Production of infectious hepatitis C virus in tissue culture from a cloned viral genome (original) (raw)

Nature Medicine volume 11, pages 791–796 (2005)Cite this article

A Corrigendum to this article was published on 01 August 2005

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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Acknowledgements

This study was supported by a Grant-in-Aid for Scientific Research from the Japan Society for the Promotion of Science, by a Grant from Toray Industries, Inc., by the Program for Promotion of Fundamental Studies in Health Sciences of the Pharmaceuticals and Medical Devices Agency (PMDA), by the Research on Health Sciences focusing on Drug Innovation from the Japan Health Sciences Foundation, by the National Heart, Lung and Blood Institute contract N01-HB-27091 for the use of chimpanzees, by a grant from the European Community (QLK2-CT-2002-01329), and by a grant from the Deutsche Forschungsgemeinschaft (SFB 638; Teilprojekt A5). T.K. was partially supported by Hepatitis Virus Research Foundation of Japan. The authors are grateful to J. Bukh for providing the pJ6CF plasmid, to S. Foung for provision of the antibody CBH5, to S. Koike for his comments, to K. Yasui, S. Sone and J.-I. Tanabe for their support, to G. Koutsoudakis and E. Steinmann, and to S. Kallis and U. Herian for technical assistance. We are also grateful to K. Takagi, K. Hiramatsu and members of Central Clinical Laboratory in Nagoya City University Hospital, R. Sapp of the Liver Diseases Branch, H. McClure of the Southwest Foundation for Biomedical Research for their technical assistance, H. Barth, B. Bartosch, T. Baumert, J. Encke and C. Sarrazin for providing human sera.

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Author notes

  1. Takaji Wakita, Thomas Pietschmann and Ralf Bartenschlager: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Microbiology, Tokyo Metropolitan Institute for Neuroscience, Tokyo, 183-8526, Japan
    Takaji Wakita, Takanobu Kato, Tomoko Date, Michiko Miyamoto & Zijiang Zhao
  2. Department of Molecular Virology, University Heidelberg, Im Neuenheimer Feld 345, Heidelberg, 69120, Germany
    Thomas Pietschmann & Ralf Bartenschlager
  3. Department of Clinical Molecular Informative Medicine, Nagoya City University Graduate School of Medical Sciences, Nagoya, 467-8601, Japan
    Takanobu Kato & Masashi Mizokami
  4. Liver Disease Branch, NIDDK, National Institute of Health, Bethesda, 20892, Maryland, USA
    Takanobu Kato & T Jake Liang
  5. Southwest Foundation for Biomedical Research, San Antonio, 78227, Texas, USA
    Krishna Murthy
  6. Department of Virology, University of Heidelberg, Im Neuenheimer Feld 324, Heidelberg, 69120, Germany
    Anja Habermann & Hans-Georg Kräusslich

Authors

  1. Takaji Wakita
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  2. Thomas Pietschmann
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  3. Takanobu Kato
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  4. Tomoko Date
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  5. Michiko Miyamoto
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  6. Zijiang Zhao
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  7. Krishna Murthy
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  8. Anja Habermann
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  9. Hans-Georg Kräusslich
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  10. Masashi Mizokami
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  11. Ralf Bartenschlager
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  12. T Jake Liang
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Correspondence toTakaji Wakita.

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Wakita, T., Pietschmann, T., Kato, T. et al. Production of infectious hepatitis C virus in tissue culture from a cloned viral genome.Nat Med 11, 791–796 (2005). https://doi.org/10.1038/nm1268

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