title: Isolation and characterization of an Huh.7.5.1-derived cell clone highly permissive to hepatitis C virus.
abstract: An efficient cell culture and infection system for hepatitis C virus (HCV) facilitates analyses of the complete virus life cycle. Human hepatic Huh7.5.1 cells and an HCV-JFH1 strain have been widely employed in infection experiments. In the present study, cultured Huh7.5.1 cells exhibited heterogeneous phenotypes of HCV infection. Using single-cell cloning of Huh7.5.1 cells, we isolated a clone highly permissive to HCV (Huh7.5.1-8) and a CD81-defective clone nonpermissive to HCV (Huh7.5.1-5). Expression of CD81 in Huh7.5.1-5 cells restored permissiveness to HCV, indicating that CD81 is essential for HCV infection and a defect in CD81 causes nonpermissiveness to HCV in Huh7.5.1-5 cells. Huh7.5.1-8 cells had approximately 10-fold higher HCV replication rates, with cellular HCV RNA copy numbers of >10(9) copies/μg of cellular RNA and viral titers of >10(6) infectious units/ml of culture supernatant. Permissiveness of Huh7.5.1-8 cells to HCV infection was phenotypically very stable because there was no difference in permissiveness after more than 100 passages (1-year culture). This efficient cell culture system for HCV using Huh7.5.1-8 cell provides a powerful tool for studying the HCV life cycle and constructing antiviral strategies.
authors.affiliation: Department of Biochemistry and Cell Biology; Research Institute for Biomedical Sciences, Tokyo University of Science, 278-0022..
authors.forename: Yoshitaka
authors.initials: Y
authors.lastname: Shirasago
authors.forename: Tsuyoshi
authors.initials: T
authors.lastname: Sekizuka
authors.forename: Kyoko
authors.initials: K
authors.lastname: Saito
authors.forename: Tetsuro
authors.initials: T
authors.lastname: Suzuki
authors.forename: Takaji
authors.initials: T
authors.lastname: Wakita
authors.forename: Kentaro
authors.initials: K
authors.lastname: Hanada
authors.forename: Makoto
authors.initials: M
authors.lastname: Kuroda
authors.forename: Ryo
authors.initials: R
authors.lastname: Abe
authors.forename: Masayoshi
authors.initials: M
authors.lastname: Fukasawa
mesh_terms: D002460:Cell Line; D015870:Gene Expression; D016174:Hepacivirus / Q000502:physiology*; D022781:Hepatocytes / Q000821:virology*; D006801:Humans; D060225:Tetraspanin 28 / Q000235:genetics / Q000378:metabolism; D019562:Viral Load; D014776:Virus Cultivation / Q000379:methods*; D014779:Virus Replication
chemical_list: C554798:CD81 protein, human; D060225:Tetraspanin 28