Abstract
The sustained entry of Ca2+ into CD4+CD8+ double-positive thymocytes is required for positive selection. Here we identified a voltage-gated Na+ channel (VGSC) that was essential for positive selection of CD4+ T cells. Pharmacological inhibition of VGSC activity inhibited the sustained Ca2+ influx induced by positively selecting ligands and the in vitro positive selection of CD4+ but not CD8+ T cells. In vivo short hairpin RNA (shRNA)-mediated knockdown of the gene encoding a regulatory β-subunit of a VGSC specifically inhibited the positive selection of CD4+ T cells. Ectopic expression of VGSC in peripheral AND CD4+ T cells bestowed the ability to respond to a positively selecting ligand, which directly demonstrated that VGSC expression was responsible for the enhanced sensitivity. Thus, active VGSCs in thymocytes provide a mechanism by which a weak positive selection signal can induce the sustained Ca2+ signals required for CD4+ T cell development.
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Acknowledgements
We thank A. Farr (University of Washington) for the ANV41.2 the cortical epithelial cell line; D. Kreamalmeyer for maintaining the mouse colony; S. Horvath for peptide synthesis and purification; M. Cella for advice on the production of the SCN4B-immunoglobulin fusion protein; M. Colonna (Washington University School of Medicine) for the Ceacam4-Ig fusion protein; S. Chan, S. Srivatsan and D. Bhattacharya for advice on shRNA-mediated knockdown experiments; R. Schreiber for mouse IFN-γ; J. Silva, C. Nichols and S. Goldstein (University of Chicago) for the human SCN5A–GFP construct; J. Nerbonne for discussions; N. Felix for initiating the gp250 project; G. Morris for critical data analysis and assistance with the manuscript; and K. Weber, C. Morley, A. Shaw, M. Vig, Y. Huang, K. Murphy and T. Egawa for critical reading of the manuscript and comments. Supported by the US National Institutes of Health (AI-24157 to P.M.A.).
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W.-L.L. and P.M.A. designed the study and wrote the manuscript; W.-L.L. did the experimental work; and D.L.D. generated the AND hybridoma cell lines.
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Lo, WL., Donermeyer, D. & Allen, P. A voltage-gated sodium channel is essential for the positive selection of CD4+ T cells. Nat Immunol 13, 880–887 (2012). https://doi.org/10.1038/ni.2379
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DOI: https://doi.org/10.1038/ni.2379
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