中文摘要：

目的探讨细粒棘球绦虫（Eg）Eg95-EgA31融合基因苜蓿叶蛋白提取液免疫小鼠被Eg原头节攻击感染后体内棘球蚴囊重减少率及细胞因子的变化。方法热絮凝法提取转基因苜蓿叶蛋白,同时提取转空质粒（pBI121）苜蓿叶蛋白和正常苜蓿叶蛋白作对照。32只雌性BALB/c小鼠随机均分4组,A和B组分别用转基因苜蓿叶蛋白提取液100μl灌胃和10μl滴鼻免疫小鼠,C组用转空质粒苜蓿叶蛋白提取液10μl滴鼻免疫小鼠,D组用正常苜蓿叶蛋白提取液100μl灌胃免疫小鼠。各组小鼠每3d免疫1次,连续免疫2个月。末次免疫后第8周,各组小鼠用Eg原头节腹腔注射攻击感染（50个/只）,感染后第24周剖杀小鼠,分离并称重细粒棘球蚴囊,计算囊重减少率;取脾,分离脾细胞,用Eg粗抗原（EgAg）、伴刀豆球蛋白A（ConA）或脂多糖（LPS）刺激培养,收集脾细胞培养上清液,常规ELISA法检测脾细胞培养上清液中白介素-12（IL-12）、白介素-10（IL-10）、干扰素-γ（IFN-γ）和肿瘤坏死因子-α（TNF-α）水平。结果A、B、C和D组小鼠的棘球蚴囊重分别为（28.0±36.0）、（41.0±33.0）、（72.0±36.0）和（78.0±57.0）mg,A组低于D组（P〈0.05）,囊重减少率为64.1%。A组的IFN-γ、1L-12和TNF-α水平高于D组（P〈0.01）,分别为（925.0±88.6）、（22.5±2.7）和（82.5±11.7）pg/ml,IL-10水平低于D组（P〈0.01）,为（125.0±26.7）pg/ml;B组小鼠脾的IFN-γ和TNF-α水平高于D组（P〈0.01）,分别为（750.0±100.0）和（80.0±13.1）pg/ml,IL-12和IL-10水平与D组相比差异无统计学意义（P〉0.05）;C组小鼠脾细胞因子水平与D组相比,差异无统计学意义（P〉0.05）。当用EgAg、ConA或LPS刺激时,刺激组的细胞因子水平高于相应的未刺激组（P〈0.05或P〈0.01）,ConA或LPS刺激组的细胞因子水平高于EgAg刺激组（P〈0.05或P〈0.01）。结论转细粒棘球绦虫Eg95-EgA31融合基因苜蓿叶蛋白提取液可诱?

英文摘要：

Objective To investigate the weight reduction of hydatid cyst and the changes of cytokine secretion in mice immunized by leaf protein extracted from Echinococcus granulosus （Eg） Eg95-EgA31 transgenic alfalfa and challenged by Eg protoscoleces. Methods Leaf protein was extracted from E. granulosus Eg95-EgA31 transgenic alfalfa by heat coagulation method. Meanwhile, leaf protein extracted from transgenic alfalfa containing pBI121 and normal alfalfa served as control. 32 female BALB/c mice were randomly divided into 4 groups. Groups A and B were immunized intragastrically （100 μl） and intranasally （10 μl） respectively by leaf protein containing Eg95-EgA31 fusion antigen, group C was vaccinated intranasally by 10 μl leaf protein with pBI121, group D was given intragastrically 100 μl normal leaf protein. All mice were immunized once per 3 days for 2 months. Mice in all groups were challenged with 50 Eg protoscoleces on the 8th week after vaccination and sacrificed on the 24th week after infection. The weight of hydatid cysts was measured and weight-reduction rate was calculated. Spleens were collected to prepare splenocytes which were cultured under stimulation with EgAg, concanavalin A （ConA） or lipopolysaccharide（LPS）. The supernatant was collected to measure the level of IL-12, IL-10, IFN-γ and TNF-α by ELISA. Results The average weight of hydatid cysts in groups A, B, C, and D was （28.0±36.0）, （41.0±33.0）, （72.0±36.0） and （78.0±57.0） mg, respectively, the cyst weight of group A was lower than that of group D （P0.05）, decreased by 64.1%. The levels of IFN-γ, IL-12 and TNF-α in group A [（925.0±88.6）, （22.5±2.7） and （82.5±11.7） pg/ml] were higher than those of group D （P0.01）, while the IL-10 level in group A [（125.0±26.7）pg/ml] was significantly lower than that of group D （P0.01）. The level of IFN-γ[（750.0±100.0） pg/ml] and TNF-α [（80.0±13.1） pg/ml] in group B was significantly higher than those of group D （P0.01）; but th