In vivo proliferation of hepadnavirus-infected hepatocytes induces loss of covalently closed circular DNA in mice

Authors

  • Marc Lutgehetmann,

    1. Department of Medicine, University Medical Center Hamburg-Eppendorf, Hamburg, Germany
    2. Department of Medical Microbiology, Virology and Hygiene, University Medical Center Hamburg-Eppendorf, Hamburg, Germany
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    • 1

      These authors contributed equally to this work.

  • Tassilo Volz,

    1. Department of Medicine, University Medical Center Hamburg-Eppendorf, Hamburg, Germany
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    • 1

      These authors contributed equally to this work.

  • Anne Köpke,

    1. Department of Medicine, University Medical Center Hamburg-Eppendorf, Hamburg, Germany
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  • Tim Broja,

    1. Department of Medicine, University Medical Center Hamburg-Eppendorf, Hamburg, Germany
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  • Eike Tigges,

    1. Department of Medicine, University Medical Center Hamburg-Eppendorf, Hamburg, Germany
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  • Ansgar W. Lohse,

    1. Department of Medicine, University Medical Center Hamburg-Eppendorf, Hamburg, Germany
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  • Eberhard Fuchs,

    1. Clinical Neurobiology Laboratory, German Primate Center, Göttingen, Germany
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  • John M. Murray,

    1. School of Mathematics and Statistics and Clinical Research, University of New South Wales, Sydney, Australia
    2. National Centre in HIV Epidemiology and Clinical Research, University of New South Wales, Sydney, Australia
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  • Joerg Petersen,

    1. Department of Medicine, University Medical Center Hamburg-Eppendorf, Hamburg, Germany
    2. Liver Centre Hamburg, Institute for Interdisciplinary Medicine, Asklepios Clinics, St. Georg, Hamburg, Germany
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    • These authors contributed equally to this work.

  • Maura Dandri

    Corresponding author
    1. Department of Medicine, University Medical Center Hamburg-Eppendorf, Hamburg, Germany
    • Medizinische Klinik und Poliklinik, Zentrum für Innere Medizin, Universitätsklinikum Hamburg-Eppendorf, Martinistr. 52, D-20246 Hamburg, Germany
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    • These authors contributed equally to this work.

    • fax: (49)-40-7410-57232


  • Potential conflict of interest: Nothing to report.

Abstract

Chronic hepatitis B virus (HBV) infection is maintained by the presence of covalently closed circular DNA (cccDNA), the template of viral transcription and replication. In quiescent hepatocytes, cccDNA is a stable molecule that can persist throughout the hepatocyte lifespan. However, in chronic HBV infection, immunomediated cell injury and compensatory hepatocyte proliferation may favor cccDNA decline and selection of cccDNA-free cells. To investigate the impact of liver regeneration on cccDNA stability and activity in vivo, we used the urokinase-type plasminogen activator (uPA)/severe combined immunodeficiency (SCID) mouse model. Primary tupaia hepatocytes (PTHs) chronically infected with woolly monkey HBV (WM-HBV) were isolated from one highly viremic uPA/SCID chimeric mouse and transplanted into 20 uPA recipients. Expansion of transplanted PTHs and viral load changes were determined by real-time polymerase chain reaction and immunohistochemistry. Transplantation of WM-HBV infected hepatocytes led to an average of 3.8 PTH doublings within 80 days, 75% reduction of virion productivity (relaxed circular DNA/cccDNA), and lower expression levels of pregenomic RNA and hepatitis B core antigen. Remarkably, a median 2-log decline of cccDNA per cell determined during PTH proliferation was due to both dilution of the cccDNA pool among daughter cells and a 0.5-log loss of intrahepatic cccDNA loads (P = 0.02). Intrahepatic viral DNA sequences persisting at the end of the study were mostly present as replicative intermediates and not as integrated virus. Conclusion: Cell division in the setting of liver regeneration and without administration of antiviral drugs induced strong destabilization of the cccDNA reservoir, resulting in cccDNA clearance in the great majority of chronically infected hepatocytes. (Hepatology 2010)

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