title: Determinants Involved in Hepatitis C Virus and GB Virus B Primate Host Restriction.
abstract: Hepatitis C virus (HCV) only infects humans and chimpanzees, while GB virus B (GBV-B), another hepatotropic hepacivirus, infects small New World primates (tamarins and marmosets). In an effort to develop an immunocompetent small primate model for HCV infection to study HCV pathogenesis and vaccine approaches, we investigated the HCV life cycle step(s) that may be restricted in small primate hepatocytes. First, we found that replication-competent, genome-length chimeric HCV RNAs encoding GBV-B structural proteins in place of equivalent HCV sequences designed to allow entry into simian hepatocytes failed to induce viremia in tamarins following intrahepatic inoculation, nor did they lead to progeny virus in permissive, transfected human Huh7.5 hepatoma cells upon serial passage. This likely reflected the disruption of interactions between distantly related structural and nonstructural proteins that are essential for virion production, whereas such cross talk could be restored in similarly designed HCV intergenotypic recombinants via adaptive mutations in NS3 protease or helicase domains. Next, HCV entry into small primate hepatocytes was examined directly using HCV-pseudotyped retroviral particles (HCV-pp). HCV-pp efficiently infected tamarin hepatic cell lines and primary marmoset hepatocyte cultures through the use of the simian CD81 ortholog as a coreceptor, indicating that HCV entry is not restricted in small New World primate hepatocytes. Furthermore, we observed genomic replication and modest virus secretion following infection of primary marmoset hepatocyte cultures with a highly cell culture-adapted HCV strain. Thus, HCV can successfully complete its life cycle in primary simian hepatocytes, suggesting the possibility of adapting some HCV strains to small primate hosts.
  
  
  OBJECTIVE
  Hepatitis C virus (HCV) is an important human pathogen that infects over 150 million individuals worldwide and leads to chronic liver disease. The lack of a small animal model for this infection impedes the development of a preventive vaccine and pathogenesis studies. In seeking to establish a small primate model for HCV, we first attempted to generate recombinants between HCV and GB virus B (GBV-B), a hepacivirus that infects small New World primates (tamarins and marmosets). This approach revealed that the genetic distance between these hepaciviruses likely prevented virus morphogenesis. We next showed that HCV pseudoparticles were able to infect tamarin or marmoset hepatocytes efficiently, demonstrating that there was no restriction in HCV entry into these simian cells. Furthermore, we found that a highly cell culture-adapted HCV strain was able to achieve a complete viral cycle in primary marmoset hepatocyte cultures, providing a promising basis for further HCV adaptation to small primate hosts.
authors.affiliation: Institut Pasteur, Molecular Genetics of RNA Viruses Unit, Paris, France CNRS UMR 3569, Paris, France Université Paris Diderot-Sorbonne Paris Cité, Paris, France.
authors.forename: Caroline
authors.initials: C
authors.lastname: Marnata
authors.affiliation: Institut Pasteur, Molecular Genetics of RNA Viruses Unit, Paris, France CNRS UMR 3569, Paris, France Université Paris Diderot-Sorbonne Paris Cité, Paris, France.
authors.forename: Aure
authors.initials: A
authors.lastname: Saulnier
authors.affiliation: CIRI-International Center for Infectiology Research, Université de Lyon, Lyon, France, Inserm, U1111, Lyon, France, Ecole Normale Supérieure de Lyon, Lyon, France, Université Lyon 1, Centre International de Recherche en Infectiologie, Lyon, France, and CNRS, UMR5308, Lyon, France LabEx Ecofect, Université de Lyon, Lyon, France.
authors.forename: Dimitri
authors.initials: D
authors.lastname: Mompelat
authors.affiliation: Institut Pasteur, Structural Virology Unit, Paris, France CNRS UMR 3569, Paris, France.
authors.forename: Thomas
authors.initials: T
authors.lastname: Krey
authors.affiliation: Institut Pasteur, Molecular Genetics of RNA Viruses Unit, Paris, France CNRS UMR 3569, Paris, France Université Paris Diderot-Sorbonne Paris Cité, Paris, France.
