EBV转化带状疱疹患者外周血B淋巴母细胞系的建立  

Establishment of immortalized B lymphoblastoid cell lines from the patients with herpes zoster by using Epstein-Barr virus

作　　者：莫小辉 宋宁静 陈佳 谈亦 余茜 胡婷婷 谭飞 

机构地区：[1]上海市皮肤病医院中心实验室,200443

出　　处：《免疫学杂志》2013年第7期615-618,共4页Immunological Journal

基　　金：上海市卫生局课题(20124y085);上海市公共卫生重点学科建设项目(12GWZX0902)

摘　　要：目的体外建立带状疱疹患者永生化的淋巴母细胞系(LCL)。方法用EB病毒感染带状疱疹患者外周血中分离的单个核细胞(PBMC),加入CpG 2006免疫调节基序以诱导B淋巴细胞增殖,环胞菌素A抑制T淋巴细胞的活性。利用流式细胞术分析LCL膜表面分子CD抗原,利用PCR技术检测淋巴母细胞中EBNA1基因。结果 20例患者PBMC经EBV感染制备淋巴母细胞,13例转化成永生化B细胞来源的LCL,成功率为65%。转化后的LCL膜表面表达CD19和CD45,并存在EB病毒基因EBNA-1基因序列。结论利用EBV感染带状疱疹患者B淋巴细胞形成的LCL保持了成熟B淋巴细胞的生物学特性,为体外研究水痘-带状疱疹病毒(VZV)感染特异性细胞免疫和人源单克隆抗体制备打下基础。Infection of B ceils with Epstein-Barr virus （EBV） leads to proliferation and subsequent immortalization. In this stury, we aimed to establish EBV transformed B lymphoblastoid cell lines （LCL） in patients with herpes zoster （HZ） in vitro. The peripheral blood mononuclear cells （PBMC） were isolated from the patients with HZ by routine method, and then infected with EBV in the presence of CpG 2006 DNA motifs and cyclosporine A （CyA）. By observing morphological characteristics with microscope, detecting expression of surface molecules with flow cytometry, and analyzing LCL DNA for EBV genomic sequences with polymerase chain reaction （PCR）, total of thirteen strains of LCL were established from twenty patients with HZ. Furthermore, CD19 and CD45 were positively detected on the surface of LCL and all LCL were positive for EBNA-1 gene. In this study, we established EBV- immortalized LCL successfully, which would provides target cells for further research on the cellular immunity of HZ and the production of human monoclonal antibodies against Variella-Zoster virus （VZV）.

关 键 词：水痘-带状疱疹病毒 EB病毒 淋巴母细胞系 带状疱疹 

分 类 号：R752.12[医药卫生—皮肤病学与性病学]