尘螨疫苗免疫治疗对过敏性哮喘小鼠气道抗铜绿假单胞菌感染能力的影响

Effect of dust mite vaccine immunotherapy on the ability of airway to resist Pseudomonas aeruginosa infection in allergic asthma mice

目的探讨尘螨疫苗免疫治疗对过敏性哮喘小鼠气道抗铜绿假单胞菌感染能力的影响。方法取24只BALB/c小鼠随机分为4组(非哮喘组、非哮喘感染组、哮喘感染组和哮喘感染治疗组),每组6只。建立粉尘螨过敏性哮喘小鼠模型,采用尘螨疫苗进行过敏原特异性免疫治疗,并经鼻感染铜绿假单胞菌。取小鼠肺组织及支气管肺泡灌洗液(BALF)分析小鼠肺部炎症状况;取小鼠肺组织匀浆进行铜绿假单胞菌选择性培养,并分析小鼠肺组织及BALF沉渣细菌载量;免疫荧光组织化学染色分析小鼠气道上皮细胞β防御素3(MBD-3)的表达水平,聚合酶链反应检测MBD-3的mRNA水平,并对以上数据进行统计学处理。结果哮喘感染治疗组小鼠肺部炎性浸润比哮喘感染组轻,哮喘感染治疗组小鼠BALF中的炎性细胞低于哮喘感染组(P<0.05)。哮喘感染治疗组小鼠肺组织及BALF细菌菌落数低于哮喘感染组(P<0.05)。哮喘感染组MBD-3表达水平低于非哮喘感染组和哮喘感染治疗组(P<0.05),而哮喘感染治疗组和非哮喘感染组MBD-3表达差异无统计学意义(P>0.05)。结论尘螨疫苗免疫治疗可能通过上调MBD-3的表达来降低哮喘小鼠气道铜绿假单胞菌感染的严重程度,从而增强过敏性哮喘小鼠气道抗铜绿假单胞菌感染能力。

Objective To explore the effect of dust mite vaccine immunotherapy on the ability of airway to resist Pseudomonas aeruginosa infection in allergic asthma mice.Methods Twenty four BALB/c mice were randomly divided into 4 groups(non-asthmatic group,non-asthmatic infection group,asthma infection group and asthma infection treatment group),six in each group.The mouse models of dust mite allergic asthma were established,and the dust mite vaccine was used for allergen-specific immunotherapy,and Pseudomonas aeruginosa was infected through the nose.The lung tissues and bronchoalveolar lavage fluid(BALF)of mice were taken to analyze the lung inflammation,and the lung tissue homogenates were taken for selective culture of Pseudomonas aeruginosa,and the bacterial load of mouse lung tissue and BALF sediment was analyzed.Immunofluorescence histochemical staining was used to analyze the expression level of mouse beta defensin-3(MBD-3)in mouse airway epithelial cells,and the mRNA level of MBD-3 was detected by polymerase chain reaction,and the above data was analyzed statistically.Results The lung inflammation infiltration in the asthma infection treatment group was less than that in the asthma infection group,and the inflammatory cells in the BALF of the asthma infection treatment group were lower than those in the asthma infection group(P<0.05).The number of bacterial colonies in lung tissue and BALF of asthma infection treatment group were lower than those in the asthma infection group(P<0.05).The expression level of MBD-3 in the asthma infection group was lower than that in the non-asthma infection group and the asthma infection treatment group(P<0.05),but there was no statistically significant difference in MBD-3 expression between the asthma infection treatment group and the non-asthma infection group(P>0.05).Conclusion Immunotherapy with dust mite vaccine may reduce the severity of Pseudomonas aeruginosa infection in airway of asthma mice by up-regulating the expression of airway MBD-3,thereby enhancing the ability of airway to resist Pseudomonas aeruginosa infection in allergic asthmatic mice.