树突状细胞疫苗联合CpG-ODN对膀胱肿瘤细胞的免疫学效应研究

Immunological effects of dendritic cells plused with tumor antigen in combination with CpG oligodeoxynucleotide on bladder tumor cells in vitro

目的探讨负载肿瘤抗原的树突状细胞(DC)疫苗联合含未甲基化胞嘧啶鸟嘌呤(CpG)序列的寡脱氧核糖核苷酸(CpG-ODN)对膀胱肿瘤细胞的免疫学效应。方法用小鼠BTT739细胞反复冻融抗原致敏培养第7天的DC,48h后分别加入CpG-ODN及肿瘤坏死因子-α(TNF-α)诱导DC成熟,制备负载肿瘤抗原的DC疫苗,灭活后用于刺激T淋巴细胞制备效应细胞(CTL),乳酸脱氢酶释放实验测定各组CTL对BTT739细胞的杀伤活性,ELISA法测定效应细胞培养上清液的干扰素-γ(IFN-γ)水平,同时检测了CTL对自体淋巴细胞的杀伤活性。结果负载肿瘤抗原的DC诱导的效应细胞分泌较高水平的IFN-γ,并对BTT739细胞具有较高的杀伤效应,CpG-ODN活化的DC疫苗诱导的效应细胞分泌IFN-γ水平及对肿瘤细胞的杀伤活性均高于TNF-α活化的DC疫苗诱导的效应细胞(P<0.01)。DC疫苗对自体淋巴细胞无明显免疫杀伤活性。结论CpG-ODN可增强负载肿瘤抗原的DC疫苗对膀胱肿瘤细胞的免疫杀伤效应,并且对自体淋巴细胞无免疫杀伤活性,为膀胱癌的免疫治疗提供了实验基础。

Objective To study the immunological effects of dendritic cells plus tumor antigen in combination with CpG oligodeoxynucleotide on bladder tumor cells in vitro. Methods After 7d culture, DCs were plused with BTT739 cell antigen and CpG- ODN,TNF-α were added. The antitumor immune responses of DCs plused with BTT739 cell antigen against BTT739 cells was assessed by determining the potent effect of cytotoxic T lymphocytes（CTL） response in vitro. The production of interferon-γ（IFN-γ） by CTL was assessed by ELISA assay. CTL-mediated cell lysis to autologous lymphocytes was also assayed. Results The production of INF 7 and specific lysis of BTT739 by effector cells stimulated by DC vaccine generated with CpG-ODN were higher than that generated with TNF-α （P〈 0. 01）, whereas DC vaccine had no killing effects to antologous lymphocytes. Conclusion CpG- ODN can enhance the cytotoxicity of DC vaccine on bladder tumor cells. DC vaccine has no killing effects to autologous lymphocytes. This founding shows that the DC vaccine has characteristics of high effects and good safety and provides the experimental foundation of immunotherapy for bladder tumor.