title: An infectious molecular clone of a Japanese genotype 1b hepatitis C virus.
abstract: We describe an infectious molecular clone of a Japanese genotype 1b strain of hepatitis C virus (HCV-N). The molecularly cloned sequence of HCV-N was compared with alignments of other HCV sequences, leading to the identification of 15 unique, nonconservative amino acid substitutions within the HCV-N open reading frame (ORF). These were repaired to the consensus genotype 1b residue, and the infectivity of RNA transcribed from the repaired clone was assessed by intrahepatic inoculation of a chimpanzee. Viral RNA was first detected in the serum of this chimpanzee 3 weeks following inoculation, and was intermittently present over the next 14 weeks. A strong and persistent anti-HCV serological response developed 13 weeks following inoculation, with seroconversion in the recombinant immunoblot assay (RIBA). A weaker, transient serological response, characterized by seroconversion in a third-generation enzyme-linked immunosorbent assay (ELISA) but not RIBA, occurred between weeks 1 and 5. This may have represented an anamnestic response to HCV antigens translated directly from the intrahepatically inoculated RNA, because the animal previously had undergone 2 unsuccessful attempts at rescue of HCV by intrahepatic RNA inoculation. There was neither biochemical nor histological evidence of liver disease. Although this is within the range of expected outcomes in an HCV-naive chimpanzee, prior immunologic priming may have modified the infection in this animal. The HCV-N clone is the first infectious molecular clone of HCV that is comprised entirely of genotype 1b sequence, and it contains an ORF sequence that is significantly divergent from that of a previously described genotype 1a/1b chimera.
authors.affiliation: Department of Microbiology and Immunology, The University of Texas Medical Branch at Galveston, Galveston, TX, USA.
authors.forename: M R
authors.initials: MR
authors.lastname: Beard
authors.forename: G
authors.initials: G
authors.lastname: Abell
authors.forename: M
authors.initials: M
authors.lastname: Honda
authors.forename: A
authors.initials: A
authors.lastname: Carroll
authors.forename: M
authors.initials: M
authors.lastname: Gartland
authors.forename: B
authors.initials: B
authors.lastname: Clarke
authors.forename: K
authors.initials: K
authors.lastname: Suzuki
authors.forename: R
authors.initials: R
authors.lastname: Lanford
authors.forename: D V
authors.initials: DV
authors.lastname: Sangar
authors.forename: S M
authors.initials: SM
authors.lastname: Lemon
mesh_terms: D020413:3' Untranslated Regions / Q000235:genetics; D020121:5' Untranslated Regions / Q000737:chemistry / Q000235:genetics; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D016384:Consensus Sequence; D004279:DNA, Viral / Q000737:chemistry / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D019143:Evolution, Molecular; D005838:Genotype; D016174:Hepacivirus / Q000145:classification* / Q000235:genetics*; D006526:Hepatitis C / Q000097:blood / Q000503:physiopathology; D018936:Hepatitis C Antigens / Q000737:chemistry / Q000235:genetics; D006801:Humans; D015151:Immunoblotting; D007564:Japan; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D002679:Pan troglodytes; D010802:Phylogeny; D012367:RNA, Viral / Q000097:blood / Q000235:genetics; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid
chemical_list: D020413:3' Untranslated Regions; D020121:5' Untranslated Regions; D004279:DNA, Viral; D018936:Hepatitis C Antigens; D012367:RNA, Viral