两种膀胱癌抗原致敏方式对树突状细胞分泌IL-12的影响和意义  

Impact and significance of two kinds of bladder-tumor antigen loading methods on DCs secreting IL-12

作　　者：杨惠祥[1] 高靖[2] 王志勇[1] 于满[1] 

机构地区：[1]承德医学院附属医院泌尿外科,河北承德067000 [2]承德医学院附属医院计算机网络中心,河北承德067000

出　　处：《中国现代医学杂志》2010年第22期3380-3382,3386,共4页China Journal of Modern Medicine

摘　　要：目的研究负载膀胱癌冻融抗原和酸洗抗原肽对树突状细胞(DCs)分泌IL-12的影响和意义。方法反复冻融法和弱酸洗脱法分别获得膀胱癌细胞株BIU-87肿瘤抗原;联合应用重组人GM-CSF、IL-4和TNF-α体外诱导健康志愿者DCs并负载膀胱癌肿瘤抗原;ELISA法检测第3、6、9d培养细胞的上清液,评价DCs分泌IL-12p70的能力。结果联合应用重组人GM-CSF、IL-4和TNF-α可在体外诱导出DCs;经ELISA方法检测,不同时期DCs分泌IL-12的量不同,负载肿瘤抗原DCs较未负载肿瘤抗原DCs有更强的分泌能力,负载酸洗抗原肽组分泌IL-12的量高于负载膀胱癌冻融抗原组(P<0.05)。结论 IL-12在DCs刺激特异性CTLs的过程中可能发挥重要作用,其分泌量受DCs的成熟状态及某些刺激信号的影响,膀胱癌肿瘤抗原是其刺激信号之一。酸洗抗原肽对DC分泌IL-12有更强的刺激能力。【Objective】 To study the impact and significance of two kinds of different bladder-tumor antigen loading methods on DCs secreting IL-12.【Methods】 BIU-87 cells tumor lysate were obtained by frozen and melt repeatedly,while the antigen peptides of BIU -87 cells were obtained by mild acid eluted technique.DCs were generated form human peripheral blood monocytes （PBMCS）,which were isolated from peripheral blood of healthy donors.To induce DCs,the PBMCS were cultured in the presence of rhGM-CS and rhIL-4 and rhTNF-αin vitro for 9 days.Then DCs were loaded with bladder-tumor lysate or peptides.Concentration of IL-12 p70 in the cell culture supernatants were quantified on every 3 days by ELISA technique.【Results】 PBMCs from healthy volunteer could be induced into DCs in vitro.Supernatants of different condition DCs were analyzed for IL-12 p70 secretion by ELISA technique.DCs loaded with tumor antigen secreted significantly higher amount of IL-12 than insensitive DCs （P 0.05）.Among them DCs loaded with tumor antigen peptides had the maximum secretion （P 0.05）.【Conclusions】IL-12 may play an important role in the process of DCs stimulating specific CTLs.The secretion of IL-12 was affected by grown status and some stimulant signals,and bladder-tumor antigen was one of the stimulant signals.DCs loaded with acid eluted bladder-tumor antigen peptides can secrete higher amount of IL-12.

关 键 词：白细胞介素-12 树突状细胞 膀胱癌 生物治疗 

分 类 号：R737.14[医药卫生—肿瘤]