Abstract
Phagocytosis is a fundamental cellular process that is pivotal for immunity as it coordinates microbial killing, innate immune activation and antigen presentation. An essential step in this process is phagosome acidification, which regulates many functions of these organelles that allow phagosomes to participate in processes that are essential to both innate and adaptive immunity. Here we report that acidification of phagosomes containing Gram-positive bacteria is regulated by the NLRP3 inflammasome and caspase-1. Active caspase-1 accumulates on phagosomes and acts locally to control the pH by modulating buffering by the NADPH oxidase NOX2. These data provide insight into a mechanism by which innate immune signals can modify cellular defenses and establish a new function for the NLRP3 inflammasome and caspase-1 in host defense.
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Acknowledgements
This work was funded by grants from US National Institutes of Health National Institute of Allergy and Infectious Diseases to L.M.S. (RO1 AI079198) and K.A.F. (RO1 AI093752), National Institutes of Health National Institute on Aging to K.J.M. (RO1 AG020255), grants from the Crohns' and Colitis and Hood foundations and National Institutes of Health National Institute of Diabetes and Digestive and Kidney Diseases to A.L.-H. (RO1 DK093695), and postdoctoral fellowships from Massachusetts General Hospital Executive Committee on Research to A.S. and New England Regional Center of Excellence–Biodefense and Emerging Infectious Diseases (U54 AI057159 to N.P.). Electron microscopy was performed by M. McKee in the Microscopy Core of the Center for Systems Biology.
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A.S., C.E.B. and W.K.E.I. designed, analyzed and performed experiments. V.A.K.R., M.B., N.P., A.T. and S.K.V. contributed to experimental design, provided discussions and assisted with experiments. K.J.M., K.A.F. and A.L.-H. contributed to data analysis, discussions and manuscript preparation. L.M.S. designed and analyzed experiments, supervised the project and prepared the manuscript with A.S. and C.E.B.
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Supplementary Table 1
List of potential phagosome-associated caspase-1 substrates. Potential caspase-1 cleavage sites in all the phagosome-associated proteins identified in ref. 26 were determined by the SitePrediction algorithm (sheet 1; http://www.dmbr.ugent.be/prx/bioit2-public/SitePrediction/). Data were filtered at 99% confidence. Potential phagosome caspase-1 substrates identified by SitePrediction were further classified by DAVID analysis to identify enriched gene functions (sheet 2). (XLS 87 kb)
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Sokolovska, A., Becker, C., Ip, W. et al. Activation of caspase-1 by the NLRP3 inflammasome regulates the NADPH oxidase NOX2 to control phagosome function. Nat Immunol 14, 543–553 (2013). https://doi.org/10.1038/ni.2595
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DOI: https://doi.org/10.1038/ni.2595
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