A strategy for the identification of HBV receptors with HBV pseudotype

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It is near a half century since hepatitis B virus(HBV) was identified. An iceberg of HBV receptor has not been elucidated, though there are some reports on infection systems and on the receptor molecules. Thus, we still have not reached finding a real HBV receptor and there have been no useful and convenient infection system in vitro and in vivo for HBV, which makes it impossible for us to understand a precise HBV life cycle and HBV involved related diseases. Here, we designed and tried to generate an HBV pseudotype, which had a viral particle containing a retrovirus capsid and a genome inside surrounded by HBV membrane proteins. We proved successful generation of this pseudotype by immunoprecipitation with anti-HBVs antibodies and by CsCl density gradient ultracentrifugation, followed by RT-PCR targeting a retroviral gene, a GFP gene in this case, respectively. Though our established system is constructed on growth dependent integration of retroviral genomes and thus was very hard to observe its infection in a primary human hepatocytes culture system, successful generation of the HBV pseudotype will make it possible for us to perform a biological assay to clone an HBV receptor based on infectivity and will facilitate its separation and identification.We have not identified an HBV receptor candidate yet. It has been suggested, however, that there could be an attachment factor in ordinary cultured cell lines derived from hepatocellular carcinomas.