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Activation of caspase-1 by the NLRP3 inflammasome regulates the NADPH oxidase NOX2 to control phagosome function

Nature Immunology volume 14, pages 543–553 (2013)Cite this article

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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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Figure 1: Active caspase-1 is recruited to S. aureus–containing phagosomes.
Figure 2: Phagocytosis of S. aureus triggers early activation of the NLRP3 inflammasome and caspase-1.
Figure 3: Inhibition of caspase-1 blocks the acidification of S. aureus–containing phagosomes.
Figure 4: The NLRP3 inflammasome regulates acidification of S. aureus–containing phagosomes.
Figure 5: Caspase-1 has phagosome-associated substrates that regulate pH.
Figure 6: NLRP3 inflammasome and caspase-1 regulate the pH of phagosomes containing Gram-positive but not Gram-negative bacteria.
Figure 7: Impact of caspase-1 on the function of phagosomes.

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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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Author notes

  1. W K Eddie Ip

    Present address: Present address: Department of Immunobiology, Yale School of Medicine, New Haven, Connecticut, USA.,

  2. Anna Sokolovska and Christine E Becker: These authors contributed equally to this work.

Authors and Affiliations

  1. Developmental Immunology and Center for Computational and Integrative Biology, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts, USA

    Anna Sokolovska, Christine E Becker, W K Eddie Ip, Matthew Brudner, Nicholas Paquette, Antoine Tanne, Adam Lacy-Hulbert & Lynda M Stuart

  2. Division of Infectious Diseases and Immunology, Department of Medicine, University of Massachusetts Medical School, Worcester, Massachusetts, USA

    Vijay A K Rathinam, Sivapriya K Vanaja & Katherine A Fitzgerald

  3. Departments of Medicine and Cell Biology, New York University Medical Center, New York, New York, USA

    Kathryn J Moore

  4. The Broad Institute of Harvard and Massachusetts Institute of Technology, Cambridge, Massachusetts, USA

    Lynda M Stuart

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Contributions

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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Correspondence to Anna Sokolovska or Lynda M Stuart.

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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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