Viperin restricts chikungunya virus replication and pathology.

Chikungunya virus (CHIKV) is a mosquito-borne arthralgia arbovirus that is reemergent in sub-Saharan Africa and Southeast Asia. CHIKV infection has been shown to be self-limiting, but the molecular mechanisms of the innate immune response that control CHIKV replication remain undefined. Here, longitudinal transcriptional analyses of PBMCs from a cohort of CHIKV-infected patients revealed that type I IFNs controlled CHIKV infection via RSAD2 (which encodes viperin), an enigmatic multifunctional IFN-stimulated gene (ISG). Viperin was highly induced in monocytes, the major target cell of CHIKV in blood. Anti-CHIKV functions of viperin were dependent on its localization in the ER, and the N-terminal amphipathic α-helical domain was crucial for its antiviral activity in controlling CHIKV replication. Furthermore, mice lacking Rsad2 had higher viremia and severe joint inflammation compared with wild-type mice. Our data demonstrate that viperin is a critical antiviral host protein that controls CHIKV infection and provide a preclinical basis for the design of effective control strategies against CHIKV and other reemerging arthrogenic alphaviruses.

[1]  Anjali Rohatgi,et al.  ISG15 Is Critical in the Control of Chikungunya Virus Infection Independent of UbE1L Mediated Conjugation , 2011, PLoS pathogens.

[2]  Alain Michault,et al.  Type I IFN controls chikungunya virus via its action on nonhematopoietic cells , 2010, The Journal of experimental medicine.

[3]  T. Decker,et al.  IFNs and STATs in innate immunity to microorganisms. , 2002, The Journal of clinical investigation.

[4]  Monica Chan,et al.  Active Infection of Human Blood Monocytes by Chikungunya Virus Triggers an Innate Immune Response , 2010, The Journal of Immunology.

[5]  D. Levy,et al.  Differential viral induction of distinct interferon‐α genes by positive feedback through interferon regulatory factor‐7 , 1998, The EMBO journal.

[6]  Larissa B. Thackray,et al.  The Interferon-Inducible Gene viperin Restricts West Nile Virus Pathogenesis , 2011, Journal of Virology.

[7]  D. Levy,et al.  The virus battles: IFN induction of the antiviral state and mechanisms of viral evasion. , 2001, Cytokine & growth factor reviews.

[8]  P. Cresswell,et al.  Viperin (cig5), an IFN-inducible antiviral protein directly induced by human cytomegalovirus , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[9]  M C ROBINSON,et al.  An epidemic of virus disease in Southern Province, Tanganyika Territory, in 1952-53. I. Clinical features. , 1955, Transactions of the Royal Society of Tropical Medicine and Hygiene.

[10]  Fabrice Chrétien,et al.  A Mouse Model for Chikungunya: Young Age and Inefficient Type-I Interferon Signaling Are Risk Factors for Severe Disease , 2008, PLoS pathogens.

[11]  J. Broderick,et al.  The antiviral protein viperin is a radical SAM enzyme , 2010, FEBS letters.

[12]  B. Williams,et al.  Functional classification of interferon‐stimulated genes identified using microarrays , 2001, Journal of leukocyte biology.

[13]  B. Williams,et al.  Identification of genes differentially regulated by interferon alpha, beta, or gamma using oligonucleotide arrays. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[14]  M. Diamond,et al.  Cutting Edge: Independent Roles for IRF-3 and IRF-7 in Hematopoietic and Nonhematopoietic Cells during Host Response to Chikungunya Infection , 2012, The Journal of Immunology.

[15]  A. Salonen,et al.  Viral RNA Replication in Association with Cellular Membranes , 2005, Current topics in microbiology and immunology.

[16]  Z. Adelman,et al.  Accurate Strand-Specific Quantification of Viral RNA , 2009, PloS one.

[17]  G. Pruijn,et al.  Viperin, a key player in the antiviral response. , 2012, Microbes and infection.

[18]  Haitao Guo,et al.  Identification of Five Interferon-Induced Cellular Proteins That Inhibit West Nile Virus and Dengue Virus Infections , 2010, Journal of Virology.

