牙周炎基因疫苗pVAX1-HA2、pVAX1-HA2/IL-15鼻腔黏膜免疫SD大鼠的实验研究  

Experimental study on gene vaccine pVAX1-HA2-FimA and pVAX1-HA2-FimA/IL-15 by intranasal immunization in rat

作　　者：白国辉 曾凤娇 杨琳 陈靖 刘建国 田源 BAI Guohui;ZENG Fengjiao;YANG Lin;CHEN Jing;LIU Jianguo;TIAN Yuan(Key Laboratory of Oral Disease of Higher Schools in Guizhou Province,Zunyi 563000,China;Guiyang Stomatological Hospital,Guiyang;Hospital/School of Stomatology,Zunyi Medical University)

机构地区：[1]贵州省普通高等学校口腔疾病研究特色重点实验室,遵义563000 [2]贵阳市口腔医院 [3]遵义医科大学附属口腔医院

出　　处：《实用口腔医学杂志》2020年第2期275-279,共5页Journal of Practical Stomatology

基　　金：国家自然科学基金项目(编号:81560186);贵州省科技计划项目(编号:黔科合基础[2019]1333号);贵州省科学技术基金项目(编号:黔科合J字LKZ[2012]15号);贵州省普通高等学校口腔疾病研究特色重点实验室开放课题(编号:SKLODR-2016-04)。

摘　　要：目的:观察牙周炎基因疫苗pVAX1-HA2、pVAX1-HA2/IL-15经鼻黏膜免疫SD大鼠后的免疫原性及其目的蛋白在免疫部位的表达。方法:32只SD大鼠分别经pVAX1-HA2、pVAX1-HA2/IL-15、pVAX1(空载体对照)和生理盐水(阴性对照)经鼻腔黏膜免疫(n=8)。ELISA法测定唾液SIgA抗体水平、免疫组织化学检测目的蛋白的表达情况以及进行动物的一般安全性检测。结果:①牙周炎疫苗经鼻黏膜免疫后可以诱导机体产生唾液SIgA抗体,且抗体水平在实验组明显高于空载组和阴性对照组(P<0.05)。②目的蛋白在鼻腔黏膜表达。③pVAX1-HA2/IL-15免疫组较pVAX1-HA2组产生更高水平的抗体。④大鼠体重无明显差异,重要脏器未见明显的病理学改变。结论:①牙周炎基因疫苗能诱导大鼠共同黏膜免疫系统产生免疫应答,具有免疫原性且具有安全性。②IL-15可作为该疫苗的免疫佐剂。Objective:To observe the immune effect of periodontitis vaccine pVAX1-HA2,pVAX1-HA2/IL-15 in rats by nasal mucosa routes and the expression of interest protein in situ.Methods:32 SD rats were immunized with 4 different vaccines,pVAX1-HA2,pVAX1-HA2/IL-15,pVAX1(blank control and normal saline(negivitive control)respectively(n=8).The level of SIgA antibody in saliva was measured by ELISA,the expression of the target protein was detected by immunohistochemistry,and the general safety test of the animal was performed.Results:①The level of salivary SIgA antibodies in the vaccine groups was significantly higher than that in the blank group and the negative control group(P<0.05).②The expression of interest protein was found in nasal mucosa.③The immunization with the IL-15 immunoadjuvant recombinant plasmid induced a higher level of antibody response.④There was no significant difference in body weight among the groups(P>0.05).No significant pathological change was observed in the major organs of the rats.Conclusion:①Periodontitis vaccine can induce immune responses in the common mucosal immune system by intranasal immunization in rats.②The cytokine IL-15 can be considered as an immune adjuvant.

关 键 词：基因疫苗 鼻腔黏膜免疫 共同黏膜免疫 免疫佐剂 

分 类 号：R781.42[医药卫生—口腔医学]