肺癌可溶性抗原与树突状细胞构建肿瘤疫苗的杀伤机制体外研究  

Preparation of tumor vaccine with soluble antigen of lung cancer and dendritic cells and study in vitro on its killing mechanism

作　　者：关晓海[1] 杨文凯[1] 伊雪[2] 李占清[1] 邬鹏宇[1] 张宇[1] 陈国生[1] 

机构地区：[1]华北煤炭医学院附属医院心胸外科,河北唐山063000 [2]华北煤炭医学院附属医院病理科,河北唐山063000

出　　处：《第四军医大学学报》2007年第9期806-808,共3页Journal of the Fourth Military Medical University

基　　金：河北省科技厅资助项目(0127611030)

摘　　要：目的:通过肺癌的可溶性抗原致敏的树突状细胞(DC)激活细胞毒性T细胞(CTL)的杀伤作用各个环节的研究,初步探讨其作用机制及影响条件.方法:从健康人外周血中提取外周血单核细胞(PBMC),加入GM-CSF和IL-4后,在第4日加入不同的促成熟因子,观察DC的表型变化及测定上清中的细胞因子情况.通过DC表面HLA-DR的表达来比较不同抗原致敏DC的优劣.应用不同浓度的肿瘤可溶性抗原(TSA),DC负载TSA的不同时间,不同活化时间的CTL及不同TAK与GLC-82细胞的效靶比来测定其中各种引起TAK最高杀伤活力的情况.结果:DC较好的促成熟因子是MCM,第7日时达到高峰;TSA是较好的致敏DC抗原;最佳效靶比为1∶50.结论:肺癌可溶性抗原是良好的DC致敏物,可在单核细胞条件培养基(MCM)协助下致敏并使DC成熟,表达更多的HLA-DR,高效激活T细胞.AIM： To explore the mechanism and conditions of activating the killing effect of cytotoxic T lymphocyte （CTL） by dendritic cells （DC） sensitized by soluble antigen of lung cancer. METHODS： Peripheral blood mononuclear cell （PBMC） was drawn from healthy person, then cultured in the medium where GM-CSF and IL-d were added, and different maturation promoting factors were added to the medium at the dth day. Observe the change of DC phenotype and measure the expressions of cytokines in the supernatant. We compared the effect of different antigens through observing the emergence of HLA-DR of DC. We applyed different concentrations of tumor soluble antigen （ TSA）, different loading time of TSA, different activated time of CTL and different ratios of GLC82 to TAK to survey the suitable conditions for activating TAK with most killing ability. RESULTS： The best maturation promoting factor for DC was MCM. DC reached the climax on the 7th day. TSA was the better antigen used to stimulate DC, and it can induce more DC with HLA-DR. The most fit effector/target cell ratio was 1：50. CONCLUSION： Soluble antigen of lung cancer is a better matter for triggering DC with more HLA- DR. It can mature DC with the aid of MCM. The T cells can be activated effectively by DC sensitized by TSA.

关 键 词：树突状细胞 可溶性抗原 抗原呈递 T淋巴细胞 细胞毒性 

分 类 号：R730.54[医药卫生—肿瘤]