重组人IL-2激活外周血干细胞移植物的抗白血病细胞毒作用

Anti-leukemia cytotoxicity of peripheral blood stem cell grafts activated by rhIL-2

目的 :观察重组人IL 2体外激活G CSF动员的健康供者外周血干细胞 ( peripheralbloodstemcell,PBSC)移植物的抗白血病细胞毒作用 ,以及rhIL 2处理对PBSC移植物造血功能的影响。方法 :细胞培养条件下 ,PBSC移植物与不同浓度rhIL 2 ( 10 0 0 ,50 0和 2 50U /ml)共培养 2 4及 72h ,采用乳酸脱氢酶 (LDH)释放法测定激活的PBSC移植物细胞毒作用 ,并应用流式细胞术分析细胞表型变化 ;采用集落形成试验测定PBSC移植物形成CFU GM、BFU E的能力。结果 :经rhIL 2体外处理的PBSC移植物 ,其细胞毒作用显著增强 ,不同rhIL 2浓度之间无显著差异 ,培养 2 4及72h则有显著差异。rhIL 2体外处理对PBSC移植物形成CFU GM、BFU E能力无抑制作用 ,且有一定的促进。结论 :提示在临床异基因外周血干细胞移植 (allo PBSCT)前 ,体外以rhIL 2处理移植物 ,可显著增强其抗白血病细胞毒作用 ;有可能在输入体内后诱导和 (或 )增强移植物抗白血病和移植物抗肿瘤效应。

Objective:To investigate the anti-leukemia cytotoxicity of peripheral blood stem cell( PBSC ) graft from normal donor activated by rhIL-2 and the effect of rhIL-2 on hematopoiesis in PBSC grafts. Methods:PBCS collection was incubated with rhIL-2 of various concentrations (1 000 U/ml,500 U/ml and 250 U/ml) at 37℃ in a humidified atmosphere of 5%CO 2 for 24 h or 72 h. Anti-leukemia cytotoxicity was tested in K562 and Raji cell lines with LDH release assay. Cell phenotypes were analyzed by flow cytometry. The ability of PBSC graft to form CFU-GM and BFU-E was evaluated in semi-solid methylcellulose cultures. Results:With rhIL-2 activation, anti-leukemia cytotoxicity of PBSC graft was significantly augmented. The effect of various rhIL-2 concentrations on cytotoxicity had no statistical difference. PBSC graft developed marked cytotoxicity after 24 h of incubation with IL-2, and significant increase in cytotoxicity was seen after 72 h incubation. rhIL-2 Did not inhibit, but,conversely, promoted PBSC to form CFU-GM and BFU-E to some extent. Conclusions:Prior to allo-PBSC transplantation, IL-2 treatment of PBSC in vitro may lead to the generation of anti-leukemia cytotoxicity,which possibly mediates the graft-versus-leukemia effect in vivo when the graft is infused to recipient. [