摘要
目的 :观察肿瘤细胞冻融裂解物致敏的树突状细胞 (dendriticcell,DC)瘤苗对颅内胶质瘤免疫治疗的效果。方法 :通过立体定向的方法接种C6胶质瘤细胞建立Wistar大鼠C6胶质瘤动物模型 ;自大鼠骨髓分离DC前体细胞 ,经重组大鼠粒细胞巨噬细胞集落刺激因子 (rrGM CSF) +白细胞介素 4 (rrIL 4 )诱导培养、扩增获得功能性DCs;DCs经反复冻融的C6胶质瘤细胞裂解物体外致敏后皮下回输荷瘤大鼠体内 ,1次 周 ,共 5次。观察荷瘤大鼠的存活期 ;以流式细胞仪检测循环血中CD8+T淋巴细胞水平及体外细胞毒反应 ;ELISA法检测血清中IL 10及IFN γ水平。结果 :经肿瘤细胞冻融裂解物致敏的树突状细胞免疫治疗的荷瘤大鼠生存期明显延长 ,外周血中CD8+T淋巴细胞比例增加 ;并且治疗组大鼠循环血中IFN γ水平明显升高 ,而IL 10水平很低 ,甚至检测不到。结论
Objective:To investigate the therapeutic efficacy of immunotherapy with dendritic cells-based vaccine against intracranial gliomas in rats.Methods:C6 glioma cells were injected into the brain of Wistar rat under stereotactic monitor to establish an animal model of glioma.The precursors of dendritic cells were isolated from bone marrow of rats,stimulated in vitro with recombinant rat granulocyte-macrophage colony-stimulating factor(rrGM-CSF)and interleukin-4(rrIL-4).Cultured cell populations were confirmed to be functional DCs.These DCs were then pulsed ex vivo with C6 tumor-lysates prepared by three cycles of freezing to -80℃ and thawing to 0℃ and subsequently injected subcutaneously into rats harboring intracranial C6 tumor.Rats from different group were treated with five weekly subcutaneous injections of either control media,unpulsed DCs,or DCs pulsed with tumor-lysates.The animals were followed for survival,the percentage of CD8 +T cells in peripheral blood and cytotoxicity assay in vitro were determined by FACS.The level of cytokine IFN-γ and IL-10 were detected by ELISA.Results:The results indicated that C6 glioma model rats treated with tumor-lysate-pulsed DCs led to prolonged survival time,increased the percentage of CD8 + T lymphocyte in peripheral blood in comparing with control group.Cytotoxicity assays suggest that vaccination with these tumor-lysate-pulsed DCs can induce specific cytotoxic T lymphocytes against C6 tumor cells compared with control group.Furthermore,significantly enhanced IFN-γ and reduced IL-10 (even undetectable)were observed in rats treated with pulsed-DCs.Conclusion:Data supported the therapeutic efficacy of systemic vaccination with DCs pulsed with tumor-lysates against intracranial glioma.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2004年第5期311-314,共4页
Chinese Journal of Immunology
