摘要
目的 :了解急性粒细胞白血病 (AML)M2a细胞诱导自体及异体T细胞受体 (TCR)Vβ亚家族T细胞克隆性增殖情况及其细胞毒性。方法 :采用混合淋巴细胞 /肿瘤细胞培养 (MLTC)方法 ,用M2a细胞体外诱导正常人和M2a患者自体的外周血T细胞 ,用RT -PCR扩增经MLTC获得的自体和异体T细胞的 2 4个TCRVβ亚家族基因的互补决定区 (CDR3) ,阳性产物进一步经基因扫描分析确定M2a细胞诱导的T细胞克隆性特点。同时用MTT法对淋巴细胞因子激活的杀伤细胞 (LAK细胞 )和MLTC获得的T细胞进行细胞毒性分析。结果 :M2a细胞诱导的自体及异体T细胞均出现了 4 - 17个TCRVβ亚家族的优势表达 ,部分表达的Vβ亚家族T细胞具有克隆性增殖的特点。MLTC的不同时期和不同个体来源对TCRVβ亚家族T细胞表达及克隆性增殖有所影响。与LAK细胞相比 ,MLTC获得的T细胞以CD8+ T细胞为主 ,对M2a细胞具有较高的特异性细胞毒性作用。结论 :MLTC获得的TCRVβ亚家族克隆性T细胞是自体和异体T细胞对M2a细胞刺激所产生的一种特异性细胞免疫反应。M2a细胞诱导后的T细胞对M2a细胞具有特异性细胞毒性作用。
AIM: To investigate clonal expansion and specific cytotoxicity of TCR Vβ subfamily T cells from acute myelogenous leukemia M_ 2a subtype (AML-M_ 2a)patients and normal individuals induced by AML-M_ 2a cells, respectively. METHODS: Complementarity determining region 3(CDR3) of TCR β with variable region genes was amplified in autologous or allogeneic T cells from mixed lymphocyte and tumor culture (MLTC) using RT-PCR. The positive products were further analyzed to identify the clonality of T cells by genescan. The specific cytotoxicity of T cells was analyzed by MTT. RESULTS: T cells from both M_ 2a patients and normal individuals after MLTC showed high response to M_ 2a cells with 4-17 TCR Vβ subfamily dominant utilization, one or two clonal expansion of T cells were identified in some predominant TCR Vβ subfamilies. Difference of distribution and clonal expansion of TCR Vβ subfamily T cells were related to source of T cells and the phase during MLTC. Compared with LAK cell, most of T cells from MLTC were CD3+CD8+T cells with higher and more specific cytoxicity to the induced cells, M_ 2a cells, but not HL60 or K562 cell line. CONCLUSION: Clonal expansion of TCR Vβ subfamily T cells stimulated selectively by M_ 2a cells may be a specific immune response of autologous and allogeneic T cells to M_ 2a cells associated antigen. The T cells induced by M_ 2a cells have the ability of specific cytoxicity to the AML-M_ 2a cells.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2002年第10期1196-1200,共5页
Chinese Journal of Pathophysiology
基金
国家自然科学基金 (No.39870 35 8)
广东省自然科学基金 (No .980 70 5 )
广州市科委重点科技攻关计划 (98-Z -0 39- 0 1)
广东省医学科研基金 (A19992 71)资助项目
