3种苦马豆素人工抗原的合成及其免疫原性研究

比较了3种苦马豆素(Swainsonine,SW)人工抗原对小鼠的免疫原性,筛选出最佳的蛋白载体,为制备疯草毒素疫苗奠定基础。活性酯法制备SW-OVA、SW-BSA、SW-HSA 3种SW人工抗原,冷冻干燥后,紫外分光光度法测定偶联率。将56只昆系小白鼠随机分为SW-OVA大剂量组(A组)、SW-OVA小剂量组(B组)、SW-BSA大剂量组(C组)、SW-BSA小剂量组(D组)、SW-HSA大剂量组(E组)、SW-HSA小剂量组(F组)和空白对照组(G组),每组8只。A~F组分别接种3种SW人工抗原,首次免疫抗原的大、小剂量分别为50μg/只和100μg/只,第15天进行第2次免疫,抗原量均为第1次免疫抗原量的1.5倍,第29天进行第3次免疫,抗原量均为第2次免疫抗原量的1.5倍,G组对照,注射等量生理盐水。第28天开始检测血清中SW抗体效价。紫外光谱测定结果显示,本试验制备的SW-OVA、SW-BSA、SW-HSA 3种SW人工抗原偶联率分别为13、10和19。ELISA测定结果显示,A和B组的SW抗体效价一直保持很高的水平,A和B组小鼠血清的SW抗体效价于第98天分别到达最高峰13(log2)和12(log2);C~F组的SW抗体效价较低。结果证实,SW-OVA作为理想的免疫原,能够刺激小鼠的免疫应答,诱导产生很高的SW抗体,在防治动物疯草中毒方面具有理想的应用前景。

Studies on Synthesis of 3 Different Artificial Antigens and Their Immunogenicity Against Swainsonine

The immunogenicity of 3 different artificial antigens against swainsonine was studied relatively on mice,acquiring the better protein for carrying,to support the foundation of vaccine against Locoweed.SW-OVA,SW-BSA,SW-HSA were obtained by active eater's method,then drying and assaying conjugate ratios of artificial antigens against swainsonine by ultraviolet spectrophotometry.56 kunming mice were randomly divided into large-dose immune group of SW-OVA(group A),mini-dose immune group of SW-OVA(group B),large-dose immune group of SW-BSA(group C),mini-dose immune group of SW-BSA(group D),large-dose immune group of SW-HSA(group E),mini-dose immune group of SW-HSA(group F),and blank control group(group G),8 mice per group.Mice of group A~F were vaccinated with 3 different artificial antigens against swainsonine.The dose of antigen of the first immunization was 0.050 mg per mouse and 0.100 mg per mouse,respectively.The second immunization was done at the day 15 and the dose of antigen was 1.5 times as the first immunization.The third immunization was done at the day 29 and the dose of antigen was 1.5 times as the second immunization.Group G was control and injected with physiologic saline.Serum SW antibody titers in each group were assayed initially at the day 28.The results of ultraviolet spectra assay showed that the conjugate ratio of SW-OVA,SW-BSA,SW-HSA were 13,10 and 19 respectively.ELISA assay showed that serum SW antibody titers of group A and B had been always keeping at a higher level,SW antibody titers of group A and B increased to the peak 13(log2)and 12(log2)respectively at the day 98.However,that of group C~F were lower.The test confirmed that,SW-OVA as the best immunogen,can stimulate immune response of mice,and induce higher level of SW antibody,that show the applied perspective for prevention and cure on Locoweed intoxication.