EBV转化的人乳头瘤病毒感染者B淋巴母细胞系的建立及应用

建立EB病毒(EBV)转化的B淋巴母细胞系(LCL),应用此细胞系作为抗原递呈细胞筛选抗原特异性的T细胞克隆。收集人乳头瘤病毒(HPV)感染者外周血20份,分离外周血单个核细胞(PBMC),利用Pan T细胞试剂盒去除PBMC中的T细胞,获得non-T淋巴细胞,EBV病毒转化non-T淋巴细胞。应用转化的LCL细胞作为抗原递呈细胞,ELISPOT筛选可能的HPV抗原特异性的T细胞克隆。成功建立18个LCLs,建株成功率为90%。转化成功的LCL细胞体积增大,聚集成团状或簇状。建株失败的两份标本,用于转化的non-T淋巴细胞数少于2×106。应用LCL细胞作为抗原递呈细胞筛选出64个HPV抗原特异性的T细胞克隆。成功建立LCLs,此细胞系细胞可作为体外研究HPV特异性免疫反应的刺激细胞。

Establishment and Application of EBV-Transformed B Lymphoblast Cell Line from HPV Infected Persons

An EBV-transformed B lymphoblast cell line（LCL） was established and applied the cell line as a presenting cells to screen antigen-specific T cell clone.Peripheral blood mononuclear cells（PBMC） were isolated from 20 blood specimens from women infected human papillomavirus（HPV） and T cells were depleted from PBMC by Pan T cell isolation kit.Non-T cells lymphocyte were obtained and transformed with EBV.HPV-specific T cell clones were screened using LCL as antigen presenting cells.The results showed that 18 LCLs were established successfully.Cell volume expansion and clustered growth were observed in LCL cells.The number of non-T cell was less than 2×106 for two specimen unable to establish LCL.64 HPV-specific T cell clones were selected.Therefore,LCLs were successfully established and they could serve as stimulating cells in specific immune response to HPV.