防龋基因疫苗pcDNA3-PAc兔鼻腔黏膜免疫防龋的实验研究  

Study on gene vaccine pcDNA3-PAc against dental caries by intranasal immunization in rabbits

作　　者：王晶[1,2] 刘建国[2] 姜晶[2] 王伟[2] 白国辉[2] 管晓燕[2] 

机构地区：[1]滨州医学院附属医院口腔内科,山东滨州256603 [2]遵义医学院附属口腔医院口腔内科,贵州遵义563003

出　　处：《上海口腔医学》2014年第2期133-137,共5页Shanghai Journal of Stomatology

基　　金：贵州省优秀青年科技人才培养计划[黔科合人字(2005)0509];贵州省高等学校重点学科建设项目[遵医院办发(2012)15号];遵义医学院优秀科研团队培育项目[遵医院办发(2012)12号]~~

摘　　要：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；400μgpcDNA3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〈0．05）。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1．5kg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。PURPOSE： To investigate the efficiency of pcDNA3-PAc on Japanese long-eared white rabbits by intranasal immunization, and observe the appreciate gene vaccine dose in rabbit immunization. METHODS： Thirty Japanese long- eared white rabbits were randomly divided into 5 groups （6 in each group） as follows： 200, 400, 600 μg pcDNA3-PAe plasmid group; 400 μg pcDNA3 group and inactivated whole-cell vaccine positive control group. The rabbits were immunized twice, and plasmid groups and whole-cell group were coupled Freund＇s adjuvant with 1：1 ratio. The specific IgG and S-IgA antibodies in serum and saliva were detected with indirect ELISA method. The data was analyzed with SPSS 17.0 software package. RESULTS： ① The peak time of the antibodies appeared between 8-10 weeks after the first immunization; ②The specific antibodies IgG and S-IgA could be detected 2 weeks after immunization; ③The level of salivary specific S-IgA and serum specific IgG of peDNA3-PAe were significantly higher than negative groups （P〈0.05）. The titer of the 200 μg was significantly lower than those of 400 μg and 600 μg group （P〈0.05）. CONCLUSIONS： ① The recombinant plasmid pcDNA3-PAc has immunogenicity, which can induce specific immune responses for 14 weeks in rabbits. ②The results of the present study show that 200 μg, 400 μg and 600 μg are effective immunizational dosage to 1.5 kg Japanese long-eared white rabbits. ③400 μg and 600 μg pcDNA3-PAe can be considered as the optimal dosage than 200 μg at present experimental condition.

关 键 词：基因疫苗 龋病 变形链球菌 表面蛋白抗原 

分 类 号：R392[医药卫生—免疫学]