细粒棘球绦虫(中国大陆株)14-3-3重组蛋白的免疫保护力  

Protective immunity of Eg14-3-3 against Echinococcus granulosus in mice

作　　者：李宗吉[1,2] 雄英[1,2] 孙俊峰[1,2] 赵巍[1,2] 

机构地区：[1]宁夏医科大学科学技术研究中心 [2]医学遗传学与细胞生物学教研室,宁夏银川750004

出　　处：《西安交通大学学报（医学版）》2012年第6期676-679,706,共5页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：国家自然科学基金资助项目(No.30260105;No.30660176)~~

摘　　要：目的探讨细粒棘球蚴Eg14-3-3重组蛋白的免疫保护性及其伴随的免疫反应。方法 ICR小鼠随机分为rEg14-3-3蛋白免疫组和PBS佐剂对照组,每隔2周背部皮下免疫1次,连续免疫3次。在第3次免疫后6周,用细粒棘球蚴活的原头蚴进行攻击感染,感染后24周杀鼠取脾,分离培养脾细胞,MTT检测淋巴细胞增殖率,用试剂盒检测脾细胞培养上清液的IL-2、IFN-γ、IL-10和IL-4水平,同时检获棘球蚴包囊数并计算减囊率,用ELISA法测定血清中IgG及其亚型和IgE水平。结果与佐剂对照组比较,rEg14-3-3蛋白免疫组小鼠的免疫保护力为84.47%,rEg14-3-3蛋白免疫组小鼠血清IgG、IgG1、IgG2a抗体水平高于对照组(P<0.05);IgE略低,但两组间差异无统计学意义;淋巴细胞增殖实验结果显示rEg14-3-3免疫组小鼠脾淋巴细胞特异抗原刺激后增殖显著,免疫组淋巴细胞培养上清中IFN-γ、IL-2分泌水平显著高于对照组,而IL-4、IL-10在两组间差异无统计学意义。结论 rEg14-3-3蛋白能诱导小鼠产生高水平的免疫保护力,是潜在的疫苗候选抗原分子。Objective To investigate the protective immunity against Echinococcus granulosus in mice immunized with rEg14-3-3.Methods ICR mice were subcutaneously immunized three times with rEg14-3-3,followed by the challenge with Echinococcus granulosus protoscoleces intraperitoneally,and then sacrificed in the sixth month post-challenge to detect the proliferation of splenocytes with MTT assay and to measure the secretion of IL-2,IL-4,IL-10 and IFN-γ with ELISA.The rate of reduced hydatid cyst and the levels of IgE,IgG and IgG subclasses in sera were examined.Results Compared with the control group,mice vaccinated with rEg14-3-3 and challenged intraperitoneally with E.granulosus protoscoleces revealed significant protective immunity of 84.47%（P0.05）.Enzyme-linked immunosorbent assay and Western blot analysis indicated that immunized mice generated specific high level of IgG against rEg14-3-3.The prevailing isotypes of IgG induced by rEg14-3-3 in mice were IgG1 and IgG2a.Spleen lymphocytes from mice immunized with rEg14-3-3 showed a significant proliferation response to rEg14-3-3.The culture of spleen cells showed that secretion of IFN-γ and IL-2 increased significantly in the vaccinated mice whereas IL-4 and IL-10 levels did not differ significantly between vaccinated and control mice.Conclusion The results indicate that rEg14-3-3 vaccination elicit significant levels of protective immunity against Echinococcus granulosus infection.Thus,rEg14-3-3 protein is a promising candidate as an effective vaccine to prevent cystic echinococcosis.

关 键 词：细粒棘球蚴 rEg14-3-3 疫苗 免疫保护力 IL-2 IFN-γ IL-10 IL-4 IgG IGE 

分 类 号：R383.33[医药卫生—医学寄生虫学]