细粒棘球绦虫重组Egmyophilin蛋白诱导免疫小鼠脾细胞增殖及细胞因子水平的变化

摘要

To investigate the immunologic mechanism in mice immunized with recombinant Egmyophilin vaccine of E. Granulosus and against challenge with Egprotoscolecs, ICR mice were subcutaneously and intranasally vaccinated by the vaccine, following by the challenge with Egprotoscolices on the 8th week of vaccination and killed on the 20th week of infection to obtai spleen. Spleen cells were separated and the proliferation was determined by MTT method. Then the cells were collected, stained by PI Annexin V-FITC. The supernatant was collected and measure the level of IL-2, IL-10, IFN-γ and TNF-α by ELISA, and the blank vector and PBS were served as control. As demonstrated by the MTT assay, the recombinant Egmyophilin vaccine could stimulate specifically the proliferation of splenic T lymphocytes in mice. ELISA assay indicated that the level of IL-2, IFN-y and TNF-a in immunity group were significantly increased after stimulus with ConA or recombinant Egmyophilin. The results indicated that the recombinant Egmyophilin vaccination induces Thl cellular response in the mice a-gainst the challenge of Egprotoscoleces.%目的 探讨细粒棘球蚴绦虫(Echinococcus granulosus,Eg)亲肌肉抗原重组疫苗(Egmyophilin)诱导ICR小鼠产生免疫保护机制.方法 将亲肌肉抗原重组疫苗皮下注射免疫ICR小鼠,同时设空质粒组及PBS对照组.末次免疫后8 w,用棘球绦虫原头节腹腔注射进行攻击感染,感染后20 w剖杀小鼠,取脾,脾细胞悬液加入EgAg、ConA(刀豆蛋白A)或脂多糖(LPS)刺激培养后,四甲基偶氮唑盐比色法(MTT法)检测免疫小鼠T淋巴细胞增殖情况;Annexin V-FITC和碘化丙啶(PI)双染色法检测脾细胞的凋亡发生率;常规ELISA法检测脾细胞培养上清液中IL-2、IL-10、IFN-γ和TNF-α水平.结果 与空质粒组及PBS对照组相比,脾细胞凋亡发生率明显降低,脾T淋巴细胞增殖水平显著升高,脾细胞培养上清液中IL-2、IFN-γ和TNF-α水平显著增高,IL-10水平明显降低.结论 棘球蚴绦虫亲肌肉抗原重组疫苗可诱导免疫鼠脾T淋巴细胞增殖,以诱导Th1型免疫应答为主,对抗Eg原头蚴攻击感染.