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

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

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-α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。

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.