EB病毒转化的丙型肝炎病人外周血B淋巴细胞中丙型肝炎Ⅱ型病毒持续存在的研究  

Persistence of hepatitis C virus type Ⅱ in patient′s peripheral blood B lymphocytes transformed by Epstein Barr virus

作　　者：程计林[1] 陈良标[2] 仝文斌[1] 陈佩兰[2] 刘宝玲[3] 高建恩[1] 杜少才 冯百芳[1] 

机构地区：[1]北京医科大学人民医院肝病研究所,100044 [2]北京军区总医院 [3]河南省焦作市人民医院

出　　处：《中华医学杂志》2000年第5期349-353,共5页National Medical Journal of China

基　　金：卫生部青年科研基金课题资助项目 !(98 1 2 6 9)

摘　　要：目的　观察丙型肝炎病人外周血B细胞长期存活状态下其中是否仍有丙型肝炎病毒(HCV)存在。方法　用EB病毒感染并使HCV阳性外周血B细胞转化 ,获得含HCV的外周血永生化B细胞 (LCL)。而后 ,每月应用套式RT PCR检测一次培养细胞和上清中的HCV RNA ,并用酶切法进行HCV基因分型 ,电镜和免疫电镜观察HCV在该细胞中存在的部位及其形态结构特征。结果　经连续 1年检测 ,HCV在传代细胞中持续存在 ,培养细胞中的HCVRNA负链和培养上清中的HCV呈间断阳性。HCV基因为Ⅱ型。电镜下病毒颗粒呈圆球型 ,双层膜结构 ,定位于胞质空泡内 ,直径多为 45～ 70nm ,个别为 110nm。结论　HCV可以在培养的EB病毒转化B细胞系中较长时间存在、复制和分泌 ,为病人外周血单个核细胞中HCV持续存在提供了又一证据 ,对于理解HCV感染慢性化机理。Objective To study the persistence and replication of hepatitis C virus (HCV) RNA in human peripheral blood B cells transformed by Epstein Barr virus (EBV) and cultured in vitro. Methods EBV was used for infecting B lymphocytes from one hepatitis C patient with HCV positive in the peripheral blood mononuclear cells(PBMC)and transforming them into lymphoblasts capable of being propagated indefinitely. Then, HCV RNA of the cultured cells and supernatants was detected by reverse transcriptase polymerase chain reaction (RT PCR) every one month. HCV gene was typed by enzyme separating method. Electron microscopy and immunoelectron microscopy were employed to locate HCV in the cells and observe the character of its configuration. Results HCV positive strand RNA was detected in the cultured cells for 1 year. Interestingly, the HCV positive strand RNA was identified in supernatants and the negative strand RNA was also observed in the cultured cells intermittently. HCV gene was typeⅡ. Electron microscopy observed HCV spherical virus like particles with a diameter of approximately 45 nm to 70 nm, individual particles 110 nm, in the LCL cytoplasmic vesicles. Conclusion HCV may exist in the cultured cell line for a longer period and reproduce in and secrete out of them. HCV locates mainly in the cytoplasm. The cell line might be useful in analyzing the mechanisms of HCV persistence in PBMC.

关 键 词：丙型肝炎 外周血 B细胞 HCV E-B病毒 

分 类 号：R512.63[医药卫生—内科学]