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

doi:10.13701/j.cnki.kqyxyj.2020.04.011

口腔医学研究 ›› 2020, Vol. 36 ›› Issue (4): 350-354.DOI: 10.13701/j.cnki.kqyxyj.2020.04.011

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

田源^1,2, 陈靖², 曾凤娇³, 于航³, 刘霞³, 刘建国^2,3, 白国辉^2,3*

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

收稿日期:2019-07-19 出版日期:2020-05-28 发布日期:2020-05-28
通讯作者: 白国辉,E-mail:baiguohui1228@126.com
作者简介:田源(1982~ ),女,山东菏泽人,副主任医师,硕士,研究方向:口腔内科常见疾病的免疫学防治。
基金资助:
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)

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

TIAN Yuan^1,2, CHEN Jing², ZENG Fengjiao³, YU Hang³, LIU Xia³, LIU Jianguo^2,3, BAI Guohui^2,3*

1. Affiliated Dental Hospital of Zunyi Medical University, Zunyi 563000, China;
2. Key Laboratory of Oral Disease of Higher Schools in Guizhou Province, Zunyi 563000, China;
3. Zunyi Medical University, Zunyi 563000, China

Received:2019-07-19 Online:2020-05-28 Published:2020-05-28

摘要/Abstract

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。

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

Abstract: 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.

Key words: gene vaccine for periodontitis, porphyromonas gingival, mucosal immunity, follicular helper T lymphocytes, secretory immunoglobulin A

田源, 陈靖, 曾凤娇, 于航, 刘霞, 刘建国, 白国辉. 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[J]. 口腔医学研究, 2020, 36(4): 350-354.

TIAN Yuan, CHEN Jing, ZENG Fengjiao, YU Hang, LIU Xia, LIU Jianguo, BAI Guohui. Changes of Tfh Cells in SD Rats Immunized with Periodontitis DNA Vaccine pVAX1-HA2-FimA-(IL-15) through Nasal Mucosa[J]. Journal of Oral Science Research, 2020, 36(4): 350-354.

参考文献

[1] 李雨阳,孟维艳.牙龈卟啉单胞菌抵抗固有免疫防御的研究进展[J].口腔医学研究,2019,35(2):12-14.
[2] 王昆,申道南,吴亚菲,等.牙龈卟啉单胞菌相关牙周炎疫苗有效免疫原及其途径和佐剂[J].国际口腔医学杂志,2015, 42(3): 328-333.
[3] Jee J,Bonnegarde-Bernard A,Duverger A,et al. Neutrophile negarively regulate induction of mucosl IgA responses after sublingual immunization [J]. Mucosal immunol, 2015, 8(4):735-745.
[4] 秦妍,管晓燕,白国辉, 等.基因疫苗黏膜免疫及其机制的研究进展[J].贵州医药,2016,40(4):426-428.
[5] Crotty S. T follicular helper cell differentiation,function,and roles in disease [J]. Immunity,2014,41(4): 529-542.
[6] 闫天萌,左亚刚.滤泡性辅助性T细胞及其相关皮肤病[J].中国麻风皮肤病杂志,2018,34(7):62-64.
[7] 黄菲,胡成进,刘晓斐.滤泡辅助性T细胞在感染性疾病中的研究进展[J].国际检验医学杂志,2018,39(13):1629-1631.
[8] Crotty S. Follicular helper CD4 T cells(Tfh) [J]. Annu RevImmunol, 2011, 29(1): 621-663.
[9] Maceiras AR , Almeida SCP , Mariotti-Ferrandiz E , et al. T follicular helper and T follicular regulatory cells have different TCR specificity [J]. Nat Commun, 2017, 8:15067.
[10] Gong F, Zhu HY, Zhu J, et al. Circulating CXCR5⁺ CD4⁺ T cells participate in the IgE accumulation in allergic asthma [J]. Immunol Lett, 2018, 197:9-14.
[11] Takebe T, Sakamoto K, Higami Y, et al. A novel mouse model for tracking the fate of CXCR5-expressing T cells [J]. Biochem Biophys Res Commun, 2018, 495(2):1642-1647.
[12] Zhao PW, Shi X, Li C, et al. IL-33 enhances humoral immunity against chronic HBV infection through activating CD4⁺CXCR5⁺ TFH cells [J]. J Interferon Cytokine Res, 2015, 35(6):454-463.
[13] Tahiliani V, Hutchinson TE, Abboud G, et al. OX40 cooperates with ICOS to amplify follicular Th cell development and germinal center reactions during infection [J]. J Immunol, 2017, 198(1):218-228.
[14] Stone EL, Pepper M, Katayama CD, et al. ICOS coreceptor signaling inactivates the transcription factor FOXO1 to promote Tfh cell differentiation [J]. Immunity, 2015, 42(2):239-251.
[15] 王波,李媛,韩晓芳.ICOS转基因小鼠肝纤维化病程中ICOS对Tfh极化的作用[J].国际免疫学杂志,2019,42(1):24-31.
[16] 曹云峰,张荣,周俊,等.康复新口服液对口腔溃疡患者疼痛缓解,唾液SIgA水平、IgG水平的影响[J].中国生化药物杂志,2017,(7):45-46.
[17] Yang F, Wei XM, Liang WW, et al. A critical role for IL-21 receptor signaling in the coxsackievirus B3-induced myocarditis [J]. Inflammation, 2017, 40(4):1428-1435.
[18] Zhang J, Ren M, Zeng H, et al. Elevated follicular helper T Cells and expression of IL-21 in thyroid tissues are involved in the pathogenesis of Graves’ disease [J]. Immunol Res, 2015, 62(2):163-174.
[19] Holm TL, Tornehave D, Søndergaard H, et al. Evaluating IL-21 as a potential therapeutic target in Crohn's disease [J]. Gastroenterol Res Pract, 2018, 2018:5962624.
[20] 徐淑琴,包韦乔,吴蓓颖,等.激素对系统性红斑狼疮患者外周血滤泡辅助性T细胞及其亚群组成的影响[J].中华微生物学和免疫学杂志, 2017, 37(11):816-821.

