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Defective survival of naive CD8+ T lymphocytes in the absence of the β3 regulatory subunit of voltage-gated calcium channels

Nature Immunology volume 10, pages 1275–1282 (2009)Cite this article

Abstract

The survival of T lymphocytes requires sustained, Ca2+ influx–dependent gene expression. The molecular mechanism that governs sustained Ca2+ influx in naive T lymphocytes is unknown. Here we report an essential role for the β3 regulatory subunit of voltage-gated calcium (Cav) channels in the maintenance of naive CD8+ T cells. Deficiency in β3 resulted in a profound survival defect of CD8+ T cells. This defect correlated with depletion of the pore-forming subunit Cav1.4 and attenuation of T cell antigen receptor (TCR)-mediated global Ca2+ entry in CD8+ T cells. Cav1.4 and β3 associated with T cell signaling machinery and Cav1.4 localized in lipid rafts. Our data demonstrate a mechanism by which Ca2+ entry is controlled by a Cav1.4-β3 channel complex in T cells.

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Figure 1: Expression analysis of pore-forming α-subunits and regulatory β-subunits of Cav channels.
Figure 2: Characterization of T cells from Cacnb3−/− mice.
Figure 3: The Ca2+-NFAT pathway is impaired in Cacnb3−/− CD8+ T cells.
Figure 4: Survival of naive CD8+ T cells requires the β3 subunit.
Figure 5: Homeostatic maintenance of naive CD8+ T cells is critically dependent on the β3 subunit.
Figure 6: Upregulation of Fas and downregulation of anti-apoptotic gene expression in Cacnb3−/− CD8+ T cells.
Figure 7: Defective CD8+ T cell–dependent in vivo effector response in Cacnb3−/− mice.
Figure 8: Lack of Cav1.4 pore-forming subunits in Cacnb3−/− naive CD8+ T cells.
Figure 9: Association of Cav1.4 and β3 with T cell signaling components and localization of Cav1.4 in lipid rafts.

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Acknowledgements

We thank E. Eynon for suggestions; F. Manzo for preparing the manuscript; C. Rathinam for critical reading of the manuscript; D. Butkus for thymectomies; T. Taylor for fluorescence-activated cell sorting; and F. Balamuth for advice on lipid raft isolation. Supported by the Arthritis Foundation (M.K.J.), the Fondation pour la Recherche Médicale (A.B.), the Arthritis National Research Foundation (A.B.), Deutsche Forschungsgemeinschaft (V.F. and M.F.), Homburger Forschungsförderungsprogramm (V.F. and M.F.), the Canadian Institutes of Health Research (J.E.M.) and the Howard Hughes Medical Institute (M.K.J., A.B. and R.A.F.).

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Authors and Affiliations

  1. Department of Immunobiology, Yale University School of Medicine, New Haven, Connecticut, USA

    Mithilesh K Jha & Richard A Flavell

  2. Université Cadi Ayyad, Faculté polydisciplinaire Safi, Safi, Morocco

    Abdallah Badou

  3. Institute for Experimental and Clinical Pharmacology and Toxicology, University of Saarland, Homburg, Germany

    Marcel Meissner, Marc Freichel & Veit Flockerzi

  4. Hotchkiss Brain Institute, University of Calgary, Calgary, Canada

    John E McRory

  5. Howard Hughes Medical Institute, Yale University School of Medicine, New Haven, Connecticut, USA

    Richard A Flavell

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Contributions

M.K.J. conceived of, designed and did all experiments, except Ca2+ imaging, and wrote the paper; A.B. did and analyzed Ca2+ imaging experiments; A.B., M.M., J.E.M., M.F. and V.F. contributed reagents and edited the manuscript and R.A.F. provided overall directions, supervised the project and wrote the paper.

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Correspondence to Richard A Flavell.

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Jha, M., Badou, A., Meissner, M. et al. Defective survival of naive CD8+ T lymphocytes in the absence of the β3 regulatory subunit of voltage-gated calcium channels. Nat Immunol 10, 1275–1282 (2009). https://doi.org/10.1038/ni.1793

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