肺癌可溶性抗原联合超抗原修饰致敏DC诱导抗瘤免疫的研究  

Study on anti-tumor immunoreaction induced by dendritic cells modified by lung cancer soluble antigen and superantigen

作　　者：马昌云[1] 吴芳[1] 孔繁义[1] 戴国光[1] 

机构地区：[1]沧州市中心医院胸外科,河北沧州061001

出　　处：《临床肿瘤学杂志》2010年第1期21-25,共5页Chinese Clinical Oncology

摘　　要：目的观察经肺癌肿瘤可溶性抗原(TSA)和超抗原金黄色葡萄球菌肠毒素A(SEA)联合修饰致敏树突状细胞(DC)体外诱导抗肺癌的免疫效应。方法3mol/L氯化钾提取法获得人肺癌细胞GLC-82的可溶性抗原;从人外周血单个核细胞(PBMC)中诱导扩增DC,并用流式细胞仪(FCM)检测表型;以肺癌TSA和SEA联合修饰致敏的DC、单纯肺癌抗原致敏的DC和未经抗原修饰的DC分别与同种异体外周血T淋巴细胞共同孵育,刺激T淋巴细胞活化增殖(作为效应细胞分别称为TSA-SEA-DCL、TSA-DCL、DCL),直接活细胞计数法观察增殖倍数;MTT法检测不同DC:T淋巴细胞比例的效应细胞对靶细胞GLC-82的体外杀伤效应。结果诱导出高表达CD1a、CD80、HLA-DR的DC,光镜下具有典型的DC特性;联合抗原修饰后的DC具有较强的免疫刺激活性,少量致敏DC即可强烈激发T细胞的增殖;TSA-SEA-DCL对靶细胞GLC-82的杀伤率明显高于TSA-DCL及DCL;联合抗原修饰的DC以1:100与T淋巴细胞共孵后的杀伤肿瘤细胞效应最强。结论经肺癌TSA和SEA联合修饰致敏的DC可强烈激发同种异体T淋巴细胞活化增殖;肺癌TSA联合超抗原SEA诱导的DC疫苗对肺癌细胞有高效杀伤作用,经肺癌TSA与超抗原SEA联合修饰DC的活性明显强于单用肺癌TSA。Objective Through the study on anti-tumor immunoreaction against lung cancer induced by dendritic cells（DC） modified by lung cancer tumor soluble antigen （TSA） and superantigen（ staphylococcal enterotoxin A, SEA） in vitro, we provide definite experiment basis for the clinical application of vaccine based on DC. Methods Through the experiment we got the soluble antigen polypeptide by 3mol/L potassium chloride. DCs were cultured and obtained from peripheral blood mononuclear cell. Phenotype of DC was identified by flow cytometer（FCM）. DCs modified by TSA and SEA, DCs modified by TSA or DCs modified by nothing were cultivated together with T lymphocyte, and the obtained cells were named TSA-SEA-DCL or TSA-DCL or DCL as effector cells. T cells were amplified and activated. Proliferation multiple of T cells was counted. The anti-tumor activity of every effector cells was assayed with MTT method. Results Induced DCs expressed more CDla, CD80 and HLA-DR, which had typical cell trait such as tree branch. The stimulating T cell activity of DC modified by SEA and TSA was strong. The kill ratio of the TSA-SEA-DCL in GLC-82 cell line was larger than TSA-DCL and DCL in vitro. DCs that modified by TSA and SEA and cultivated together with T lymphocyte with a proportion of 1：100 had the strongest anti-tumor effects. Conclusion DCs modified by TSA and SEA can activate T cells strongly. High potency anti-lung caner effect can be prepared in virtue of DC bacterin induced by lung caner TSA and SEA. The activity of DC modified by TSA and SEA is stronger than by TSA alone.

关 键 词：树突状细胞 肿瘤可溶性抗原 金黄色葡萄球菌肠毒素A 肺癌 

分 类 号：R734.2[医药卫生—肿瘤]