Immune evasion by hepatitis C virus NS3/4A protease-mediated cleavage of the Toll-like receptor 3 adaptor protein TRIF.

Toll-like receptors (TLRs) bind pathogen-specific ligands early in infection, initiating signaling pathways that lead to expression of multiple protective cellular genes. Many viruses have evolved strategies that block the effector mechanisms induced through these signaling pathways, but viral interference with critical proximal receptor interactions has not been described. We show here that the NS3/4A serine protease of hepatitis C virus (HCV), a virus notorious for its ability to establish persistent intrahepatic infection, causes specific proteolysis of Toll-IL-1 receptor domain-containing adaptor inducing IFN-beta (TRIF or TICAM-1), an adaptor protein linking TLR3 to kinases responsible for activating IFN regulatory factor 3 (IRF-3) and NF-kappaB, transcription factors controlling a multiplicity of antiviral defenses. NS3/4A-mediated cleavage of TRIF reduces its abundance and inhibits polyI:C-activated signaling through the TLR3 pathway before its bifurcation to IRF-3 and NF-kappaB. This uniquely broad mechanism of immune evasion potentially limits expression of multiple host defense genes, thereby promoting persistent infections with this medically important virus.

[1]  Stanley M. Lemon,et al.  Regulation of Interferon Regulatory Factor-3 by the Hepatitis C Virus Serine Protease , 2003, Science.

[2]  B. Beutler,et al.  Inferences, questions and possibilities in Toll-like receptor signalling , 2004, Nature.

[3]  R. Bartenschlager,et al.  Identification of the Hepatitis C Virus RNA Replication Complex in Huh-7 Cells Harboring Subgenomic Replicons , 2003, Journal of Virology.

[4]  A. Urbani,et al.  Activity of purified hepatitis C virus protease NS3 on peptide substrates , 1996, Journal of virology.

[5]  M. Yoneyama,et al.  Review: Control of IRF-3 Activation by Phosphorylation , 2002 .

[6]  R. Lanford,et al.  DNA Microarray Analysis of Chimpanzee Liver during Acute Resolving Hepatitis C Virus Infection , 2001, Journal of Virology.

[7]  C. Rice,et al.  Specificity of the hepatitis C virus NS3 serine protease: effects of substitutions at the 3/4A, 4A/4B, 4B/5A, and 5A/5B cleavage sites on polyprotein processing , 1994, Journal of virology.

[8]  A. Iwasaki,et al.  Toll-like receptor control of the adaptive immune responses , 2004, Nature Immunology.

[9]  G. Stark,et al.  IRF-3-dependent, NFκB- and JNK-independent activation of the 561 and IFN-β genes in response to double-stranded RNA , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[10]  Shizuo Akira,et al.  The RNA helicase RIG-I has an essential function in double-stranded RNA-induced innate antiviral responses , 2004, Nature Immunology.

[11]  Jiahuai Han,et al.  Identification of Lps2 as a key transducer of MyD88-independent TIR signalling , 2003, Nature.

[12]  T. Maniatis,et al.  IKKε and TBK1 are essential components of the IRF3 signaling pathway , 2003, Nature Immunology.

[13]  J. Pawlotsky.  Pathophysiology of hepatitis C virus infection and related liver disease. , 2004, Trends in microbiology.

[14]  S. Lemon,et al.  Control of antiviral defenses through hepatitis C virus disruption of retinoic acid-inducible gene-I signaling. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[15]  S. Lemon,et al.  Virus-Host Cell Interactions during Hepatitis C Virus RNA Replication: Impact of Polyprotein Expression on the Cellular Transcriptome and Cell Cycle Association with Viral RNA Synthesis , 2004, Journal of Virology.

[16]  C. Rice,et al.  Hepatitis C virus NS3 serine proteinase: trans-cleavage requirements and processing kinetics , 1994, Journal of virology.

[17]  I. Haga,et al.  The Poxvirus Protein A52R Targets Toll-like Receptor Signaling Complexes to Suppress Host Defense , 2003, The Journal of experimental medicine.

[18]  R. De Francesco,et al.  Structure and function of the hepatitis C virus NS3-NS4A serine proteinase. , 2000, Current topics in microbiology and immunology.

