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

• Shirasago Yoshitaka

  Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases Research Institute for Biomedical Sciences, Tokyo University of Science

• Sekizuka Tsuyoshi

  Pathogen Genomics Center, National Institute of Infectious Diseases

• 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 >10⁹ copies/μg of cellular RNA and viral titers of >10⁶ 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

Keywords

• hepatitis C virus
• hepatocytes
• replication

Details

• CRID: 1390282681218530304
• NII Article ID: 40020410899
• NII Article ID: 130004716751
• NII Book ID: AA1132885X
• DOI: 10.7883/yoken.jjid.2014.231
• ISSN: 18842836
• ISSN: 13446304
• NDL BIB ID: 026280891
• PubMed: 25420655
• Web Site: https://ndlsearch.ndl.go.jp/books/R000000004-I026280891
• Web Site: https://search.jamas.or.jp/link/ui/2016013571
• Text Lang: en
• Data Source:
  • JaLC
  • NDL
  • Crossref
  • PubMed
  • CiNii Articles
  • KAKEN
• Abstract License Flag: Disallowed