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Immunocytochemistic Study of HCV Antigens in Human Peripheral Blood Mononuclear Cell Transformed by Epstein-barr Virus.
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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。Abstract: 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 wasnt done in the nucleolus.HCV may exist in the cultured cell line for a longer period and reproduce in and secrete out of them.
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Key words:
- HCV /
- Blymphocyte
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