白介素-13受体α2抗原肽致敏的DC-CIK细胞对人胶质瘤U_(251)细胞的杀伤效应

Killing effect of interleukin-13 receptor alpha 2 (IL-13Rα2) sensitized DC-CIK cells on human glioblastoma U_(251) cells

目的:观察人脑胶质母细胞瘤特异性抗原(IL-13Rα2)致敏的同源细胞因子诱导的杀伤细胞(cytokine induced killer,CIK)在体外对U251的细胞毒效应。方法:用密度梯度分离法获取HLA-A*0201+健康志愿者的人外周血树突状细胞(dendritic cells,DC)的前体细胞与淋巴细胞,并在体外诱导而获得成熟的DC细胞和CIK细胞。收获IL-13Rα2抗原负载的DC与CIK细胞共培养,使DC递呈肿瘤抗原予CIK细胞,并激活CIK细胞。激活的CIK细胞与U251细胞在96孔板内混合培养24 h后,以CCK-8试剂间接检测其对U251细胞杀伤率。结果:IL-13Rα2抗原肽成功负载DC,CIK细胞被负载抗原的DC激活并增殖;特异性抗原肽激活的CIK细胞对人胶质瘤U251细胞有杀伤效应(P<0.01)。结论:IL-13Rα2抗原肽激活的CIK细胞对人胶质瘤U251细胞有杀伤效应。

Objective： To determine the cytokine induced killer（CIK） cells＇ cytot oxic effect in human U251 in vitro, after co-culturing with dendritic cells （DC） which was pulsed with IL-13Rα2 antigen that is highly and specifically express in human giiobiastoma. Methods： The lymphocyte and the monocytederived DC generated from healthy HLA.A＊0201＊ positive donors peripherai biood and were separated using standard Ficoli-Hypaque gradient density centrifugation. DC were pulsed with interieukin-13 receptor alpha 2 peptide in vitro and then co-cultured with lymphocyte. After induced CIK ceils and U251 cells were cocuitured 24 hours in 96 welis, CCK-8 were added every well equally. 2 hours passed and then, the OD value were detected and recorded. ResuIts： On IL-13Rα2 antigen uptake, DC efficient presentation of the antigen to CIK cells, resulting in CIK cells activation and proiiferation. The induced CIK cells showed specific lysis against human U251 cells（P〈 0.01）. Conclusion： The results suggest that IL-13Rα2 antigen specific induced CIK ceils can kill human glioma U251 cells.