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  

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

作　　者：郭萌[1] 王效远[1] 宋文哲[1] 李向农[1] 梁骁[1] 李莹[1] 黄智龙[1] 

机构地区：[1]徐州医学院附属医院普通外科,江苏徐州221002

出　　处：《细胞与分子免疫学杂志》2011年第11期1180-1183,1187,共5页Chinese Journal of Cellular and Molecular Immunology

摘　　要：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。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 lymphocytes group The expression of CD83 and CD86 on the surface of DCs, the expression of CD4 and CD8a on the surface of T lymphocytes, the apoptotic rates of MCF-Уcells after treated by DC vaccine were measured by flow cytometry; IFN-y levels of all groups were detected by ELISA and the growth inhibition of MCF-7cells after being treated with DC vaccine was tested by MTT- colorimetry. RESULTS： The results of flow cytometry revealed that high levels CD83 and CD86 were ex- pressed on the surface of DCs; high levels CD4 and CD8a were expressed on the surface of T lymphocytes; the IFN-y levels in survivin（4）/Th group [ （66.50 ±3.34）ng/Ll were significantly higher than that in survivin （4） group [ （46.10 ±1.35） ng/L], empty plasmid group[ （25.17 ±0.32） ng/L], untransfected group [ （25.47 ±0.95 ） ng/L ] or T lympho- cytes group [ （23. 73 ±0.50） ng/L] （ P 〈 0.05）. The inhibition rate of MCF-7 cells in survivin（4）/Th group was significantly higher than that in survivin（4）group, empty plasmid group, untransfected group or T lymphocytes group （ P 〈 0.05 ）. The apoptotic rate of MCF-7 cells in survivin （4）/Th clroup was （10.63 ±0.29） % after treated by DC vaccine, which was significantly higher than that in in survivin （4）group, empty plasmid group, untransfected group or T lymphocytes group（ P 〈 0. 05）. CONCLUSION： The DCs vaccine loaded with multi- CD8＋ CTL epitopes of survivin has strong antitumor eff

关 键 词：生存素 表位 树突状细胞 肿瘤疫苗 

分 类 号：R730.51[医药卫生—肿瘤]