论文部分内容阅读
目的研究AML-M5b患者外周血T细胞TCR Vβ亚家族表达及克隆性增殖特点,并分析分选的患者T细胞经体外诱导、活化及短期培养后其TCR Vβ亚家族的利用及克隆性增殖情况。方法运用RT-PCR-基因扫描技术分析初治AML-M5b患者外周血T细胞TCR Vβ亚家族利用和克隆性增殖,免疫磁珠分选患者T淋巴细胞,体外利用自体白血病DC细胞联合CD3单抗、CD28对T细胞进行诱导、活化,并对诱导后T细胞进行TCR Vβ亚家族表达和克隆性增殖分析。结果9例患者T细胞在体外获得增殖,TCR Vβ分析显示患者外周血T细胞均表达部分Vβ亚家族,体外诱导前后的T细胞TCR Vβ亚家族表达和克隆性增殖特点不完全相同,有部分出现新的Vβ亚家族表达及克隆性增殖,有部分Vβ亚家族在诱导前后始终表现为克隆性增殖,体外杀伤性分析发现诱导后的T细胞对自体白血病细胞具有一定的识别作用。结论AML-M5患者TCR Vβ亚家族T细胞的倾斜性分布和克隆性增殖,部分Vβ亚家族T细胞在诱导前后始终保持克隆性增殖状态,可能是机体针对白血病相关抗原的特异性CTL,体外诱导活化及短期培养后患者T细胞TCR Vβ亚家族表达数量增加,并出现新的克隆性增殖Vβ亚家族T细胞,提示体外诱导活化能够产生白血病患者自体的白血病特异性CTL。
Objective To investigate the TCR Vβ subfamily expression and clonal proliferation in peripheral blood T lymphocyte subsets of AML-M5b patients and to analyze the TCR Vβ subfamily utilization and clonal proliferation in vitro after being induced, activated and cultured in vitro . Methods TCR Vβ subfamilies and clonal proliferation of peripheral blood mononuclear cells (PBMCs) from patients with untreated AML-M5b were analyzed by RT-PCR-gene scanning. T lymphocytes were sorted by immunomagnetic beads. , CD28 on T cell induction, activation, and T cells after induction of TCR Vβ subfamily expression and clonal proliferation analysis. Results T cells from 9 patients proliferated in vitro. TCR Vβ analysis showed that some T cells expressed Vβ subfamilies in peripheral blood. The TCR Vβ subfamilies and clonal proliferative characteristics of T cells before and after T cells were not completely the same The new Vβ subfamily expression and clonal proliferation, some Vβ subfamilies showed clonality proliferation before and after induction, in vitro cytotoxicity analysis showed that the induced T cells have certain recognition of autologous leukemia cells. Conclusions The TCR Vβ subfamily T cells in AML-M5 have a skewed distribution and clonal proliferation. Some Vβ subfamilies T cells maintained clonal proliferation before and after induction, which may be the specific CTLs for leukemia-associated antigens in vitro. After activation and short-term culture, TCR Vβ subfamily T cells increased in number and a new clonal proliferation of Vβ subfamily T cells, suggesting that induction of activation in vitro can produce leukemia-specific leukemia-specific CTL.