多抗原肽免疫血清体外阻断丙型肝炎病毒感染外周血单个核细胞的研究

摘要

目的 探讨多抗原肽(MAP)免疫兔血清体外阻断丙型肝炎病毒(HCV)感染人外周血单个核细胞.方法 用兔抗MAP血清按一定比例混合后接种于HCV感染的人外周血单个核细胞,采用逆转录聚合酶链反应、免疫组化和原位杂交技术分别检测细胞内或上清中HCV RNA、细胞内HCV NS3和HCV NS5抗原的表达.结果 兔抗MAP血清按1:1混合后,外周血单个核细胞仍可检出HCV RNA,细胞有HCV NS3抗原表达;而按5:1和10:1混合后,单个核细胞内未检测出HCV正负链RNA,未见特异性抗原表达,原位杂交也未检测出HCV RNA存在.结论 兔抗MAP血清在体外能部分阻断HCV感染.%Objective To investigate whether an anti-multiple antigen peptide (MAP)rahbit serum against hypervariable region 1 of hepatitis C virus(HCV)can block viral infection of peripheral blood mononuclear cells in vitro. Methods An anti-MAP rabbit serum was mixed with positive sera of HCV RNA respectively in some series of proportions. The cells and supernatant were harvested at various phases during the cultures. Both the plus and minus strands of HCV RNA in infected cells or supernatants were examined by reverse transcription polymerase chain reaction (RT-PCR). The HCV NS3 and HCV NS5 antigens in infected cells were respectively detected with the monoclonal antihodies to them by immunohistochemical assay.The minus strand of HCV RNA in infected cells were localized by in situ hyhridization. Results When the anti-MAP rabbit serum and inf'ectious serum (1:1)were added into peripheral blood mononuclear cells,HCV RNA could be detected in culture cells,and the positive signals of NS3 and NS5 antigens could he detected within cytoplasm of infected cells. The minus strand of HCV RNA was located within cytoplasm. But when the anti-MAP rahbit serum and infectious serum (5:1and 10:1)were added into peripheral blood mononuclear cells,the HCV RNA,NS3 and NS5 antigens were negative in the infected cells. The minus strand of HCV RNA was not found in the cytoplasm of the cells. Conclusions The anti-MAP rahbit serum could partly prevent the cultured cells from being infected hy HCV.