Isolation and Characterization of an Huh.7.5.1-Derived Cell Clone Highly Permissive to Hepatitis C Virus

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Author(s)

    • Shirasago Yoshitaka
    • Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases|Research Institute for Biomedical Sciences, Tokyo University of Science
    • Saito Kyoko
    • Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases
    • Suzuki Tetsuro
    • Department of Infectious Diseases, Hamamatsu University School of Medicine
    • Wakita Takaji
    • Department of Virology II, National Institute of Infectious Diseases
    • Hanada Kentaro
    • Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases
    • Kuroda Makoto
    • Pathogen Genomics Center, National Institute of Infectious Diseases
    • Abe Ryo
    • Research Institute for Biomedical Sciences, Tokyo University of Science
    • Fukasawa Masayoshi
    • Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases

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<sup>9</sup> copies/μg of cellular RNA and viral titers of >10<sup>6</sup> 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.

Journal

  • Japanese Journal of Infectious Diseases

    Japanese Journal of Infectious Diseases 68(2), 81-88, 2015

    National Institute of Infectious Diseases, Japanese Journal of Infectious Diseases Editorial Committee

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