SD大鼠经鼻黏膜免疫牙周炎基因疫苗pVAX1-HA2-FimA-(IL-15)后Tfh细胞的变化  

Changes of Tfh Cells in SD Rats Immunized with Periodontitis DNA Vaccine pVAX1-HA2-FimA-(IL-15)through Nasal Mucosa

作　　者：田源 陈靖 曾凤娇 于航 刘霞 刘建国 白国辉 TIAN Yuan;CHEN Jing;ZENG Fengjiao;YU Hang;LIU Xia;LIU Jianguo;BAI Guohui(Affiliated Dental Hospital of Zunyi Medical University, Zunyi 563000, China;Key Laboratory of Oral Disease of Higher Schools in Guizhou Province, Zunyi 563000, China;Zunyi Medical University, Zunyi 563000, China)

机构地区：[1]遵义医科大学附属口腔医院牙体牙髓科,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]遵义医科大学,贵州遵义563000

出　　处：《口腔医学研究》2020年第4期350-354,共5页Journal of Oral Science Research

基　　金：国家自然科学基金(编号:81560186);贵州省科技计划项目(黔科合基础[2019]1333号);遵义市科技计划课题[遵市科合HZ字(2019)21号];贵州省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SD大鼠经鼻黏膜免疫牙周炎基因疫苗后Tfh细胞的变化,探讨Tfh细胞在黏膜免疫应答产生特异性sIgA抗体中的作用。方法:采用鼻滴方式免疫SD大鼠,A组为生理盐水组,B组为空载体(pVAX1)组,C组为牙周炎疫苗pVAX1-HA2-FimA-(IL-15)组,ELISA法检测大鼠唾液中特异性sIgA抗体的水平;流式细胞技术检测鼻黏膜Tfh细胞及CD4+CXCR5+ICOS+Tfh细胞的表达。结果:第3~7周C组大鼠唾液中特异性sIgA抗体表达水平高于A组和B组(P<0.05);C组FITC(CD4)和Aleax flour 594(CXCR5)融合后呈黄色荧光,A组和B组的免疫荧光双染结果中未见双阳性荧光信号;C组大鼠鼻黏膜细胞中CD4+CXCR5+Tfh及CD4+CXCR5+ICOS+Tfh细胞的表达均高于A组和B组(P<0.05);且发现大鼠唾液中特异性sIgA与大鼠鼻黏膜CD4+CXCR5+Tfh及CD4+CXCR5+ICOS+Tfh细胞均呈正相关关系。结论:牙周炎基因疫苗pVAX1-HA2-FimA-(IL-15)经鼻黏膜免疫大鼠后可激活Tfh细胞在鼻黏膜的表达,表达水平与特异性sIgA抗体水平呈正相关关系,说明Tfh细胞可能在促进机体产生特异性sIgA抗体的过程中起到重要作用。Objective:To investigate the changes of Tfh cells in SD rats immunized with pVAX1-HA2-FimA-(IL-15)eriodontitis gene vaccine,and to investigate the role of Tfh cells in mucosal immune response to produce specific SIgA antibody.Methods:SD rats were randomly divided into three groups.Group A was nasal saline,group B was nasal drop vector(pVAX1),and group C was nasal drop periodontitis vaccine pVAX1-HA2-FimA-(IL-15).ELISA was used to detect the level of specific SIgA antibody in rat saliva,immunofluorescence technique was used to detect the localization of Tfh cells in nasal mucosa,and flow cytometry was used to detect Tfh cells in nasal mucosa and CD4+CXCR5+ICOS+Tfh Expression of cells.Results:The expression level of specific SIgA antibody in saliva of group C was always higher than that of group A and group B(P<0.05).The fusion of FITC(CD4)and Aleax flour 594(CXCR5)in group C showed yellow fluorescence,and the double immunofluorescence staining results of group A and group B showed no double positive fluorescence signal.The expression of CD4+CXCR5+Tfh and CD4+CXCR5+ICOS+Tfh in nasal mucosa of group C rats was higher than those of group A and group B(P<0.05).The specific SIgA in rat saliva and rat nasal mucosa CD4 were found.There were correlations between CD4+CXCR5+Tfh and CD4+CXCR5+ICOS+Tfh cells.Conclusion:pVAX1-HA2-FimA-(IL-15),a genetic vaccine for periodontitis,can activate the expression of Tfh cells in nasal mucosa after immunizing rats with specific SIgA antibody,and the expression level is positively correlated with the level of specific SIgA antibody,suggesting that Tfh cells may play an important role in promoting the production of specific SIgA antibody in the body.

关 键 词：牙周炎基因疫苗 牙龈卟啉单胞菌 黏膜免疫 滤泡性辅助性T淋巴细胞 分泌型免疫球蛋白 

分 类 号：R78[医药卫生—口腔医学]