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Clonal deletion and the fate of autoreactive thymocytes that survive negative selection

Nature Immunology volume 13, pages 569–578 (2012)Cite this article

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Abstract

Clonal deletion of autoreactive thymocytes is important for self-tolerance, but the intrathymic signals that induce clonal deletion have not been clearly identified. We now report that clonal deletion during negative selection required CD28-mediated costimulation of autoreactive thymocytes at the CD4+CD8lo intermediate stage of differentiation. Autoreactive thymocytes were prevented from undergoing clonal deletion by either a lack of CD28 costimulation or transgenic overexpression of the antiapoptotic factors Bcl-2 or Mcl-1, with surviving thymocytes differentiating into anergic CD4−CD8− double-negative thymocytes positive for the T cell antigen receptor αβ subtype (TCRαβ) that 'preferentially' migrated to the intestine, where they re-expressed CD8α and were sequestered as CD8αα+ intraepithelial lymphocytes (IELs). Our study identifies costimulation by CD28 as the intrathymic signal required for clonal deletion and identifies CD8αα+ IELs as the developmental fate of autoreactive thymocytes that survive negative selection.

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Figure 1: CD28 costimulation–deficient mice have more TCRαβ+ DN thymocytes.
Figure 2: Effect of transgenes encoding Bcl-2 and Mcl-1 on the appearance of TCRαβ+ DN thymocytes.
Figure 3: Expression of Mtv-reactive Vβ+ TCRs in pre- and post-selection thymocyte subsets.
Figure 4: TCRαβ+ DN thymocytes are the progeny of DP thymocytes.
Figure 5: Effect of thymic selection on the expression of Mtv-reactive Vβ+ TCRs.
Figure 6: Developmentally diverted TCRαβ+ DN thymocytes migrate to the intestine, where they become CD8αα+ IELs.
Figure 7: Developmentally diverted TCRαβ+ DN thymocytes become CD8αα+ IELs.
Figure 8: Runx3 is required for the differentiation of TCRαβ+ DN thymocytes into developmentally diverted CD8αα+ IELs.

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Acknowledgements

We thank N. Taylor, J. DiSanto and R. Hodes for critical reading of the manuscript; A. Sharpe (Harvard University) for B7-deficient mice, T. Honjo (Kyoto University) for PD-1-deficient mice; D. Littman (New York University) for Runx3-YFP reporter mice; and S. Sharrow, A. Adams and L. Granger for flow cytometry. Supported by the Intramural Research Program of the National Cancer Institute Center for Cancer Research of the US National Institutes of Health.

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Authors and Affiliations

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

    Leonid A Pobezinsky, Xuguang Tai, Susanna Jeurling, François Van Laethem, Jung-Hyun Park & Alfred Singer

  2. Ludwig Center for Cancer Research, University of Lausanne, Epalinges, Switzerland

    Georgi S Angelov

  3. Science Applications International Corporation-Frederick, National Cancer Institute-Frederick Cancer Research and Development Center, Frederick, Maryland, USA

    Lionel Feigenbaum

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Contributions

L.A.P. designed the study, did experiments, analyzed data and contributed to the writing of the manuscript; G.S.A., X.T., S.J. and F.V.L. did experiments and analyzed data; J.-H.P. and L.F. generated transgenic mice, and A.S. designed the study, analyzed data and wrote the manuscript.

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Correspondence to Alfred Singer.

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Pobezinsky, L., Angelov, G., Tai, X. et al. Clonal deletion and the fate of autoreactive thymocytes that survive negative selection. Nat Immunol 13, 569–578 (2012). https://doi.org/10.1038/ni.2292

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