防龋基因疫苗pcDNA3-PAc兔鼻腔黏膜免疫防龋的实验研究

上海口腔医学 ›› 2014, Vol. 23 ›› Issue (2): 133-137.

防龋基因疫苗pcDNA3-PAc兔鼻腔黏膜免疫防龋的实验研究

王晶^{1, 2}, 刘建国², 姜晶², 王伟², 白国辉², 管晓燕²

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

收稿日期:2013-04-22 出版日期:2014-04-20 发布日期:2014-05-21
通讯作者: 刘建国, E-mail:13087891001@163.com
作者简介:王晶(1971-), 女, 硕士, 主治医师, E-mail:tree1668@163.com
基金资助:
贵州省优秀青年科技人才培养计划;贵州省高等学校重点学科建设项目;遵义医学院优秀科研团队培育项目

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

WANG Jing^1,2, LIU Jian-guo², JIANG Jing², WANG Wei², BAI Guo-hui², GUAN Xiao-yan²

1.Department of Oral Medicine, Affiliated Hospital of Binzhou Medical University. Binzhou 256603, Shandong Province;
2.Department of Oral Medicine, Affiliated Stomatology Hospital, Zunyi Medical University. Zunyi 563003, Guizhou Province, China

Received:2013-04-22 Online:2014-04-20 Published:2014-05-21

摘要/Abstract

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、600 μg pcDNA3-PAc质粒组;400 μg 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、600 μg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);③200 μg组与400 μg及600 μg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、600 μg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、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, 表面蛋白抗原

Abstract: 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-PAc 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 pcDNA3-PAc 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: ①The 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-PAc can be considered as the optimal dosage than 200 μg at present experimental condition.

Key words: Gene vaccine, Dental caries, Streptococcus mutans, PAc

中图分类号:

R781

王晶, 刘建国, 姜晶, 王伟, 白国辉, 管晓燕. 防龋基因疫苗pcDNA3-PAc兔鼻腔黏膜免疫防龋的实验研究[J]. 上海口腔医学, 2014, 23(2): 133-137.

WANG Jing, LIU Jian-guo, JIANG Jing, WANG Wei, BAI Guo-hui, GUAN Xiao-yan. Study on gene vaccine pcDNA3-PAc against dental caries by intranasal immunization in rabbits[J]. Shanghai Journal of Stomatology, 2014, 23(2): 133-137.

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