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Recognition of RNA virus by RIG-I results in activation of CARD9 and inflammasome signaling for interleukin 1β production

Nature Immunology volume 11, pages 63–69 (2010)Cite this article

An Addendum to this article was published on 18 December 2013

This article has been updated

Abstract

Interleukin 1β (IL-1β) is a potent proinflammatory factor during viral infection. Its production is tightly controlled by transcription of Il1b dependent on the transcription factor NF-κB and subsequent processing of pro-IL-1β by an inflammasome. However, the sensors and mechanisms that facilitate RNA virus–induced production of IL-1β are not well defined. Here we report a dual role for the RNA helicase RIG-I in RNA virus–induced proinflammatory responses. Whereas RIG-I-mediated activation of NF-κB required the signaling adaptor MAVS and a complex of the adaptors CARD9 and Bcl-10, RIG-I also bound to the adaptor ASC to trigger caspase-1-dependent inflammasome activation by a mechanism independent of MAVS, CARD9 and the Nod-like receptor protein NLRP3. Our results identify the CARD9–Bcl-10 module as an essential component of the RIG-I-dependent proinflammatory response and establish RIG-I as a sensor able to activate the inflammasome in response to certain RNA viruses.

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Figure 1: RIG-I signaling is required and sufficient for IL-1β production after infection with an RNA virus.
Figure 2: RIG-I controls IL-1β production and caspase-1 activation after detecting RNA viruses.
Figure 3: MAVS and CARD9 are essential for RIG-I-induced production of IL-1β but are dispensable for inflammasome activation.
Figure 4: MAVS, CARD9 and Bcl-10 control pro-IL-1β production after RIG-I ligation.
Figure 5: RIG-I engages ASC to induce inflammasome activation.
Figure 6: RIG-I-induced inflammasome activation is independent of NLRP3.
Figure 7: Mda5 requires NLRP3 for inflammasome activation.

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  • 15 May 2013

    In 2010 we reported in Nature Immunology how the sensing of cytosolic RNA triggers the generation of mature interleukin 1β (IL-1β; Poeck et al., Nat. Immunol. 11, 63–69 (2010)). We demonstrated that ligation of the RNA helicase RIG-I triggered the adaptor CARD9 downstream of the signaling adaptor MAVS (IPS-1) to induce the transcription factor NF-κB for the generation of pro-IL-1β and in parallel activated caspase-1 for the generation of mature IL-1β. We presented data that indicated similar generation of pro-IL-1β and IL-1β in wild-type bone marrow–derived dendritic cells (BMDCs) and BMDCs deficient in CARD9 or the adaptor Bcl-10 after transfection of the synthetic DNA poly(dA:dT) (Fig. 3c (right) and Fig. 4c,e,f). However, additional experiments in our laboratory have shown that CARD9- or Bcl-10-deficient BMDCs have impaired production of pro-IL-1β and IL-1β not only after sensing of RNA but also after the transfection of poly(dA:dT). We speculate that a particular aliquot of poly(dA:dT) used during the preparation of our study published in 2010 might have been contaminated with lipopolysaccharide or some other CARD9-independent trigger of the innate immune system. Still, the central conclusions of that manuscript (that RIG-I signals via CARD9 and the inflammasome to control IL-1β) remain unchanged and have been confirmed by multiple independent laboratories. Nevertheless, we would like to correct the idea that CARD9- or Bcl-10-deficient cells have normal IL-1β responses to the transfection of poly(dA:dT).

References

  1. Kawai, T. & Akira, S. Toll-like receptor and RIG-I-like receptor signaling. Ann. NY Acad. Sci. 1143, 1–20 (2008).

    Article  CAS  Google Scholar 

  2. Alexopoulou, L., Holt, A.C., Medzhitov, R. & Flavell, R.A. Recognition of double-stranded RNA and activation of NF-κB by Toll-like receptor 3. Nature 413, 732–738 (2001).

    Article  CAS  Google Scholar 

  3. Heil, F. et al. Species-specific recognition of single-stranded RNA via toll-like receptor 7 and 8. Science 303, 1526–1529 (2004).

    Article  CAS  Google Scholar 

  4. Hemmi, H. et al. Toll-like receptor recognizes bacterial DNA. Nature 408, 740–745 (2000).

    Article  CAS  Google Scholar 

  5. Burckstummer, T. et al. An orthogonal proteomic-genomic screen identifies AIM2 as a cytoplasmic DNA sensor for the inflammasome. Nat. Immunol. 10, 266–272 (2009).

