IL-13Rα2致敏的DC—CTL对人胶质瘤干细胞的体外杀伤效应

Killing effect of interleukin - 13 receptor alpha 2 （ IL-13Rα2） sensitized DC - CTL cells on human glioblastoma stem cells in vitro

目的比较IL-13Rα2致敏的DC—CTL对人胶质瘤干细胞和普通胶质瘤细胞的体外杀伤效应。方法用神经干细胞无血清培养法培养出U251胶质瘤干细胞并进行免疫荧光鉴定，GM—CSF、IL-4体外诱导成树突状细胞（DCs），与人工合成的IL-13Rα2（345-354）多肽孵育后再激活T淋巴细胞，CFDA—SE／PI双荧光染料标记的流式细胞计数法检测其对U251胶质瘤干细胞和普通U251胶质瘤细胞的体外杀伤作用。结果IL-13Rα2（345-354）抗原肽致敏的Dc—CTL对U251胶质瘤干细胞的杀伤率为（18．59±1．42）％，高于未用抗原肽致敏的DC—CTL及CIK对胶质瘤干细胞的杀伤率，其杀伤率分别为（10．35±0．98）％和（7．15±0．58）％，差异具有统计学意义（P〈0．001）。同时，IL-13Rα2（345-354）抗原肽致敏的DC—CTL对U251胶质瘤干细胞的杀伤率也高于其对普通U251细胞（11．53±0．65）％的杀伤率（P=0．001）。结论IL-13Rα2（345-354）致敏的Dc—CTL在体外对胶质瘤干细胞具有杀伤作用，且杀伤率高于对普通的胶质瘤细胞。

Objective To compare the killing effect of IL-13Rα2 sensitized DC - CTL cells on human glioblastoma stem cells and ordinary glioblastoma cells in vitro. Methods Tumor sphere cells were isolated from glioma cell line U251 with serum -free incubation techniques which used to isolated normal neural stem cells, the immunofluorescence staining was employed to identify the glioma stem cells. Mononuclear cells generated from healthy HLA ＊ A＊ 0201 ＋ positive donors peripheral blood were separated using standard Ficoll -Hypaque gradient density centrifugation. Corresponsive DCS from PBMC were co - cultured in RPMI1640 with GM - CSF and IL -4, and then matured by the artificial IL-13Rα2（345-354） to stimulate autologous T cells. CFSE and PI iluorochrome stain assay was used for detecting the killing effect in vitro. Results The killing effect of IL-13Rα2 sensitized DC - CTL cells on human glioblastoma stem cells was （ 18. 59 ± 1.42） % , which was much higher than that of uselessIL-13Rα2（345-354） sensitized DC - CTL cells and CIK on glioblastoma stem cells. The killing effect was （ 10. 35 ± 0. 98 ） % and （7. 15 ± 0. 58 ） %, recepecively. There were statistically differences among them Meanwhile , the killing effect of IL-13Rα2 sensitized DC - CTL cells on human glioblastoma stem cells was also higher than that on the ordinary U251 cells （ 11.53 ± 0. 65） % （ P = 0. 001 ）. Conclusions The IL-13Rα2（345-354）induced CTLs exhibited specific cytotoxic against U251 glioma stem cells and the killing rate was much higher than that of U251 ceils in vitro.