CAJ | 학술논문

目的应用热诱导凋亡U251细胞致敏树突状细胞,观察其诱导特异性细胞毒性T细胞的能力,探讨其对肿瘤细胞的杀伤机制.方法应用改良Inaba法,从小鼠骨髓细胞中诱导出树突状细胞,应用电镜和流式细胞技术行表型鉴定,然后和经热诱导凋亡的胶质瘤细胞共培养获得树突状细胞疫苗,细胞计数试剂盒( CCK-8)检测疫苗在体外促T细胞增殖和杀伤肿瘤细胞作用,将疫苗经尾静脉注入荷瘤裸鼠体内,检测体内肿瘤生长抑制作用.结果 44℃孵育3h能有效诱导胶质瘤细胞凋亡,凋亡率达(40.10±1.08)％.负载凋亡细胞抗原的树突状细胞在体外能有效促进T细胞增殖,增加干扰素(IFN)-γ的分泌,并随效靶比增加,对胶质瘤细胞的杀伤率增大.体内注射疫苗4周后,治疗组肿瘤体积(301.704±21.659) mm3,空白对照组为(487.116 ±65.975) mm3,差异有统计学意义(P＜0.01).结论热诱导凋亡胶质瘤细胞致敏的树突状细胞能在体外有效促进T细胞增殖,诱导特异性细胞毒性T细胞杀伤胶质瘤细胞,有效抑制体内肿瘤生长.
목적 응용열유도조망U251세포치민수돌상세포,관찰기유도특이성세포독성T세포적능력,탐토기대종류세포적살상궤제.방법 응용개량Inaba법,종소서골수세포중유도출수돌상세포,응용전경화류식세포기술행표형감정,연후화경열유도조망적효질류세포공배양획득수돌상세포역묘,세포계수시제합( CCK-8)검측역묘재체외촉T세포증식화살상종류세포작용,장역묘경미정맥주입하류라서체내,검측체내종류생장억제작용.결과 44℃부육3h능유효유도효질류세포조망,조망솔체(40.10±1.08)％.부재조망세포항원적수돌상세포재체외능유효촉진T세포증식,증가간우소(IFN)-γ적분비,병수효파비증가,대효질류세포적살상솔증대.체내주사역묘4주후,치료조종류체적(301.704±21.659) mm3,공백대조조위(487.116 ±65.975) mm3,차이유통계학의의(P＜0.01).결론 열유도조망효질류세포치민적수돌상세포능재체외유효촉진T세포증식,유도특이성세포독성T세포살상효질류세포,유효억제체내종류생장.
Objective To investigate the ability and mechanism of dendritic cells (DCs) inducing specific cytotoxicity T cells to kill giloma cells.Methods DCs were induced and cultured from the murine bone marrow cells by using modified Inaba method in vitro.The biological characteristics of DCs were detected by an electron microscope and flow cytometry.The DCs vaccine was obtained by mixing DCs with apoptotic glioma cells induced by heating treatment.The capacity of DCs vaccine promoting proliferation and killing glioma cells was tested by the method of cell counting kit-8 (CCK-8).DCs vaccine was injected into the tumor-bearing mice via the tail vein to examine the tumor growth inhibition in vivo.Results The flow cytometry confirmed that DCs vaccine highly expressed the typical surface markers CD11C,CD80,CD86 and MHC Ⅱ,and that the optimal thermal condition for inducing apoptosis of glioma cells was 44 ℃for 3 h with the maximum apoptosis rate being (40.10 ± 1.08)％.DCs loaded with apoptotic glioma cells antigen could effectively promote T cell proliferation,and increase the interferon-γ (IFN-γ) secretion.And increased effector/target ratio enhanced the cytotoxity activity.Four weeks after vaccine treatment,tumor size was (301.704 ±21.659) mm3 in treatment group,and (487.116 ±65.975) mm3 in blank control group (P ＜0.01 ).Conclusion DCs,sensitized by heating treatment apoptotic glioma cells,can effectively promote T cell proliferation and induce specific cytotoxic T cells to kill glioma cellsin vitro,and inhibit tumor growth in vivo.