Abstract
Voltage-gated proton currents regulate generation of reactive oxygen species (ROS) in phagocytic cells. In B cells, stimulation of the B cell antigen receptor (BCR) results in the production of ROS that participate in B cell activation, but the involvement of proton channels is unknown. We report here that the voltage-gated proton channel HVCN1 associated with the BCR complex and was internalized together with the BCR after activation. BCR-induced generation of ROS was lower in HVCN1-deficient B cells, which resulted in attenuated BCR signaling via impaired BCR-dependent oxidation of the tyrosine phosphatase SHP-1. This resulted in less activation of the kinases Syk and Akt, impaired mitochondrial respiration and glycolysis and diminished antibody responses in vivo. Our findings identify unanticipated functions for proton channels in B cells and demonstrate the importance of ROS in BCR signaling and downstream metabolism.
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Acknowledgements
We thank U. Klein for discussions; S.-D. Zhang for assistance with statistical analysis; the laboratories of P. Nicotera, G. Cohen, P. Salomoni (Medical Research Council Toxicology Unit) for reagents and discussions; K. Young for help in the acquisition and analysis of data; A. Tobin, I. Forsythe, E. Tulchinsky, M. Kriajevska, E. Raven, J. Basran and K. Brown (University of Leicester) for reagents and use of equipment; F. Batista (Cancer Research UK) and A. Knight (University of Newcastle) for the HEL-specific B cell lines A20 D1.3 and LK35.2 HyHEL10; R. Snowden and K. Topping for assistance with flow cytometry and sorting; and the Medical Research Council (G0802524) for sorter funding. Supported by the Medical Research Council, the Lymphoma Research Foundation (USA), Philip Morris and the US National Institutes of Health (HL-61437).
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M.C. and M.J.S.D. designed research and wrote the manuscript; E.V., T.E.D., I.C.M.M., K.C. and R.D.G. designed some aspects of research; M.C., M.K.B., T.H., R.S.B., K.C., K.P., B.M., V.V.C., D.M., D.D. and M.K. did experiments; C.L. provided some technical support; E.V., T.E.D., I.C.M.M. and K.C. edited the manuscript; and all authors reviewed and approved the manuscript.
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Capasso, M., Bhamrah, M., Henley, T. et al. HVCN1 modulates BCR signal strength via regulation of BCR-dependent generation of reactive oxygen species. Nat Immunol 11, 265–272 (2010). https://doi.org/10.1038/ni.1843
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DOI: https://doi.org/10.1038/ni.1843
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