[19]  Zhenghong Yuan,et al.  Viperin inhibits hepatitis C virus replication by interfering with binding of NS5A to host protein hVAP-33. , 2012, The Journal of general virology.

[20]  B. Autran,et al.  Persistent Chronic Inflammation and Infection by Chikungunya Arthritogenic Alphavirus in Spite of a Robust Host Immune Response , 2010, The Journal of Immunology.

[21]  S. Lemon,et al.  The antiviral protein viperin inhibits hepatitis C virus replication via interaction with nonstructural protein 5A , 2011, Hepatology.

[22]  Bryan R. G. Williams,et al.  Interferon-inducible antiviral effectors , 2008, Nature Reviews Immunology.

[23]  M. Albert,et al.  Biology and pathogenesis of chikungunya virus , 2010, Nature Reviews Microbiology.

[24]  N. Reich,et al.  The Interferon Stimulated Gene 54 Promotes Apoptosis* , 2010, The Journal of Biological Chemistry.

[25]  A. Powers,et al.  Changing patterns of chikungunya virus: re-emergence of a zoonotic arbovirus. , 2007, The Journal of general virology.

[26]  P. Vidalain,et al.  Mapping of Chikungunya Virus Interactions with Host Proteins Identified nsP2 as a Highly Connected Viral Component , 2012, Journal of Virology.

[27]  Y. Leo,et al.  Chikungunya fever in Singapore: acute clinical and laboratory features, and factors associated with persistent arthralgia. , 2010, Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology.

[28]  Baohua Gu,et al.  Identification of Three Interferon-Inducible Cellular Enzymes That Inhibit the Replication of Hepatitis C Virus , 2007, Journal of Virology.

[29]  Esther G. L. Koh,et al.  Chikungunya Virus Neutralization Antigens and Direct Cell-to-Cell Transmission Are Revealed by Human Antibody-Escape Mutants , 2011, PLoS pathogens.

[30]  X. de Lamballerie,et al.  Infectious clones of Chikungunya virus (La Réunion isolate) for vector competence studies. , 2006, Vector borne and zoonotic diseases.

[31]  R. Medzhitov,et al.  Type I interferons in host defense. , 2006, Immunity.

[32]  K. Honda,et al.  Type I Inteferon Gene Induction by the Interferon Regulatory Factor Family of Transcription Factors , 2006 .

[33]  P. Cresswell,et al.  Viperin: a multifunctional, interferon-inducible protein that regulates virus replication. , 2011, Cell host & microbe.

[34]  J. Tong,et al.  Immuno-biology of Chikungunya and implications for disease intervention. , 2009, Microbes and infection.

[35]  P. Cresswell,et al.  The antiviral protein, viperin, localizes to lipid droplets via its N-terminal amphipathic α-helix , 2009, Proceedings of the National Academy of Sciences.

[36]  S. Cole,et al.  Adrenergic inhibition of innate anti-viral response: PKA blockade of Type I interferon gene transcription mediates catecholamine support for HIV-1 replication , 2006, Brain, Behavior, and Immunity.

[37]  W. Lumsden.  An epidemic of virus disease in Southern Province, Tanganyika Territory, in 1952-53. II. General description and epidemiology. , 1955, Transactions of the Royal Society of Tropical Medicine and Hygiene.

[38]  K. Choo,et al.  IL-1β, IL-6, and RANTES as Biomarkers of Chikungunya Severity , 2009, PloS one.

[39]  Frank Kunst,et al.  Genome Microevolution of Chikungunya Viruses Causing the Indian Ocean Outbreak , 2006, PLoS medicine.

[40]  M. Colonna,et al.  Viral sensors: diversity in pathogen recognition , 2008, Immunological reviews.

[41]  B. Williams,et al.  Identification of genes differentially regulated by interferon α, β, or γ using oligonucleotide arrays , 1998 .

[42]  Wan Ariffin Bin Abdullah.  Singapore Med J , 1993 .

[43]  L. Ng,et al.  Chikungunya: a bending reality. , 2009, Microbes and infection.

[44]  P. Cresswell,et al.  Human Cytomegalovirus Directly Induces the Antiviral Protein Viperin to Enhance Infectivity , 2011, Science.

