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

Experiment of the periodontitis gene vaccine pVAX1-HA2-FimA and pVAX1-HA2-FimA/IL-15 by intranasal immunization in SD rats

作　　者：曾凤娇 杨琳 刘建国 于航 陈靖 白国辉 ZENG Fengjiao;YANG Lin;LIU Jianguo;YU Hang;CHEN Jing;BAI Guohui(School of Stomatology,Zunyi Medical University,Zunyi,Guizhou 563000,China;Key Laboratory of Oral Disease of Higher Schools in Guizhou Province,Zunyi,Guizhou 563000,China;Department of General,Guiyang Stomatological Hospital,Guiyang,Guizhou 550000,China)

机构地区：[1]遵义医科大学口腔学院,贵州遵义563000 [2]贵州省普通高等学校口腔疾病研究特色重点实验室,贵州遵义563000 [3]贵阳市口腔医院综合科,550000

出　　处：《重庆医学》2019年第10期1621-1624,1629,共5页Chongqing medicine

基　　金：国家自然科学基金项目(81560186);贵州省科技厅社发攻关项目(黔科合SY字[2013]3043号)

摘　　要：目的观察牙周炎基因疫苗pVAX1-血凝集素黏附结构域(HA2)-菌毛蛋白(FimA)、pVAX1-HA2-FimA/白细胞介素(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大鼠分为4组,A组:pVAX1-HA2-FimA组;B组:pVAX1-HA2-FimA/IL-15组;C组:pVAX1组;D组:生理盐水组,共免疫3次。利用酶联免疫吸附实验(ELISA)测定免疫后大鼠唾液分泌型IgA(sIgA)抗体水平,通过免疫组织化学检测鼻黏膜目的蛋白的表达。结果 A、B组基因疫苗免疫SD大鼠,唾液标本中sIgA型抗HA2抗体和抗FimA抗体水平在免疫第1周开始上升,2～5周逐步上升,于第5周达到高峰,自第6周开始缓慢下降,第9周抗体基本降到免疫前水平。C组及D组的抗体水平较低且无明显起伏变化。含有IL-15免疫佐剂重组质粒免疫组可诱导更高水平的抗体反应。免疫组织化学观察到鼻黏膜组织黏膜固有层存在目的蛋白的表达;HA2蛋白主要在细胞质中表达,FimA蛋白则分布在细胞质及细胞间质中。结论牙周炎基因疫苗能够成功诱导实验大鼠共同黏膜免疫系统产生免疫应答,具有免疫原性。IL-15作为牙龈卟啉单胞菌基因疫苗的佐剂可增强免疫效果。Objective To observe the immune effect of periodontitis vaccine pVAX1-HA2-FimA and pVAX1-HA2-FimA/IL-15 in rats by intranasal immunization,and investigate the expression of interest protein in situ.Methods 32 SD rats were divided into 4 groups,group A:the pVAX1-HA2-FimA group;group B:the pVAX1-HA2-FimA/IL-15 group;group C:the pVAX1 group;group D:the normal saline group.All rats were immunized three times.The secretory IgA(sIgA)antibodies were detected by ELISA and the expression of interest protein were detected by immunohistochemistry technique.Results The levels of sIgA anti-HA2 antibody and anti-FimA antibody in saliva of SD rats in group A and B began to rise at the first week of immunity,and gradually increased during the second to fifth weeks,reached the peak at the fifth week,and slowly decreased from the sixth week,reduced to the pre-immunity level at the ninth week.Antibody levels were low and did not fluctuate significantly.The antibody contained IL-15 immune adjuvant recombinant plasmid induced a higher level of antibody response.The expression of interest protein in the inherent mucosa of nasal mucosa was observed by immunohistochemistry.HA2 protein is mainly expressed in the cytoplasm,while FimA protein is distributed in the cytoplasm and interstitium.Conclusion Periodontitis gene vaccine can successfully induce immune responses in the common mucosal immune system by intranasal immunization in rats.IL-15 can be used as adjuvant of porphyromonas gingival gene vaccine to enhance the immune effect.

关 键 词：牙髓卟啉单胞菌 鼻黏膜 佐剂 免疫 基因疫苗 

分 类 号：R781.42[医药卫生—口腔医学]