应用热诱导凋亡法制备靶向胶质瘤细胞的树突状细胞疫苗  

Dendritic cells pulsed with heat-treated U251 cells induce antitumor immunity against malignant glioma

作　　者：冀保卫[1] 陈谦学[1] 刘宝辉[1] 吴立权[1] 田道锋[1] 郭振涛[1] 纪振刚[1] 

机构地区：[1]武汉大学人民医院神经外科,430060

出　　处：《中华实验外科杂志》2012年第3期452-455,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（30772223）

摘　　要：目的应用热诱导凋亡U251细胞致敏树突状细胞，观察其诱导特异性细胞毒性T细胞的能力，探讨其对肿瘤细胞的杀伤机制。方法应用改良Inaba法，从小鼠骨髓细胞中诱导出树突状细胞，应用电镜和流式细胞技术行表型鉴定，然后和经热诱导凋亡的胶质瘤细胞共培养获得树突状细胞疫苗，细胞计数试剂盒（CCK-8）检测疫苗在体外促T细胞增殖和杀伤肿瘤细胞作用，将疫苗经尾静脉注入荷瘤裸鼠体内，检测体内肿瘤生长抑制作用。结果44℃孵育3h能有效诱导胶质瘤细胞凋亡，凋亡率达（40．10±1．08）％。负载凋亡细胞抗原的树突状细胞在体外能有效促进T细胞增殖，增加干扰素（IFN）-γ的分泌，并随效靶比增加，对胶质瘤细胞的杀伤率增大。体内注射疫苗4周后，治疗组肿瘤体积（301．704±21．659）mm2，空白对照组为（487．116±65．975）mm2，差异有统计学意义（P〈0．01）。结论热诱导凋亡胶质瘤细胞致敏的树突状细胞能在体外有效促进T细胞增殖，诱导特异性细胞毒性T细胞杀伤胶质瘤细胞，有效抑制体内肿瘤生长。Objective To investigate the ability and mechanism of dendritic cells （DCs） inducing specific cytotoxicity T cells to kill giloma cells. Methods DCs were induced and cultured from the murine bone marrow cells by using modified Inaba method in vitro. The biological characteristics of DCs were de- tected by an electron microscope and flow cytometry. The DCs vaccine was obtained by mixing DCs with apoptotic glioma ceils induced by heating treatment. The capacity of DCs vaccine promoting proliferation and killing glioma cells was tested by the method of cell counting kit-8 （CCK-8）. DCs vaccine was injected into the tumor-bearing mice via the tail vein to examine the tumor growth inhibition in vivo. Results The flow cytometry confirmed that DCs vaccine highly expressed the typical surface markers CD11 C, CD80, CD86 and MHC H , and that the optimal thermal condition for inducing apoptosis of glioma cells was 44℃ for 3 h with the maximum apoptosis rate being （40. 10 ±1.08）%. DCs loaded with apoptotic glioma cells antigen could effectively promote T cell proliferation, and increase the interferon-3, （IFN-T） secretion. And increased effector/target ratio enhanced the cytotoxity activity. Four weeks after vaccine treatment, tumor size was （301. 704 ±21. 659） mm3 in treatment group, and （487. 116 ±65. 975） mm3 in blank con- trol group （ P 〈 0.01 ）. Conclusion DCs, sensitized by heating treatment apoptotie glioma cells, can effectively promote T cell proliferation and induce specific eytotoxic T cells to kill glioma cells in vitro, and inhibit tumor growth in vivo.

关 键 词：脑胶质瘤 树突状细胞 肿瘤疫苗 免疫疗法 

分 类 号：R73[医药卫生—肿瘤]