变形链球菌基因疫苗pcDNA3gtfB鼻黏膜免疫家兔实验探究.docVIP

变形链球菌基因疫苗pcDNA3gtfB鼻黏膜免疫家兔实验探究 　【摘要】 目的 观察变形链球菌防龋基因疫苗pcDNA3gtfB经鼻腔黏膜免疫日本长耳大白兔后的免疫反应性，初步观察不同剂量质粒pcDNA3gtfB免疫机体后的抗体效价。方法 实验分两部分，第一部分：30只日本长耳大白兔平均分为5组，每组6只，分别为：200 μg、400 μg 、600 μg pcDNA3gtfB质粒组，400 μg pcDNA3组，灭活全菌疫苗阳性对照组。质粒组及全菌组以1：1比例添加弗氏佐剂后进行免疫，共免疫两次。第二部分：采用间接ELISA法检测血液、唾液中特异性IgG、SIgA。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1～2周，各质粒组兔血液、唾液中特异性抗GTF的IgG、SIgA抗体水平同阴性对照组比较差异有统计学意义 (Plt;0.05);③200 μg组与400 μg及600 μg组比较多时点差异有统计学意义(Plt;0.05)。结论 ①基因疫苗pcDNA3gtfB具有免疫反应性;②200 μg 、400 μg、600 μg的基因疫苗对于体重1.5 kg左右的兔都是有效的免疫剂量;③在本研究的条件下，400 μg、600 μg疫苗组效价优于200 μg组。 【关键词】 基因疫苗;龋病;变形链球菌;葡糖基转移酶 　　【Abstract】 Objective To investigate the efficiency of pcDNA3gtfB on Japans Longeared rabbits by intranasal immunization, and observe the appreciate gene vaccine dose in rabbit immunization.Methods The study includes two parts. Part one: 30 Japan’s Longeared rabbits were randomly divided into 5 groups(each group 6) as follows: 200 μg, 400 μg, 600 μg pcDNA3gtfB plasmid group，400 μg pcDNA3 group，inactivated wholecell vaccine positive control group.In the process of experiment the rabbits were immunized twice,and plasmid groups and wholecell group were coupled Freund’s adjuvant with 1∶1 ratio.Part two:The specific IgG and SIgA in blood and saliva were detected by indirect ELISA method.Results ①The peak time of the antibodies appeared between 810 weeks after the first immunization.②The specific antibodies IgG and SIgA could be detected 12 week after immunization.③The level of salivary specific SIgA and serum specific IgG of pcDNA3gtfB were significantly higher than negative groups (Plt;0.05). The titer of the 200 μg is significantly lower than those of 400 μg and 600 μg groups (Plt;0.05).Conclusion ①The recombinant plasmid pcDNA3gtfB has immunogenicity, which can induce specific immune responses in rabbits.②The results of the present study showed that 200 μg, 400 μg and 600 μg were effective immunizational dosage to 1.5 kilograms Japan’s Longeared rabbits. ③400 μg and 600 μg pcDNA3g