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目的:观察防龋基因疫苗pcDNA3-PAc经鼻腔黏膜免疫日本长耳大白兔后的免疫反应性,确定不同剂量质粒pcDNA3-PAc免疫机体后的抗体效价。方法:30只日本长耳大白兔随机分为5组,每组6只,分别为200、400、600μg pcDNA3-PAc质粒组;400μg pcDNA3质粒组(阴性对照组);灭活全菌疫苗组(阳性对照组)。质粒组及全菌组以1:1比例添加弗氏佐剂后进行免疫,共免疫2次。采用间接ELISA法检测血液、唾液中特异性IgG、S-IgA抗体。采用SPSS17.0软件包对数据进行统计学分析。结果:①血液、唾液中抗体高峰时间为初始免疫后8~10周左右:②自免疫后第3周开始,200、400、600μg组兔血清、唾液中特异性抗PAc的IgG、S-IgA抗体水平与阴性对照组相比,差异有显著性(P<0.05);③200μg组与400μg及600μg组相比,多时间点差异有显著性(P<05)。结论:①防龋基因疫苗pcDNA3-PAc具有免疫反应性,可诱导免疫反应长达14周;②200、400、600μg的防龋基因疫苗pcDNA3-PAc对体重1.5 kg左右的兔都是有效的免疫剂量;③在本研究条件下,400、600μg疫苗组效价优于200μg组。
OBJECTIVE: To observe the immunoreactivity of anti-caries gene vaccine pcDNA3-PAc in nasal mucosa of Japanese white-eared rabbits and determine the antibody titer after immunization with different doses of pcDNA3-PAc. Methods: Thirty Japanese white-bellied rabbits were randomly divided into 5 groups with 6 mice in each group: 200, 400 and 600 μg pcDNA3-PAc plasmid group; 400 μg pcDNA3 plasmid group (negative control group); inactivated whole-cell vaccine group Positive control group). The plasmid group and the whole bacteria group were immunized with Freund’s adjuvant in a ratio of 1: 1, and were co-immunized twice. Indirect ELISA was used to detect specific IgG and S-IgA antibodies in blood and saliva. SPSS17.0 software package for statistical analysis of the data. Results: ① The peak time of antibody in blood and saliva was about 8 to 10 weeks after initial immunization: ② From the 3rd week after immunization, IgG, S-IgA specific anti-PAc in serum, saliva of 200, 400 and 600μg rabbits Compared with the negative control group, the antibody level was significantly different (P <0.05). (3) Compared with the 400μg and 600μg groups, the difference was significant (P <0.05). Conclusion: ①Anti-cariogenic vaccine pcDNA3-PAc is immunoreactive and can induce an immune response for up to 14 weeks. ② 200,400,600μg of cariogenic vaccine pcDNA3-PAc is an effective immunization dose for rabbits weighing about 1.5 kg ; ③ Under the conditions of this study, 400 and 600μg vaccine group titers better than the 200μg group.