CAJ | 학술논문

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 - 13 Rα2(345-354)致敏的DC - CTL在体外对胶质瘤于细胞具有杀伤作用,且杀伤率高于对普通的胶质瘤细胞.
목적 비교IL - 13Rα2치민적DC - CTL대인효질류간세포화보통효질류세포적체외살상효응.방법 용신경간세포무혈청배양법배양출U251효질류간세포병진행면역형광감정,GM- CSF、IL-4체외유도성수돌상세포(DCs),여인공합성적IL - 13Rα2(345-354)다태부육후재격활T림파세포,CFDA - SE/PI쌍형광염료표기적류식세포계수법검측기대U251효질류간세포화보통U251효질류세포적체외살상작용.결과IL - 13Rα2(345-354)항원태치민적DC - CTL대U251효질류간세포적살상솔위(18.59±1.42)％,고우미용항원태치민적DC - CTL급CIK대효질류간세포적살상솔,기살상솔분별위(10.35±0.98)％화(7.15±0.58)％,차이구유통계학의의(P＜0.001).동시,IL- 13Rα2(345-354)항원태치민적DC - CTL대U251효질류간세포적살상솔야고우기대보통U251세포(11.53±0.65)％적살상솔(P=0.001).결론 IL - 13 Rα2(345-354)치민적DC - CTL재체외대효질류우세포구유살상작용,차살상솔고우대보통적효질류세포.
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 teehniques 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 Fieoll -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-13α2(345-354)to stimulate autologous T cells.CFSE and PI fluorochrome 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 useless IL - 13α(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 cells in vitro.