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。
Objectives This study sought to investigate immunoprotection provided by recombinant Echinococcus granulosus myophilin(rEg.myophilin) and tentatively analyze the mechanism of this protection.Methods One hundred and eleven male ICR mice were randomly divided into 3 groups,control groups A and B and experimental group C.Mice were subcutaneously injected with PBS;PBS and adjuvant;or PBS,adjuvant,and rEg.myophilin.After two boosters,mice were intraperitoneally challenged with Eg protoscoleces(PSCs),and then their humoral immune response was determined based on specific IgG,IgG1,IgG2a,IgG2b,IgG3,and IgE.Cellular immunity was also determined using IFN-γ,IL-4,and IL-10.Results Mice immunized with rEg.myophilin had effective immunoprotection(reduction in cyst load of more than 86.11%).Mice immunized with rEg.myophilin had specific IgG,IgG1,IgG2a and IgE.IFN-γ and IL-4 significantly increased after immunization one or more times,but neither IgG2b nor IgG3 increased.Nevertheless,IL-10 significantly increased in the control groups but tended to decline in the rEg.myophilin group.Conclusion rEg.myophilin effectively provided protective immunity.(1) This may have led to an IgG and IgE-mediated humoral immune response,as well as cell-mediated immunity,as indicated by IFN-γ.(2) IL-4 helped to stimulate a change in the subtype of IgE,which plays an important role in eosinophil-related anti-parasite immunity.