Strain-dependent viral dynamics and virus-cell interactions in a novel in vitro system supporting the life cycle of blood-borne hepatitis C virus

Authors

  • Hussein Hassan Aly,

    1. Laboratory of Human Tumor Viruses, Institute for Virus Research, Kyoto University, Kyoto, Japan
    2. Hepatology Department, National Hepatology and Tropical Medicine Research Institute, Cairo, Egypt
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  • Yue Qi,

    1. Laboratory of Viral Oncology, Graduate School of Biostudies, Kyoto University, Kyoto, Japan
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  • Kimie Atsuzawa,

    1. Department of Anatomy, Fujita Health University School of Medicine, Toyoake, Japan
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  • Nobuteru Usuda,

    1. Department of Anatomy, Fujita Health University School of Medicine, Toyoake, Japan
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  • Yasutsugu Takada,

    1. Department of Surgery, Division of Hepato-Pancreato-Biliary and Transplant Surgery, Graduate School of Medicine, Kyoto University, Kyoto, Japan
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  • Masashi Mizokami,

    1. Research Center for Hepatitis and Immunology, International Medical Center of Japan Kounodai Hospital, Ichikawa, Japan
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  • Kunitada Shimotohno,

    Corresponding author
    1. Center for Human Metabolomic Systems Biology, Keio University, Tokyo, Japan
    • Center for Human Metabolomic Systems Biology, Keio University, 35, Shinano-machi, Shinjuku-ku, Tokyo, 160-8582, Japan
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    • fax: 81-3-5363-3592

  • Makoto Hijikata

    Corresponding author
    1. Laboratory of Human Tumor Viruses, Institute for Virus Research, Kyoto University, Kyoto, Japan
    2. Laboratory of Viral Oncology, Graduate School of Biostudies, Kyoto University, Kyoto, Japan
    • Laboratory of Human Tumor Viruses, Institute for Virus Research, Kyoto University, 53, Kawaharacho, Shogoin, Sakyoku, Kyoto, 606-8507, Japan
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    • fax: 81-75-751-3998


  • Potential conflict of interest: Nothing to report.

Abstract

We developed an in vitro system that can be used for the study of the life cycle of a wide variety of blood-borne hepatitis C viruses (HCV) from various patients using a three-dimensional hollow fiber culture system and an immortalized primary human hepatocyte (HuS-E/2) cell line. Unlike the conventional two-dimensional culture, this system not only enhanced the infectivity of blood-borne HCV but also supported its long-term proliferation and the production of infectious virus particles. Both sucrose gradient fractionation and electron microscopy examination showed that the produced virus-like particles are within a similar fraction and size range to those previously reported. Infection with different HCV strains showed strain-dependent different patterns of HCV proliferation and particle production. Fluctuation of virus proliferation and particle production was found during prolonged culture and was found to be associated with change in the major replicating virus strain. Induction of cellular apoptosis was only found when strains of HCV-2a genotype were used for infection. Interferon-alpha stimulation also varied among different strains of HCV-1b genotypes tested in this study. Conclusion: These results suggest that this in vitro infection system can reproduce strain-dependent events reflecting viral dynamics and virus-cell interactions at the early phase of blood-borne HCV infection, and that this system can allow the development of new anti-HCV strategies specific to various HCV strains. (HEPATOLOGY 2009.)

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