选择性去除表达CD25^+、CD69^+的同种异体反应T细胞的实验研究

A study on selective depletion of alloreactive donor T cells based on expression of CD25^+ and CD69^+

目的探讨体外选择性去除表达CD25+、CD69+的同种异体反应T细胞协调移植物抗宿主病和移植物抗肿瘤的可能性。方法流式细胞仪检测初次单向混合淋巴细胞反应CD25+、CD69+表达情况,磁性细胞分离系统在细胞表达高峰去除CD25+、CD69+同种异体反应T细胞。MTT法在各时间点检测不同抗原组再次混合淋巴细胞反应的细胞增殖情况。结果再次单向混合淋巴细胞反应显示明显降低了对相同刺激原的反应性(48·82±13·71)%,保留大部分对无关刺激原的反应性(65·47±9·84)%和肿瘤刺激原的反应性(64·83±3·65)%。结论体外选择性去除表达CD25+、CD69+的同种异体反应T细胞可以协调移植物抗宿主病和移植物抗肿瘤。

Objective To study the possibility of selective depletion of alloreactive donor T cells based on expression of CD25^＋ and CD69^＋ to coordinate graft-versus-host disease（GVHD） and graftversus-tumor（GVT）. Methods Eexpressions of both CD25^＋ and CD69^＋ in first mixed lymphocyte culture（MLC） samples were analyzed by flow cytometry. Alloreactive lymphocytes were identified by expression of both CD25^＋ and CD69^＋ , and selectively removed by magnetic cell sorting（MACS）. Cells prolification in secondary MLC samples was assayed at the indicated time points by MTT, Results Mixed lymphocyte culture assays demonstrated that this technique led to a significant decrease in alloreactivity of donor cells 48. 82±13.71%, which at the same time preserved reactivity against third party cells 65.47±9.84% and anti-tumor cells 64.83±3.65%. Conclusion The selective depletion of alloreactive donor T cells based on expression of two activation-induced antigens CD25^＋ and CD69^＋ can coordinate GVHD and GVT.