    Article  Google Scholar 

  6. Fernandes-Alnemri, T., Yu, J.W., Datta, P., Wu, J. & Alnemri, E.S. AIM2 activates the inflammasome and cell death in response to cytoplasmic DNA. Nature 458, 509–513 (2009).

    Article  CAS  Google Scholar 

  7. Hornung, V. et al. AIM2 recognizes cytosolic dsDNA and forms a caspase-1-activating inflammasome with ASC. Nature 458, 514–518 (2009).

    Article  CAS  Google Scholar 

  8. Roberts, T.L. et al. HIN-200 proteins regulate caspase activation in response to foreign cytoplasmic DNA. Science 323, 1057–1060 (2009).

    Article  CAS  Google Scholar 

  9. Andrejeva, J. et al. The V proteins of paramyxoviruses bind the IFN-inducible RNA helicase, mda-5, and inhibit its activation of the IFN-β promoter. Proc. Natl. Acad. Sci. USA 101, 17264–17269 (2004).

    Article  CAS  Google Scholar 

  10. Yoneyama, M. et al. The RNA helicase RIG-I has an essential function in double-stranded RNA-induced innate antiviral responses. Nat. Immunol. 5, 730–737 (2004).

    Article  CAS  Google Scholar 

  11. Pichlmair, A. & Reis e Sousa, C. Innate recognition of viruses. Immunity 27, 370–383 (2007).

    Article  CAS  Google Scholar 

  12. Hornung, V. et al. 5′-Triphosphate RNA is the ligand for RIG-I. Science 314, 994–997 (2006).

    Article  Google Scholar 

  13. Pichlmair, A. et al. RIG-I-mediated antiviral responses to single-stranded RNA bearing 5′-phosphates. Science 314, 997–1001 (2006).

    Article  CAS  Google Scholar 

  14. Schlee, M. et al. Recognition of 5′ triphosphate by RIG-I helicase requires short blunt double-stranded RNA as contained in panhandle of negative-strand virus. Immunity 31, 25–34 (2009).

    Article  CAS  Google Scholar 

  15. Schmidt, A. et al. 5′-triphosphate RNA requires base-paired structures to activate antiviral signaling via RIG-I. Proc. Natl. Acad. Sci. USA 106, 12067–12072 (2009).

    Article  CAS  Google Scholar 

  16. Kato, H. et al. Differential roles of MDA5 and RIG-I helicases in the recognition of RNA viruses. Nature 441, 101–105 (2006).

    Article  CAS  Google Scholar 

  17. Kawai, T. et al. IPS-1, an adaptor triggering RIG-I- and Mda5-mediated type I interferon induction. Nat. Immunol. 6, 981–988 (2005).

    Article  CAS  Google Scholar 

  18. Meylan, E. et al. Cardif is an adaptor protein in the RIG-I antiviral pathway and is targeted by hepatitis C virus. Nature 437, 1167–1172 (2005).

    Article  CAS  Google Scholar 

  19. Seth, R.B., Sun, L., Ea, C.K. & Chen, Z.J. Identification and characterization of MAVS, a mitochondrial antiviral signaling protein that activates NF-κB and IRF 3. Cell 122, 669–682 (2005).

    Article  CAS  Google Scholar 

  20. Nakhaei, P., Genin, P., Civas, A. & Hiscott, J. RIG-I-like receptors: Sensing and responding to RNA virus infection. Semin. Immunol. 21, 215–222 (2009).

    Article  CAS  Google Scholar 

  21. Yu, H.B. & Finlay, B.B. The caspase-1 inflammasome: a pilot of innate immune responses. Cell Host Microbe 4, 198–208 (2008).

    Article  CAS  Google Scholar 

  22. Allen, I.C. et al. The NLRP3 inflammasome mediates in vivo innate immunity to influenza a virus through recognition of viral RNA. Immunity 30, 556–565 (2009).

    Article  CAS  Google Scholar 

  23. Gross, O. et al. Syk kinase signalling couples to the Nlrp3 inflammasome for anti-fungal host defence. Nature 459, 433–436 (2009).

    Article  CAS  Google Scholar 

  24. Ichinohe, T., Lee, H.K., Ogura, Y., Flavell, R. & Iwasaki, A. Inflammasome recognition of influenza virus is essential for adaptive immune responses. J. Exp. Med. 206, 79–87 (2009).

    Article  CAS  Google Scholar 

  25. Thomas, P.G. et al. The intracellular sensor NLRP3 mediates key innate and healing responses to influenza A virus via the regulation of caspase-1. Immunity 30, 566–575 (2009).

