乙型肝炎病毒核心抗原特异性细胞毒性T淋巴细胞作用的靶细胞的建立

Establishment of target cells of cytotoxic T-lymphocyte response with specifity of HBcAg

目的建立ＨＢｃＡｇ特异性ＣＴＬ作用的靶细胞系统，为进一步研究ＨＢＶ基因变异对ＣＴＬ细胞毒效应的影响奠定基础。方法采用两种质粒ｐＸＴ１和ＥＢＯ－ｐｌｐｐ构建ＨＢＶＣ基因真核细胞表达载体并转染水生化Ｂ细胞，ＤＮＡ分析和流式细胞仪检测ＨＢＶＣ基因在宿主细胞中的复制、表达。结果在转染重组质植ｐＸＴ１－ＨＢＶｃ和ＥＢ０－ｐｌｐｐ－ＨＢＶｃ的Ｂ细胞中，均能检测到目的基因；分另有８９．８１％和８８．５１％的宿主细胞能检测到ＨＢｃＡｇ；连续培养３周后，分别有８８．３０％和７２．１９％的宿主细胞表达ＨＢｃＡｇ。结论重组逆转录病毒表达载体ｐＸＴ１－ＨＢＶｃ和ＥＢ病毒表达载体ＥＢＯ－ｐｌｐｐ－ＨＢＶｃ转染的永生化人外周血Ｂ细胞能有效地表达靶抗原（ＨＢｃＡｇ），可作为较为理想的ＨＢＶ特异性ＣＴＬ作用的靶细胞。

Objective To establish target cell system of CTL response with specificity of HBV for studying the effect of CTLcytotoxicity caused by HBV gene mutation. Methods Two recombinant plasmids pXT1-HBVc and EBO-plpP-HBVc weretransferred into immortalized B cells, replication and expression of HBV C gene in the host cells were detected by DNA analysis andflow cytometry. Results DNA fragments of the same size with the goal gene can be detected in the host cells, and the expressedHBcAg was detected in 89.81 % and 88.51 % of the host cells with transfection of recombinant plasmid pXT1-HBVc and EBO-plppHBVc respectively; and in 88.30% and 72.19% after 3 weeks of culture. Conclusion The immortalized B cells transfected with therecombinant plasmid pXT1-HBVc and EBO-plpp-HBVc can expressed target antigen(HBcAg) effectively, which can serve assuitable target cells of CTL response with specifity of HBV.