论文部分内容阅读
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 f 1T epitope vaccine of dust mite allergen on specific immunotherapy in asthmatic mice. Methods Thirty mice in each group were randomly divided into three groups: PBS group, asthma group and immunotherapy group. The asthma group and the immunotherapy group were intraperitoneally injected with 100 μL of a sensitizing solution containing 100 μg / mL Der f1 [PBS solution containing 2% Al (OH) 3] on days 0, 7 and 14, respectively. The PBS group was treated with PBS solution [2% Al (OH) 3] instead. The asthma group and immunotherapy group inhaled the above sensitized solution from day 21 onwards, and the PBS solution was inhaled into the above PBS solution atomized for 30 min every day for 7 days. In the immunotherapy group, 200μL of der f1T cell epitope protein (100μg / mL) was injected intraperitoneally 30min before the atomization on the 25th to 27th days for specific immunotherapy. The PBS group was replaced by PBS solution. Mice were sacrificed 24 hours after the last inhalation and mice were killed. Bronchoalveolar lavage fluid (BALF) and serum from each group of mice were collected and eosinophils were counted in BALF. Lung tissue was taken to make lung histopathology. Spleen cells were isolated Join Der f1 co-culture made of spleen cell culture supernatant. The levels of IL-13 and IFN-γ in the culture supernatant of BALF and spleen cells, and the levels of serum specific IgE (sIgE) and IgG2a were detected by ELISA. Results Microscopic examination of lung tissue showed that inflammation in immunotherapy group was significantly better than that in asthma group, and infiltration of inflammatory cells was decreased. The number of eosinophils in BALF of mice was significantly lower than that in asthma group (P <0.01). The level of IL-13 in BALF in immunotherapy group was significantly lower than that in asthma group (P <0.01). However, the content of IL-13 in BALF was opposite to that in asthma group (P <0.01). The levels of IL-13 in the spleen cell culture supernatants in each group were significantly lower than those in the asthma group (P <0.01), while the levels of IFN-γ and IL-13 in the immunized group were significantly higher than those in the asthma group (P < 0.01). Serum specific IgE levels in the immunotherapy group were significantly lower than those in the asthma group (P <0.01). The level of IgG2a was the opposite of that of the IgE group. The immunotherapy group was significantly higher than the asthma group (P <0.01). Conclusion Der f1 T cell epitope vaccine can effectively relieve inflammation in asthmatic mice and correct the imbalance of Th1 / Th2.