    Article  CAS  Google Scholar 

  26. Poeck, H. et al. 5′-Triphosphate-siRNA: turning gene silencing and Rig-I activation against melanoma. Nat. Med. 14, 1256–1263 (2008).

    Article  CAS  Google Scholar 

  27. Gack, M.U. et al. TRIM25 RING-finger E3 ubiquitin ligase is essential for RIG-I-mediated antiviral activity. Nature 446, 916–920 (2007).

    Article  CAS  Google Scholar 

  28. Hara, H. & Saito, T. CARD9 versus CARMA1 in innate and adaptive immunity. Trends Immunol. 30, 234–242 (2009).

    Article  CAS  Google Scholar 

  29. Goodridge, H.S. et al. Differential use of CARD9 by dectin-1 in macrophages and dendritic cells. J. Immunol. 182, 1146–1154 (2009).

    Article  CAS  Google Scholar 

  30. Gross, O. et al. Card9 controls a non-TLR signalling pathway for innate anti-fungal immunity. Nature 442, 651–656 (2006).

    Article  CAS  Google Scholar 

  31. Wang, L. et al. Card10 is a novel caspase recruitment domain/membrane-associated guanylate kinase family member that interacts with BCL10 and activates NF-κB. J. Biol. Chem. 276, 21405–21409 (2001).

    Article  CAS  Google Scholar 

  32. Rawlings, D.J., Sommer, K. & Moreno-Garcia, M.E. The CARMA1 signalosome links the signalling machinery of adaptive and innate immunity in lymphocytes. Nat. Rev. Immunol. 6, 799–812 (2006).

    Article  CAS  Google Scholar 

  33. Henry, T., Brotcke, A., Weiss, D.S., Thompson, L.J. & Monack, D.M. Type I interferon signaling is required for activation of the inflammasome during Francisella infection. J. Exp. Med. 204, 987–994 (2007).

    Article  CAS  Google Scholar 

  34. Steinberg, C. et al. The IFN regulatory factor 7-dependent type I IFN response is not essential for early resistance against murine cytomegalovirus infection. Eur. J. Immunol. 39, 1007–1018 (2009).

    Article  CAS  Google Scholar 

  35. Schroder, K., Muruve, D.A. & Tschopp, J. Innate immunity: cytoplasmic DNA sensing by the AIM2 inflammasome. Curr. Biol. 19, R262–R265 (2009).

    Article  CAS  Google Scholar 

  36. Colonna, M. All roads lead to CARD9. Nat. Immunol. 8, 554–555 (2007).

    Article  CAS  Google Scholar 

  37. Geijtenbeek, T.B. & Gringhuis, S. I. Signalling through C-type lectin receptors: shaping immune responses. Nat. Rev. Immunol. 9, 465–479 (2009).

    Article  CAS  Google Scholar 

  38. Werninghaus, K. et al. Adjuvanticity of a synthetic cord factor analogue for subunit Mycobacterium tuberculosis vaccination requires FcRgamma-Syk-Card9-dependent innate immune activation. J. Exp. Med. 206, 89–97 (2009).

    Article  CAS  Google Scholar 

  39. Hsu, Y.M. et al. The adaptor protein CARD9 is required for innate immune responses to intracellular pathogens. Nat. Immunol. 8, 198–205 (2007).

    Article  CAS  Google Scholar 

  40. Hara, H. et al. The adaptor protein CARD9 is essential for the activation of myeloid cells through ITAM-associated and Toll-like receptors. Nat. Immunol. 8, 619–629 (2007).

    Article  CAS  Google Scholar 

  41. Su, H. et al. Requirement for caspase-8 in NF-κB activation by antigen receptor. Science 307, 1465–1468 (2005).

    Article  CAS  Google Scholar 

  42. Takahashi, K. et al. Roles of caspase-8 and caspase-10 in innate immune responses to double-stranded RNA. J. Immunol. 176, 4520–4524 (2006).

    Article  CAS  Google Scholar 

  43. Bauernfeind, F.G. et al. Cutting edge: NF-κB activating pattern recognition and cytokine receptors license NLRP3 inflammasome activation by regulating NLRP3 expression. J. Immunol. 183, 787–791 (2009).

    Article  CAS  Google Scholar 

  44. Franchi, L., Eigenbrod, T. & Nunez, G. Cutting edge: TNF-α mediates sensitization to ATP and silica via the NLRP3 inflammasome in the absence of microbial stimulation. J. Immunol. 183, 792–796 (2009).

    Article  CAS  Google Scholar 

  45. Dostert, C. et al. Innate immune activation through Nalp3 inflammasome sensing of asbestos and silica. Science 320, 674–677 (2008).

