title: Production of infectious hepatitis C virus in tissue culture from a cloned viral genome.
abstract: Hepatitis C virus (HCV) infection causes chronic liver diseases and is a global public health problem. Detailed analyses of HCV have been hampered by the lack of viral culture systems. Subgenomic replicons of the JFH1 genotype 2a strain cloned from an individual with fulminant hepatitis replicate efficiently in cell culture. Here we show that the JFH1 genome replicates efficiently and supports secretion of viral particles after transfection into a human hepatoma cell line (Huh7). Particles have a density of about 1.15-1.17 g/ml and a spherical morphology with an average diameter of about 55 nm. Secreted virus is infectious for Huh7 cells and infectivity can be neutralized by CD81-specific antibodies and by immunoglobulins from chronically infected individuals. The cell culture-generated HCV is infectious for chimpanzee. This system provides a powerful tool for studying the viral life cycle and developing antiviral strategies.
authors.affiliation: Department of Microbiology, Tokyo Metropolitan Institute for Neuroscience, Tokyo 183-8526, Japan. wakita@tmin.ac.jp
authors.forename: Takaji
authors.initials: T
authors.lastname: Wakita
authors.forename: Thomas
authors.initials: T
authors.lastname: Pietschmann
authors.forename: Takanobu
authors.initials: T
authors.lastname: Kato
authors.forename: Tomoko
authors.initials: T
authors.lastname: Date
authors.forename: Michiko
authors.initials: M
authors.lastname: Miyamoto
authors.forename: Zijiang
authors.initials: Z
authors.lastname: Zhao
authors.forename: Krishna
authors.initials: K
authors.lastname: Murthy
authors.forename: Anja
authors.initials: A
authors.lastname: Habermann
authors.forename: Hans-Georg
authors.initials: HG
authors.lastname: Kräusslich
authors.forename: Masashi
authors.initials: M
authors.lastname: Mizokami
authors.forename: Ralf
authors.initials: R
authors.lastname: Bartenschlager
authors.forename: T Jake
authors.initials: TJ
authors.lastname: Liang
mesh_terms: D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D015703:Antigens, CD / Q000276:immunology; D055592:Biophysical Phenomena; D001703:Biophysics; D006528:Carcinoma, Hepatocellular / Q000821:virology; D045744:Cell Line, Tumor; D003001:Cloning, Molecular; D016679:Genome, Viral; D016174:Hepacivirus / Q000254:growth & development* / Q000302:isolation & purification / Q000472:pathogenicity*; D006526:Hepatitis C / Q000188:drug therapy / Q000276:immunology; D006801:Humans; D007106:Immune Sera; D008113:Liver Neoplasms / Q000821:virology; D008854:Microscopy, Electron; D002679:Pan troglodytes; D012367:RNA, Viral; D060225:Tetraspanin 28; D014162:Transfection; D014776:Virus Cultivation / Q000379:methods
chemical_list: D000906:Antibodies; D015703:Antigens, CD; C554798:CD81 protein, human; D007106:Immune Sera; D012367:RNA, Viral; D060225:Tetraspanin 28