树突状细胞疫苗的免疫学效应实验研究

摘要

Objective To explore the immunological effects and mechanisms of dendritic cell vaccine.Methods DC vaccine against MB 49 cells were prepared by loading MB 49 cells, and the model was established in mice. After making the model, the 7 days, 14 days, the vaccine were injected in right anterior axillary of vaccine group mice, and the mice of PBS control group were injected with PBS, normal feeding to normal group.21 days after modeling, lfow cytometry was used to detect the number of CD 4+ and CD 4+CD 69+T cells in spleen of mice. The T cells were cultured for 48 hours, and IFN-γ was detected by ELISA. The number of CD 4+ and CD 8+cells in tumor tissues was detected by immunohistochemical method.Results Mouse spleen, CD 4+andCD 4+CD 69+ cells count, DC vaccine group was higher than the PBS control group and normal group,P<0.01, had difference statistically signiifcance. ( one of themP<0.05 ). The level of IFN-γof T cell culture supernatant , DC vaccine group was higher than that of PBS control group and the normal group,P<0.01, had difference statistically signiifcance. The number of CD 4+, CD 8+ T cells in tumor tissue, DC vaccine group was higher than that of PBS control group,P<0.01, had difference statistically significance. ConclusionDC vaccine stimulated T cell activation and proliferation, increased the number of CD 4+, CD 4+ CD 69+ cells in the spleen,increased IFN-γsecretion, and increased the number of CD 4+, CD 8+T cells in the tumor tissues. DC 2.4 vaccine can inhibit the growth of tumor cells in mice by killing the tumor cells.%目的：探讨树突状细胞疫苗的免疫学效应和抑瘤机制。方法制备负载MB 49细胞粗提全抗原的DC疫苗；建立小鼠MB 49膀胱癌皮下移植瘤模型。造模后第7 d、14 d，疫苗组小鼠于右前腋皮下注射疫苗，PBS对照组相应注射PBS，正常组正常饲养。造模第21 d，流式细胞术检测小鼠脾CD 4+、CD 4+ CD 69+T细胞数；取分离后的T细胞培养48 h，取上清，ELISA法检测IFN-γ水平。应用免疫组化方法检测瘤组织内CD 4+、CD 8+细胞数。结果小鼠脾CD 4+、CD 4+CD 69+细胞数，DC疫苗组高于PBS对照组和正常组，P＜0.01，差异具有统计学意义。（其中1组P＜0.05）。T细胞培养上清IFN-γ水平，DC疫苗组高于PBS对照组和正常组，P＜0.01，差异具有统计学意义。瘤组织内CD 4+、CD 8+T细胞数，DC疫苗组高于PBS对照组，P＜0.01，差异具有统计学意义。结论 DC 2.4疫苗刺激T细胞活化和增殖，使脾内CD 4+、CD 4+CD 69+细胞数增加，IFN-γ分泌增多，同时趋化迁移至瘤组织的CD 4+、CD 8+T细胞数增多，使模型小鼠自身杀伤清除肿瘤细胞能力增强，从而抑制了模型小鼠肿瘤的生长。