Intrinsic Phenotypic Differences of Asthmatic Epithelium and Its Inflammatory Responses to Respiratory Syncytial Virus and Air Pollution
Intrinsic Phenotypic Differences of Asthmatic Epithelium and Its Inflammatory Responses to Respiratory Syncytial Virus and Air Pollution
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DOI：
10.1165/rcmb.2011-0031oc
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发表时间：
2011-11-01
影响因子：
6.4
通讯作者：
Knight, Darryl A.
中科院分区：
文献类型：
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作者：
Hackett, Tillie-Louise;Singhera, Gurpreet K.;Knight, Darryl A.
与哮喘相关的医疗保健费用的很大一部分可归因于疾病的恶化。在气道内，上皮形成粘膜免疫屏障，这是抵抗常见环境损伤（例如呼吸道合胞病毒（RSV）和颗粒物）的第一结构细胞防御。我们试图描述分化的哮喘源性气道上皮细胞培养物的表型及其对环境挑战的内在炎症反应。气液界面（ALI）培养物由哮喘（n = 6）和非哮喘（n = 6）气道上皮细胞产生。通过免疫组化分析气道组织和ALI培养物的细胞角蛋白-5、E-钙粘蛋白、Ki 67、Muc 5AC、NF-κ B、p38活化和细胞凋亡。将ALI培养物暴露于RSV（4 × 10（6）空斑形成单位/ml）、加拿大环境卫生部收集的颗粒物质（EHC-93，100 μ g/ml）或机械损伤24、48和96小时，并使用Luminex和ELISA分析基底外侧上清液的炎性细胞因子。哮喘患者气道切片和体外培养的ALI患者气道上皮细胞分化程度较低，其特征为基底细胞数量增加，细胞角蛋白-5表达，p38丝裂原活化蛋白激酶磷酸化增加，粘附连接蛋白E-钙粘蛋白减少。哮喘和非哮喘培养物之间的跨上皮电阻没有差异。在RSV感染、EHC-93暴露或机械创伤后，哮喘ALI培养物释放的IL-6、IL-8和粒细胞巨噬细胞集落刺激因子浓度高于非哮喘培养物（P < 0.05）。这种平行的离体和体外研究表明，与非哮喘上皮细胞相比，哮喘上皮细胞对常见环境挑战的表型和异常炎症反应发生了本质性改变。
A substantial proportion of healthcare cost associated with asthma is attributable to exacerbations of the disease. Within the airway, the epithelium forms the mucosal immune barrier, the first structural cell defense against common environmental insults such as respiratory syncytial virus (RSV) and particulate matter. We sought to characterize the phenotype of differentiated asthmatic-derived airway epithelial cultures and their intrinsic inflammatory responses to environmental challenges. Air-liquid interface (ALI) cultures were generated from asthmatic (n = 6) and nonasthmatic (n = 6) airway epithelial cells. Airway tissue and ALI cultures were analyzed by immunohistochemistry for cytokeratin-5, E-cadherin, Ki67, Muc5AC, NF-kappa B, the activation of p38, and apoptosis. ALI cultures were exposed to RSV (4 x 10(6) plaque forming unit/ml), particulate matter collected by Environmental Health Canada (EHC-93, 100 mu g/ml), or mechanically wounded for 24, 48, and 96 hours and basolateral supernatants analyzed for inflammatory cytokines, using Luminex and ELISA. The airway epithelium in airway sections of patients with asthma as well as in vitro ALI cultures demonstrated a less differentiated epithelium, characterized by elevated numbers of basal cells marked by the expression of cytokeratin-5, increased phosphorylation of p38 mitogen-activated protein kinase, and less adherens junction protein E-cadherin. Transepithelial resistance was not different between asthmatic and nonasthmatic cultures. In response to infection with RSV, exposure to EHC-93, or mechanical wounding, asthmatic ALI cultures released greater concentrations of IL-6, IL-8, and granulocyte macrophage colony-stimulating factor, compared with nonasthmatic cultures (P < 0.05). This parallel ex vivo and in vitro study of the asthmatic epithelium demonstrates an intrinsically altered phenotype and aberrant inflammatory response to common environmental challenges, compared with nonasthmatic epithelium.