水凝胶负载pVAX1-SpaP/P防龋DNA疫苗以4种途径产生免疫效应  

Hydrogel loaded anti-caries DNA vaccine pVAX1-SpaP/P produces immune effects in four ways

作　　者：李虎 管晓燕 李敏 董竞男 肖茜文 白国辉 王铭蔚 刘建国 Li Hu;Guan Xiaoyan;Li Min;Dong Jingnan;Xiao Qianwen;Bai Guohui;Wang Mingwei;Liu Jianguo(School of Stomatology,Zunyi Medical University,Zunyi 563000,Guizhou Province,China;Zunyi Medical University,Zunyi 563000,Guizhou Province,China;Special Key Laboratory of Oral Diseases Research,Higher Education Institution in Guizhou Province,Zunyi 563000,Guizhou Province,China)

机构地区：[1]遵义医科大学口腔医学院,贵州省遵义市563000 [2]遵义医科大学,贵州省遵义市563000 [3]贵州省普通高校口腔疾病研究特色重点实验室,贵州省遵义市563000

出　　处：《中国组织工程研究》2022年第27期4340-4345,共6页Chinese Journal of Tissue Engineering Research

基　　金：贵州省暨遵义市医用生物材料研发人才基地建设项目(黔人领发[2018]3号,遵委[2019]69号),项目负责人:刘建国;遵义市科技支撑计划项目[遵市科合HZ字(2020)297号],项目负责人:白国辉;遵义医科大学学术新苗培养及创新探索专项项目(黔科合平台人才[2017]5733-037),项目负责人:白国辉;珠海市医学科研基金项目(ZH3310200018PJL),项目负责人:王铭蔚。

摘　　要：背景:防龋DNA疫苗价格低廉、安全有效,有望成为人类抵抗龋病的重要手段,因而学者在寻找一种能高效诱导抗体产生的免疫途径或保护疫苗效价的缓释系统及佐剂,成为近年研究的热点。目的:观察聚乳酸乙醇酸-聚乙二醇-聚乳酸乙醇酸水凝胶负载pVAX1-SpaP/P防龋基因疫苗的免疫效应。方法:以聚乳酸乙醇酸-聚乙二醇-聚乳酸乙醇酸水凝胶分别负载重组质粒pVAX1-SpaP/P与空载质粒pVAX1。随机将28只新西兰大白兔分为7组,其中4组分别为股四头肌注射、颌下腺区皮下注射、口服、鼻腔滴注负载重组质粒pVAX1-SpaP/P的水凝胶,另3组为对照组,阴性对照组1股四头肌注射负载空载质粒pVAX1的水凝胶,阴性对照组2口服负载空载质粒pVAX1的水凝胶,空白对照组股四头肌注射生理盐水,初次免疫后1周增强免疫1次,间隔2周后加强免疫第2次,共免疫3次。应用ELISA法检测免疫前后唾液与血清中IgA型抗SpaP抗体、血清中IgG型抗SpaP抗体水平;免疫结束后第3天,免疫组化SP法检测免疫部位pVAX1-SpaP/P蛋白的表达。结果与结论:①经不同途径免疫负载重组质粒pVAX1-SpaP/P的水凝胶,免疫后2周兔血清中IgG型抗SpaP抗体与IgA型抗SpaP抗体水平开始升高,IgG型抗SpaP抗体水平在第10周达到峰值,IgA型抗SpaP抗体水平在第6周达到峰值,口服组6-12周的IgG型抗SpaP抗体水平高于其他6组(P<0.05),颌下腺组4-8周的IgA型抗SpaP抗体水平高于其他6组(P<0.05);②经不同途径免疫负载重组质粒pVAX1-SpaP/P的水凝胶,免疫后1周兔唾液中IgA型抗SpaP抗体水平开始升高,第8周达到峰值,颌下腺组1-6周的抗体水平高于其他6组(P<0.05),口服组8-12周的抗体水平高于其他6组(P<0.05);免疫后第2周兔唾液中IgG型抗SpaP抗体水平开始升高,第10周达到峰值,口服组第2,3,6周的抗体水平高于其他6组(P<0.05),股四头肌组第10周的抗体水平高于其他6组(P<0.05);③经不同途径免疫�BACKGROUND:Due to low price,safety and effectiveness,anti caries DNA vaccine is expected to become an important means of human anti caries.Scholars are looking for an effective immune pathway to induce antibody production or a sustained-release system and adjuvant to protect vaccine titer,which has become a research focus in recent years.OBJECTIVE:To observe the immune effect of the sustained release hydrogel poly(lactic/glycolic acid)-polyethylene glycol-poly(lactic/glycolic acid)loaded with anti-caries gene vaccine pVAX1-SpaP/P.METHODS:The recombinant plasmid pVAX1-SpaP/P and the empty plasmid pVAX1 were loaded with poly(lactic/glycolic acid)-polyethylene glycol-poly(lactic/glycolic acid)hydrogel,respectively.Twenty-eight New Zealand white rabbits were randomly divided into seven groups.Quadriceps femoris injection,submandibular gland subcutaneous injection,oral administration,and nasal drip injection of hydrogel loaded with recombinant plasmid pVAX1-SpaP/P were performed in four groups.The remaining groups were the control groups.In the negative control group 1,quadriceps muscle was injected with hydrogel loaded with empty plasmid pVAX1.In the negative control group 2,hydrogel loaded with empty plasmid pVAX1 was orally taken.The quadriceps femoris of the blank control group was injected with saline.The immunization was enhanced once a week after the initial immunization.The second immunization was conducted one week apart,and immunized for the third time after an interval of two weeks.ELISA was used to detect anti-SpaP IgA antibody in saliva and anti-SpaP IgG antibody in serum before and after immunization.At 3 days after the end of the three immunizations,the expression of pVAX1-SpaP/P protein was observed by immunohistochemistry technique.RESULTS AND CONCLUSION:(1)Two weeks after immunization with hydrogels loaded with recombinant plasmid pVAX1-SpaP/P via different pathways,the levels of anti-SpaP IgA antibody in saliva and anti-SpaP IgG antibody in serum were significantly increased in rabbits.Anti-SpaP IgG

关 键 词：龋病 细胞表面抗原SpaP 基因疫苗 水凝胶 疫苗 免疫应答 变异链球菌 目的蛋白 

分 类 号：R459.9[医药卫生—治疗学] R392.9[医药卫生—临床医学]