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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 protective effect of Eggs-derived Eg95-EgA31 fusion gene alfalfa vaccine on challenge of Eg protoscoleces in BALB / c mice. Methods The leaf protein of transgenic alfalfa was extracted by hot-flocculation method, and the concentration was 20μg / μl. BALB / c mice were inoculated intranasally with 100μl (containing about 1μg of fusion antigen) orally and 10μl (containing 0.1μg of fusion antigen) intranasally to immunize BALB / c mice respectively. pBI121) alfalfa leaf protein and normal alfalfa leaf protein control group. At 8 weeks after the last immunization, challenge was performed with Eg protoscoleces (50 Eg protoscoleces per mouse per mouse), and the mice were sacrificed at 24 weeks after infection, the cysts were seized and the cystic masses were reduced The blood samples were collected for eyeball blood and IgG and its subclasses and IgE levels were detected by enzyme-linked immunosorbent assay (ELISA). Results The weight of cysts in the mice inoculated with the vaccine was significantly lower than that of the normal control group (P <0.05), and the serum IgG, IgG2b and IgE levels were significantly lower The levels were significantly higher than that of the normal alfalfa leaf protein control group, and their values were 0.175 ± 0.013, 0.096 ± 0.028 and 0.096 ± 0.028, respectively. Conclusions Oral administration of Eg95-EgA31 fusion gene alfalfa vaccine can protect the immunized mice against infection by Eg protoscoleces, and IgG, IgG2b and IgE play an important role in vaccine-induced protection.