CAJ | 학술논문

目的研究应用脑胶质瘤干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MHC Ⅱ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.
목적 연구응용뇌효질류간세포항원제비적수돌상세포(DC)역묘대효질류세포적살상작용.방법 반복동융법획득원우효질류세포계U251중적종류간세포적포용물,화원우소서간충질간세포적DC공배양,획득효질류간세포역묘.류식세포의검측역묘표면표지변화；CCK-8법검측기촉T세포증식능력이급대효질류세포적파향살상작용；ELISA법검측배양기중간우소-γ적함량.용열처리U251세포제비적DC역묘작위대조조.결과 여대조조상비,뇌효질류간세포역묘적CD80、CD86、CD11c화MHC Ⅱ등표면표지표체현저제고,구유경강적촉T세포증식능력(P＜0.01),대U251세포특이성파향살상솔수효파비적제고이증가,최고위(78.508±4.156)％,상응적간우소-γ분비수평야체도최고.결론 효질류간세포역묘능경유효지유도특이성세포독성T세포,표현출경강적항종류특성.
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 cells 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-γ.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.