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1×10 7FBL3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5 d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10 d后加入小剂量的IL-2。 结果： 共培养至21 d， T细胞可扩增100倍以上，其中CD8+细胞比率明显增高。乳酸脱氢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 1×10 7 DCs pulsed with 5×10 7 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.

国家863计划（2002AA205051）资助项目