摘要
目的利用表达结核分枝杆菌蛋白Ag85B、ESAT-6、Rv2031c和Rv2626c的重组腺病毒(Ad-AE-R2)滴鼻黏膜免疫小鼠,探究其对哮喘小鼠气道炎症的免疫预防作用。方法本研究为实验研究,采用单纯随机抽样方法。将18只6~8周龄BALB/c雌性小鼠随机分为3组:空白对照组、哮喘模型组和免疫预防组,每组6只。免疫预防组滴鼻免疫108 PFU Ad-AE-R2,空白对照组和哮喘模型组滴鼻等量生理盐水。小鼠免疫3周后,利用卵清蛋白诱导免疫预防组和哮喘模型组小鼠哮喘。空白对照组用生理盐水雾化作为对照。末次激发小鼠哮喘24 h后,麻醉处死各组小鼠,检测小鼠支气管肺泡灌洗液(BALF)和外周血中IgE和γ干扰素(IFN-γ)、白细胞介素5(IL-5)、IL-13等细胞因子的含量,HE染色观察肺组织病理学改变。结果与空白对照组相比,哮喘模型组BALF和血清中的IgE含量增加(均P<0.05),BALF中IFN-γ/IL-5下降(P<0.01)。与哮喘模型组相比,免疫预防组BALF中IL-13含量降低(P<0.01),BALF和血清中IFN-γ/IL-5升高(均P<0.05)。小鼠肺组织病理切片HE染色结果表明,与空白对照组相比,哮喘模型组小鼠肺组织结构出现明显损伤,气道黏膜上皮细胞局部脱落,管壁及基底膜显著增厚,气道及血管周围可见大量炎症细胞浸润;与哮喘模型组相比,免疫预防组小鼠肺组织中的炎症性病理改变明显减轻。结论利用卵清蛋白成功诱导小鼠气道炎症,建立小鼠哮喘模型。Ad-AE-R2黏膜免疫可以提升小鼠气道Th1型免疫反应,促进Th1/Th2型细胞因子平衡,对小鼠哮喘具有一定的预防保护效果,为哮喘的免疫预防提供研究参考。
Objective To explore the immunoprophylaxis effect of a recombinant adenovirus expressing Mycobacterium tuberculosis proteins Ag85B,ESAT-6,Rv2031c,and Rv2626c(Ad-AE-R2)on the airway inflammation in asthmatic mice by intranasal mucosal immunization.Methods This was an experimental study.A simple random sampling method was used.Eighteen female BALB/c mice aged 6-8 weeks were randomly divided into 3 groups,immunoprophylaxis group,asthma model group,and blank control group,with 6 mice in each group.The immunoprophylaxis group was immunized with 108 PFU recombinant adenovirus of Ad-AE-R2 per mice intranasally,while the blank control group and the asthma model group were intranasally administered the same amount of normal saline.Three weeks after immunization,OVA was used to induce asthma in the immunoprophylaxis group and the asthma model group.The blank control group was nebulized with normal saline as a control.Twenty-four hours after the last challenge of asthma,the mice in each group were anesthetized and sacrificed,and the content of IgE,the cytokines of IFN-γ,IL-5,and IL-13 in bronchoalveolar lavage fluid(BALF)and in peripheral blood were detected.Pathological changes of the lung tissue in mice were observed by HE staining.Results Compared with the blank control group,the contents of IgE in both BALF and sera of mice in asthma model group were increased(P<0.05),with the ratio of IFN-γ/IL-5 in BALF decreased(P<0.0001).Compared with asthma model group,the content of IL-13 in BALF of mice in immunoprophylaxis group was decreased(P<0.01),with the ratios of IFN-γ/IL-5 in BALF(P<0.0001)and sera(P<0.05)of mice both increased.Compared with the blank control group,the HE staining for histopathological changes of the lung tissue in asthma model group showed that the lung tissue structure was significantly damaged,the airway mucosal epithelial cells were partially shed,the tube wall and basement membrane were significantly thickened,and the airways and blood vessels showed extensive infiltration of inflammatory cells.Com
作者
向光宇
陈海沣
靳芊芊
王丽梅
Xiang Guangyu;Chen Haifeng;Jin Qianqian;Wang Limei(The Third Team of Students,School of Basic Medicine,Air Force Medical University,Xi′an 710032,China;Department of Microbiology and Pathogen Biology,School of Basic Medicine,Air Force Medical University,Xi′an 710032,China)
出处
《国际呼吸杂志》
2023年第6期646-651,共6页
International Journal of Respiration
基金
陕西省重点研发计划(2019SF-090)。
关键词
结核分枝杆菌
重组腺病毒
哮喘
黏膜免疫
免疫预防
Mycobacterium tuberculosis
Recombinant adenovirus
Asthma
Mucosal immunity
Immunoprophylaxis
