Antiviral strategies in chronic hepatitis B virus infection: I. Establishment of an in vitro system using the duck hepatitis B virus model

Authors

  • Naomi Bishop,

    1. Hepatitis Research Unit, Macfarlane Burnet Centre for Medical Research, Fairfield Hospital, Fairfield, Victoria, Australia
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  • Gilda Civitico,

    1. Hepatitis Research Unit, Macfarlane Burnet Centre for Medical Research, Fairfield Hospital, Fairfield, Victoria, Australia
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  • Yanyan Wang,

    1. Hepatitis Research Unit, Macfarlane Burnet Centre for Medical Research, Fairfield Hospital, Fairfield, Victoria, Australia
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  • Kejian Guo,

    1. Hepatitis Research Unit, Macfarlane Burnet Centre for Medical Research, Fairfield Hospital, Fairfield, Victoria, Australia
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  • Chris Birch,

    1. Hepatitis Research Unit, Macfarlane Burnet Centre for Medical Research, Fairfield Hospital, Fairfield, Victoria, Australia
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  • Ian Gust,

    1. Hepatitis Research Unit, Macfarlane Burnet Centre for Medical Research, Fairfield Hospital, Fairfield, Victoria, Australia
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  • Dr. Stephen Locarnini

    Corresponding author
    1. Hepatitis Research Unit, Macfarlane Burnet Centre for Medical Research, Fairfield Hospital, Fairfield, Victoria, Australia
    • Hepatitis Research Unit, Macfarlane Burnet Centre for Medical Research, Fairfield Hospital, Fairfield, Victoria 3078, Australia
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Abstract

Primary duck hepatocyte (PDH) cultures were established from ducklings congenitally infected with the duck hepatitis B virus (DHBV), plated onto feeder cell layers of irradiated human embryonic lung fibroblasts, and observed for 2 to 3 weeks. This system permitted the survival of the PDH in a differentiated form free of fibroblastic overgrowth for at least 3 weeks. The hepatocytes were shown to contain all the replicative DNA intermediates found during DHBV replication as well as the DHBV structural proteins PRE-S1, PRE-S2, and S of duck hepatitis B surface antigen (DHBsAg). The pool of supercoiled (SC) DHBV DNA increased dramatically from days 10 to 14 postplating. This PDH-feeder cell layer cell culture model provides a convenient system to study the effects of conventional inhibitors of DHBV replication and compounds targeted at the supercoiled form of DHBV DNA. This approach should allow the evaluation of a variety of strategies for treating chronic carriers of hepadnaviruses.

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