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

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

[1]	张慧彦, 李保胜, 张震阳, 韩春雨, 李尊泰, 齐晓爽, 付齐月, 孟维艳. D/L-缬氨酸对牙龈卟啉单胞菌及其生物膜的作用[J]. 口腔医学研究, 2020, 36(7): 648-653.
[2]	邬扬绚, 向黎, 黄志强, 褚金海. 釉基质蛋白衍生物调节牙龈卟啉单胞菌感染牙周膜干细胞成骨分化的作用及机制[J]. 口腔医学研究, 2020, 36(6): 534-538.
[3]	王腾飞, 徐燕, 徐涵颖, 韩旭, 沈继龙, 程婷. 卵黄抗体缓释型纳米羟基磷灰石的制备及其抗菌性能的研究[J]. 口腔医学研究, 2020, 36(6): 591-594.
[4]	张源, 李新月. 脂多糖抑制牙周膜细胞骨唾液酸蛋白表达中微小RNA的筛选与验证[J]. 口腔医学研究, 2020, 36(5): 477-480.
[5]	许志强, 曾秀峰, 洪少楠, 曾秀霞, 贺于奇, 黄俊徽. 不同管径二氧化钛纳米管的抗菌性能研究[J]. 口腔医学研究, 2020, 36(3): 230-234.
[6]	王素文, 赖思煜, 习利军, 王利宏. 黄芪甲苷调节人牙周膜细胞炎症反应及成骨分化的作用及机制研究[J]. 口腔医学研究, 2020, 36(2): 121-125.
[7]	孙天祥, 苗棣, 王宝彦, 陈悦. 高糖对牙龈卟啉单胞菌LPS引发牙周膜细胞炎症反应的影响[J]. 口腔医学研究, 2020, 36(12): 1123-1127.
[8]	左源, 李维善. 牙龈卟啉单胞菌脂多糖对人脐静脉内皮细胞表达小凹蛋白-1的影响[J]. 口腔医学研究, 2020, 36(1): 75-78.
[9]	聂然, 周延民. 慢性牙周炎与阿尔茨海默病相关性的研究进展[J]. 口腔医学研究, 2019, 35(5): 419-422.
[10]	李雨阳,孟维艳. 牙龈卟啉单胞菌抵抗固有免疫防御的研究进展[J]. 口腔医学研究, 2019, 35(2): 113-115.
[11]	叶兴如, 徐燕, 庞罡, 王莹, 周莉莉, 蒋鹏, 沈继龙, 王荣海. RT-qPCR检测CPC对中重度慢性牙周炎患者口内环境中红色复合体的影响[J]. 口腔医学研究, 2018, 34(8): 852-856.
[12]	梁珊珊, 邢文燕, 季耀庭, 江汉, 杜民权. 牙龈卟啉单胞菌对大鼠胎盘及仔鼠出生体重的影响[J]. 口腔医学研究, 2018, 34(5): 495-499.
[13]	郭海盈, 任洪芋, 季耀庭, 江汉, 杜民权. p-Akt对牙龈卟啉单胞菌诱导的人胚胎滋养层细胞凋亡的影响[J]. 口腔医学研究, 2018, 34(5): 500-504.
[14]	唐路, 薛栋, 白雨豪, 王鹏程, 周维, 李文婷, 董岩, 曾红燕, 赵颖. 牙龈卟啉单胞菌对血管平滑肌细胞钙化作用的研究[J]. 口腔医学研究, 2018, 34(2): 130-133.
[15]	黄琳砚,赖仁发,黄怡. 牙周炎与动脉粥样硬化信号相互作用的关联影响[J]. 口腔医学研究, 2018, 34(12): 1271-1273.