NB4和HL-60细胞诱导的脐血T细胞TCR Vβ基因谱系分析

Analysis of TCR Vβ repertoire in cord blood T cells induced by NB4 cells and HL-60 cells

目的:了解经NB4细胞和HL-60细胞体外诱导后脐血T细胞的TCR Vβ亚家族限制性利用和克隆性增殖情况。方法:采用混合淋巴细胞/肿瘤细胞培养(MLTC)方法,用经丝裂霉素灭活的NB4细胞和HL-60细胞体外诱导脐血T细胞增殖,利用RT-PCR分析经MLTC前后T细胞的24个TCR Vβ亚家族的利用情况,并用基因扫描分析其克隆性特点。结果:诱导前脐血T细胞表达20个TCR Vβ亚家族,诱导后的T细胞所表达的TCR Vβ亚家族随培养时间增加而逐渐减少,且诱导组呈现出克隆趋势或寡克隆增殖特点。两种细胞诱导后10 d均可见Vβ1的克隆增殖出现变化,此外,NB4诱导的T细胞出现Vβ15,而HL-60诱导的T细胞则出现Vβ23的克隆增殖情况。结论:NB4细胞和HL-60细胞可诱导脐血TCR VβT细胞克隆性增殖,此优势表达的克隆性T细胞可能与特异性抗原介导的细胞免疫反应有关。

Aim： To investigate the restricted usage and clonal expansion of TCR Vβ subfamily T ceils in cord blood induced by NB4 ceils and HL- 60 cells. Methods： Mixed lymphocyte tumor cells culture （MLTC） was performed with mixing cord blood mononuelear cells and NB4 cells or HL - 60 ceils treated with mitomycin. The usage of 24 TCR Vβ subfamilies were analyzed in T cells before and after MLTC by RT- PCR. The PCR products were further analyzed by genescan to identify the clonality of T cells. Results：20 TCR Vβ subfamilies were detected in cord blood T ceils before MLTC, while the number of expressed TCR Vβ subfamilies after MLTC were reduced with increasing culture time, and clonal tendency or oligoclonal expansion of TCR Vβ subfamily T cells were identified. The clonal expansion of TCR Vβ subfamily was detected in T cells induced by both NB4 cells and HL - 60 cellls at 10th day of MLTC. Moreover, clonal expanded Vβ15 and Vβ23 T ceils were identified in T ceils induced by NB4 cells or HL- 60 respectively. Condusions： Oligoclonal and clonal tendency TCR Vfl subfamily T cells could be identified in cord blood T cells induced by both NB4 and HL- 60 cell line, and the clonal expanded T cells may be related to antigen specific cell inmmne response.