肺癌可溶性抗原联合超抗原诱导的DC疫苗对肺癌细胞杀伤作用的研究  

Study On Effect of Dendritic Cells Bacterin Induced by Superantigen and Lung Cancer Soluble Antigen Against Lung Cancer Cells

作　　者：马昌云[1] 吴芳[1] 孔繁义[1] 韩丽荣[1] 

机构地区：[1]沧州市中心医院胸外科,沧州061001

出　　处：《中国肺癌杂志》2009年第6期539-542,共4页Chinese Journal of Lung Cancer

摘　　要：背景与目的通过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)体外诱导对肺癌细胞杀伤作用的研究,为以DC为基础的肺癌免疫瘤苗的临床应用提供一定的实验依据。方法3MKCl法提取人肺癌可溶性抗原;从人外周血单个核细胞(PBMC)中诱导扩增DC并鉴定;肺癌TSA和不同质量浓度的SEA联合修饰致敏DC;以联合抗原修饰后的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T淋巴细胞共同孵育,MTT法测定混合淋巴细胞反应中DC的免疫刺激活性,诱导产生具有识别肺癌抗原的特异性CTL(作为效应细胞分别称为TSA-SEA-DCL、TSA-DCL、DCL);用流式细胞仪FCS及免疫染色法分析鉴定DC和效应细胞表型;MTT法检测各效应细胞对靶细胞体外杀伤效应。结果诱导出表达CD1a,CD80,HLA-DR分子的DC;经肺癌TSA和SEA联合修饰致敏后,上述分子表达上调;联合修饰后的DC具有较强的免疫刺激活性,DC/T比例1:10可能为最适比例;TSA-SEA-DCL中CD3+CD8+细胞的比例大幅度增加;TSA-SEA-DCL对靶细胞GLC-82的杀伤率明显高于TSA-DCL及DCL,亦明显高于对肺癌CALU-6和人红白血病K562细胞的杀伤率。结论经肺癌TSA和SEA联合修饰致敏的DC可诱导同种异体T淋巴细胞活化增殖产生CD8+表达增加的CTL;联合抗原诱导的DC疫苗对肺癌细胞有高效特异性的杀伤作用,肺癌TSA和超抗原SEA联合修饰的DC的活性明显强于单用肺癌TSA修饰。Background and objective Through the study on Effect of Dendritic Cells Bacterin Induced by Superantigen （staphylococcal enterotox in A, SEA） and Lung Cancer Soluble Antigen （TSA） Against Lung Cancer Cells in vitro, we provide definite experiment basis for the clinic application of vaccine based on DC. The experiment got the solul^le antigen polypeptide through 3M kcl. Lung Cancer TSA and SEA was used to trigger the DC extracting from people peripheral blood and induced ,We produced the DC loaded with antigen polypeptide and SEA, made the T lymphocyte from healthy person＇s peripheral blood, amplified and activated（as initial T lymphocyte）, induced CTL which can recognize lung cancer antigen. DC modified by TSA and SEA, DC modified by TSA or DC modified by nothing is incubated together with T lymphocyte and the obtained cellswere named TSA-SEA-DCL or TSA-DCL or DCL as effector cells. The activity of T lymphocyte excited by DC was assayed with MTT method in Mixed lymphocyte response. Phenotypes of DC and effector cells were analysed by FCS and immunity coloration. MTr assay was used to determine cytotoxicity t of every effector cells. Results Induced DC expressed CD 1a, CD80 and HLA-DR cell. Through modification of SEA and lung cancer TSA, its expression upregulated. The stimulating T cell activity of DC modified by SEB and TSA is strong. The DC/T, 1：10 is the most suitable proportion. The ratio of CD^3＋ and CD^8＋ T cell is rising. The killing ratio of the TSA-SEA-DCL in vitro to GLC-82 is larger than TSA-DCL and DCL, also larger than in vitro to CALU-6 and K562. Conclusion DC modified by SEB and TSA can induce CTL expressed more CD8＋. High potency and relatively specific anti-lung caner immunocyte can be prepared in virtue ofDC Bacterin Induced by TSA and SEA. The activity of DC modified by TSA and SEA is stronger than by only TSA of lung caner.

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]