树突状细胞疫苗治疗G422胶质母细胞瘤的实验研究

Experimental Study of Treatment of Intracranial G422 Glioblastoma with Bone Marrow-derived Dendritic Cells Vaccine

目的研究G422肿瘤细胞RNA体外冲击致敏的树突状细胞(DC)回输诱导体内产生保护性免疫反应及对颅内荷瘤小鼠的免疫治疗效应,探讨以树突状细胞为基础的疫苗对脑胶质瘤治疗的可行性。方法提取G422胶质母细胞瘤RNA或肿瘤提取物冲击致敏DC制成疫苗,分别进行免疫保护和免疫治疗的体内实验,以及细胞毒性T淋巴细胞(CTL)体外诱导及活性检测,并与PBS、未经致敏的DC、G422肿瘤细胞RNA组进行对照。疫苗能诱导产生针对G422肿瘤抗原特异性CTL,差异具有非常显著性(P<0.01),接种疫苗后的小鼠能抵抗G422的再次攻击(P<0.01),荷瘤小鼠接种疫苗后生存期较对照组延长(P<0.05)。结果疫苗能诱导产生针对G422肿瘤抗原特异性CTL,差异具有非常显著性(P<0.01),接种疫苗后的小鼠能抵抗G422的再次攻击(P<0.01),荷瘤小鼠接种疫苗后生存期较对照组延长(P<0.05)。结论肿瘤RNA或肿瘤提取物冲击致敏DC能诱导小鼠产生抗原特异性CTL,激活抗肿瘤T细胞,具有免疫保护和免疫治疗作用。该实验为DC疫苗免疫治疗胶质瘤提供了实验基础。

Objectives To investigate the anti-tumor efficacy of dendritic cell (DC)-based vaccines pulsed with tumor RNA or extracts in mice with intracranial G422 glioblastoma and to discuss the feasibility of this active immunotherapy strategy for gliomas. Methods DC were isolated from mouse bone marrow precursors stimulated in vitro with granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4.These DC were then pulsed with tumor RNA or extracts.The curative effect of unpulsed DC,G422 tumor RNA or extracts,or DC pulsed with G422 tumor RNA or extracts on the intracranial G422 glioblastomas were studied in the mice.CLT activity induced by DC pulsed with G422 tumor RNA or extracts was determined.The survival time of the mice was observe and analyzed.The brains were removed and examined pathologically in the mice when they died.Results Immunization using DC pulsed with tumor RNA or extracts induced G422-specific CTL responses that were significantly better than that in the mice treated with PBS,unpulsed DC,G422 tumor RNA or extracts (P<0.01).The survival time of mice with intracranial glioblastomas was significandy longer in the tumor RNA or extract-pulsed DC-treated group than that in the other groups (P<0.01).Conclusion DC pulsed with tumor RNA or extracts derived from autologous tumors represent a promising approach to the immunotherapy of gliomas, which can specifically activate antitumor T cells and lead to significantly prolonged survival in tumor-bearing animals.The experiment provides scientific basis for clinical use of DC vaccine.