细粒棘球绦虫转Eg95-EgA31融合基因苜蓿疫苗诱导的保护力观察  

Immunoprotection of Transgenic Alfalfa(Medicago sativa) Containing Eg95-EgA31 Fusion Gene of Echinococcus granulosus against Eg Protoscoleces

作　　者：叶艳菊[1] 李文桂[1] 

机构地区：[1]重庆医科大学附属第一医院传染病寄生虫病研究所,重庆400016

出　　处：《热带医学杂志》2010年第3期235-237,共3页Journal of Tropical Medicine

基　　金：国家自然科学基金(No.30671835)

摘　　要：目的探讨细粒棘球绦虫(Eg)转Eg95-EgA31融合基因苜蓿疫苗免疫BALB/c小鼠后对Eg原头节攻击感染的保护性作用。方法热絮凝法提取转基因苜蓿的叶蛋白,配成浓度为20μg/μl。分别用100μl(约含1μg融合抗原)口服灌胃和10μl(约含0.1μg融合抗原)滴鼻接种免疫BALB/c小鼠,每3d免疫1次,连续免疫2月,同时设转空质粒(pBI121)苜蓿叶蛋白及正常苜蓿叶蛋白对照组。末次免疫后8周,用Eg原头节进行攻击感染(腹腔注射50个Eg原头节/每只小鼠),感染后24周剖杀各组小鼠,检获包囊质量,计算囊重减少率;采眼球血,常规酶联免疫吸附试验法(enzyme-linked immunosorbent assay,ELISA)检测血清中IgG及其亚类和IgE水平。结果疫苗口服灌胃接种组小鼠检获包囊质量明显降低,囊重减少率为64.1%,与正常蛋白对照组相比,差异有统计学意义(P<0.05),血清中IgG、IgG2b和IgE水平显著高于正常苜蓿叶蛋白对照组,其值分别为0.175±0.013、0.096±0.028和0.096±0.028。结论细粒棘球绦虫转Eg95-EgA31融合基因苜蓿疫苗口服接种能使免疫鼠获得保护力以抵抗Eg原头节攻击感染,IgG、IgG2b和IgE在疫苗诱导的保护力中起重要作用。Objective To investigate the immunoprotection of the transgenic Alfalfa（Medicago sativa） containing Eg95-EgA31 fusion gene of Echinococcus granulosus against Eg protoscoleces.Methods The leaf protein was extracted from the transgenic alfalfa by heat-coagulation method.BALB /c mice were immunized intranasally or orally with the leaf protein once per 3 days for 2 months.At the same time,the leaf protein from transgenic plant transformed with pBI-121 blank vector and the normal leaf protein without foreign antigen was served as control.The mice were then challenged intraperitoneally with Eg protoscoleces（50 protoscoleces per mouse） on week 8 after the last vaccination and sacrificed on week 24 post-infection to count the rate of reduced hydatid cyst.The antigen specific antibody（IgE,IgG and its subclasses） in the sera was evaluated by ELISA.Results In the oral immunization group,the hydatid cyst weight significant decreased by 64.1%（P〈0.05）,as compared with the normal protein control,and the antibody titers of IgG（0.175±0.013）,IgG2b（0.096±0.028） and IgE（0.096±0.028） were obviously higher than that in the control group.Conclusion The protection against Eg protoscoleces can be induced in mice immunized orally with the transgenic alfalfa containing Eg95-EgA31 fusion gene of Echinococcus granulosus,and IgG,IgG2b and IgE may play important roles in the protection.

关 键 词：细粒棘球绦虫 转基因植物 苜蓿 保护力 

分 类 号：R383.33[医药卫生—医学寄生虫学]