慢性粒细胞白血病相关的TCR VβT细胞的诱导及其限制性和克隆性分析

The inducement, restricted usage and clonal expansion of TCR Vβsubfamily T cells related to chronic myelogenous leukemia

目的:了解体外诱导CML细胞相关TCR Vβ亚家族T细胞克隆性增殖及其杀伤性的情况。方法:采用混合淋巴细胞/白血病细胞培养法(MLTC)体外将CML细胞、K562细胞和bcr-abl多肽与供者外周血单个核细胞混合培养和扩增,利用RT-PCR-基因扫描技术分析培养后T细胞的TCR Vβ谱系的限制性利用和克隆性增殖情况,并经LDH法分析诱导增殖T细胞的杀伤性。结果:经bcr-abl多肽、CML细胞和K562细胞诱导扩增1-2周后,供者外周血T细胞表达10-13个Vβ亚家族,在Vβ16和Vβ21出现克隆性增殖T细胞,在Vβ5和Vβ13出现寡克隆生长趋势T细胞。诱导扩增后的T细胞对CML和K562细胞具有特异性杀伤作用。结论:利用CML细胞、K562细胞和bcr-abl多肽可在体外诱导出CML细胞特异性CTL,该CTL可能为优势表达的Vβ亚家族克隆性T细胞。

AIM: To investigate the anti-leukemia effect, the restricted usage and clonal expansion of TCR Vβ subfamily T cells from donor peripheral blood induced by chronic myelogenous leukemia(CML) cells, K562 cells and bcr-abl peptide, respectively. METHODS: T cells in donor's peripheral blood were stimulated with CML cells, K562 cells and bcr3-abl2 peptide and amplified by MLTC, to induce the CML specific cytotoxic T lymphocytes. The induced T cells were further analyzed for the restricted usage and clonal expansion of TCR Vβ subfamilies by using RT-PCR and genescan analysis, and the detection of specific cytotoxicity in CML by LDH release assay. RESULTS: 10-13 Vβ subfamilies were expressed in T cells from donor peripheral blood which were induced with CML cells, K562 cells and bcr-abl peptide in 1-2 weeks by MLTC. Oligoclonal T cell in Vβ16, Vβ21 and oligoclonal tendency T cells in Vβ5, Vβ13 subfamilies were identified in induced T cells, which have the ability of specific cytoxicity to CML cells and K562 cells. CONCLUSION: The anti-CML cytotocity T cells were induced by CML cells, K562 cells and bcr-abl peptide. These induced T cells with specific cytoxicity effect may come from the clonal expansion TCR Vβ subfamily T cells.