初步探讨丙型肝炎病毒体外感染的致病机理

Preliminary Study on Pathogenesis of Hepatitis C Virus Infection of Normal Tupaia Hepatocytes in Vitro

目的:初步探讨原代培养肝细胞对HCV的易感性,并在此基础上观察HCV感染后肝细胞形态学的变化。方法:用HCV RNA阳性血清与培养肝细胞共同孵育后,收集不同时相点标本,以逆转录聚合酶链反应(RT-PCR)、分别细胞内或上清中正负链RNA,免疫组化检测细胞内HCV NS3、NS5抗原的表达以及原位杂交检测细胞内HCV负链RNA,同时在电镜下观察细胞形态学变化。结果:接种感染血清后第3～5d,即可在细胞内或培养上清中检出HCV正负链RNA;感染肝细胞内可见HCV NS3、NS5抗原表达,阳性物质位于胞浆中;原位杂交法证实细胞内存在负链RNA,也位于胞浆中;光镜下未发现感染细胞形态学变化,电镜下发现胞浆内含有大量的脂肪空泡、多泡体,并可见疑似病毒样结构。结论:HCV不但能感染树鼩肝细胞,而且在体外能直接致细胞病变。

Objective: To investigate the susceptibility of normal Tupaia hepatocytes in primary culture by hepatitis C virus in vitro and observe their morphologic changes. Methods s Both the plus and minus strands of HCV RNA in infected cells or supernatant were examined by reverse transcription polymerase chain reaction (RT-PCR). The HCV NS3, NS5 antigens in infected cells were respectively detected with the monoclonal antibodies response to the antigens by immuno-histochemical assay. The minus strand of HCV RNA in ...