MUC1/Y基因疫苗治疗膀胱肿瘤的实验研究

MUC1/Y plasmids as tumor vaccine in the experimental study of anti-tumor roles in the bladder cancer

目的探讨人黏蛋白MUC1/Y基因DNA疫苗诱导小鼠特异性细胞毒性T细胞(cytotoxic Tlym-phocytes,CTL)及体液免疫应答的作用,为人膀胱癌疫苗的应用提供基础实验研究资料。方法通过pEGFP-C3-MUC1/Y质粒免疫雄性BALB/c小鼠,采用ELISA法检测小鼠抗体生成情况和CTL分泌IL-2和IFN-γ的量,并用LDH释放法测定CTL对膀胱癌细胞株BIU-87的特异性杀伤率。结果在接种疫苗的小鼠体内特异性抗体明显升高,CTL分泌的IL-2和IFN-γ较对照组高。在效靶比为100∶1、50∶1、25∶1、12.5∶1时,实验组特异性CTL对BIU-87杀伤率分别为50.9%、45.9%、47.5%和18.6%,而对照组(空载体pEGFP-C3和灭菌生理盐水)分别为18.3%、10.7%、13.8%、6.3%;16.5%、11.9%、8.6%和10.3%。实验组与对照组两者之间差异具有显著性(P<0.01)。结论MUC1/YDNA疫苗能够诱导小鼠体内产生抗人膀胱癌黏蛋白的特异性抗体,并能诱导产生特异杀伤表达MUC1/Y的靶细胞的CTL,为MUC1/Y基因疫苗用于膀胱肿瘤生物免疫提供一种新的治疗策略。

[Objective] To study the roles of the DNA vaccine containing human mucins gene in the fields of inducing specific cytotoxic T lymphocytes （CTL） and humoral immune response, so that the basic experimental data can be provided for the clinic application of human bladder cancer vaccine. [Methods] The pEGFP-C3-MUC1/Y plasmicls was used to immunize male BALB/c mice. Subsequently, the production of anti-MUC1/Y antibody, IL-2 and IFN-γ were examined by ELISA. Also, the specific killing of CTL targeting to BIU-87 cell was examined by LDH release method. [Results] The production of antibodies against MUC1/Y rose and the levels of IL-2 and IFN-γ increased after immunization with the plasmids as compared with those of control groups. When the rate of effector cells to target cells were 100：1, 50：1, 25：1 and 12.5：1, the specific lysis in pEGFP-C3-MUC1/Y groups reached 50.9%, 45.9%, 47.5%, 18.6%, while control groups 18.3%, 10.7% 13.8%, 6.3%; 16.5%, 11.9%, 8.6%, 10.3%, respectively. The former group was different significantly from the later two groups （P 〈0.01）. [Conclusions] The constructed DNA vaccines can induce the productivity of antibody against human bladder cancer mucins, as well as the CTL capable of killing the target cells expressing MUC1/Y. The study supports the application of MUC1/Y vaccine in the therapy of human bladder cancer.