DLI治疗CML病人的克隆性增殖TCR Vβ细胞分析

摘要

Objective: To investigate the clonal expansion of TCR Vβsubfamily T c ells which were considered as GVL effective cells after donor lymphocytes infusi on (DLI) in patients with relapse CML after allo-BMT. Methods: The CDR3 of TCR Vβ 24 subfamily genes were amplified in samples of peripheral blood mononuclear c ells at different time points before and after DLI, which were drawn from 2 case s with relapse CML treated by allo-BMT, to observe the usage of TCR Vβ reperto ir e, the PCR products were further labeled with fluorescent and analyzed by genesc an technique for the CDR3 size, to evaluating clonality of the detectable TCR V β T cells.Results:Only 4-11 Vβ subfamily T cells could be identified in CML ca ses before DLI, and 12-21 Vβ subfamily T cells could be detected in samples fr om CML which display remission after DLI. Genescan analysis showed that new clona l expansion TCR Vβ subfamily T cells could be found in samples after DLI. Conclusion: The skew distribution of TCR Vβ subfamily T cells could be found on patie n ts with relapse CML after allo-BMT, and this skewing pattern may stage to stage to normal pattern during the complete remission. The GVL effect may exert throu g h some clonal expansion TCR Vβ subfamily T cells, during the treatment of DLI i n relapse CML.%目的：了解供者淋巴细胞输注（DLI）后，产生GVL效应的克隆性TCRVβ亚家族T细胞的情况。方法：利用RT-PCR分别扩增2例CML经allo-BMT后复发病人DIL治疗前后不同时间外周血单个核细胞的TCRVβ24个亚家族基因的CDR3，了解病人各Vβ亚家族的利用情况。阳性的PCR产物进一步经荧光素标记和基因扫描分析产物的CDR3长度，了解T细胞克隆性。结果：DLI治疗前病人外周血仅表达4～11个Vβ亚家族，而DLI治疗后缓解期则可检测到12～21个Vβ亚家族。基因扫描显示DLI后病人外周血出现某些新的Vβ亚家族克隆性增殖T细胞。结论：病人的TCRVβ亚家族T细胞存在倾斜性分布现象，该倾斜现象在完全缓解时逐步恢复正常。DLI治疗病人时，可能通过某些克隆性增殖TCRVβ亚家族T细胞而发挥GVL作用。