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目的 研究肿瘤抗原致敏树突状细胞瘤苗治疗胶质瘤的疗效。方法 从大鼠的骨髓中加入GM CSF和IL 4培养树突状细胞 ,采用相差显微镜、扫描电镜观察了树突状细胞的形态学特点 ;免疫组织化学双标技术、流式细胞免疫学技术观察了其特异性抗体OX6和OX62 的表达 ;微酸洗脱方法抽提C6肿瘤抗原体外致敏树突状细胞 ,建立C6胶质瘤脑内动物模型 ,体内应用抗原致敏的树突状细胞 ,观察脑内肿瘤的疗效 ,并取致敏的动物的脾脏T细胞体外加入C6肿瘤抗原 ,加入IL 2培养进行CTL活性检测。结果 培养第 8天可见有典型的树突状细胞 ;免疫组织化学双标可见有OX6和OX62 阳性细胞 ,流式细胞免疫学分析发现OX62 阳性率为 93 0 3% ,OX6的阳性率为 92 0 1% ;体外细胞毒试验发现对相应的C6肿瘤细胞有明显的杀伤作用 ;动物实验发现实验组可见肿瘤组织内有大量的坏死 ,而对照组仅有少量的坏死 ,两者相比P <0 0 1。结论 体内注射采用微酸洗脱的抗原肽致敏树突状细胞瘤苗能够引起荷瘤动物脑肿瘤大面积的坏死 ,该方法为将来临床免疫治疗胶质瘤充分调动机体的免疫系统奠定基础。
Objective To study the therapeutic effect of tumor antigen-sensitized dendritic cell tumor vaccine on glioma. Methods Dendritic cells were cultured with GM CSF and IL-4 from bone marrow of rats. Morphological characteristics of dendritic cells were observed by phase-contrast microscopy and scanning electron microscopy. Immunohistochemical double-labeled technique and flow cytometry The specific antibodies OX6 and OX62 were expressed. The acidic extraction of C6 tumor antigen was used to sensitize dendritic cells in vitro. The animal model of C6 glioma was established in vivo. Antigen-sensitized dendritic cells (DCs) Observation of the efficacy of brain tumors, and take the sensitized animal spleen T cells in vitro addition of C6 tumor antigen, added IL2 culture CTL activity detection. Results The typical dendritic cells were seen on the 8th day of culture. OX6 and OX62 positive cells were observed by immunohistochemistry. The immunophenotype of OX62 was 93 0 3% and the positive rate of OX6 was 92 0 1%; in vitro cytotoxicity test found that the corresponding C6 tumor cells have obvious killing effect; animal experiments found that the experimental group showed a large number of tumor necrosis, while the control group only a small amount of necrosis, both P <0 0 1. Conclusion In vivo injection of microdialysis-eluting peptide antigen-dendritic cell vaccine can cause large-scale necrosis of brain tumor in tumor-bearing animals, which lays the foundation for the future clinical immunotherapy of glioma to fully mobilize the body’s immune system.