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目的研究细粒棘球绦虫(中国大陆株)重组亲肌肉抗原(rEg.myophilin)的免疫保护作用及其机制,为包虫病分子疫苗的研制提供新的候选分子。方法 (1)选取111只ICR小鼠随机分为A、B对照组和C实验组,分别于皮下注射PBS、PBS+佐剂和PBS+佐剂+候选疫苗,之后分别用同样的试剂进行两次加强免疫;(2)用Eg原头蚴进行攻击感染,感染后25周剖杀小鼠,检获棘球蚴包囊,计算免疫保护力;(3)按不同的时间点分批次对小鼠采血、取脾,用ELISA法测定血清中IgG及其亚型和IgE水平,并检测其细胞因子IFN-γ、IL-4和IL-10水平。结果 rEg.myophilin能诱导小鼠产生≥86.11%的免疫保护力;免疫小鼠血清IgG、IgG1、IgG2a、IgE和细胞因子IFN-γ、IL-4水平均增高,对照组攻击感染后IL-10增高,实验组未增高。结论 rEg.myophilin免疫后IgG及其亚类IgG1和IgE介导的体液免疫在抗细粒棘球蚴感染过程中发挥重要作用。IL-4在IgE亚型类别转换中起关键作用,这一机制可能直接影响IgE参与的嗜酸性粒细胞相关的抗寄生虫感染。
Objective To study the immunoprotection and its mechanism of recombinant e antigen of muscle (rEg.myophilin) of Echinococcus granulosus (Chinese mainland strain), and to provide a new candidate molecule for the development of molecular vaccine against hydatid disease. Methods (1) 111 ICR mice were randomly divided into A, B control group and C experimental group, and were subcutaneously injected with PBS, PBS + adjuvant and PBS + adjuvant + candidate vaccine respectively, and then twice with the same reagent Immunization; (2) challenge with Eg protothecae infection, mice were killed 25 weeks after infection, echinococcosis cysts were collected to calculate the immune protection; (3) according to different time points in batches of mice blood The spleen was removed and the levels of IgG and its subtypes and IgE in serum were determined by ELISA. The levels of IFN-γ, IL-4 and IL-10 were measured. Results The mice immunized with rEg.myophilin showed an immunopotency of ≥86.11%. The levels of IgG, IgG1, IgG2a, IgE and IFN-γ, IL-4 in the immunized mice were increased. The levels of IL-10 Increased, the experimental group did not increase. Conclusion IgG and IgGI and IgE-mediated humoral immunity after immunization with rEg.myophilin play an important role in the anti-Echinococcus granulosus infection. IL-4 plays a pivotal role in the IgE subtype shift, a mechanism that may directly affect eosinophil-associated anti-parasitic infections involving IgE.