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目的探讨害嗜鳞螨Ⅱ类变应原Lep d2对尘螨致敏哮喘小鼠的特异性免疫治疗效果。方法将30只SPF级BALB/c小鼠随机分为PBS组、哮喘组、Lep d2治疗组,于实验开始时及实验第7、14天用尘螨提取液对哮喘组及Lep d2治疗组小鼠腹腔注射致敏,第21天开始雾化吸入激发,连续7 d,第25~27天开始,Lep d2免疫治疗组于雾化前30 min用Lep d2变应原进行免疫治疗,PBS组及哮喘组用PBS进行腹腔注射及雾化吸入。最后1次雾化激发24 h后,收集支气管肺泡灌洗液(BALF)和血清,并进行脾细胞培养。采用HE染色观察各组小鼠肺组织病理变化;采用ELISA法测定BALF和脾细胞培养液上清中IFN-γ、IL-5、IL-13及血清中特异性Ig E(s Ig E)、s Ig G2a抗体水平。结果哮喘组及Lep d2治疗组小鼠出现哮喘急性发作症状,PBS组未出现该症状。哮喘组肺部炎性细胞浸润明显,PBS组及Lep d2治疗组肺部炎症较轻。Lep d2治疗组BALF和脾细胞培养液上清中IFN-γ浓度均高于哮喘组(P均<0.01),但IL-5和IL-13浓度则均低于哮喘组(P均<0.01)。Lep d2治疗组s Ig E抗体水平低于哮喘组(P<0.01);但s Ig G2a抗体水平高于哮喘组(P均<0.01)。结论害嗜鳞螨Ⅱ类变应原Lep d2疫苗可有效减轻哮喘小鼠肺部炎症。
Objective To investigate the immunotherapeutic effect of Lep d2, a key allergen of Acaroid pest scale Ⅱ, on dust mite-sensitized asthmatic mice. Methods Thirty SPF BALB / c mice were randomly divided into PBS group, asthma group and Lep d2 treatment group. At the beginning of the experiment and on the 7th and 14th day of the experiment, the dust mite extract was used to treat asthma and Lep d2 treatment groups Rats were sensitized by intraperitoneal injection and started inhalation on day 21 for 7 d. From the 25th to 27th day, Lep d2 immunotherapy group was immunized with Lep d2 allergen at 30 min before aerosolization. PBS and The asthma group was intraperitoneally injected with PBS and inhaled by nebulization. After the last aerosol challenge, bronchoalveolar lavage fluid (BALF) and serum were collected and cultured in spleen cells. The pathological changes of lung tissue in each group were observed by HE staining. The levels of IFN-γ, IL-5 and IL-13 in the culture supernatant of BALF and splenocytes were detected by ELISA. The serum specific IgE, s Ig G2a antibody levels. Results Asthma and Lep d2-treated mice developed acute exacerbation of asthma, but no symptoms were observed in PBS group. Inflammatory cells in the lungs of asthmatic group were significantly infiltrated, and pulmonary inflammation in PBS group and Lep d2-treated group was lighter. The concentrations of IFN-γ in BALF and spleen cell supernatant of Lep d2 group were higher than those in asthma group (all P <0.01), but the concentrations of IL-5 and IL-13 in Lep d2 group were lower than those in asthma group (all P <0.01) . The level of s Ig E antibody in Lep d2 treatment group was lower than that in asthma group (P <0.01), but s Ig G2a antibody level was higher than that in asthma group (all P <0.01). Conclusions Lep d2, an allergen-attenuated Acaroidid class II allergen, can effectively reduce lung inflammation in asthmatic mice.