    Article  CAS  Google Scholar 

  46. Gitlin, L. et al. Essential role of mda-5 in type I IFN responses to polyriboinosinic:polyribocytidylic acid and encephalomyocarditis picornavirus. Proc. Natl. Acad. Sci. USA 103, 8459–8464 (2006).

    Article  CAS  Google Scholar 

  47. Mariathasan, S. et al. Differential activation of the inflammasome by caspase-1 adaptors ASC and Ipaf. Nature 430, 213–218 (2004).

    Article  CAS  Google Scholar 

  48. Michallet, M.C. et al. TRADD protein is an essential component of the RIG-like helicase antiviral pathway. Immunity 28, 651–661 (2008).

    Article  CAS  Google Scholar 

  49. Petrilli, V. et al. Activation of the NALP3 inflammasome is triggered by low intracellular potassium concentration. Cell Death Differ. 14, 1583–1589 (2007).

    Article  CAS  Google Scholar 

  50. LeibundGut-Landmann, S. et al. Syk- and CARD9-dependent coupling of innate immunity to the induction of T helper cells that produce interleukin 17. Nat. Immunol. 8, 630–638 (2007).

    Article  CAS  Google Scholar 

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Acknowledgements

We thank J. Tschopp (University of Lausanne) for critical reading of the manuscript, discussions and NLRP3-deficient, ASC-deficient and MAVS-deficient mice and plasmids; and A. Krug (Technical University of Munich) for EMCV. This work includes parts of a thesis by M.B. at the University of Munich. Supported by Bundesministerium für Bildung und Forschung Biofuture (G.H.), Deutsche Forschungsgemeinschaft (SFB704, SFB670, SFB832 and KFO177 to G.H.; Sonderforschungsbereiche to S.E., V.H. and J.R.; Graduiertenkolleg 1202 to M.B.; and RO 2525/3–1 to S.R.), the Center for Integrated Protein Science Munich (S.E.), the European Research Council (V.H.) and Deutsche Krebshilfe (J.R.).

Author information

Author notes

  1. Olaf Gross

    Present address: Present address: Department of Biochemistry, University of Lausanne, Epalinges, Switzerland.,

  2. Hendrik Poeck, Michael Bscheider, Olaf Gross, Gunther Hartmann, Veit Hornung and Jürgen Ruland: These authors contributed equally to this work.

Authors and Affiliations

  1. III. Medizinische Klinik, Klinikum rechts der Isar, Technische Universität München, Munich, Germany

    Hendrik Poeck, Olaf Gross, Katrin Finger, Susanne Roth, Nicole Hannesschläger, Christian Peschel & Jürgen Ruland

  2. Institute of Clinical Chemistry and Pharmacology, Universitätsklinikum Bonn, Bonn, Germany

    Martin Schlee, Winfried Barchet, Gunther Hartmann & Veit Hornung

  3. Center of Integrated Protein Science Munich and Division of Clinical Pharmacology, Department of Internal Medicine, University of Munich, Munich, Germany

    Hendrik Poeck, Michael Bscheider, Simon Rothenfusser & Stefan Endres

  4. Laboratory of Signaling in the Immune System, Helmholtz-Zentrum München–Germany, Research Center for Environmental Health, Neuherberg, Germany

    Susanne Roth & Jürgen Ruland

  5. Department of Biochemistry, University of Lausanne, Epalinges, Switzerland

    Manuele Rebsamen

  6. Department of Host Defense, Research Institute for Microbial Diseases, Osaka University, Japan

    Hiroki Kato & Shizuo Akira

  7. Department of Geriatric Medicine, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan

    Satoshi Inoue

  8. Research Center for Genomic Medicine, Saitama Medical School, Saitama, Japan

    Satoshi Inoue

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Contributions

H.P., M.B., O.G., G.H., V.H. and J.R. designed the research; H.P., M.B., O.G., K.F., S.R., N.H., M.R. and M.S. did experiments; W.B., H.K., S.A. and S.I. contributed critical reagents; H.P., M.B., O.G., S.R., S.E., C.P., V.H., G.H. and J.R. analyzed results; H.P. and M.B. prepared the figures; and H.P., M.B., O.G., G.H. and J.R. wrote the paper.

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Correspondence to Jürgen Ruland.

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Poeck, H., Bscheider, M., Gross, O. et al. Recognition of RNA virus by RIG-I results in activation of CARD9 and inflammasome signaling for interleukin 1β production. Nat Immunol 11, 63–69 (2010). https://doi.org/10.1038/ni.1824

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