摘要
目的:观察重组疫苗E.coli LLO/WT1诱导人外周血树突状细胞(dendritic cell,DC)成熟及其致敏的T细胞对WT1+肿瘤细胞的杀伤作用.方法:以E.coli为对照,采用E.coliLLO/WT1刺激健康人外周血树突状细胞后,流式细胞术分析DC的CD83,CD86和HLA-DR的表达,MTT法观察活化DC促进T细胞增殖和杀伤WT1+人胃癌细胞SGC-7901的效果,ELISA检测DC及DC和T细胞混合培养上清液内IL-12和IFN-γ的含量.结果:E.coliLLO/WT1较E.coli刺激的DC表达更高水平的CD83,CD86和HLA-DR,其阳性率分别为78.1%,83.4%,93.2%和65.8%,61.2%,75.8%;两组DC培养上清内IL-12的含量分别为(270.42±3.85)ng/L和(204.17±4.23)ng/L(P<0.05);E.coli LLO/WT1活化的DC较E.coli活化的DC更明显地促进了同源T细胞增殖,混合培养细胞上清液内IFN-γ的含量也更高,分别为(86.32±2.00)ng/L和(44.71±1.45)ng/L(P<0.05);活化的T细胞发挥了更强的溶解WT1+SGC-7901的作用,其杀伤率分别为68.44±3.78和36.76±2.63(P<0.05).结论:重组疫苗E.coliLLO/WT1有效地活化了人外周血树突状细胞,并促进了WT1+特异性抗肿瘤T细胞免疫.
AIM：To investigate the role of recombinant E.coli LLO/WT1 in inducing the maturation of human peripheral blood dendritic cells（DC）and promoting WT1＋ tumor resolving effect of the activated T cells.METHODS：After stimulated by E.coli LLO/WT1 and E.coli respectively,the surface molecules expression of DC was determined by flow cytometry;the proliferation and WT1＋ SGC-7901 cells resolving effect of the activated T cells were determined by MTT.The concentration of IL-12 and IFN-γ in the supernatant of DC and co-cultured DC and T cells were determined by ELISA respectively.RESULTS：Compared with E.coli,E.coli LLO/WT1 stimulated DC expressing higher level of CD83,CD86 and HLA-DR,the positive rates were 78.1%,83.4%,93.2% and 65.8%,61.2%,75.8%;the activated DC secreted higher level of IL-12 also,the concentration of IL-12 in the supernatant was（270.42±3.85）and（204.17±4.23）ng/L（P〈0.05）.E.coli LLO/WT1 activated DC promoted more autogenic T cell proliferation and secreted higher level IFN-γ than.E.coli activated DC,the concentration of IFN-γ in the supernatant was（86.32±2.00）and（44.71±1.45）ng/L（P〈0.05）;and the proliferated T cells resolved more WT1＋ SGC-7901 cells,the killing rates were 68.44±3.78 and 36.76±2.63（P〈0.05）.CONCLUSION：Recombinant E.coli LLO/WT1 effectively stimulates human peripheral blood DC maturation and promote WT1＋ specific anti-tumor T cell immunity.
出处
《第四军医大学学报》
北大核心
2009年第19期1887-1890,共4页
Journal of the Fourth Military Medical University
基金
重庆市教委基金(KJ080319)
