荷载人生存素的多表位树突状细胞疫苗的抗肿瘤活性研究

目的:观察荷载人生存素(survivin)的多表位树突状细胞(DC)疫苗的抗肿瘤活性.方法:分别将含4个sur-vivin的HLA-A2类限制性CD8+ CTL表位和1个CD4+ Th细胞表位的重组真核表达质粒pPIRESneo3.0-survivin(4)/Th,以及含4个CD8+ CTL表位的重组质粒pPIRESneo3.0-survivin (4),转染人DC并制备DC疫苗.实验分为survivin(4)/Th组、survivin (4)组、空质粒组、未转染DC与T细胞共培养组和单独T淋巴细胞组.DC疫苗作用后,MCF-7细胞的凋亡率明显高于survivin(4)组(P＜0.05),亦明显高于空质粒组、未转染DC与T细胞共培养组和单独T淋巴细胞组(P＜0.05).运用流式细胞术(FCM)分别检测DC表面CD83、CD86、T淋巴细胞表面CD4、CD8a的表达以及DC疫苗作用后MCF-7乳腺癌细胞的凋亡.用ELISA法检测上清中IFN-γ的含量.用四甲基偶氮唑蓝(MTT)比色法检测DC疫苗诱导的CTL对MCF-7细胞的抑制率.结果:流式细胞术检测显示,人DC高表达CD83、CD86;人外周血T淋巴细胞高表达CD4、CD8a;survivin (4)/Th组IFN-γ的含量[(66.50±3.34) ng/L]明显高于survivin(4)组[(46.10±1.35)ng/L]、空质粒组[(25.17±0.32) ng/L]、未转染DC与T细胞共培养组[(25.47±0.95)ng/L]和单独T淋巴细胞组[(23.73 +0.50)ng/L],P＜0.05.survivin (4)/Th组中MCF-7细胞的抑制率明显高于survivin (4)组、空质粒组、未转染DC与T细胞共培养组和单独T淋巴细胞组(P＜0.05).DC疫苗作用后,MCF-7细胞的凋亡率[ (10.63±0.29)％]明显高于survivin(4)组(P＜0.05),亦明显高于空质粒组、未转染DC与T细胞共培养组和单独T淋巴细胞组(P＜0.05).结论:荷载多个survivin的CD8+ CTL表位的树突状细胞肿瘤疫苗具有很强的抗肿瘤活性,CD4+ Th细胞对CD8+ CTL的抗肿瘤方面有明显的促进作用.

The study of anti-tumor activities of DC vaccine loaded with multi-epotipes of survivin

AIM: To observe the anti-tumor activity of dendritic cell(DC)vaccine loaded with multi-epitopes of survivin.METHODS: The recombinant plasmid pPIRESneo3.0-survivin(4)/Th which include four survivin HLA-A2-restricted CD8+ CTL epitopes and a CD4+Th epitope,pPIRESneo3.0-survivin(4) which include four survivin CD8+ CTL epitopes,were transfected into human dendritic cells respectively.There were five groups,which included survivin(4)/Th group,survivin(4)group,empty plasmid group,untransfected group and T lymphocy...