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Receptor interacting protein kinase 2–mediated mitophagy regulates inflammasome activation during virus infection

Nature Immunology volume 14, pages 480–488 (2013)Cite this article

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Abstract

NOD2 receptor and the cytosolic protein kinase RIPK2 regulate NF-κB and MAP kinase signaling during bacterial infections, but the role of this immune axis during viral infections has not been addressed. We demonstrate that Nod2−/− and Ripk2−/− mice are hypersusceptible to infection with influenza A virus. Ripk2−/− cells exhibited defective autophagy of mitochondria (mitophagy), leading to enhanced mitochondrial production of superoxide and accumulation of damaged mitochondria, which resulted in greater activation of the NLRP3 inflammasome and production of IL-18. RIPK2 regulated mitophagy in a kinase-dependent manner by phosphorylating the mitophagy inducer ULK1. Accordingly, Ulk1−/− cells exhibited enhanced mitochondrial production of superoxide and activation of caspase-1. These results demonstrate a role for NOD2-RIPK2 signaling in protection against virally triggered immunopathology by negatively regulating activation of the NLRP3 inflammasome and production of IL-18 via ULK1-dependent mitophagy.

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Figure 1: RIPK2 deficiency leads to hyperinflammation.
Figure 2: RIPK2 modulates cytokine and chemokine production.
Figure 3: IL-18 mediates hyperinflammation in Ripk2−/− mice.
Figure 4: Both hematopoietic and lung epithelial cells contribute to hypercytokinemia.
Figure 5: Elevated IL-18 in Ripk2−/− cells is NLRP3 inflammasome-dependent.
Figure 6: RIPK2 modulates inflammasome activation through autophagy.
Figure 7: RIPK2 specifically regulates mitophagy and accumulation of damaged mitochondria to modulate inflammasome activation.
Figure 8: RIPK2 regulates activation of the critical autophagy protein ULK1 in response to IAV infection.

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Acknowledgements

We thank members of the Veterinary Pathology Core lab at St. Jude for their work in processing of hematoxylin and eosin–stained and immunohistochemistry slides, members of the Cell and Tissue Imaging Core facility for their help in preparing and imaging transmission electron microscopy samples, and A. Coyle, E. Grant, J. Bertin (Millennium Pharmaceuticals), T. Mak (University of Toronto) and R. Flavell (Yale University) for providing mutant mice. M.L. is supported by grants from the EU Framework Program 7 (Marie-Curie grant 256432), European Research Council (grant 281600) and the Fund for Scientific Research Flanders (G030212N, 1.2.201.10.N.00 and 1.5.122.11.N.00). This work was supported by US National Institutes of Health grants (AR056296, AI101935 and CA163507) and the American Lebanese Syrian Associated Charities to T.-D.K.

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Authors and Affiliations

  1. Department of Immunology, St. Jude Children's Research Hospital, Memphis, Tennessee, USA

    Christopher Lupfer, Paul G Thomas, Paras K Anand, Sandra Milasta, Jennifer Martinez, Gonghua Huang, Maggie Green, Hongbo Chi, Douglas R Green, Peter C Doherty & Thirumala-Devi Kanneganti

  2. Veterinary Pathology Core, St. Jude Children's Research Hospital, Memphis, Tennessee, USA

    Peter Vogel

  3. Department of Pathology, St. Jude Children's Research Hospital, Memphis, Tennessee, USA

    Mondira Kundu

  4. Gastrointestinal Unit and Center for the Study of Inflammatory Bowel Disease, Harvard Medical School, Boston, Massachusetts, USA

    Ramnik J Xavier

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

    Ramnik J Xavier

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

    Ramnik J Xavier

  7. Department of Biochemistry, Ghent University, Ghent, Belgium

    Mohamed Lamkanfi

  8. Department of Medical Protein Research, Vlaams Instituut voor Biotechnologie, Ghent, Belgium

    Mohamed Lamkanfi

  9. Department of Medicine, University of Colorado Denver, Aurora, Colorado, USA

    Charles A Dinarello

  10. Department of Microbiology and Immunology, University of Melbourne, Parkville, Victoria, Australia

    Peter C Doherty

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  1. Christopher Lupfer
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Contributions

C.L. designed and conducted experiments, and wrote the manuscript. P.G.T. helped conduct the initial experiments in vivo in Ripk2−/− mice and helped design T cell experiments. P.K.A. conducted experiments with L. monocytogenes and helped write the manuscript. P.V. helped design experiments and interpret histopathology data. S.M. helped design and conduct experiments related to mitophagy and mitochondrial damage. J.M. helped design experiments related to mitophagy and mitochondrial damage. G.H. helped design and conduct experiments with Mapk14 conditional knockout mice. M.G. conducted experiments for IL-18, NF-κB and MAPK signaling. M.K. helped design experiments, and provided Ulk1−/− mice and other reagents for ULK1 experiments. H.C. helped design experiments and provided Mapk14 conditional knockout mice. R.J.X. helped design experiments and provided reagents for ATG16-related experiments and autophagy in general. D.R.G. helped design experiments and provided reagents for mitophagy-related experiments. M.L. helped design experiments and provided reagents for caspase-1 activation and interaction studies. C.A.D. helped design experiments and provided reagents for IL-18 neutralization studies. P.C.D. helped design experiments, provided reagents for T cell experiments and helped write the manuscript. T.-D.K. conceived project, designed experiments, analyzed data, helped write the manuscript and oversaw the project.

Corresponding author

Correspondence to Thirumala-Devi Kanneganti.

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Lupfer, C., Thomas, P., Anand, P. et al. Receptor interacting protein kinase 2–mediated mitophagy regulates inflammasome activation during virus infection. Nat Immunol 14, 480–488 (2013). https://doi.org/10.1038/ni.2563

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