树突状细胞的培养及其诱导的神经胶质瘤体外杀伤作用研究  

Generation of dendritic cells from human peripheral blood for the study of antitumor efficacy 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：《中国神经精神疾病杂志》2004年第3期182-185,共4页Chinese Journal of Nervous and Mental Diseases

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 12例胶质瘤细胞 ,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对胶质瘤细胞有明显的杀伤作用 ,显著高于K5 6 2细胞对照组 ,并且杀伤率随着效靶比的增加而增加。结论　从人外周血中诱导的DCs负载胶质瘤抗原后 ,激活的CTLs在体外对胶质瘤能产生高效的杀伤作用 。Objective To investigate the antitumor efficiency of the special cytotoxic T lymphocytes activated by dendritic cells(DCs) loaded with glioma antigens in vitro, and to study the feasibility of immunotherapy for 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. DCs with specitic tumor antigen were obtained by inducing the cultured peripheral blood monocytes with combining application of human colony stimulating factors for granulocyte and macrophage, recombinant human interleukin 4 and tumor necrosis factor α. After cultured in vitro with glioma lysates, DCs were used to transform T lymphocytes into cytotoxic T lymphocytes(CTLs) for glioma cells. The cytotoxicity of CTLs to human glioma cells was assayed by MTT. Results The CTLs could kill glioma cells markedly in vitro and the killing rate was much higher than that in K562 control, which increased with the augmentation of the proportion of effector cells to target cells. Conclusions The antigen specific CTLs killed glioma cells effectively and specifically ,which might play a role in clinical immunotherapy.

关 键 词：树突状细胞 胶质瘤 肿瘤免疫 

分 类 号：R739.4[医药卫生—肿瘤]