key: cord-0860241-9jninw8x authors: Moriyama, Miyu; Nagai, Minami; Maruzuru, Yuhei; Koshiba, Takumi; Kawaguchi, Yasushi; Ichinohe, Takeshi title: Influenza virus-induced oxidized DNA activates inflammasomes date: 2020-06-14 journal: iScience DOI: 10.1016/j.isci.2020.101270 sha: ca187d299e0a6978b241c9779044b32ff75fa3a0 doc_id: 860241 cord_uid: 9jninw8x Summary Influenza virus M2 and PB1-F2 proteins have been proposed to activate the Nod-like receptor family, pyrin domain-containing 3 (NLRP3) inflammasome in macrophages by altering intracellular ionic balance or mitochondrial reactive oxygen species (ROS) production. However, the precise mechanism by which these viral proteins trigger the NLRP3 inflammasome activation remains unclear. Here we show that influenza virus stimulates oxidized DNA release from macrophages. Ion channel activity of the M2 protein or mitochondrial localization of the PB1-F2 protein was required for oxidized DNA release. While the oxidized DNA enhanced influenza virus-induced IL-1β secretion, inhibition of mitochondrial ROS production by antioxidant Mito-TEMPO decreased the virus-induced IL-1β secretion. In addition, we show that influenza virus stimulates IL-1β secretion from macrophages in an AIM2-dependent manner. These results provide a missing link between influenza viral proteins and the NLRP3 inflammaseome activation, and reveal the importance of influenza virus-induced oxidized DNA in inflammasomes activation. DNase I (D). Cell-free supernatants were collected and analyzed for IL-1β by ELISA. 581 These data are from three independent experiments (mean ± s.e.m.). **P < 0.01, 582 ***P < 0.001; (one-way ANOVA and Tukey's test). See also Figure S3 . 583 584 The NLRP3 inflammasome 314 mediates in vivo innate immunity to influenza A virus through recognition of viral 315 Response of host inflammasomes to viral infection Severe Acute 323 Respiratory Syndrome Coronavirus Viroporin 3a Activates the NLRP3 Inflammasome PtdIns4P on dispersed trans-Golgi network mediates 326 NLRP3 inflammasome activation A novel influenza A virus mitochondrial protein 330 that induces cell death Correction: Influenza A Virus NS1 Protein Inhibits the NLRP3 Inflammasome A 335 single mutation in the PB1-F2 of H5N1 (HK/97) and 1918 influenza A viruses 336 contributes to increased virulence Oxysterol 339 Restraint of Cholesterol Synthesis Prevents AIM2 Inflammasome Activation Innate 342 antiviral responses by means of TLR7-mediated recognition of single-stranded RNA Pore-Forming Protein Gasdermin D Regulates Interleukin-1 Secretion from Living 348 5'-Triphosphate RNA is the ligand for 351 RIG-I Inflammasome 354 recognition of influenza virus is essential for adaptive immune responses Influenza virus activates 358 inflammasomes via its intracellular M2 ion channel Microbiota regulates immune defense against respiratory tract 363 influenza A virus infection Mitochondrial protein mitofusin 2 is required for NLRP3 inflammasome activation after RNA virus infection Encephalomyocarditis virus viroporin 2B 372 activates NLRP3 inflammasome Innate immunity to influenza virus infection Caspase-11 cleaves 379 gasdermin D for non-canonical inflammasome signalling BAK/BAX 400 macropores facilitate mitochondrial herniation and mtDNA efflux during apoptosis PB1-F2 proteins from H5N1 and 20 century pandemic 404 influenza viruses cause immunopathology Expression of the 1918 influenza A virus 408 PB1-F2 enhances the pathogenesis of viral and secondary bacterial pneumonia Activation of the NLRP3 inflammasome 412 by IAV virulence protein PB1-F2 contributes to severe pathophysiology and disease The RNA-and TRIM25-Binding Domains of 416 Influenza Virus NS1 Protein Are Essential for Suppression of NLRP3 Inflammasome-Mediated Interleukin-1beta Secretion High ambient temperature dampens adaptive 421 immune responses to influenza A virus infection Influenza A virus M2 protein 424 triggers mitochondrial DNA-mediated antiviral immune responses Autophagy proteins 428 regulate innate immune responses by inhibiting the release of mitochondrial DNA 429 mediated by the NALP3 inflammasome Severe acute respiratory syndrome coronavirus E protein transports 434 calcium ions and activates the NLRP3 inflammasome IL-1R signaling in dendritic cells 437 replaces pattern-recognition receptors in promoting CD8(+) T cell responses to 438 influenza A virus RIG-I-mediated antiviral responses to single-stranded RNA bearing 443 5'-phosphates Peptide Derived from Avian Influenza A Virus H7N9 Induces Inflammation via 447 Activation of the NLRP3 Inflammasome RIG-I detects viral genomic RNA 451 during negative-strand RNA virus infection Cutting Edge: 455 DNA in the Lung Microenvironment during Influenza Virus Infection Tempers 456 Inflammation by Engaging the DNA Sensor AIM2 Cleavage of GSDMD by inflammatory caspases determines 460 pyroptotic cell death Oxidized mitochondrial DNA 463 activates the NLRP3 inflammasome during apoptosis Influenza A mutant viruses with altered NS1 protein function 467 provoke caspase-1 activation in primary human macrophages, resulting in fast apoptosis 468 and release of high levels of interleukins 1beta and 18 The adaptor MAVS promotes NLRP3 mitochondrial localization and 473 inflammasome activation The NLRP3 inflammasome: molecular 477 activation and regulation to therapeutics The intracellular 481 sensor NLRP3 mediates key innate and healing responses to influenza A virus via the 482 regulation of caspase-1 Incoming RNA virus 487 nucleocapsids containing a 5'-triphosphorylated genome activate RIG-I and antiviral Influenza A virus protein PB1-F2 translocates into mitochondria via 493 Tom40 channels and impairs innate immunity AIM2 Inflammasome Is Critical for Lung Injury and Mortality New mitochondrial DNA synthesis enables 502 NLRP3 inflammasome activation NF-kappaB Restricts Inflammasome 506 Activation via Elimination of Damaged Mitochondria A role for mitochondria in 509 NLRP3 inflammasome activation The authors declare no competing interests. 311