Scavenger receptor class B type I is a key host factor for hepatitis C virus infection required for an entry step closely linked to CD81

Hepatitis C virus (HCV) is a major cause of chronic hepatitis worldwide. Scavenger receptor class B type I (SR‐BI) has been shown to bind HCV envelope glycoprotein E2, participate in entry of HCV pseudotype particles, and modulate HCV infection. However, the functional role of SR‐BI for productive HCV infection remains unclear. In this study, we investigated the role of SR‐BI as an entry factor for infection of human hepatoma cells using cell culture–derived HCV (HCVcc). Anti–SR‐BI antibodies directed against epitopes of the human SR‐BI extracellular loop specifically inhibited HCVcc infection in a dose‐dependent manner. Down‐regulation of SR‐BI expression by SR‐BI–specific short interfering RNAs (siRNAs) markedly reduced the susceptibility of human hepatoma cells to HCVcc infection. Kinetic studies demonstrated that SR‐BI acts predominately after binding of HCV at an entry step occurring at a similar time point as CD81–HCV interaction. Although the addition of high‐density lipoprotein (HDL) enhanced the efficiency of HCVcc infection, anti–SR‐BI antibodies and SR‐BI–specific siRNA efficiently inhibited HCV infection independent of lipoprotein. Conclusion: Our data suggest that SR‐BI (i) represents a key host factor for HCV entry, (ii) is implicated in the same HCV entry pathway as CD81, and (iii) targets an entry step closely linked to HCV–CD81 interaction. (HEPATOLOGY 2007.)

[1]  Seth M. Klein,et al.  Comparison of Class B Scavenger Receptors, CD36 and Scavenger Receptor BI (SR-BI), Shows That Both Receptors Mediate High Density Lipoprotein-Cholesteryl Ester Selective Uptake but SR-BI Exhibits a Unique Enhancement of Cholesteryl Ester Uptake* , 1999, The Journal of Biological Chemistry.

[2]  Norbert Perrimon,et al.  Drosophila RNAi Screen Reveals CD36 Family Member Required for Mycobacterial Infection , 2005, Science.

[3]  Robert J. Linhardt,et al.  Cellular Binding of Hepatitis C Virus Envelope Glycoprotein E2 Requires Cell Surface Heparan Sulfate* , 2003, Journal of Biological Chemistry.

[4]  M. Krieger,et al.  The Efficient Cellular Uptake of High Density Lipoprotein Lipids via Scavenger Receptor Class B Type I Requires Not Only Receptor-mediated Surface Binding but Also Receptor-specific Lipid Transfer Mediated by Its Extracellular Domain* , 1998, The Journal of Biological Chemistry.

[5]  Toshiaki Maruyama,et al.  Complete Replication of Hepatitis C Virus in Cell Culture , 2005, Science.

[6]  N. Webb,et al.  High Density Lipoprotein Uptake by Scavenger Receptor SR-BII* , 2004, Journal of Biological Chemistry.

[7]  M. Brown,et al.  A receptor-mediated pathway for cholesterol homeostasis. , 1986, Science.

[8]  C. Cheng‐Mayer,et al.  Hepatitis C virus glycoproteins mediate pH-dependent cell entry of pseudotyped retroviral particles , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[9]  D. Steinberg,et al.  Dissociation of tissue uptake of cholesterol ester from that of apoprotein A-I of rat plasma high density lipoprotein: selective delivery of cholesterol ester to liver, adrenal, and gonad. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[10]  A. Shavinskaya,et al.  Construction and characterization of infectious intragenotypic and intergenotypic hepatitis C virus chimeras. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[11]  J. Auwerx,et al.  Liver receptor homolog 1 controls the expression of the scavenger receptor class B type I , 2002, EMBO reports.

[12]  B. Bartosch,et al.  Infectious Hepatitis C Virus Pseudo-particles Containing Functional E1–E2 Envelope Protein Complexes , 2003, The Journal of experimental medicine.

[13]  R. Bartenschlager,et al.  Production of infectious hepatitis C virus in tissue culture from a cloned viral genome , 2005, Nature Medicine.

[14]  Charles M. Rice,et al.  Highly Permissive Cell Lines for Subgenomic and Genomic Hepatitis C Virus RNA Replication , 2002, Journal of Virology.

[15]  D. Malide,et al.  CLA-1 and its splicing variant CLA-2 mediate bacterial adhesion and cytosolic bacterial invasion in mammalian cells , 2006, Proceedings of the National Academy of Sciences.

[16]  D. Lavillette,et al.  Characterization of host‐range and cell entry properties of the major genotypes and subtypes of hepatitis C virus , 2005, Hepatology.

[17]  P. Borrow,et al.  Analysis of the binding of hepatitis C virus genotype 1a and 1b E2 glycoproteins to peripheral blood mononuclear cell subsets. , 2005, The Journal of general virology.

[18]  R. Bartenschlager,et al.  Characterization of the Early Steps of Hepatitis C Virus Infection by Using Luciferase Reporter Viruses , 2006, Journal of Virology.