[19]  M. Yoneyama,et al.  Induction of IRF‐3/‐7 kinase and NF‐κB in response to double‐stranded RNA and virus infection: common and unique pathways , 2001, Genes to cells : devoted to molecular & cellular mechanisms.

[20]  C. Rice,et al.  Subcellular Localization, Stability, andtrans-Cleavage Competence of the Hepatitis C Virus NS3-NS4A Complex Expressed in Tetracycline-Regulated Cell Lines , 2000, Journal of Virology.

[21]  C. Amici,et al.  NF‐κB and virus infection: who controls whom , 2003, The EMBO journal.

[22]  S. Akira,et al.  Role of Adaptor TRIF in the MyD88-Independent Toll-Like Receptor Signaling Pathway , 2003, Science.

[23]  Steven R. LaPlante,et al.  An NS3 protease inhibitor with antiviral effects in humans infected with hepatitis C virus , 2003, Nature.

[24]  B. Beutler,et al.  LPS, dsRNA and the interferon bridge to adaptive immune responses: Trif, Tram, and other TIR adaptor proteins. , 2004, Journal of endotoxin research.

[25]  C. Seeger,et al.  Replication of Hepatitis C Virus Subgenomes in Nonhepatic Epithelial and Mouse Hepatoma Cells , 2003, Journal of Virology.

[26]  Shizuo Akira,et al.  Toll/IL-1 Receptor Domain-Containing Adaptor Inducing IFN-β (TRIF) Associates with TNF Receptor-Associated Factor 6 and TANK-Binding Kinase 1, and Activates Two Distinct Transcription Factors, NF-κB and IFN-Regulatory Factor-3, in the Toll-Like Receptor Signaling 1 , 2003, The Journal of Immunology.

[27]  A. Tramontano,et al.  Rational design and functional expression of a constitutively active single-chain NS4A-NS3 proteinase. , 1998, Folding & design.

[28]  W. McKeon.  Sources of health care data for social work leaders. , 1999, Continuum.

[29]  S. Lemon,et al.  Regulating Intracellular Antiviral Defense and Permissiveness to Hepatitis C Virus RNA Replication through a Cellular RNA Helicase, RIG-I , 2005, Journal of Virology.

[30]  L. Seeff.  Natural history of hepatitis C , 1997, Clinics in liver disease.

[31]  J. Hiscott,et al.  The interferon antiviral response: from viral invasion to evasion , 2002, Current opinion in infectious diseases.

[32]  Michael G. Katze,et al.  Viruses and interferon: a fight for supremacy , 2002, Nature Reviews Immunology.

[33]  T. Akazawa,et al.  TICAM-1, an adaptor molecule that participates in Toll-like receptor 3–mediated interferon-β induction , 2003, Nature Immunology.

[34]  Guo-Ping Zhou,et al.  Triggering the Interferon Antiviral Response Through an IKK-Related Pathway , 2003, Science.

[35]  S. Akira,et al.  Cutting Edge: A Novel Toll/IL-1 Receptor Domain-Containing Adapter That Preferentially Activates the IFN-β Promoter in the Toll-Like Receptor Signaling1 , 2002, The Journal of Immunology.

[36]  J. Silver,et al.  Replication of Subgenomic Hepatitis C Virus Rnas in a Hepatoma Cell Line , 1999 .

[37]  J. Hiscott,et al.  Triggering the interferon response: the role of IRF-3 transcription factor. , 1999, Journal of interferon & cytokine research : the official journal of the International Society for Interferon and Cytokine Research.

[38]  Tak W. Mak,et al.  Toll-like receptor 3-mediated activation of NF-κB and IRF3 diverges at Toll-IL-1 receptor domain-containing adapter inducing IFN-β , 2004 .

[39]  Daniel R. Caffrey,et al.  LPS-TLR4 Signaling to IRF-3/7 and NF-κB Involves the Toll Adapters TRAM and TRIF , 2003, The Journal of experimental medicine.

[40]  R. Flavell,et al.  Recognition of double-stranded RNA and activation of NF-κB by Toll-like receptor 3 , 2001, Nature.

[41]  L. Wodicka,et al.  Genomic analysis of the host response to hepatitis C virus infection , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[42]  B. Rehermann,et al.  Hepatitis C virus infection: when silence is deception. , 2003, Trends in immunology.