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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ＨＣＶＲＮＡ 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。
Objective To establish an infected cell model that is close to the body’s natural state of infection and can stably support long-term replication of hepatitis C virus (HCV) in vitro. Methods Human hepatocellular carcinoma cell line 7721 was co-incubated with serum of patients with chronic hepatitis C for 8 hours. HCV RNA in cells and/or culture supernatants was detected by reverse transcription polymerase chain reaction, immunohistochemistry, and in situ hybridization, respectively. , HCV antigen expression and localization of HCV RNA in infected cells. Results From the 2nd to the 66th day after co-incubation with infected serum and cells, HCV positive and negative strand RNAs were detected intermittently from intracellular and culture supernatants. Even if the cells were passaged 6 times between them, the HCV NS3 and NS5 antigens were The cells can be stably expressed. In situ hybridization shows that HCV RNA positive substances are located in the cytoplasm. Conclusion 7721 cells are not only susceptible to HCV, but also can stably support the long-term replication of HCV in vitro. This model can be used for the study of HCV infection, replication mechanism, evaluation of antiviral drugs, and preliminary screening of protective antibodies/vaccines.