Fas/FasL途径在造血干细胞移植免疫中的实验研究  

Apoptosis of the allo-antigen specific T cells and leukemic cells induced by Fas/FasL pathway

作　　者：肖娟[1] 邹萍[1] 刘忠文[1] 马艳萍[1] 胡中波[1] 刘凌波[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院血液病研究所,武汉430022

出　　处：《中华器官移植杂志》2003年第6期351-353,共3页Chinese Journal of Organ Transplantation

基　　金：国家自然科学基金资助项目 ( 39770 76 7)

摘　　要：目的　探讨转染FasL的CD34+ 细胞对同种抗原特异性T细胞和白血病细胞凋亡的影响。方法　丝裂霉素C预处理表达外源FasL的骨髓CD34+ 细胞 ,分别与经过富集的T细胞 (刺激细胞 /效应细胞比例为 5∶1)和白血病细胞U937混合培养 ,借助原位末端标记法 (TUNEL)和流式细胞仪 (FCM )检测T细胞和白血病细胞的凋亡率。结果　分别以未转染和转染外源FasL的CD34+ 细胞为刺激细胞 ,5d后 ,T细胞凋亡率分别为 :3.2 %± 1.1%、12 .1%± 1.5 % (P <0 .0 1) ,经IL 2作用后 ,凋亡率上升至 17.6 %± 1.3% (P <0 .0 1) ;与白血病细胞U937混合培养 18h后 ,白血病细胞凋亡率分别为 :5 .0 %± 1.3%、10 .8%± 0 .6 % (P <0 .0 1) ,加入阿糖胞苷后 ,凋亡率上升至 17.9%±1.3% (P <0 .0 1)。结论　转染外源FasL的骨髓CD34+ 细胞 ,可诱导同种抗原特异性T细胞和白血病细胞凋亡。提示通过Fas/FasL途径可选择性清除移植物中同种抗原特异性T细胞 ,有望为造血干细胞移植中防治移植物抗宿主病 (GVHD) ,保留移植物抗白血病效应 (GVL)提供新的策略。Objective To assess the value of CD34 + cells transferred exogenous Fas ligand (FasL) in inducing apoptosis of the allo-antigen specific T cells and leukemic cells.Methods The CD34 + cells were transfected with FasL or not, treated with mitomycin C, either mixed with allo-antigen specific T cells in the presence or absence of IL-2 for 5 days, or mixed with leukaemic cells U937 in the presence or absence of cytosine arabinoside (Ara-C) for 18 h. Apoptosis of T cells and U937 cells were detected by TdT nick end mediated dUTP labeling (TUNEL) and flow cytometry (FCM).Results After being induced by CD34 + cells transfected with FasL or not, the ratio of apoptosis of T cells was 12.1?% ± 1.5?% , 3.2?% ± 1.1?% respectively ( P < 0.01 ); In the presence of IL-2, the ratio reached to 17.6?% ± 1.3?% ( P < 0.01 ); The ratio of apoptosis of U937 was 10.8?% ± 0.6?% , 5.0?% ± 1.3?% respectively ( P < 0.01 ). When being induced by FasL +CD34 ++Ara-C, the ration of apoptosis of T cells was 17.9?% ± 1.3?% ( P < 0.01 ).Conclusion CD34 + cells transfected with exogenous FasL could induce apoptosis of allo-antigen specific T cells and leukaemic cells, suggesting it was possible to provide a new method to prevent GVHD and preserve GVL in hematopoietic stem cell transplantation.

关 键 词：造血干细胞移植 免疫 实验研究 FAS配体 移植物抗宿主病 T淋巴细胞 

分 类 号：R457.7[医药卫生—治疗学]