PRODUCTION OF HEPATITIS-B VIRUS-PARTICLES IN HEP-G2 CELLS TRANSFECTED WITH CLONED HEPATITIS-B VIRUS-DNA
PRODUCTION OF HEPATITIS-B VIRUS-PARTICLES IN HEP-G2 CELLS TRANSFECTED WITH CLONED HEPATITIS-B VIRUS-DNA
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DOI：
10.1073/pnas.84.4.1005
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发表时间：
1987-02-01
影响因子：
11.1
通讯作者：
ACS, G
中科院分区：
文献类型：
--
作者：
SELLS, MA;CHEN, ML;ACS, G
将携带G418抗性基因的质粒和四个5‘’-3‘’串联的乙肝病毒基因组拷贝导入肝母细胞瘤细胞系Hep G2，使基因组DNA的两个二聚体相互之间为3‘’-3‘’。在G418存在下生长的一个克隆细胞产生高水平的乙肝e抗原和乙肝表面抗原。HBVDNA由这些细胞以染色体整合序列的形式携带，在外周以松弛的环状、共价闭合的和不完整的HBV基因组拷贝的形式携带。在条件培养基中也检测到了病毒DNA，其浮力密度是感染性丹麦人和未成熟核心颗粒的特征。最后，免疫电子显微镜分析显示了与22 nm球形和丝状乙肝表面抗原颗粒以及42 nm Dane颗粒形态相同的乙肝病毒特异性成分。因此，我们已经证明，Hep G2细胞不仅可以支持几种复制的HBVDNA中间体的组装和分泌，还可以支持Dane样颗粒的组装和分泌。这个体外系统现在可以用来研究乙肝病毒的生命周期以及免疫活性细胞与携带乙肝病毒的细胞的反应。
The hepatoblastoma cell line Hep G2 was transfected with a plasmid carrying the gene that confers resistance to G418 and four 5''-3'' tandem copies of the hepatitis B virus (HBV) genome positioned such that two dimers of the genomic DNA are 3''-3'' with respect to one another. Cells of one clone that grew in the presence of G418 produce high levels of hepatitis B e antigen and of hepatitis B surface antigen. HBV DNA is carried by these cells as chromosomally integrated sequences and episomally as relaxed circular, covalently closed, and incomplete copies of the HBV genome. Viral DNA was detected also in conditioned growth medium at the buoyant densities characteristic for infectious Dane and immature core particles. Finally, HBV-specific components morphologically identical to the 22-nm spherical and filamentous hepatitis B surface antigen particles as well as 42-nm Dane particles were visualized by immunoelectron microscopic analysis. Therefore, we have demonstrated that the Hep G2 cell line can support the assembly and secretion not only of several of the replicative intermediates of HBV DNA but also of Dane-like particles. This in vitro system can now be used to study the life cycle of HBV and the reaction of immunocompetent cells with cells carrying HBV.