title: Evidence for in vitro replication of hepatitis C virus genome in a human T-cell line.
abstract: A human T-cell line, MOLT-4, either uninfected or infected with murine retroviruses, was tested for its susceptibility to hepatitis C virus (HCV) infection. The cell cultures were inoculated with a serum containing HCV and then examined for the presence of viral sequences by cDNA/PCR. In murine retrovirus-infected MOLT-4 (MOLT-4 Ma) cells, intracellular minus-strand viral RNA, a putative replication intermediate, was first detected 3 days after inoculation, and the maximum signal was seen on day 7. When the cells were continuously subcultured in fresh medium, HCV sequences were intermittently detected in cells over a period of 3 weeks. In MOLT-4 cells free of retroviruses, replication of minus-strand HCV RNA appeared less efficient than in MOLT-4 Ma cells. The presence of minus-strand viral RNA in MOLT-4 Ma cells inoculated with HCV was confirmed by in situ hybridization with a strand-specific RNA probe. Immunofluorescence tests with antibodies specific for HCV core and NS4 antigens showed that MOLT-4 Ma cells were positive for viral antigen 7 days after inoculation. Thus, it appears likely that the HCV genome can replicate in the human T-cell line MOLT-4.
authors.affiliation: Department of Enteroviruses, National Institute of Health, Tokyo, Japan.
authors.forename: Y K
authors.initials: YK
authors.lastname: Shimizu
authors.forename: A
authors.initials: A
authors.lastname: Iwamoto
authors.forename: M
authors.initials: M
authors.lastname: Hijikata
authors.forename: R H
authors.initials: RH
authors.lastname: Purcell
authors.forename: H
authors.initials: H
authors.lastname: Yoshikura
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D001483:Base Sequence; D002460:Cell Line; D016679:Genome, Viral; D016174:Hepacivirus / Q000235:genetics*; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D002679:Pan troglodytes; D016133:Polymerase Chain Reaction / Q000379:methods; D012367:RNA, Viral / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics*; D013601:T-Lymphocytes; D014779:Virus Replication
chemical_list: D000911:Antibodies, Monoclonal; D009838:Oligodeoxyribonucleotides; D012367:RNA, Viral