中文摘要：

目的研究应用脑胶质瘤于细胞抗原制备的树突状细胞（DC）疫苗对胶质瘤细胞的杀伤作用。方法反复冻融法获得源于胶质瘤细胞系U251中的肿瘤干细胞的胞溶物，和源于小鼠间充质干细胞的DC共培养，获得胶质瘤干细胞疫苗。流式细胞仪检测疫苗表面标志变化；CCK-8法检测其促T细胞增殖能力以及对胶质瘤细胞的靶向杀伤作用；ELISA法检测培养基中干扰素-γ的含量。用热处理U251细胞制备的DC疫苗作为对照组。结果与对照组相比，脑胶质瘤干细胞疫苗的CD80、CD86、CD11c和MHCU等表面标志表达显著提高，具有更强的促T细胞增殖能力（P〈0．01），对U251细胞特异性靶向杀伤率随效靶比的提高而增加，最高为（78．508±4．156）％，相应的干扰素-γ分泌水平也达到最高。结论胶质瘤干细胞疫苗能更有效地诱导特异性细胞毒性T细胞，表现出更强的抗肿瘤特性。

英文摘要：

Objective To explore the ability of anti - glioma cells and the mechanism of specific T cells induced by dendritic cells loaded with the antigen of glioma stem cells. Methods The cancer stem cells were cultured from the U251 cell line. The lysate of glioma stem cells was obtained through the repeated freezing and thawing method. Dendritic cells （DCs） were prepared from mouse mesenchymal stem cells. The glioma stem cell vaccine was produced by mixing tumor stem cell lysate with DCs. Then, the surface markers of DCs were checked by flowcytometry. CCK - 8 method was used to detect the DC vaccine＇ s ability of promoting T cells proliferation and killing U251 cells. The level of IFN - γ in the supernatant was checked by ELISA. Dendritic ceils pulsed with heat -treated U251 were used as the control group. Results After the stimulation glioma stem cell lysate, the expression of surface molecules of DCs was up - regulated, including CD80 （95.10 ± 0. 44 ） %, CD86 （ 92. 90 ± 4. 85 ） %, CD11C （ 75.53 ± 0. 50 ） %, and MHC II （94. 27 ±2. 30）%. Compared with control groups, glioma stem cell vaccines could stimulate the proliferation of T cells more strongly, induce more effective immunoresponse against glioma cells （the highest killing rate was （78. 508 ± 4. 156）% at effector： target ratio 80：1 ）, and strongly boost the secretion of interferon- ±/. Conclusions Dendritic cells pulsed with glioma stem cells＇ lysate can effectively induce specific cytotoxic T cells, show strong anti - tumor properties, and provide a new potent way for the immunotherapy of human brain malignant gliomas with dendritic cell vaccines.