乙型肝炎病毒(HBV)假型颗粒的成功生成一种用于HBV受体鉴定和HBV感染性研究的通用工具

K. Ueda, H. Omori
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引用次数: 4

摘要

乙型肝炎病毒(HBV)被发现已有近半个世纪。HBV受体分子和HBV进入肝细胞的机制尚未完全阐明,尽管有一些关于感染系统和受体分子的报道。因此,我们仍然没有找到真正的HBV受体,也没有有效方便的体外和体内HBV感染系统,这使得我们无法准确了解HBV的生命周期和HBV相关疾病。也需要一个HBV感染系统来探索基于证据的HBV相关疾病的治疗方法和手段。在这里,我们设计并试图产生一种HBV假型,它具有一个含有逆转录病毒衣壳的病毒颗粒和一个被HBV膜蛋白包围的基因组。我们通过抗乙肝病毒抗体的免疫沉淀和CsCl密度梯度超离心,然后分别针对逆转录病毒基因和EGFP基因进行RT-PCR,证明了这种假型的成功产生。虽然我们建立的系统是建立在逆转录病毒基因组的生长依赖性整合上,因此很难在原代人肝细胞培养系统中观察到它的感染情况,但HBV假型的成功产生将使我们有可能进行基于感染性克隆HBV受体的生物学试验,并将促进其分离和鉴定
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Successful Generation of Hepatitis B virus (HBV) Pseudotype Particle; A Versatile Tool for Identification of the HBV Receptor and Investigation of HBV Infectivity
It is near a half century since hepatitis B virus (HBV) was identified. HBV receptor molecules and the entry mechanism of HBV into hepatocytes have not been elucidated completely, 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. An HBV infection system is really needed to explore ways and means of treatment of HBV related diseases based on evidence as well. Here, we designed and tried to generate an HBV pseudotype, which has 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, an EGFP 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
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