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目的研究不同抗原剂量的亚单位疫苗对小鼠的免疫效果。方法将重组LTB-VP1和BSA蛋白分别用氢氧化铝吸附后,制备成不同浓度的亚单位疫苗,以100、50、20、10、5和1μg/只的剂量免疫昆明小鼠,共免疫3次。于初次免疫后第14、28、42、56天断尾采血,分离血清,采用间接ELISA法检测小鼠血清中特异性IgG抗体水平和特异性IgG1/IgG2a比率,并采用尿素变性法检测小鼠血清中抗体相对亲合力指数(abidity index,AI)。结果 LTB-VP1和BSA抗原激发的抗体水平随免疫次数的增加而不断升高,与抗原剂量呈非线性关系,不同抗原的合适使用剂量也不相同。LTB-VP1抗原以20μg/只剂量产生的抗体水平和抗体的AI值最高;IgG1/IgG2a平均值为1.04,各剂量组Th1/Th2型免疫反应处在一种平衡状态。BSA抗原在5和100μg/只剂量时激发的抗体水平相近,抗体的AI值差异有统计学意义(P<0.05);IgG1/IgG2a平均值为2.4,各剂量组以Th2型免疫反应为主。结论抗体水平不随抗原剂量的增加而增加,低剂量抗原即可产生较好的免疫效果,为兽用疫苗的临床使用提供了数据参考。
Objective To study the immune effect of subunit vaccines with different antigen doses on mice. Methods Recombinant LTB-VP1 and BSA protein were respectively adsorbed on aluminum hydroxide to prepare different concentrations of subunit vaccines. Kunming mice were immunized with 100, 50, 20, 10, 5 and 1 μg / Times. At the 14th, 28th, 42th and 56th day after the initial immunization, the serum was collected from the tail and the serum was separated. The specific IgG antibody level and the specific IgG1 / IgG2a ratio in the serum of the mice were detected by indirect ELISA, and the mice were detected by urea denaturation Serum antibody relative affinity index (abidity index, AI). Results The levels of antibodies raised by LTB-VP1 and BSA antigen increased with the increase of the number of immunizations, showing a non-linear relationship with the dose of antigen. The appropriate dosage of different antigens was also different. The antibody level and antibody AI value of LTB-VP1 antigen was the highest at 20μg / dose; the average IgG1 / IgG2a value was 1.04, and the Th1 / Th2 immune responses of each dose group were in an equilibrium state. The antibody levels of BSA antigen at 5 and 100 μg / dose were similar, and the antibody AI values were significantly different (P <0.05). The mean of IgG1 / IgG2a was 2.4. The Th2 type immunoreaction was the major factor in each dose group. Conclusion The antibody level does not increase with the increase of antigen dosage, low dose of antigen can produce better immune effect, and provide reference for the clinical use of veterinary vaccine.