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Inducible Major Vault Protein Plays a Pivotal Role in Double-Stranded RNA− Or Virus-Induced Proinflammatory Response

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Inducible Major Vault Protein Plays a Pivotal Role in Double-Stranded RNA− Or Virus-Induced Proinflammatory Response Inducible Major Vault Protein Plays a Pivotal Role in Double-Stranded RNA− or Virus-Induced Proinflammatory Response This information is current as Nanfang Peng, Shi Liu, Zhangchuan Xia, Sheng Ren, Jian of September 28, 2021. Feng, Mingzhen Jing, Xin Gao, Erik A. C. Wiemer and Ying Zhu J Immunol 2016; 196:2753-2766; Prepublished online 3 February 2016; doi: 10.4049/jimmunol.1501481 Downloaded from http://www.jimmunol.org/content/196/6/2753 Supplementary http://www.jimmunol.org/content/suppl/2016/02/02/jimmunol.150148 Material 1.DCSupplemental http://www.jimmunol.org/ References This article cites 53 articles, 22 of which you can access for free at: http://www.jimmunol.org/content/196/6/2753.full#ref-list-1 Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision by guest on September 28, 2021 • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2016 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology Inducible Major Vault Protein Plays a Pivotal Role in Double-Stranded RNA– or Virus-Induced Proinflammatory Response Nanfang Peng,*,1 Shi Liu,*,1 Zhangchuan Xia,* Sheng Ren,* Jian Feng,* Mingzhen Jing,* Xin Gao,* Erik A. C. Wiemer,† and Ying Zhu* Pathogen invasion triggers robust antiviral cytokine production via different transcription factor signaling pathways. We have previously demonstrated that major vault protein (MVP) induces type I IFN production during viral infection; however, little is known about the role of MVP in proinflammatory responses. In this study, we found in vitro that expression of MVP, IL-6, and IL-8 was inducible upon dsRNA stimulation or viral infection. Moreover, MVP was essential for the induction of IL-6 and IL-8, as impaired expression of IL-6 and IL-8 in MVP-deficient human PBMCs, human lung epithelial cells (A549), and THP- Downloaded from 1 monocytes, as well as in murine splenocytes, peritoneal macrophages, and PBMCs from MVP-knockout (MVP2/2) mice, was observed. Upon investigation of the underlying mechanisms, we demonstrated that MVP acted in synergy with AP-1 (c-Fos) and CCAAT/enhancer binding protein (C/EBP)b–liver-enriched transcriptional activating protein to activate the IL6 and IL8 pro- moters. Introduction of mutations into the AP-1 and C/EBPb binding sites on the IL6 and IL8 promoters resulted in the loss of synergistic activation with MVP. Furthermore, we found that MVP interacted with both c-Fos and C/EBPb. The interactions 2/2 promoted nuclear translocation and recruitment of these transcription factors to IL6 and IL8 promoter regions. In the MVP http://www.jimmunol.org/ mouse model, significantly decreased expression of early antiviral cytokines resulted in higher viral titer in the lung, higher mortality, and heavier lung damage after infection with lethal influenza A virus. Taken together, our findings help to delineate a novel role of MVP in host proinflammatory response. The Journal of Immunology, 2016, 196: 2753–2766. ecognition of pathogen-associated molecular patterns whereas TLRs 1–13, except TLR10, are expressed in mice (2). by pattern recognition receptors (PRRs) initiates a local TLRs are expressed on both immune cells, including macro- R inflammatory response characterized by production of phages, dendritic cells, and B cells, and nonimmune cells, such as diverse proinflammatory cytokines and chemokines, and this re- fibroblasts and epithelial cells (3). The RLRs consist of RIG-I, by guest on September 28, 2021 sponse acts as a first-line defense, protecting the host from invading melanoma differentiation–associated protein (MDA)5, and LGP2. microbial pathogens. The main PRRs include membrane-bound TLR3 along with RIG-I and MDA5 have been shown to rec- TLRs and cytosolic retinoic acid–inducible gene (RIG)-I–like ognize viral dsRNA generated as a replicative intermediate receptors (RLRs) (1). To date, 13 mammalian TLR paralogs during ssRNA virus infection and the synthetic dsRNA analog have been identified. TLRs 1–10 are expressed in humans, polyinosinic-polycytidylic acid [poly(I:C)] (3–5). Although re- cently challenged by some reports, the notion that influenza A *State Key Laboratory of Virology, College of Life Sciences, Wuhan University, virus (IAV) produces replicative intermediate dsRNA after infec- Wuhan, Hubei 430072, China; and †Department of Medical Oncology, Erasmus tion is widely accepted (4). As the first sensor of viral dsRNA, University Medical Center Cancer Institute, Rotterdam 3000 CA, the Netherlands TLR3 recognizes dsRNA and signals through NF-kB and MAPKs 1 N.P. and S.L. are cofirst authors. to produce proinflammatory cytokines and chemokines via Toll/ Received