粉尘螨1类变应原Der f1 T细胞表位疫苗对哮喘小鼠特异性免疫治疗的实验研究

Der f1 T cell epitope vaccine from Dermatophagoides farinae as the specific immunotherapy in asthma mice

目的探讨粉尘螨1类变应原Der f1T细胞表位疫苗对哮喘小鼠特异性免疫治疗的效果。方法30只小鼠按每组10只随机分为三组,PBS组、哮喘组和免疫治疗组。哮喘组和免疫治疗组分别在第0、7、14天经小鼠腹腔注射100μL含100μg/mL Der f1的致敏液[含2%Al（OH）3的PBS液],PBS组以PBS液[含2%Al（OH）3]代替。哮喘组和免疫治疗组自第21天起,雾化吸入上述致敏液,PBS组雾化吸入上述PBS液,每天雾化30min,连续雾化7d。免疫治疗组在第25~27天雾化前30min,经腹腔注射100μg/mL的Der f1T细胞表位蛋白200μL,进行特异性免疫治疗,PBS组以PBS液代替。最后一次雾化吸入24h后处死小鼠,收集每组小鼠的支气管肺泡灌洗液（BALF）和血清并计数BALF中嗜酸性粒细胞,取肺组织制作肺组织病理切片,取脾脏分离脾细胞加入Der f1共培养制成脾细胞培养上清。ELISA检测BALF和脾细胞培养上清中IL-13和IFN-γ含量及血清中特异性IgE（sIgE）和IgG2a水平。结果肺组织病理切片镜检说明,免疫治疗组比哮喘组炎症明显好转,炎症细胞浸润减少。小鼠BALF中嗜酸性粒细胞数结果为免疫治疗组嗜酸性粒细胞数明显低于哮喘组（P〈0.01）。BALF中IL-13含量免疫治疗组明显低于哮喘组（P〈0.01）,而IFN-γ含量与IL-13含量变化相反,免疫治疗组明显高于哮喘组（P〈0.01）。各组脾细胞培养上清中IL-13,免疫治疗组明显低于哮喘组（P〈0.01）,而IFN-γ含量与IL-13含量变化相反,免疫治疗组明显高于哮喘组（P〈0.01）。血清中特异性IgE含量结果为免疫治疗组明显低于哮喘组（P〈0.01）,IgG2a变化水平与IgE变化正好相反,免疫治疗组明显高于哮喘组（P〈0.01）。结论 Der f1T细胞表位疫苗能有效缓解哮喘小鼠的炎症反应并纠正Th1/Th2失衡。

Objective To investigate the effect of Der f1 Tcell epitope vaccine in asthma mice as a specific immunotherapy.Methods 30 mice were randomly divided into three groups：PBS group,asthma group and SIT group.On days 0,7and 14,the mice in asthma group and SIT group were intraperitoneally injected 100μg of Der f1,while PBS group received PBS injection.From day 21,the mice in asthma group and SIT group were challenged with Der f1（100μg/mL）inhalation for 7successive days,and the mice in SIT group were injected subcutaneously 200μg of Der f1 Tcell epitope vaccine before inhalation.during Day 25 and Day 27,while the mice in PBS group were treated with PBS.24 hours after the last inhalation,the mice were sacrificed,and the levels of IFN-γand IL-13 in bronchoalveolar lavage fluid（BALF）and spleen cell culture supernatants as well as the serum levels of specific IgE（sIgE）and IgG2 awere detected with ELISA.The number of eosinophils in BALF were counted.Results The inflammation in the SIT group was significantly improved than the asthma group,with inflammatory cells reduced.The counts of eosinophils in BALF and spleen cell culture supernatant in the SIT group were significantly lower than in asthma group（P〈0.01）,while those of IFN-γwere significantly higher than in the asthma group（P〈0.01）.The content of specific IgE in serum in the SIT group was significantly lower than in the asthma group（P〈0.01）,while that of IgG2 awas significantly higher than in the asthma group（P〈0.01）.Conclusion Der f1 Tcell epitope vaccine can effectively improve the inflammatory reaction and the Th1/Th2 imbalance in asthma mice.