PC-3冻融抗原致敏DC诱导CTL杀伤PC-3细胞体外实验研究  

Study on Cytotoxic T Lymphocyte Activation by PC-3 Antigen-loaded Dendritic cells for Killing of PC-3 Cells in Vitro

作　　者：周海滨[1] 付强[1] 汪亮亮[1] 唐青[1] 廖鑫鑫[1] 

机构地区：[1]九江学院附属医院泌尿外科,江西九江332000

出　　处：《赣南医学院学报》2015年第4期510-513,共4页JOURNAL OF GANNAN MEDICAL UNIVERSITY

基　　金：九江市科技支撑计划项目(20135011)

摘　　要：目的:研究PC-3冻融抗原致敏的树突状细胞(DC)体外诱导的细胞毒性T淋巴细胞(CTL)对PC-3细胞的杀伤效应。方法:冻融法制备PC-3细胞裂解物抗原成分;联合应用rh GM-CSF、rh IL-4和rh TNF-α体外诱导人外周血单个核细胞(PBMC)分化出成熟DC,并用PC-3细胞冻融抗原致敏DC;用LDH释放法检测,对比PC-3细胞冻融抗原致敏DC组、未致敏的DC组、淋巴细胞组、K562冻融抗原致敏DC组分别激活CTL对PC-3细胞的杀伤作用。结果:PBMC在细胞因子rh GM-CSF、rh IL-4和rh TNF-α的联合诱导下分化出成熟DC;在靶效比为10∶1及20∶1时,PC-3冻融抗原致敏的DC组体外诱导的CTL对PC-3细胞的杀伤率分别为(58.2±16.0)%、(92.1±23.3)%,均高于其它对照组,差异有统计学意义(P<0.05)。结论:PC-3冻融抗原致敏的DC在可诱导激活的CTL,在体外对PC-3细胞具有明显的特异性杀伤作用。Objective： To study the effect of dendritic cells（ DC） stimulated with PC-3 cells lysate in inducing specific cytotoxic T lymphocytes（ CTL） against PC-3 cells in vitro. Methods： Tumor antigens were extracted from the prostate cancer line PC-3 with the repeated freeze-thaw method. After separation from heparinized venous blood of healthy donors by Ficoll-Hypaque density-gradient centrifugation,peripheral blood mononuclear cells（ PBMC） were cultured and induced by rh GM-CSF and rh IL-4 and rh TNF-α to differentiate into DCs. The T cells were stimulated by DCs loaded with freeze-thawed PC-3 cells and T-cell cytotoxicities were measured by lactate dehydrogenase（ LDH） assay. Results： The DCs could be successfully obtained from peripheral blood monocytes after the culture with rh GM-CSF,rh IL-4,and rh TNF-α. Mixed lymphocyte reactions induced by the antigen-loaded DCs were significantly stronger than those induced by antigen-unloaded DCs（ P 〈0. 05）. At the effector to target ratio of 10∶ 1 and 20∶ 1,cytotoxicities against PC-3 cells by CTLs derived from culture with the antigen-loaded DCs were the strongest（ P 〈0. 05）. Conclusion： CTLs derived from DCs pulsed with PC-3 cells lysate show effective and specific cytotoxicity against PC-3 cells.

关 键 词：树突状细胞 前列腺癌 免疫治疗 

分 类 号：R730.3[医药卫生—肿瘤]