人肝癌细胞系7721丙型肝炎病毒体外感染模型的建立
作者：宋志强 郝飞
单位：400038 重庆，第三军医大学西南医院全军传染病专科中心
关键词：肝炎病毒，丙型；7721细胞系；细胞培养；细胞模型；感染
中华传染病杂志990402 【摘要】 目的 建立接近体内自然感染状态并能稳定支持丙型肝炎病毒(HCV)体外长期复制的感染细胞模型。方法 将人肝癌细胞系7721与慢性丙型肝炎患者血清共温育8小时后，分别以逆转录-聚合酶链反应、免疫组化、原位杂交检测细胞和/或培养上清中的HCV RNA、HCV抗原表达及HCV RNA在感染细胞内的定位。结果 感染血清和细胞共同温育后的第2～66天，从细胞内和培养上清中均可间断地检测出HCV正、负链RNA，即使其间细胞传代6次，HCV NS3、NS5抗原在细胞内能稳定表达，原位杂交显示HCV RNA阳性物质多位于细胞浆。结论 7721细胞不但对HCV易感，而且可以稳定地支持HCV的体外长期复制，此模型可以用于HCV感染、复制机制的研究、抗病毒药物的评价及保护性抗体/疫苗的初步筛选。
Establishment of HCV infection model in vitro using a human liver cell line 7721
SONG Zhiqiang, HAO Fei.
Center for Infectious Disease of Chinese PLA, Southwest Hospital, Third Military Medical University, Chongqing 400038
【Abstract】 Objective To establish a cell model that is close to the HCV replication in vivo and can support long-term HCV replication in vitro.Methods A human liver cell line 7721 was inoculated with serum from a chronic hepatitis C patient for 8 hours. After incubation, the presence of HCV RNA, the expression of HCV antigen and the location of HCV RNA in cells and/or supernatant were detected by RT-PCR, immunohistochemistry and in situ hybridization respectively. Results It was found that plus-and minus-strand of HCV RNA could be discontinuously detected in both cells and supernatant as late as 66 days after inoculation even if cells had been subcultured for 6 times. HCV NS3、NS5 antigens could be, http://www.100md.com
单位：400038 重庆，第三军医大学西南医院全军传染病专科中心
关键词：肝炎病毒，丙型；7721细胞系；细胞培养；细胞模型；感染
中华传染病杂志990402 【摘要】 目的 建立接近体内自然感染状态并能稳定支持丙型肝炎病毒(HCV)体外长期复制的感染细胞模型。方法 将人肝癌细胞系7721与慢性丙型肝炎患者血清共温育8小时后，分别以逆转录-聚合酶链反应、免疫组化、原位杂交检测细胞和/或培养上清中的HCV RNA、HCV抗原表达及HCV RNA在感染细胞内的定位。结果 感染血清和细胞共同温育后的第2～66天，从细胞内和培养上清中均可间断地检测出HCV正、负链RNA，即使其间细胞传代6次，HCV NS3、NS5抗原在细胞内能稳定表达，原位杂交显示HCV RNA阳性物质多位于细胞浆。结论 7721细胞不但对HCV易感，而且可以稳定地支持HCV的体外长期复制，此模型可以用于HCV感染、复制机制的研究、抗病毒药物的评价及保护性抗体/疫苗的初步筛选。
Establishment of HCV infection model in vitro using a human liver cell line 7721
SONG Zhiqiang, HAO Fei.
Center for Infectious Disease of Chinese PLA, Southwest Hospital, Third Military Medical University, Chongqing 400038
【Abstract】 Objective To establish a cell model that is close to the HCV replication in vivo and can support long-term HCV replication in vitro.Methods A human liver cell line 7721 was inoculated with serum from a chronic hepatitis C patient for 8 hours. After incubation, the presence of HCV RNA, the expression of HCV antigen and the location of HCV RNA in cells and/or supernatant were detected by RT-PCR, immunohistochemistry and in situ hybridization respectively. Results It was found that plus-and minus-strand of HCV RNA could be discontinuously detected in both cells and supernatant as late as 66 days after inoculation even if cells had been subcultured for 6 times. HCV NS3、NS5 antigens could be, http://www.100md.com