Production of infectious hepatitis C virus in tissue culture from a cloned viral genome.
Journal: 2005/August - Nature Medicine
ISSN: 1078-8956
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.
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Nat Med 11(7): 791-796

Production of infectious hepatitis C virus in tissue culture from a cloned viral genome

+3 authors
Department of Microbiology, Tokyo Metropolitan Institute for Neuroscience, Tokyo 183-8526, Japan
Department of Molecular Virology, University Heidelberg, Im Neuenheimer Feld 345, 69120 Heidelberg, Germany
Department of Clinical Molecular Informative Medicine, Nagoya City University Graduate School of Medical Sciences, Nagoya 467-8601, Japan
Liver Disease Branch, NIDDK, National Institute of Health, Bethesda, Maryland 20892, USA
Southwest Foundation for Biomedical Research, San Antonio, Texas, 78227, USA
Department of Virology, University of Heidelberg, Im Neuenheimer Feld 324, 69120 Heidelberg, Germany
Correspondence should be addressed to T.W. (pj.ca.nimt@atikaw)
These authors contributed equally to this work.

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.

Abstract

HCV is a major cause of chronic liver diseases12. Development of selectable drugs and efficient vaccines has been hampered by poor virus growth in cell culture3. Although subgenomic replicons replicate efficiently in cultured cells4, for unknown reasons infectious viral particles are not produced56. Subgenomic replicons of the HCV genotype 2a JFH1 strain cloned from an individual with fulminant hepatitis7 replicate efficiently in Huh7 cells and do not require cell culture–adaptive mutations810. In this study, we show that transfection of in vitro–transcribed full-length JFH1 RNA into Huh7 cells results in secretion of viral particles that are infectious for cultured cells and a chimpanzee.

Footnotes

Accession numbers. The Genbank accession numbers for the consensus sequence of JFH1 is {"type":"entrez-nucleotide","attrs":{"text":"AB047639","term_id":"13122261","term_text":"AB047639"}}AB047639 and for pJFH1 it is {"type":"entrez-nucleotide","attrs":{"text":"AB047640","term_id":"13122263","term_text":"AB047640"}}AB047640.

Competing Interests Statement: The authors declare that they have no competing financial interests.

Footnotes

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