草分支杆菌疫苗治疗哮喘模型小鼠的实验研究

Effects of Mycobacteria Vaccine on Asthma Model in Mice

目的 研究草分支杆菌疫苗对哮喘模型小鼠的疗效及其作用机制。方法 将 18只BALB/c小鼠分为3组 ,每组各 6只 ,其中卵蛋白致敏哮喘组 (OVA组 )和草分支杆菌疫苗治疗组 (Utilin组 )皮下注射卵蛋白致敏制作哮喘模型 ,然后用卵蛋白激发 2次 ;阴性对照组 (NS组 )皮下注射生理盐水 (NS) ,然后NS激发 2次。Utilin组在激发前后分别给予草分支杆菌疫苗 0 .5 μg腹腔注射 3次 ,其他两组不作干预。 3组分别在第 2次激发后第 1、2、3、4周眼眶后静脉丛采血测OVA特异性免疫球蛋白IgE ,并于激发后第 4周处死小鼠测肺泡灌洗液 (BALF)中的细胞总数及嗜酸性粒细胞 (EOS)计数 ,肺组织病理切片观察形态学改变 ,并测定脾细胞培养上清液中OVA特异性IFN γ。结果 Utilin组BALF中细胞总数为 (2 9.5 1± 5 .81)× 10 4 /mL、EOS为 (2 .88± 0 .96 )× 10 4 /mL ,明显低于OVA组 [分别为 (4 0 .15± 6 .12 )× 10 4 /mL和 (6 .91± 1.92 )× 10 4 /mL],P <0 .0 5 ;Utilin组肺组织炎症反应较OVA组明显减轻 ;Utilin组脾细胞培养上清液中OVA特异性IFN γ的浓度为 (4 6 9± 86 )pg/mL ,明显高于OVA组 (193± 80 ) pg/mL ,P <0 .0 5 ;Utilin组激发后第 3周和第 4周OVA特异性IgE分别为 (0 .2 99± 0 .0 92 )(OD值 ) ,(0 .2 6 7± 0 .

Purpose: To study the efficacy and its mechanism of mycobacteria vaccine for asthma model in mice. Methods: 18 female BALB/c mice were divided into 3 groups. Both group OVA and group Utilin were immunized subcutaneously with OVA/alum and challenged with OVA. Group NS was injected and challenged with NS. Group Utilin was injected with mycobacteria vaccine 0.5 μg intraperitioneally before and after challenged three times. Thereafter blood samples were obtained every week for four times. OVA-specific IgE were measured by ELISA. BALF, lung tissues and spleen cells were collected after fourth blood collection. Total cell numbers and its classification of BALF were counted. IFN-γ produced by OVA-stimulated spleen cells were determined by ELISA. Results: Pathological examination showed that in group utilin airway inflammation and eosinophil infiltration in airway and lung parenchyma were inhibited as compared with group OVA. In group utilin total cell numbers and its classification of BALF were decreased, serum level of IgE was decreased, and the Th1 cytokine IFN-γ in spleen cell supernatants was increased as compared with group OVA. Conclusions: Mycobacteria vaccine is an effective immune-modulation, it could treat asthma by the means of adjustment of the balance of Th1/Th2.