Production of infectious hepatitis C virus in tissue culture from a cloned viral genome.
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.
Wakita
Takaji
T
Department of Microbiology, Tokyo Metropolitan Institute for Neuroscience, Tokyo 183-8526, Japan. wakita@tmin.ac.jp
Pietschmann
Thomas
T
Kato
Takanobu
T
Date
Tomoko
T
Miyamoto
Michiko
M
Zhao
Zijiang
Z
Murthy
Krishna
K
Habermann
Anja
A
Kräusslich
Hans-Georg
HG
Mizokami
Masashi
M
Bartenschlager
Ralf
R
Liang
T Jake
TJ
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
D000906:Antibodies; D015703:Antigens, CD; C554798:CD81 protein, human; D007106:Immune Sera; D012367:RNA, Viral; D060225:Tetraspanin 28