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目的了解急性髓系白血病KG1a细胞体外诱导人外周血T细胞的特异性细胞毒作用和T细胞受体(TCR)Vβ谱系表达和克隆情况。方法分别以Jurkat和Raji细胞作为阳性和阴性对照,建立TCRVβ谱基因扫描技术,并用其检测5例健康个体T细胞克隆表型。采用混合淋巴/肿瘤细胞培养方法,以照射后的KG1a细胞作为刺激原体外诱导外周血单个核细胞增殖,应用LDH释放法检测不同效靶比时诱导前后T细胞杀伤KG1a细胞的活性,同时利用基因扫描技术分析诱导后TCRVβ亚家族的克隆性增殖特点。结果基因扫描显示5例健康人全部TCRVβ谱型均呈高斯(钟型)分布,各亚家族表达频率接近,但呈现不同的多态性和长度分布。KG1a细胞可以诱导出呈单克隆、寡克隆或寡克隆趋势生长的亚家族T细胞,并具有特异性识别和杀伤KG1a细胞的功能。结论KG1a细胞在体外可诱导T细胞呈克隆性增殖,此优势增殖的克隆性T细胞具有特异性细胞毒作用,对KG1a细胞具有选择性杀伤作用。
Objective To investigate the specific cytotoxicity and the expression and cloning of T cell receptor (TCR) Vβ in vitro induced by KG1a cells of acute myeloid leukemia in vitro. Methods Jurkat and Raji cells were used as positive and negative controls to establish TCRVβ gene scanning technique and to detect T cell clone phenotype in 5 healthy individuals. The mixed lymphoid / tumor cell culture method was used to induce the proliferation of peripheral blood mononuclear cells (PBMCs) from irradiated KG1a cells. LDH release assay was used to detect the killing effect of KG1a cells before and after treatment with different target ratios. At the same time, Analysis of clonal proliferation of TCRVβ subfamily induced by scanning technique. Results The results of gene scan showed that all TCRVβ profiles of five healthy individuals showed a Gaussian distribution, and the frequency of each subfamily was close but showed different polymorphism and length distribution. KG1a cells can induce sub-family T cells that grow in the tendency of monoclonal, oligoclonal or oligoclonal growth and have the function of specifically recognizing and killing KG1a cells. Conclusion KG1a cells can induce clonal proliferation of T cells in vitro. The dominant clonal T cells proliferate in a specific cytotoxicity and selectively kill KG1a cells.