for publication June 30, 2015. Accepted for publication January 4, 2016. IL-1R domain–containing adaptor inducing IFN-b–dependent This work was supported by Major State Basic Research Development Program of pathways (6). The interaction between Toll/IL-1R domain– China Grant 2013CB911102; National Natural Science Foundation of China Grants 81461130019, 81271821, 31500149, and 31570870; and by National Mega Project containing adaptor inducing IFN-b and TNFR-associated factor on Major Infectious Diseases Prevention Grant 2012ZX10004503-004. The funding (TRAF)6 activates TGF-b–activated protein kinase (TAK1), agencies had no role in study design, data collection, or analysis, decision to publish, which activates both the NF-kB signaling pathway by mediating or preparation of the manuscript. IkB kinase activation and the MAPK signaling pathway by si- Address correspondence and reprint requests to Prof. Ying Zhu, State Key Laboratory of Virology, College of Life Sciences, Wuhan University, 16 Luojia Hill Road, Wuhan, multaneously phosphorylating MAPK kinases 3 and 6 (7–9). RIG-I Hubei 430072, China. E-mail address: [email protected] and MDA5 sense the intracellular dsRNA and signal through The online version of this article contains supplemental material. mitochondrial antiviral signaling protein–dependent pathways Abbreviations used in this article: ATF, activating transcription factor; C/EPB, (10). Mitochondrial antiviral signaling protein contains multiple CCAAT/enhancer binding protein; ChIP, chromatin immunoprecipitation; h, human; TRAF-interacting motifs, which interact with TRAF family IAV, influenza A virus; IRF, IFN regulatory factor; LAP, liver-enriched transcriptional activating protein; LIP, liver-enriched transcriptional inhibitory protein; m, mouse; members, including TRAF2, TRAF3, and TRAF6 and down- MDA, melanoma differentiation–associated protein; MOI, multiplicity of infection; stream with MAP3K and TAK1. Similar to the TLR3 signaling MVP, major vault protein; PM, peritoneal macrophage; poly(I:C), polyinosinic- polycytidylic acid; PRR, pattern recognition receptor; RIG, retinoic acid–inducible pathway, TRAF6 and TAK1 play critical roles in the activation of gene; RLR, retinoic acid–inducible gene-I–like receptor; shRNA, short hairpin RNA; NF-kB and MAPK signaling pathways (10–12). TRAF6 can also TAK1, TGF-b–activated protein kinase 1; TCID50, 50% tissue culture infective dose; interact with MAPK kinase 1, which in turn contributes to the TRAF, TNFR-associated factor. activation of IkB kinase and MAPKK, leading to activation of the Copyright Ó 2016 by The American Association of Immunologists, Inc. 0022-1767/16/$30.00 NF-kB and MAPK signaling pathways (13). www.jimmunol.org/cgi/doi/10.4049/jimmunol.1501481 2754 MVP REGULATES THE EXPRESSION OF IL-6 AND IL-8 MAPKs such as JNK, ERK, and p38 MAPK (14) target the medium. All media were supplemented with 10% heat-inactivated FBS, downstream transcription factors such as AP-1 and CCAAT/ 100 U/ml penicillin, and 100 mg/ml streptomycin sulfate and all cell enhancer binding protein (C/EBP)b (15–17). AP-1 is a hetero- cultures were maintained at 37˚C in a 5% CO2 incubator. All cell lines above were purchased from American Type Culture Collection. dimeric basic region leucine zipper (bZIP) protein comprised of members of the Jun, Fos, activating transcription factors (ATF), Virus and the Maf subfamily (18). The Jun proteins (c-Jun, JunB, and The influenza virus A/Hong Kong/498/97 (H3N2) strain used in this study JunD) both homodimerize and heterodimerize with Fos, whereas was provided by the China Center for Type Culture Collection. The final the Fos proteins (c-Fos, FosB, Fra-1, and Fra-2) only hetero- concentration of H3N2 virus infection used in this study was a multiplicity dimerize with members of the Jun family (14). C/EBPb, which is a of infection (MOI) of 1. The influenza virus A/FM/1/47 (H1N1) used for member of the C/EBP transcription factor family, is widely in- in vivo experiments was adapted for lethality in mice through serial lung passage. volved in cytokine production in response to a variety of stimuli Stock virus was propagated in 10-d-old embryonated chicken eggs (19). C/EBPb binding motifs are found in the functional regula- (Shijun Li laboratory at Central China Agricultural University) for 36–48 h tory region of genes encoding IL-6, IL-8, and TNF-a (20). The at 37˚C. The allantoic fluid was then harvested, and aliquots were stored at 2 C/ebpb gene is translated into at least three different protein 80˚C before being used. isoforms, namely the liver-enriched transcriptional activating Reagent proteins (LAPs), the 38 kDa (full) and the 34 kDa (LAP), and the 20-kDa liver-enriched transcriptional inhibitory proteins Synthetic poly(I:C) (InvivoGen, San Diego, CA), recombinant human (h) IFN-g (PeproTech, London, U.K.), and recombinant mouse (m)IFN-g (LIPs).
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