AML-M_(2a)细胞体内外诱导TCR Vβ亚家族T细胞克隆性增殖情况的研究

Study on Clonal Proliferation of TCR Vβ Subfamily T Cells Induced by AML-M_(2a) Cells In Vivo and In Vitro

为了解AML-M_(2a)细胞体外诱导TCR Vβ亚家族T细胞的表达和克隆性增殖情况,利用混合淋巴细胞/肿瘤细胞培养(MLTC)体外获取AML-M_(2a)细胞诱导的T细胞,并以RT-PCR扩增4例M_(2a)患者外周血单个核细胞和4例AML-M_(2a)细胞诱导的T细胞中的24个TCR Vβ亚家族基因的互补决定区3(CDR3)。阳性产物进一步经基因扫描分析从而确定AML-M_(2a)细胞体内外诱导的TCR Vβ亚家族T细胞的克隆性增殖情况。结果显示:4例AML-M_(2a)细胞体内外访导的Vβ亚家族T细胞表达情况相似,其中3例AML-M_(2a)细胞体内外诱导的一个或两个Vβ亚家族T细胞里克隆性增殖特点。结论:AML-M_(2a)细胞体内外选择性诱导不同的TCR Vβ亚家族T细胞克隆性增殖可能是患者T细胞对AML-M_(2a)细胞相关抗原刺激所产生的一种特异性细胞免疫反应。体内外环境的不同对Vβ亚家族T细胞的克隆性增殖情况有一定的影响。

To observe the expression and clonal expansion of TCR V(5 subfamily T ceils induced by AML-Ma.cells in vivo and in vitro,complementary determining region 3 ( CDR3) of TCR B with variable region genes was amplified by using RT-PCR in both peripheral blood mononuclear cells(PBMNC) and T cells from mixed lymphocyte and tumor culture (MLTC) from four AML-M2a patients.The positive products were further analyzed to identify the clonality of T cells by genescan.The results showed that the similarity distribution of TCR Vp subfamily T cells was found in PBMNC and MLTC.One or two clonality expansion of T cells could be found in predominant TCR Vfi subfamily T cells induced by AML-M2,cells from 3 cases in vivo and in vitro.It was concluded that clonal expansion of TCR VB subfamily T cells stimulated selectively by AML-M2,cells may be a specific immune response of patient s T cells to AML-M2,cells associated antigen.The clonal proliferation of TCR VB subfamily T cells were affected somewhat by environmental difference in vivo and in vitro.