EBV转化B淋巴细胞中丙型肝炎病毒抗原表达的免疫组化研究  

Immunocytochemistic Study of HCV Antigens in Human Peripheral Blood Mononuclear Cell Transformed by Epstein-barr Virus.

作　　者：程计林[1] 耿直[2] 仝文斌[1] 范涛[1] 高建恩[1] 马文彬[1] 冯百芳[1] 

机构地区：[1]北京医科大学人民医院肝病研究所,北京100044 [2]河南省焦作市人民医院,河南焦作454100

出　　处：《临床肝胆病杂志》2001年第2期92-94,共3页Journal of Clinical Hepatology

摘　　要：观察丙型肝炎患者外周血单个核细胞 (PBMC)体外转化培养后其中是否仍有丙型肝炎病毒 (HCV)存在。用EB病毒感染患者HCV阳性PBMC并使其转化 ,获得永生化B细胞 (EBVTB)。而后 ,用套式RT -PCR检测培养细胞和上清中的HCVRNA ,用免疫组化SP法观察HCV抗原在细胞中的表达。HCV在传代细胞中持续存在 ,而培养细胞中的HCVRNA负链和培养上清中的HCV间断阳性。免疫组化发现HCVNS3、NS5和C抗原分布于胞浆、细胞膜和核膜周围也可见阳性着色 ,细胞核阴性。HCV可以在体外EB病毒转化B细胞系中较长时间存在和复制。To study persistence and replication of hepatitis C virus(HCV)in human peripheral blood mononuclear cells(PBMC)cultured in vitro.EBV was used to infect B lymphocytes from three hepatitis C patients with HCV positive in PBMC and transformed into lymphoblasts capable of being propagated indefinitely.Then,HCV RNA in the cultured cells and supernatants were detected by lestle reverse transcriptase-polymerase chain reaction(RT-PCR)and immunocytochemistric SP method was used to identify the expression of antigens of HCV in the cells.HCV plus-strand RNA could be detected in the cells for more than six months.Whereas the minus-strand RNA was also observed in the cells.lnterestingly,the HCV positive-strand RNA could be identified in supernatants intermittently.The positive brown granules of NS3,NS5,and C antigens of HCV were found and they scattered or clustered or conglomerated in the cytoplasm.The positive staining was observed on the cell membrane and karyotheca too,but wasnt done in the nucleolus.HCV may exist in the cultured cell line for a longer period and reproduce in and secrete out of them.

关 键 词：丙型肝炎病毒 B淋巴细胞 EBV 抗原 免疫组织化学 

分 类 号：R512.63[医药卫生—内科学]