APL细胞诱导脐血TCR Vα和Vβ亚家族T细胞增殖和细胞毒性研究

Clonal expansion and specific cytotoxicity of cord blood T cells induced by APL cells

目的了解急性早幼粒细胞性白血病(APL)细胞体外诱导脐血T细胞的TCR Vα和Vβ亚家族克隆性增殖及针对APL细胞特异性杀伤情况。方法采用混合淋巴细胞和肿瘤细胞培养(MLTC)方法,利用APL细胞体外诱导脐血T细胞增殖,用RT-PCR和基因扫描分析MLTC后T细胞的29个TCR Vα亚家族基因和24个TCR Vβ亚家族基因的互补决定区(CDR3),了解各亚家族的T细胞克隆性增殖特点,并应用LDH法分析诱导增殖的T细胞针对APL细胞特异性杀伤作用。结果经APL细胞刺激后增殖的脐血T细胞表达部分Vα和Vβ谱系基因,并在Vα10、Vβ5和Vβ15出现有克隆增殖趋势,诱导后T细胞对APL细胞的杀伤性均高于未诱导组的T细胞。结论APL细胞体外诱导脐血T细胞出现抗原相关的TCR Vα和Vβ亚家族优势利用和克隆性增殖;诱导后的T细胞对APL细胞具有特异性细胞毒性作用。

Objective To investigate the clonal expansion and specific cytotoxcity of TCR Vet subfamily and Vβ subfamily T cells from cord blood induced by APL cells. Methods T cells from cord blood were induced and amplified by APL cell in vitro by MLTC （mixed lymphocyte - tumor culture）. The complementary determining region 3 （CDR3） of 29 TCR Vet subfamily and 24 TCR Vβ subfamily genes from these T cells were analyzed using RT - PCR and genescan techniques, to detect the usage and clonality feature of TCR subfamilies. Induced cells at different time points were detected for the specifc cytotoxicity by LDH release assay. Results A part of TCR Vet and Vβ repertoires were detected in cord blood T cells after cultured with APL cells. There were oligoclonal tendency T cells in TCR Vet 10, TCR Vβ5 and TCR Vβ15 subfamilies. Cytotoxicity on APL cells of induced cells was significantly higher than that of the uninducded cells from CB. Conclusion Restricted usage and clonal expansion of TCR Vα and Vβ subfamily T cells from CB can be induced by APL cells in vitro; these induced cells have specific cytotoxity on APL cells.