title: HVR1-mediated antibody evasion of highly infectious in vivo adapted HCV in humanised mice.
abstract: HCV is a major cause of chronic liver disease worldwide, but the role of neutralising antibodies (nAbs) in its natural history remains poorly defined. We analysed the in vivo role of hypervariable region 1 (HVR1) for HCV virion properties, including nAb susceptibility.
  
  
  
  Analysis of HCV from human liver chimeric mice infected with cell-culture-derived prototype genotype 2a recombinant J6/JFH1 or HVR1-deleted variant J6/JFH1ΔHVR1identified adaptive mutations, which were analysed by reverse genetics in Huh7.5 and CD81-deficient S29 cells. The increased in vivo genomic stability of the adapted viruses facilitated ex vivo density analysis by ultracentrifugation and in vivo neutralisation experiments addressing the role of HVR1.
  
  
  
  In vivo, J6/JFH1 and J6/JFH1ΔHVR1depended on single substitutions within amino acids 867-876 in non-structural protein, NS2. The identified A876P-substitution resulted in a 4.7-fold increase in genomic stability. In vitro, NS2 substitutions enhanced infectivity 5-10-fold by increasing virus assembly. Mouse-derived mJ6/JFH1A876Pand mJ6/JFH1ΔHVR1/A876Pviruses displayed similar heterogeneous densities of 1.02-1.1 g/mL. Human liver chimeric mice loaded with heterologous patient H (genotype 1a) immunoglobulin had partial protection against mJ6/JFH1A876Pand complete protection against mJ6/JFH1ΔHVR1/A876P. Interestingly, we identified a putative escape mutation, D476G, in mJ6/JFH1A876P. This mutation in hypervariable region 2 conferred 6.6-fold resistance against H06 IgG in vitro.
  
  
  
  The A876P-substitution bridges in vitro and in vivo studies using J6/JFH1-based recombinants. We provide the first in vivo evidence that HVR1 protects cross-genotype conserved HCV neutralisation epitopes, which advocates the possibility of using HVR1-deleted viruses as vaccine antigens to boost broadly reactive protective nAb responses.
authors.affiliation: Copenhagen Hepatitis C Program (CO-HEP), Department of Infectious Diseases and Clinical Research Centre, Hvidovre Hospital and Department of Immunology and Microbiology, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Jannick
authors.initials: J
authors.lastname: Prentoe
authors.affiliation: Center for Vaccinology, Ghent University, Ghent, Belgium.
authors.forename: Lieven
authors.initials: L
authors.lastname: Verhoye
authors.affiliation: Copenhagen Hepatitis C Program (CO-HEP), Department of Infectious Diseases and Clinical Research Centre, Hvidovre Hospital and Department of Immunology and Microbiology, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Rodrigo
authors.initials: R
authors.lastname: Velázquez Moctezuma
authors.affiliation: Center for Vaccinology, Ghent University, Ghent, Belgium.
authors.forename: Caroline
authors.initials: C
authors.lastname: Buysschaert
authors.affiliation: Center for Vaccinology, Ghent University, Ghent, Belgium.
authors.forename: Ali
authors.initials: A
authors.lastname: Farhoudi
authors.affiliation: Department of Transfusion Medicine, Warren Grant Magnuson Clinical Center, National Institutes of Health, Bethesda, Maryland, USA.
authors.forename: Richard
authors.initials: R
authors.lastname: Wang
authors.affiliation: Department of Transfusion Medicine, Warren Grant Magnuson Clinical Center, National Institutes of Health, Bethesda, Maryland, USA.
authors.forename: Harvey
authors.initials: H
authors.lastname: Alter
authors.affiliation: Center for Vaccinology, Ghent University, Ghent, Belgium.
authors.forename: Philip
authors.initials: P
authors.lastname: Meuleman
authors.affiliation: Copenhagen Hepatitis C Program (CO-HEP), Department of Infectious Diseases and Clinical Research Centre, Hvidovre Hospital and Department of Immunology and Microbiology, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Jens
authors.initials: J
authors.lastname: Bukh
mesh_terms: D019943:Amino Acid Substitution / Q000235:genetics; D000818:Animals; D057134:Antibodies, Neutralizing / Q000097:blood; D004195:Disease Models, Animal; D005838:Genotype; D016174:Hepacivirus / Q000235:genetics* / Q000472:pathogenicity; D006526:Hepatitis C / Q000235:genetics / Q000276:immunology / Q000517:prevention & control / Q000821:virology; D066298:In Vitro Techniques; D051379:Mice; D009154:Mutation; D014761:Viral Hepatitis Vaccines / Q000235:genetics; D017361:Viral Nonstructural Proteins / Q000235:genetics; D014764:Viral Proteins / Q000235:genetics*
chemical_list: D057134:Antibodies, Neutralizing; C400562:HVR1 protein, Hepatitis C virus; D014761:Viral Hepatitis Vaccines; D017361:Viral Nonstructural Proteins; D014764:Viral Proteins
keywords: HCV; HEPATITIS C