AP-HM Department of Respiratory diseases, Université Aix-Marseille, Marseille, France.
J Allergy Clin Immunol. 2012 May;129(5):1259-1266.e1. doi: 10.1016/j.jaci.2012.01.073. Epub 2012 Mar 10.
Structural changes to the airways are features of severe asthma. The bronchial epithelium facilitates this remodeling process. Learning about the changes that develop in the airway epithelium could improve our understanding of asthma pathogenesis and lead to new therapeutic approaches.
We sought to determine the feasibility and relevance of air-liquid interface cultures of bronchial epithelium derived from endobronchial biopsy specimens of patients with different severities of asthma for studying the airway epithelium.
Human bronchial epithelial cells derived from endobronchial biopsy specimens of patients with mild and severe asthma were maintained in culture for 21 days in an air-liquid interface to reproduce a fully differentiated airway epithelium. Initially, features of remodeling that included epithelial and subepithelial layers, as well as mucus production, were assessed in paraffin-embedded endobronchial biopsy specimens to evaluate morphologic characteristics of asthmatic patients' epithelia. Ex vivo differentiated epithelia were then analyzed for morphology and function based on ultrastructural analysis, IL-8 release, lipoxin A(4) generation, mucin production, and lipoxygenase gene expression.
Morphologic and inflammatory imbalances initially observed in endobronchial biopsy specimens obtained from patients with severe or mild asthma persisted in the air-liquid interface reconstituted epithelium throughout the differentiation process to 21 days. Epithelium from patients with severe asthma produced greater levels of mucin, released more IL-8, and produced lower levels of lipoxin A(4) than that from patients with mild asthma. Expression of 15-lipoxygenase 2 was increased in epithelium from patients with severe asthma, whereas expression levels of MUC5AC, MUC5B, 5-lipoxygenase, and 15-lipoxygeanse 1 were similar to those of patients with mild asthma.
Ex vivo cultures of fully differentiated bronchial epithelium from endobronchial biopsy specimens maintain inherent phenotypic differences specifically related to the severity of asthma.
气道结构的改变是严重哮喘的特征。支气管上皮促进了这种重塑过程。了解气道上皮发生的变化可以增进我们对哮喘发病机制的理解，并为新的治疗方法提供依据。
我们旨在确定源自不同严重程度哮喘患者支气管内活检标本的气道上皮的气液界面培养的可行性和相关性，以用于气道上皮的研究。
通过将源自轻、重度哮喘患者支气管内活检标本的人支气管上皮细胞在气液界面中培养 21 天，以重现完全分化的气道上皮，从而建立气道上皮的气液界面培养。最初，我们通过石蜡包埋的支气管内活检标本评估重塑特征，包括上皮和上皮下层以及黏液产生，以评估哮喘患者上皮的形态特征。然后，根据超微结构分析、IL-8 释放、脂氧素 A4 的生成、黏蛋白产生和脂氧合酶基因表达，对体外分化的上皮进行形态和功能分析。
在严重或轻度哮喘患者的支气管内活检标本中最初观察到的形态和炎症失衡，在整个 21 天的分化过程中，在重建的气液界面上皮中持续存在。与轻度哮喘患者相比，重度哮喘患者的上皮产生更多的黏蛋白，释放更多的 IL-8，并产生更少的脂氧素 A4。15-脂氧合酶 2 在重度哮喘患者的上皮中表达增加，而 MUC5AC、MUC5B、5-脂氧合酶和 15-脂氧合酶 1 的表达水平与轻度哮喘患者相似。
源自支气管内活检标本的完全分化的支气管上皮的体外培养保留了与哮喘严重程度相关的固有表型差异。
