An in situ hybridization, molecular biological and immunohistochemical study of hepatitis delta virus in woodchucks

Authors

  • Spyros Dourakis,

    1. Liver Unit, Academic Department of Medicine, St. Mary's Hospital Medical School, Imperial College of Science, Technology and Medicine, London W2, United Kingdom
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  • Peter Karayiannis M.D.,

    Corresponding author
    1. Liver Unit, Academic Department of Medicine, St. Mary's Hospital Medical School, Imperial College of Science, Technology and Medicine, London W2, United Kingdom
    • Liver Unit, Academic Department of Medicine, St. Mary's Hospital Medical School, South Wharf Road, London W2 1NY, United Kingdom
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  • Robert Goldin,

    1. Liver Unit, Academic Department of Histopathology, St. Mary's Hospital Medical School, Imperial College of Science, Technology and Medicine, London W2, United Kingdom
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  • Michael Taylor,

    1. Liver Unit, Academic Department of Histopathology, St. Mary's Hospital Medical School, Imperial College of Science, Technology and Medicine, London W2, United Kingdom
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  • John Monjardino,

    1. Liver Unit, Academic Department of Medicine, St. Mary's Hospital Medical School, Imperial College of Science, Technology and Medicine, London W2, United Kingdom
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  • Howard C. Thomas

    1. Liver Unit, Academic Department of Medicine, St. Mary's Hospital Medical School, Imperial College of Science, Technology and Medicine, London W2, United Kingdom
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Abstract

The presence of hepatitis delta virus genomic RNA and hepatitis delta antigen was investigated in woodchuck liver and extrahepatic tissues by in situ hybridization using synthetic radiolabeled probes, Northern-blot analysis and immunohistochemical staining for hepatitis delta antigen. Hepatitis D virus RNA and hepatitis delta antigen were detected in the nuclei of infected hepatocytes but in none of the other tissues examined. Northern-blot analysis of total cell RNA confirmed these findings and revealed a series of hepatitis D virus transcripts, including full-length genomic RNA and dimers and trimers of hepatitis D virus RNA that may represent replicative intermediates. Use of single-stranded probes showed genome-size monomers and dimers to be both of genomic and antigenomic polarity, although dimers were found to be predominantly antigenomic. These findings document the strict hepatotropism of hepatitis D virus and support the rolling-circle model of genome replication for this unique, defective RNA virus.

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