[19]  F. Chisari.  Unscrambling hepatitis C virus–host interactions , 2005, Nature.

[20]  J. Dubuisson,et al.  Binding of Hepatitis C Virus-Like Particles Derived from Infectious Clone H77C to Defined Human Cell Lines , 2002, Journal of Virology.

[21]  R. Kawaguchi,et al.  Real-time detection system for quantification of hepatitis C virus genome. , 1999, Gastroenterology.

[22]  A. Szántó,et al.  High Density Lipoprotein Endocytosis by Scavenger Receptor SR-BII Is Clathrin-dependent and Requires a Carboxyl-terminal Dileucine Motif* , 2006, Journal of Biological Chemistry.

[23]  M Nauck,et al.  Quantitative determination of high-, low-, and very-low-density lipoproteins and lipoprotein(a) by agarose gel electrophoresis and enzymatic cholesterol staining. , 1995, Clinical chemistry.

[24]  J. Pawlotsky,et al.  Human Serum Facilitates Hepatitis C Virus Infection, and Neutralizing Responses Inversely Correlate with Viral Replication Kinetics at the Acute Phase of Hepatitis C Virus Infection , 2005, Journal of Virology.

[25]  James S. Owen,et al.  Scavenger Receptor BI and BII Expression Levels Modulate Hepatitis C Virus Infectivity , 2007, Journal of Virology.

[26]  Jean Dubuisson,et al.  High Density Lipoproteins Facilitate Hepatitis C Virus Entry through the Scavenger Receptor Class B Type I* , 2005, Journal of Biological Chemistry.

[27]  R. Cortese,et al.  The human scavenger receptor class B type I is a novel candidate receptor for the hepatitis C virus , 2002, The EMBO journal.

[28]  M. Krieger,et al.  Scavenger receptor class B type I is a multiligand HDL receptor that influences diverse physiologic systems. , 2001, The Journal of clinical investigation.

[29]  M. Mizokami,et al.  Efficient replication of the genotype 2a hepatitis C virus subgenomic replicon. , 2003, Gastroenterology.

[30]  Thomas Baumert,et al.  Cooperativity between Cd81 and Scavenger Dependent on Cholesterol and Initiation of Hepatitis C Virus Infection Is , 2006 .

[31]  P. Connell,et al.  Alternative forms of the scavenger receptor BI (SR-BI). , 1997, Journal of lipid research.

[32]  T. Dragic,et al.  Different Domains of CD81 Mediate Distinct Stages of Hepatitis C Virus Pseudoparticle Entry , 2006, Journal of Virology.

[33]  V. Agnello,et al.  Hepatitis C virus and other flaviviridae viruses enter cells via low density lipoprotein receptor. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[34]  Ralf Bartenschlager,et al.  High Density Lipoprotein Inhibits Hepatitis C Virus-neutralizing Antibodies by Stimulating Cell Entry via Activation of the Scavenger Receptor BI* , 2006, Journal of Biological Chemistry.

[35]  M. Kieny,et al.  Entry of hepatitis C virus pseudotypes into primary human hepatocytes by clathrin-dependent endocytosis. , 2006, The Journal of general virology.

[36]  Charles M. Rice,et al.  Claudin-1 is a hepatitis C virus co-receptor required for a late step in entry , 2007, Nature.

[37]  F. Penin,et al.  An Interplay between Hypervariable Region 1 of the Hepatitis C Virus E2 Glycoprotein, the Scavenger Receptor BI, and High-Density Lipoprotein Promotes both Enhancement of Infection and Protection against Neutralizing Antibodies , 2005, Journal of Virology.

[38]  C. Rice,et al.  Oxidized low‐density lipoprotein inhibits hepatitis C virus cell entry in human hepatoma cells , 2006, Hepatology.

[39]  D. Burton,et al.  Robust hepatitis C virus infection in vitro. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[40]  Mark Marsh,et al.  Virus Entry: Open Sesame , 2006, Cell.

[41]  P. Schürmann,et al.  Scavenger Receptor Class B Type I and Hepatitis C Virus Infection of Primary Tupaia Hepatocytes , 2005, Journal of Virology.

[42]  R. Cortese,et al.  Cell Entry of Hepatitis C Virus Requires a Set of Co-receptors That Include the CD81 Tetraspanin and the SR-B1 Scavenger Receptor* , 2003, Journal of Biological Chemistry.

[43]  M. Houghton,et al.  Binding of hepatitis C virus to CD81. , 1998, Science.

[44]  Y. D. Chen,et al.  Morphological evidence that high density lipoproteins are not internalized by steroid-producing cells during in situ organ perfusion. , 1984, The Journal of clinical investigation.

[45]  Helen H. Hobbs,et al.  Identification of Scavenger Receptor SR-BI as a High Density Lipoprotein Receptor , 1996, Science.

[46]  D. Steinberg,et al.  Uptake of high-density lipoprotein-associated apoprotein A-I and cholesterol esters by 16 tissues of the rat in vivo and by adrenal cells and hepatocytes in vitro. , 1985, The Journal of biological chemistry.