DC联合小剂量IL-2大量扩增白血病特异性CTL的实验研究  

Large-Scale Expansion of Leukemia Specific CTLs Using Lysates-Pulsed Dendritic Cell and Low-Dose IL-2 for Adoptive Immunotherapy

作　　者：楼敬伟[1] 杨建民[1] 贾新颜[1] 陈莉[1] 章卫平[1] 周红[1] 王健民[1] 

机构地区：[1]第二军医大学附属长海医院血液科,上海200433

出　　处：《中国肿瘤生物治疗杂志》2005年第2期134-137,共4页Chinese Journal of Cancer Biotherapy

基　　金：国家863计划(2002AA205051)资助项目

摘　　要：目的:建立和优化体外大量扩增白血病特异性CTL的方法。方法:用1×107FBL3细胞冻融产物(FBL3-LY)冲击的DC每10天1次对C57BL/6小鼠反复接种,在第3次接种5d后处死小鼠制备脾T细胞,每周用FBL3-LY冲击的DC刺激,体外培养10d后加入小剂量的IL-2。结果:共培养至21d,T细胞可扩增100倍以上,其中CD8+细胞比率明显增高。乳酸脱氧酶释放试验证实扩增所得CTL对FBL3细胞具有高效的杀伤作用(81.84±8.68%),而对K562细胞无特异性杀伤作用。结论:FBL3-LY冲击的DC联合小剂量IL-2可大量扩增FBL3白血病特异性CTL,该方法的建立对提高白血病特异性CTL免疫治疗的临床疗效具有重要意义。Objective:To determine the parameters settings for in vivo induction and in vitro expansion of leukemia-specific CTL. Methods:C57BL/6 mice were vaccinated repeatedly with 1×107 DCs pulsed with 5×107 FBL3- lysates (FBL3-LY) every 10 days, mice were sacrificed and the spleen T cells were collected and purified. Five days after the third immunization,T cells were restimulated weekly with FBL3-HS pulsed DC(30 Gy irradiated) , and low-dose IL-2 was added 10 days following. Results:Three weeks' co-culture of splenocytes T cells from the vaccinated mice with FBL3-LY pulsed DC in the presence of low dose IL-2 resulted in more than 100-folds' expansion of leukemia specific CTLs, which had a higher percentage of CD8+ T cell. The expanded CTLs had potent cytotoxic activity against the parental FBL3 cells (81. 84±8.68%) tested in standard LDH release assay, but no cytolytic activity above background was observed against the k562 leukemia targets. Conclusion:This regime could be considered as practical alternatives to the existing clinical immunotherapy strategies.

关 键 词：树突状细胞 免疫治疗 白血病细胞冻融产物 CTL 

分 类 号：R392.12[医药卫生—免疫学]