IN-VITRO EXPERIMENTAL-INFECTION OF PRIMARY HUMAN HEPATOCYTES WITH HEPATITIS-B VIRUS
IN-VITRO EXPERIMENTAL-INFECTION OF PRIMARY HUMAN HEPATOCYTES WITH HEPATITIS-B VIRUS
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DOI：
10.1016/0016-5085(94)90700-5
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发表时间：
1994-03-01
期刊：
影响因子：
29.4
通讯作者：
ZENTGRAF, H
中科院分区：
文献类型：
--
作者：
GALLE, PR;HAGELSTEIN, J;ZENTGRAF, H
背景/目的：研究B型肝炎病毒（HBV）与宿主细胞的相互作用需要合适的组织培养系统。本研究采用健康人肝组织的原代肝细胞建立体外生产性感染。病毒蛋白的生产进行了评估，放射免疫分析和[35 S]蛋氨酸标记，和病毒DNA的生产进行了评估，Southern印迹和内源性polymerase assay.Results：分泌高水平的B肝炎表面抗原（HBsAg）和低水平的B肝炎病毒e抗原（HBeAg）进入培养基感染后6天可检测到，并在12天后达到最大值。代谢标记显示病毒蛋白的产生是从头合成的结果。单链HBV DNA出现在感染细胞的细胞质中，通常存在于未成熟的核心中，表明病毒复制。感染后12天，可从培养基中免疫沉淀出含有HBV DNA的颗粒，该颗粒具有活性病毒DNA聚合酶。特异性针对病毒包膜的preS 1区域的抗血清能够阻断感染。二甲基亚砜的存在下，在培养基中大大提高了病毒protein.Conclusions的产量：原代成人肝细胞是有能力感染HBV。
Background/Aims:Studies on the interaction of hepatitis B virus (HBV) with its host cell require a suitable tissue culture system. This study used primary adult hepatocytes from healthy human liver tissue to establish productive infection in vitro.Methods:Hepatocytes were inoculated overnight with HBV. Production of viral proteins was assessed by radioimmunoassay and by [35S]methionine labeling, and production of viral DNA was assessed by Southern blotting and endogenous polymerase assay.Results:Secretion of high levels of hepatitis B surface antigen (HBsAg) and low levels of hepatitis B virus e antigen (HBeAg) into the medium was detectable 6 days after infection and reached maximum values after 12 days. Metabolic labeling showed production of viral proteins to be a result of de novo synthesis. The appearance of single-stranded HBV DNA in the cytoplasm of infected cells, typically present in immature cores, indicated viral replication. HBV DNA containing particles possessing an active viral DNA polymerase could be immunoprecipitated from the medium 12 days after infection. An antiserum specific for the preS1 region of the viral envelope was capable to block infection. Presence of dimethyl sulfoxide in the medium greatly improved the yield of viral proteins.Conclusions:Primary adult human liver cells are competent for infection with HBV.