HCV体外感染正常成人肝细胞的研究

目的 为进一步研究丙型肝炎免疫发病机制，临床治疗和疫苗研制提供最接近自然感染状态的理想细胞模型。方法 体外二步灌流法分离正常成人肝细胞并维持培养1月以上，丙型肝炎患者血清感染上述肝细胞并用免疫组化和逆转录-聚合酶链反应法（RT-PCR法）检测被感染肝细胞HCV抗原表达及HCVRNA。结果 正常成人肝细胞可在体外存活1月以上，加丙肝患者血清4-36d后免疫组化法可检测到肝细胞内HCVNS3、NS5抗原表达，感染后2-36d，RT-PCR法可在感染肝细胞培养上清和细胞悬液中间断检测出HCVRNA。结论 正常成人肝细胞可在体外存活1月余并被HCV感染者血清感染。

Study on the intection of nomal adult hepatocytes with HCV in vitro

Objective To provide a better cell model of closely nature infectious state for further understanding the immunopathogenesis, clinical treatment and vaccine development of hepatitis C. Methods Normal adult hepatocytes were isolated according to the two step perfusion method and cultured in vitro more than 1 month. The hepatocytes were then infected with serum of patients with hepatitis C. The expression of HCV antigen and HCV RNA in cells were detected with immunohistochemical method and RT PCR. Results Normal adult hepatocytes surrived more than 1 month in vitro . In this experiment the normal adult hepatocytes could be infected by serum containing HCV from patients with hepatitis C. The expression of HCV NS3, NS5 antigen in cells could be detected by immunohistochemical method, HCV RNA was discontinuously detected in cells and supernatant with RT PCR. Conclusion Normal adult hepatocytes may survive more than one month in vitro and can be infected by serum of patients with hepatitis C.