恶性胶质瘤相关抗原表位肽激活的树突状细胞致敏的细胞毒性T淋巴细胞靶向治疗恶性胶质瘤的实验研究
发布时间：2018-07-29 06:17
【摘要】：目的研究多种恶性胶质瘤相关抗原表位肽激活的树突状细胞(GDC)致敏的细胞毒性T淋巴细胞(GDC-CTL)对人脑胶质瘤细胞系U87的体外细胞毒活性,以及其对BALB/c nude裸小鼠神经胶质瘤模型的体内抗肿瘤作用。方法制备多种恶性胶质瘤相关抗原致敏的GDC和GDC-CTL细胞,用流式细胞仪对细胞表型进行分析。设立不同的效靶比(5∶1,10∶1,20∶1),用CCK8法测定GDC-CTL细胞体外对U87细胞的杀伤率,用酶联免疫吸附实验(ELISA)法检测GDC-CTL与U87细胞共培养48 h后培养上清液中γ干扰素(IFN-γ)水平,设立未经GDC激活的T淋巴细胞为对照组。用BALB/c Nude裸小鼠U87细胞皮下肿瘤模型观察GDC-CTL细胞体内抑瘤作用,实验分为空白对照组、模型组、静脉治疗组和局部治疗组,空白对照组于背部皮下注射0.9%Na Cl,其他3组于背部皮下注射1×107U 87细胞,在肿瘤长径达3 mm后模型组于肿瘤局部注射0.9%Na Cl,静脉治疗组和局部治疗组分别于尾静脉和肿瘤局部注射2×107GDC-CTL,每周注射3次,共2周,注射体积均为0.2 m L。观察4组小鼠的肿瘤体积生长情况,取肿瘤组织进行病理学检测。结果 GDC高表达CD83、CD1a和HLA-DR,表明成功诱导GDC成熟。CD3+T淋巴细胞比例为93.00%,CD3+CD8+T淋巴细胞为69.00%,表明GDC-CTL成功激活。效靶比为5∶1,10∶1,20∶1时:GDC-CTL组和对照组对U87细胞的杀伤率分别为(24.35±1.12)%vs(15.21±0.91)%,(38.57±2.10)%vs(23.35±1.30)%,(59.44±3.79)%vs(35.23±2.33)%;GDC-CTL组和对照组与U87细胞共培养48 h后培养上清液中IFN-γ水平分别为(405.36±27.65)vs(371.11±23.23)pg·m L~(-1),(1509.22±97.16)vs(913.54±48.35)pg·m L~(-1),(2429.57±183.18)vs(1814.97±123.24)pg·m L~(-1),在效靶比为10∶1和20∶1时,2组间差异有统计学意义(P0.05)。静脉治疗组和局部治疗组对BALB/c nude裸小鼠皮下神经胶质瘤的生长抑制率分别为34.83%和45.37%,与模型组的100.00%比较,差异均有统计学意义(P0.05),与模型组相比,局部治疗组和静脉治疗组均可见肿瘤细胞显著减少。结论 GDC-CTL是一种高效的免疫细胞,具有较强的体内外抑制胶质瘤生长的作用。
[Abstract]:Objective to study the cytotoxic activity of cytotoxic T lymphocytes (GDC-CTL) sensitized by dendritic cells (GDC) activated by epitope peptides associated with malignant glioma on human glioma cell line U87 in vitro. And its anti-tumor effect on BALB/c nude mouse glioma model in vivo. Methods GDC and GDC-CTL cells sensitized with various malignant glioma associated antigens were prepared and their phenotypes were analyzed by flow cytometry. Different effective / target ratios (5: 1 10: 1 20: 1) were established. The cytotoxicity of GDC-CTL cells to U87 cells was measured by CCK8 assay. The levels of IFN- 纬 in the supernatant of GDC-CTL and U87 cells co-cultured for 48 h were detected by enzyme-linked immunosorbent assay (ELISA). T lymphocytes without GDC activation were established as control group. BALB/c Nude nude mice U87 cell tumor model was used to observe the anti-tumor effect of GDC-CTL cells in vivo. The experiment was divided into three groups: blank control group, model group, intravenous treatment group and local treatment group. 0.9%Na Cl1 was injected subcutaneously into the back of the control group, and 1 脳 107 U87 cells were injected into the back in the other three groups. After the tumor reached 3 mm in length, the model group was injected with 0.9%Na Cl locally, and the intravenous treatment group and the local treatment group were injected with 2 脳 107 GDC-CTL in the tail vein and tumor respectively, three times a week for 2 weeks. The injection volume was 0.2mL. Tumor volume growth was observed in 4 groups of mice and tumor tissues were taken for pathological examination. Results the overexpression of CD83a CD1a and HLA-DR1 by GDC indicated that the ratio of T lymphocytes induced to GDC maturation. CD3 T lymphocytes was 93.00 and CD3 CD8 T lymphocytes were 69.00, indicating that GDC-CTL was successfully activated. 鏁堥澏姣斾负5鈭，
本文编号：2151796
[Abstract]:Objective to study the cytotoxic activity of cytotoxic T lymphocytes (GDC-CTL) sensitized by dendritic cells (GDC) activated by epitope peptides associated with malignant glioma on human glioma cell line U87 in vitro. And its anti-tumor effect on BALB/c nude mouse glioma model in vivo. Methods GDC and GDC-CTL cells sensitized with various malignant glioma associated antigens were prepared and their phenotypes were analyzed by flow cytometry. Different effective / target ratios (5: 1 10: 1 20: 1) were established. The cytotoxicity of GDC-CTL cells to U87 cells was measured by CCK8 assay. The levels of IFN- 纬 in the supernatant of GDC-CTL and U87 cells co-cultured for 48 h were detected by enzyme-linked immunosorbent assay (ELISA). T lymphocytes without GDC activation were established as control group. BALB/c Nude nude mice U87 cell tumor model was used to observe the anti-tumor effect of GDC-CTL cells in vivo. The experiment was divided into three groups: blank control group, model group, intravenous treatment group and local treatment group. 0.9%Na Cl1 was injected subcutaneously into the back of the control group, and 1 脳 107 U87 cells were injected into the back in the other three groups. After the tumor reached 3 mm in length, the model group was injected with 0.9%Na Cl locally, and the intravenous treatment group and the local treatment group were injected with 2 脳 107 GDC-CTL in the tail vein and tumor respectively, three times a week for 2 weeks. The injection volume was 0.2mL. Tumor volume growth was observed in 4 groups of mice and tumor tissues were taken for pathological examination. Results the overexpression of CD83a CD1a and HLA-DR1 by GDC indicated that the ratio of T lymphocytes induced to GDC maturation. CD3 T lymphocytes was 93.00 and CD3 CD8 T lymphocytes were 69.00, indicating that GDC-CTL was successfully activated. 鏁堥澏姣斾负5鈭，
本文编号：2151796
本文链接：https://www.wllwen.com/yixuelunwen/shenjingyixue/2151796.html
最近更新
教材专著
