特异性体外扩增人γδT淋巴细胞及其生物学特性的研究  

Establishemnt of a novel culture system for specific expansion of human γδ T cell and study of its biological properties

作　　者：李新燕[1,2] 张学光[1,2] 张毅 李明[1,2] M.M.Hallet B.Klein[1,2] 

机构地区：[1]苏州医学院免疫教研室 [2]法国科学中心分子遗传所

出　　处：《中华医学杂志》1997年第2期111-114,共4页National Medical Journal of China

基　　金：卫生部"八五"科技攻关基金

摘　　要：探讨人γδＴ淋巴细胞体外特异性激发的方法，及其杀伤肿瘤细胞的分子机理。方法采用人多发性骨髓瘤细胞株ＸＧ－７细胞作为刺激细胞，与人外周血单个核细胞体外共同培养，激发和扩增人γδＴ淋巴细胞。以流式细胞仪或间接免疫荧光技术分析细胞表型；再以５１Ｃｒ４小时释放试验检测γδＴ淋巴细胞对肿瘤细胞的杀伤效应。结果（１）采用本培养系统，可以使来自健康人或肿瘤患者的外周血γδＴ淋巴细胞得以选择性激发和扩增，由４．８％±３．４％增加至４６．０％±８．３％，在白细胞介素２（ＩＬ－２）的作用下，能迅速、大量地增殖，３周内可达１０１０以上数量级。（２）所扩增的γδＴ淋巴细胞，对肿瘤细胞株（天然杀伤细胞敏感株Ｋ５６２和淋巴因子激活的杀伤细胞敏感株Ｄａｕｄｉ）皆可介导高效杀伤活性，而对正常淋巴母细胞则无明显细胞毒作用。（３）抗热休克蛋白（ＨＳＰ）－７０单克隆抗体可阻断γδＴ淋巴细胞对ＸＧ－７细胞的反应。结论本室建立的人γδＴ淋巴细胞体外激发、扩增的方法，具有简便、迅速、重复性好的特点，对肿瘤细胞具有广谱的细胞毒活性，在肿瘤的过继免疫治疗中具有潜在的临床应用价值。Objective To study the specific expansion method of human γδ T cell and the molecular mechanism against tumor cells. Methods The peripheral blood mononuclear cells were co cultured with XG 7 cells as stimulating cells and activated for the expansion of γδ T cells. Immunophonotype was analysed by IIFA and FACS. The cytotoxicity of γδT cell against tumor cells was measured by 51 Cr four hour release assay. Results When this novel culture system was used, γδ T cells mainly expressing V γ9/Vδ 2 encoded TCR were selected and expanded rapidly from 4.8±3.4% to 46.2±8.3%. With exogenous interleukin 2, γδ T cells were expanded in large quantity 10 10 within 3 weeks. γδT cells mediated strong cytotoxic activity to different tumor cells including K562, Daudi, XG 7 cell lines, but no cytotoxic activity to normal cells was observed. XG 7 cells expressed HSP molecules and anti HSP mAB blocked the proliferation of γδT cells. Conclusion The method shows unique biological properties: brief, rapid, well repeating and wide cytotoxic activity against tumor cells. So γδT cells are of value in tumor adoptive immunotherapy.

关 键 词：T淋巴细胞.细胞毒性 抗原.肿瘤 热休克蛋白 

分 类 号：R730.51[医药卫生—肿瘤] R392.12[医药卫生—临床医学]