论文部分内容阅读
目的 探讨多抗原肽 (MAP)免疫血清在体外能否阻断丙型肝炎病毒 (HCV)感染树鼠句肝细胞 ,为研制HCV分子疫苗提供实验基础。方法 用兔抗MAP血清分别与两份感染血清按一定比例混合后接种于原代培养的树鼠句肝细胞中 ,用逆转录聚合酶链反应 (RT PCR)、免疫组化、原位杂交分别检测细胞内或上清中正负链RNA ,细胞内HCVNS3抗原的表达情况 ,以及原位检测细胞内HCV负链RNA。结果 1份HCVRNA阳性血清与兔抗MAP血清按 1∶1混合后接种于培养的树鼠句肝细胞中 ,细胞内可检出HCV正负链RNA ,并有HCVNS3抗原的表达 ,而另外 1份HCVRNA阳性血清与兔抗MAP血清按 1∶1、5∶1、10∶1混合后接种于培养的树鼠句肝细胞中 ,细胞内未检测出HCV正负链RNA ,也未见特异性抗原表达。结论 兔抗MAP血清在体外能部分阻断HCV感染。
Objective To investigate whether multi-antigen peptide (MAP) immune sera can block hepatitis C virus (HCV) -based hepatocytes in vitro and provide the experimental basis for the development of HCV molecular vaccine. Methods Rabbit anti-MAP sera were separately mixed with two infected sera at a certain ratio and then inoculated into primary cultured rat hepatocytes. RT-PCR, immunohistochemistry and in situ hybridization Detection of intracellular or supernatant positive and negative strand RNA, intracellular HCV NS3 antigen expression, and in situ detection of intracellular HCV negative strand RNA. Results 1 HCV RNA positive serum and rabbit anti-MAP serum were mixed at a ratio of 1: 1 and then inoculated into cultured rat hepatocytes. HCV positive and negative RNAs were detected in the cells with the expression of HCV NS3 antigen, while the other 1 HCV RNA positive serum and rabbit anti-MAP serum were mixed at 1: 1, 5: 1, 10: 1 and then inoculated into cultured rat hepatocytes. No positive or negative HCV RNA was detected in the cells, and no specific antigen expression. Conclusion Rabbit anti-MAP serum can partially block HCV infection in vitro.