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

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 3M KCI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;M1T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sE^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.

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

Background and objective Through the study on E ect 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 soluble 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...