Ag85B DNA vaccine suppresses airway inflammation in a murine model of asthma.
Ag85B DNA vaccine suppresses airway inflammation in a murine model of asthma.
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DOI：
10.1186/1465-9921-10-51
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发表时间：
2009-06-16
影响因子：
5.8
通讯作者：
Zhong N
中科院分区：
文献类型：
--
作者：
Wu J;Xu J;Cai C;Gao X;Li L;Zhong N
在过敏性哮喘中，Th2 淋巴细胞被认为在协调气道嗜酸性粒细胞增多和炎症方面发挥着重要作用。重置 Th1/Th2 失衡可能对哮喘有治疗作用。结核分枝杆菌 30 千道尔顿主要分泌蛋白（抗原 85B、Ag85B）可以通过诱导 Th1 主导反应来保护动物免受结核分枝杆菌感染。在本研究中，Ag85B基因被克隆到pMG质粒中以产生pMG-Ag85B质粒。重组pMG-Ag85B质粒鼻内免疫后，通过Western blotting和免疫组化染色检测小鼠支气管上皮细胞中Ag85B基因的表达。通过组织学检查和支气管肺泡灌洗（BAL）评估pMG-Ag85B质粒免疫对气道炎症的保护作用。使用 ELISA 试剂盒测定 BAL 和脾细胞培养物上清液中的 IL-4 和 IFN-γ 水平。通过用重建的pMG-Ag85B质粒进行鼻内免疫，Ag85B基因在小鼠支气管上皮细胞中成功表达。使用卵清蛋白 (OVA) 诱导的小鼠哮喘模型，pMG-Ag85B 免疫显着抑制气道上皮细胞浸润，细胞总数减少 37%（9.6 ± 2.6 × 105/ml 对比 15.2 ± 3.0 × 105/ml，p < 0.05），嗜酸性粒细胞数量减少 74% (1.4 ± 0.2 × 105/ml vs. 5.4 ± 1.1 × 105/ml, p < 0.01) 与 OVA 致敏对照组相比。 pMG-Ag85B组、OVA致敏对照组和空pMG组之间BAL液中中性粒细胞的数量没有差异。与 pMG-Ag85B 组相比，pMG-Ag85B 组 BAL 液中的 IL-4 产量显着降低（32.0 ± 7.6 pg/ml 对比 130.8 ± 32.6 pg/ml，p < 0.01）和脾细胞上清液中（5.1 ± 1.6 pg/ml 对比 10.1 ± 2.3 pg/ml，p < 0.05） OVA 致敏对照组，而 BAL 液中的 IFN-γ 产量增加（137.9 ± 25.6 pg/ml 与 68.4 ± 15.3 pg/ml，p < 0.05）和脾细胞上清液中（20.1 ± 5.4 pg/ml 与 11.3 ± 3.2 pg/ml，p < 0.05）。在 OVA 诱导的哮喘小鼠模型中，pMG-Ag85B 鼻内免疫可显着减少过敏性气道炎症，并减少嗜酸性粒细胞浸润。这种保护作用与 BAL 液和培养的脾细胞上清液中 IL-4 的减少和 IFN-γ 产量的增加有关。
In allergic asthma, Th2 lymphocytes are believed to play important roles in orchestrating airway eosinophilia and inflammation. Resetting the Th1/Th2 imbalance may have a therapeutic role in asthma. The mycobacterium tuberculosis 30-kilodalton major secretory protein (antigen 85B, Ag85B) can protect animals from M. tuberculosis infection by inducing a Th1-dominant response. In this study, the Ag85B gene was cloned into pMG plasmids to yield the pMG-Ag85B plasmid. The expression of Ag85B gene in murine bronchial epithelia cells was detected by Western blotting and immunohistochemical staining after intranasal immunization with reconstructed pMG-Ag85B plasmids. The protective effect of pMG-Ag85B plasmids immunization in airway inflammation was evaluated by histological examination and bronchoalveolar lavage (BAL). IL-4 and IFN-γ levels in the BAL and supernatant from splenocyte culture were determined using ELISA kits. The Ag85B gene was successfully expressed in murine bronchial epithelia cells by intranasal immunization with reconstructed pMG-Ag85B plasmids. Using a murine model of asthma induced by ovalbumin (OVA), pMG-Ag85B immunization significantly inhibited cellular infiltration across the airway epithelium with a 37% decrease in the total number of cells (9.6 ± 2.6 × 105/ml vs. 15.2 ± 3.0 × 105/ml, p < 0.05) and a 74% decrease in the number of eosinophils (1.4 ± 0.2 × 105/ml vs. 5.4 ± 1.1 × 105/ml, p < 0.01) compared with the OVA-sensitized control group. There was no difference in the number of neutrophils in BAL fluid between the pMG-Ag85B group, the OVA-sensitized control group and the empty pMG group. IL-4 production was significantly decreased in the BAL fluid (32.0 ± 7.6 pg/ml vs. 130.8 ± 32.6 pg/ml, p < 0.01) and in the splenocyte supernatant (5.1 ± 1.6 pg/ml vs. 10.1 ± 2.3 pg/ml, p < 0.05) in the pMG-Ag85B group compared with the OVA-sensitized control group, while IFN-γ production was increased in the BAL fluid (137.9 ± 25.6 pg/ml vs. 68.4 ± 15.3 pg/ml, p < 0.05) and in the splenocyte supernatant (20.1 ± 5.4 pg/ml vs. 11.3 ± 3.2 pg/ml, p < 0.05). In a murine model of asthma induced by OVA, intranasal immunization with pMG-Ag85B significantly reduced allergic airway inflammation with less eosinophil infiltration. This protective effect was associated with decreased IL-4 and increased IFN-γ production in the BAL fluid and in the supernatant of cultured splenocytes.
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