半相合供者淋巴细胞输注抗急性非淋巴细胞性白血病的实验研究

Study on haploidentical related donor lymphocyte infusionversus acute nonlymphocytic leukemia in vitro

目的 :阐明半相合供者淋巴细胞 (HRDLs)体外抗急性非淋巴细胞白血病 (ANLL)的机制。方法 :通过预实验 ,确定白血病细胞培养过程中加入HRDLs产生移植物抗白血病效应 (GVL)的最佳时间及最佳浓度 ,并在此条件下检测白血病细胞培养过程加入去CD8+ 和去CD4 + HRDLs后白血病细胞凋亡率及培养液中白细胞介素 2(IL 2 )浓度。结果 :在白血病细胞培养 2 4h(最佳时间 )后 ,加入浓度为 2 .5× 10 8/L(最佳浓度 )的去CD8+ HRDLs混合培养 ,其白血病细胞凋亡率 (17.0± 2 .5 ) %与对照组 (4 .4± 1.1) % (未加入HRDLs)及加入去CD4 + HRDLs组的白血病细胞凋亡率 (4 .8± 1.6 ) %相比较差异有统计学意义 ,但与加入未去CD8+ 和CD4 + HRDLs组的白血病细胞凋亡率 (17.4± 0 .6 ) %比较则差异无统计学意义。同时 ,去CD8+ HRDLs组培养液中IL 2浓度显著高于对照组和去CD4 + HRDLs组。结论 :CD4 + HRDLs在抗白血病效应中可能起着比CD8+ HRDLs更重要的作用 ,临床使用中 ,选择性去除CD8+ HRDLs,可以减少或减轻移植物抗宿主病 (GVHD) ,而不影响GVL效应 ,同时IL

Objective:To elucidate the mechanism of haploidentical related donor lymphocytes (HRDLs) anti acute nonlymphocytic leukemia(ANLL) in vitro.Method:By beforehand experiment, we found the optimal time and infusing dose under which HRDLs could generate graft-versus-leukemia, and by them, we measured the apoptosis rate of leukemic cells when infusing CD8^+/CD4^+ T cell deletion HRDLs. IL-2 concentration was measured in culture medium also.Result:The optimal time is 24 hours, the optimal dose of HRDLs is (2.5)×10~8/L.The difference of leukemic cells apoptosis rate between CD8^+ T cell deletiog HRDLs((17.6)±(2.5))% and control((4.4)±(1.1))%, CD4^+ T cell deletion-CD4^+ HRDLs((4.8)±(1.6))% is significant, but the difference between CD8^+ T cell deletion HRDLs and non-deleting HRDLs is little in ANLL NR period. IL-2 concentration of CD8^+ T cell deletion HRDLs is higher than that of CD4^+ T cell deletion.Conclusion:CD4^+ HRDLs may play more important role in GVL than CD8^+ HRDLs,so in clinical application, selective CD8^+ T cell deletion HRDLs may reduce GVHD ,not interfere with its GVL.IL-2 may be the major mechanism during GVL.