树突状细胞活化的CTLs对神经胶质瘤的体外杀伤作用研究  

Antitumor efficacy of the specific cytotoxic T lymphocytes activated by dendritic cells to glioma in vitro

作　　者：孟庆海[1] 宫安静[1] 李洛[1] 

机构地区：[1]青岛大学医学院附属医院神经外科,青岛266003

出　　处：《中华神经外科杂志》2005年第7期427-431,共5页Chinese Journal of Neurosurgery

摘　　要：目的研究负载肿瘤抗原的树突状细胞(DCs)活化的特异性细胞毒性T淋巴细胞(CTLs)对神经胶质瘤细胞的体外杀伤效应,探讨其用于临床治疗的可行性。方法体外原代培养胶质瘤细胞,冻融法获取胶质瘤细胞抗原,联合应用粒细胞/巨噬细胞集落刺激因子、白细胞介素-4和肿瘤坏死因子-α等,对人外周血单核细胞进行体外诱导来获取DCs并负载肿瘤抗原,激活自体T淋巴细胞,制备特异性CTLs,MTT法检测对胶质瘤细胞的体外杀伤效应。结果负载胶质瘤抗原的DCs激活的CTLs对胶质瘤的杀伤作用显著高于对K562细胞的杀伤作用(P<0.01),并且杀伤活性随着效靶比的增加而增加;负载胶质瘤抗原的DCs激活的CTLs对胶质瘤的杀伤活性明显高于未经胶质瘤抗原致敏的DCs刺激的CTLs对胶质瘤的杀伤作用(P<0.01)。结论联合应用细胞因子从人外周血中诱导出的DCs负载胶质瘤抗原后,激活的CTLs在体外对胶质瘤细胞能产生高效而特异的杀伤作用,为临床应用DCs瘤苗奠定基础。Objective To investigate the antitumor efficiency of the special cytotoxic T lymphocytes activated by dendritic cells loaded with glioma antigens in vitro and the feasibility of immunotherapy of glioma with dendritic cells for clinical application. Methods Glioma cells were prepared by primary culture in vitro and tumour antigens were extracted by thawing and freezing.Peripheral blood monocytes were cultured to produce DCs with recombinant human granulocyte-macrophage colony-stimulating factor, recombinant human interleukin-4, recombinant human tumor necrosis factor-α. After pulsed in vitro with glioma lysates,DCs wer used to induce T lymphocytes into cytotoxic T lymphocytes for glioma cells.The cytotoxicity of CTLs to patient's glioma cells was assayed by MTT. Results The CTLs could kill glioma cells markedly in vitro and the killing rate of glioma cells was much higher than that of K562. The CTLs were more effective to glioma than that cultured without glioma antigen.The ratio increased with the improvement of the proportion of effector cells to target cells. Conclusions The antigen specific CTLs killed glioma cells effectively and specifically ,which might play a great role in clinical therapy.

关 键 词：CTLS 树突状细胞活化 粒细胞/巨噬细胞集落刺激因子 作用研究 特异性细胞毒性T淋巴细胞 肿瘤坏死因子-α 人外周血单核细胞 体外杀伤效应 神经胶质瘤细胞 白细胞介素-4 杀伤作用 体外原代培养 MTT法检测 K562细胞 肿瘤抗原 

分 类 号：R739.4[医药卫生—肿瘤]