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Themis is a member of a new metazoan gene family and is required for the completion of thymocyte positive selection

Nature Immunology volume 10, pages 831–839 (2009)Cite this article

A Corrigendum to this article was published on 01 January 2010

This article has been updated

Abstract

T cell antigen receptor (TCR) signaling in CD4+CD8+ double-positive thymocytes determines cell survival and lineage commitment, but the genetic and molecular basis of this process is poorly defined. To address this issue, we used ethylnitrosourea mutagenesis to identify a previously unknown T lineage–specific gene, Themis, which is critical for the completion of positive selection. Themis contains a tandem repeat of a unique globular domain (called 'CABIT' here) that includes a cysteine motif that defines a family of five uncharacterized vertebrate proteins with orthologs in most animal species. Themis-deficient thymocytes showed no substantial impairment in early TCR signaling but did show altered expression of genes involved in the cell cycle and survival before and during positive selection. Our data suggest a unique function for Themis in sustaining positive selection.

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Figure 1: Lower production of CD4+ and CD8+ T cells in 5AT161 mutant mice.
Figure 2: Impaired positive selection in TCR-transgenic 5AT161 mice and normal superantigen negative selection in B10.BR 5AT161 mice.
Figure 3: Induced premature stop codon encoded by Themis in 5AT161 mice.
Figure 4: Domain architectures and evolution of the CABIT domain.
Figure 5: Themis expression.
Figure 6: Normal TCR-driven upregulation of activation markers and lower expression of development markers in Themis-mutant DP thymocytes.
Figure 7: Altered expression of survival, cell cycle and lipid metabolism genes in Themis(Y489X) thymocytes.

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  • 19 August 2009

    NOTE: In the version of this article initially published, the top right graph in Fig. 7b is incorrect. The error has been corrected in the HTML and PDF versions of the article.

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Acknowledgements

We thank K. Holmes, E. Stregevsky and B. Hague for flow sorting; D.E. Anderson for mass spectrometry; T. Myers and Q. Su for assistance with microarrays; and the staffs of the Australian Phenomics Facility, Biomedical Services Oxford, and the US National Institutes of Health Comparative Medicine Branch for animal husbandry. Supported by the US National Institutes of Health (A.L.J.); Oxford University (A.L.J.); The Division of Intramural Research, National Institute of Allergy and Infectious Diseases (R.H.S. and M.J.L.) and National Library of Medicine (L.A.), US National Institutes of Health; the Medical Research Council (R.J.C.); the Wellcome Trust (R.J.C. and C.C.G.); the Australian Research Council Federation (C.C.G.); and the National Institute for Health Research Biomedical Research Centre Programme (R.J.C.).

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Author notes

  1. Richard J Cornall and Ronald H Schwartz: These authors contributed equally to this work.

Authors and Affiliations

  1. Nuffield Department of Clinical Medicine, Oxford University, Oxford, UK

    Andy L Johnson, Tanya L Crockford, Teresa Lambe & Richard J Cornall

  2. Laboratory of Cellular and Molecular Immunology, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland, USA

    Andy L Johnson, Natalia Shulzhenko, Andre Morgun, See-Young Choi & Ronald H Schwartz

  3. National Library of Medicine, National Institutes of Health, Bethesda, Maryland, USA

    L Aravind

  4. Australian Cancer Research Foundation Genetics Laboratory, John Curtin School of Medical Research, Australian National University, and Australian Phenomics Facility, Canberra, Australia

    Heather Domaschenz, Edyta M Kucharska, Carola G Vinuesa & Christopher C Goodnow

  5. Laboratory of Immunology, National Institutes of Health, Bethesda, Maryland, USA

    Lixin Zheng & Michael J Lenardo

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Contributions

A.L.J. designed and did experiments, analyzed and interpreted results and wrote the manuscript; L.A. did the bioinformatics analysis and designed the related figures and text; N.S. and A.M. assisted with microarray experimental design and analysis; S.-Y.C. did the GST precipitation and some of the biochemical analyses by immunoblot; T.L.C. and T.L. contributed to ex vivo cellular analyses, tissue culture and gene mapping; H.D. and E.M.K. did the immunization and ANA screens; L.Z. contributed to cloning, confocal microscopy and immunoprecipitation studies; C.G.V., M.J.L. and C.C.G. helped with designing experiments and writing the manuscript; and R.J.C. and R.H.S. directed the study, analyzed and interpreted results and wrote the manuscript.

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Correspondence to Richard J Cornall or Ronald H Schwartz.

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Johnson, A., Aravind, L., Shulzhenko, N. et al. Themis is a member of a new metazoan gene family and is required for the completion of thymocyte positive selection. Nat Immunol 10, 831–839 (2009). https://doi.org/10.1038/ni.1769

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