title: Construction and characterization of infectious intragenotypic and intergenotypic hepatitis C virus chimeras.
abstract: Chronic liver disease caused by infection with hepatitis C virus (HCV) is an important global health problem that currently affects 170 million people. A major impediment in HCV research and drug development has been the lack of culture systems supporting virus production. This obstacle was recently overcome by using JFH1-based full-length genomes that allow production of viruses infectious both in vitro and in vivo. Although this improvement was important, because of the restriction to the JFH1 isolate and a single chimera consisting of J6CF and JFH1-derived sequences, broadly based comparative studies between different HCV strains were not possible. Therefore, in this study we created a series of further chimeric genomes allowing production of infectious genotype (GT) 1a, 1b, 2a, and 3a particles. With the exception of the GT3a/JFH1 chimera, efficient virus production was obtained when the genome fragments were fused via a site located right after the first transmembrane domain of NS2. The most efficient construct is a GT2a/2a chimera consisting of J6CF- and JFH1-derived sequences connected via this junction. This hybrid, designated Jc1, yielded infectious titers 100- to 1,000-fold higher than the parental isolate and all other chimeras, suggesting that determinants within the structural proteins govern kinetic and efficiency of virus assembly and release. Finally, we describe an E1-specific antiserum capable of neutralizing infectivity of all HCV chimeras.
authors.affiliation: *Department of Molecular Virology, University of Heidelberg, Im Neuenheimer Feld 345, 69120 Heidelberg, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Pietschmann
authors.forename: Artur
authors.initials: A
authors.lastname: Kaul
authors.forename: George
authors.initials: G
authors.lastname: Koutsoudakis
authors.forename: Anna
authors.initials: A
authors.lastname: Shavinskaya
authors.forename: Stephanie
authors.initials: S
authors.lastname: Kallis
authors.forename: Eike
authors.initials: E
authors.lastname: Steinmann
authors.forename: Karim
authors.initials: K
authors.lastname: Abid
authors.forename: Francesco
authors.initials: F
authors.lastname: Negro
authors.forename: Marlene
authors.initials: M
authors.lastname: Dreux
authors.forename: Francois-Loic
authors.initials: FL
authors.lastname: Cosset
authors.forename: Ralf
authors.initials: R
authors.lastname: Bartenschlager
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology / Q000494:pharmacology; D045744:Cell Line, Tumor; D046528:Chimerism; D016679:Genome, Viral / Q000235:genetics; D005838:Genotype; D016174:Hepacivirus / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D012367:RNA, Viral / Q000235:genetics; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D012367:RNA, Viral; D014759:Viral Envelope Proteins