[45]  J. Gorvel,et al.  Pathogen–endoplasmic-reticulum interactions: in through the out door , 2006, Nature Reviews Immunology.

[46]  D. Ron,et al.  Feedback Inhibition of the Unfolded Protein Response by GADD34-Mediated Dephosphorylation of eIF2α , 2001, The Journal of cell biology.

[47]  M. Strobel,et al.  Chikungunya, an epidemic arbovirosis. , 2007, The Lancet. Infectious diseases.

[48]  B. He.  Viruses, endoplasmic reticulum stress, and interferon responses , 2006, Cell Death and Differentiation.

[49]  R. Kuhn,et al.  A structural and functional perspective of alphavirus replication and assembly. , 2009, Future microbiology.

[50]  A. Salmon,et al.  Fibroblast cell lines from young adult mice of long-lived mutant strains are resistant to multiple forms of stress. , 2005, American journal of physiology. Endocrinology and metabolism.

[51]  P. Cresswell,et al.  The N-terminal Amphipathic α-Helix of Viperin Mediates Localization to the Cytosolic Face of the Endoplasmic Reticulum and Inhibits Protein Secretion , 2009, Journal of Biological Chemistry.

[52]  V. Chow,et al.  Structural characterization reveals that viperin is a radical S-adenosyl-L-methionine (SAM) enzyme. , 2010, Biochemical and biophysical research communications.

[53]  A. Dasgupta,et al.  The Presence of the Iron-Sulfur Motif Is Important for the Conformational Stability of the Antiviral Protein, Viperin , 2012, PloS one.

[54]  G. Gifford,et al.  Effect of Actinomycin D on Interferon Production by ‘Active’ and ‘Inactive’ Chikungunya Virus in Chick Cells , 1963, Nature.

[55]  Y. Leo,et al.  Clinical features and epidemiology of chikungunya infection in Singapore. , 2009, Singapore medical journal.

[56]  V. Defilippis,et al.  Chikungunya Virus Induces IPS-1-Dependent Innate Immune Activation and Protein Kinase R-Independent Translational Shutoff , 2010, Journal of Virology.

[57]  Crystal W Burke,et al.  Identification and Characterization of Interferon-Induced Proteins That Inhibit Alphavirus Replication , 2007, Journal of Virology.

[58]  N. Nasr,et al.  HIV-1 infection of human macrophages directly induces viperin which inhibits viral production. , 2012, Blood.

[59]  Philippe Pierre,et al.  Induction of GADD34 Is Necessary for dsRNA-Dependent Interferon-β Production and Participates in the Control of Chikungunya Virus Infection , 2012, PLoS pathogens.

[60]  G. Sen,et al.  Interferon‐inducible protein, P56, inhibits HPV DNA replication by binding to the viral protein E1 , 2008, The EMBO journal.

[61]  Kumanan Wilson,et al.  The New International Health Regulations and the Federalism Dilemma , 2005, PLoS medicine.

[62]  T. Ahola,et al.  Inhibitors of Alphavirus Entry and Replication Identified with a Stable Chikungunya Replicon Cell Line and Virus-Based Assays , 2011, PloS one.

[63]  Peter Cresswell,et al.  The interferon-inducible protein viperin inhibits influenza virus release by perturbing lipid rafts. , 2007, Cell host & microbe.

[64]  L. Rénia,et al.  Persistent Arthralgia Induced by Chikungunya Virus Infection is Associated with Interleukin-6 and Granulocyte Macrophage Colony-Stimulating Factor , 2011, The Journal of infectious diseases.

[65]  Karine Labadie,et al.  Chikungunya disease in nonhuman primates involves long-term viral persistence in macrophages. , 2010, The Journal of clinical investigation.

[66]  J. Vlak,et al.  Chikungunya Virus Nonstructural Protein 2 Inhibits Type I/II Interferon-Stimulated JAK-STAT Signaling , 2010, Journal of Virology.

[67]  C. Rice,et al.  Interferon-stimulated genes and their antiviral effector functions , 2011, Current Opinion in Virology.