负载冻融抗原的树突状细胞诱导特异性抗膀胱癌作用的研究

目的研究负载膀胱癌冻融抗原的树突状细胞（DC）诱导的对膀胱癌细胞的特异性杀伤效应。方法反复冻融法获得BIU-87细胞抗原；人单个核细胞在含重组人GM—CSF、IL-4和TNF-a的RPMI1640培养基中体外诱导出人DC并负载肿瘤抗原；9d后成熟DC与自体T细胞共孵育诱导膀胱癌抗原特异性CTLs； 用MTT法检测其对BIU-87体外杀伤效应；用ELISA法检测DC分泌IL-12的能力。结果负载膀胱癌抗原的DC诱导的特异性CTLs可明显杀伤BIU-87细胞，在效靶比为40：1时杀伤率为（65．5±6．4）％，显著高于各对照组（P〈0．01）；负载抗原DCs较未负载抗原DCs有更强的分泌IL-12能力（P〈0．05），而且不同时期的DC分泌的量不同。结论负载膀胱癌抗原的DC在体外可诱导出高效而特异的抗膀胱癌效应，提示以DC为中心的肿瘤生物治疗有望提高膀胱癌综合治疗水平。

Experimental study of specific cytotoxicity induced by dendrtic cells pulsed with tumor lysate towards bladder cancer cell line

Objective To observe specific cytotoxicity induced by dendrtic cells pulsed with bladder tumor lysate towards bladder cancer cell line. Methods BIU-87 ceils （human bladder transitional cell carcinoma cell line） were frozen and melt repeatedly, then tumor lysate was obtained. Human PBMCs were cultured with RPMl1640 contained rhGM-CS＋ rhIL-4＋ rhTNF-αin vitro, in the meantime pulsed with bladder tumor lysate. In the nineth day matured DCs were harvested, which were then cultured with T Cells, and induced tumor specific CTLs. Specific cytotoxicity caused by the CTLs towards bladder cancer ceils could be detected through MTT technique. DC could also excrete IL-12, which were able to be detected by ELISA technique. Results Tumor specific CTLs induced by DCs pulsed with bladder tumor lysate could kill about （65.5± 6.4）% of BIU-87 cell line when the ratio was 40 ： 1, which was significantly stronger than control groups （P〈0. 01）. The excreting IL-12 capacity of DCs sensitized by bladder tumor cell antigen was stronger than that of insensitive DCs, and the quantity was different in different stages （P 〈 0. 05）. Conclusion DC loaded bladder tumor lysate can induce highly effective and remarkably specific cytotoxicity in vitro, which indicates that anti-tumor biological therapy will be expected to improve the overall therapeutic level of bladder tumor.