF 80 kb)

Supplementary Table 3

Differential gene expression between Foxp3ΔEGFP/+ and Foxp3EGFP/+ cells. (PDF 145 kb)

Supplemental Video 1

The Foxp3-EGFP fusion protein is not found in the nucleus. A frozen lymph node section from a Foxp3ΔEGFP male was stained with anti-GFP/Alexa488 (green) and the DNA stain TO-PRO-3™ (red) (both were obtained from Molecular Probes), then scanned by confocal microscopy (1200X magnification) at 122nm intervals. Images from a representative EGFP+ cell were compiled into a video using Quicktime 7 Pro software (31 images, 6 frames/second). EGFP staining did not co-localize with TO-PRO-3™ (nuclear) staining in any of the images. (MOV 3055 kb)

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Lin, W., Haribhai, D., Relland, L. et al. Regulatory T cell development in the absence of functional Foxp3. Nat Immunol 8, 359–368 (2007). https://doi.org/10.1038/ni1445

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