authors.forename: Lisette
authors.initials: L
authors.lastname: Cohen
authors.affiliation: Institut Pasteur, Molecular Genetics of RNA Viruses Unit, Paris, France CNRS UMR 3569, Paris, France Université Paris Diderot-Sorbonne Paris Cité, Paris, France.
authors.forename: Célia
authors.initials: C
authors.lastname: Boukadida
authors.affiliation: Institut Pasteur, Molecular Genetics of RNA Viruses Unit, Paris, France CNRS UMR 3569, Paris, France Université Paris Diderot-Sorbonne Paris Cité, Paris, France.
authors.forename: Lucile
authors.initials: L
authors.lastname: Warter
authors.affiliation: CIRI-International Center for Infectiology Research, Université de Lyon, Lyon, France, Inserm, U1111, Lyon, France, Ecole Normale Supérieure de Lyon, Lyon, France, Université Lyon 1, Centre International de Recherche en Infectiologie, Lyon, France, and CNRS, UMR5308, Lyon, France LabEx Ecofect, Université de Lyon, Lyon, France.
authors.forename: Judith
authors.initials: J
authors.lastname: Fresquet
authors.affiliation: Institut Pasteur, Structural Virology Unit, Paris, France CNRS UMR 3569, Paris, France.
authors.forename: Ieva
authors.initials: I
authors.lastname: Vasiliauskaite
authors.affiliation: Institut Pasteur, Viral Genomics and Vaccination Unit, Paris, France CNRS UMR 3569, Paris, France.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Escriou
authors.affiliation: CIRI-International Center for Infectiology Research, Université de Lyon, Lyon, France, Inserm, U1111, Lyon, France, Ecole Normale Supérieure de Lyon, Lyon, France, Université Lyon 1, Centre International de Recherche en Infectiologie, Lyon, France, and CNRS, UMR5308, Lyon, France LabEx Ecofect, Université de Lyon, Lyon, France.
authors.forename: François-Loïc
authors.initials: FL
authors.lastname: Cosset
authors.affiliation: Institut Pasteur, Structural Virology Unit, Paris, France CNRS UMR 3569, Paris, France.
authors.forename: Felix A
authors.initials: FA
authors.lastname: Rey
authors.affiliation: Southwest National Primate Research Center at Texas Biomedical Research Institute, San Antonio, Texas, USA.
authors.forename: Robert E
authors.initials: RE
authors.lastname: Lanford
authors.affiliation: Imperial College, Department of Medicine, London, United Kingdom.
authors.forename: Peter
authors.initials: P
authors.lastname: Karayiannis
authors.affiliation: National Institute for Biological Standards and Control, Division of Virology, South Mimms, United Kingdom.
authors.forename: Nicola J
authors.initials: NJ
authors.lastname: Rose
authors.affiliation: CIRI-International Center for Infectiology Research, Université de Lyon, Lyon, France, Inserm, U1111, Lyon, France, Ecole Normale Supérieure de Lyon, Lyon, France, Université Lyon 1, Centre International de Recherche en Infectiologie, Lyon, France, and CNRS, UMR5308, Lyon, France.
authors.forename: Dimitri
authors.initials: D
authors.lastname: Lavillette
authors.affiliation: Institut Pasteur, Molecular Genetics of RNA Viruses Unit, Paris, France CNRS UMR 3569, Paris, France Université Paris Diderot-Sorbonne Paris Cité, Paris, France annette.martin@pasteur.fr.
authors.forename: Annette
authors.initials: A
authors.lastname: Martin
mesh_terms: D000818:Animals; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D005455:Fluorescent Antibody Technique; D029406:GB virus B / Q000502:physiology*; D057809:HEK293 Cells; D016174:Hepacivirus / Q000235:genetics / Q000502:physiology*; D022781:Hepatocytes / Q000821:virology; D058507:Host Specificity; D006801:Humans; D015151:Immunoblotting; D008018:Life Cycle Stages / Q000502:physiology*; D023421:Models, Animal; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D011323:Primates / Q000821:virology*; D017422:Sequence Analysis, DNA; D014766:Viremia; D053586:Virus Internalization
chemical_list: D017931:DNA Primers