Tracheobronchial air-liquid interface cell culture: a model for innate mucosal defense of the upper airways?
Tracheobronchial air-liquid interface cell culture: a model for innate mucosal defense of the upper airways?
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DOI：
10.1152/ajplung.90388.2008
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发表时间：
2009-01
影响因子：
4.9
通讯作者：
Sheehan, John K.
中科院分区：
文献类型：
--
作者：
Kesimer, Mehmet;Kirkham, Sara;Pickles, Raymond J.;Henderson, Ashley G.;Alexis, Neil E.;DeMaria, Genevieve;Knight, David;Thornton, David J.;Sheehan, John K.
人气管支气管上皮细胞气液界面培养已成为气道生物学研究的有力工具。在这项研究中，我们已经调查了这种培养系统是否产生“粘液”的蛋白质组成类似于在体内，诱导气道分泌物。以前的粘液分泌物的成分研究大大低估了粘蛋白的贡献，粘蛋白是正常粘液的主要结构成分。为了克服这一局限性，我们使用了一种基于质谱的方法，该方法集中在粘蛋白与大多数其他蛋白质的预先分离上。使用这种方法，我们已经比较了顶端分泌物（AS）从分化良好的原代人气管支气管细胞生长在气液界面和人气管支气管正常诱导痰（IS）的蛋白质组成。共鉴定出186种蛋白质，134种来自AS，136种来自IS; 84种蛋白质是两种分泌物共有的，其中宿主防御蛋白占主导地位。上皮粘蛋白MUC 1，MUC 4和MUC 16和凝胶形成粘蛋白MUC 5 B和MUC 5AC被确定在两种分泌物中。折射率测定表明，凝胶形成粘蛋白的主要贡献者的质量两种分泌物。当IS的组成被校正为最可能来自唾液、血清和迁移细胞的蛋白质时，两种分泌物之间存在相当大的相似性，特别是在宿主防御蛋白的类别中，其中包括粘蛋白。这表明原代细胞培养系统是研究上呼吸道先天防御方面的重要模型，特别是与仅由呼吸道细胞产物组成的粘液相关的方面。
Human tracheobronchial epithelial cells grown in air-liquid interface culture have emerged as a powerful tool for the study of airway biology. In this study, we have investigated whether this culture system produces “mucus” with a protein composition similar to that of in vivo, induced airway secretions. Previous compositional studies of mucous secretions have greatly underrepresented the contribution of mucins, which are major structural components of normal mucus. To overcome this limitation, we have used a mass spectrometry-based approach centered on prior separation of the mucins from the majority of the other proteins. Using this approach, we have compared the protein composition of apical secretions (AS) from well-differentiated primary human tracheobronchial cells grown at air-liquid interface and human tracheobronchial normal induced sputum (IS). A total of 186 proteins were identified, 134 from AS and 136 from IS; 84 proteins were common to both secretions, with host defense proteins being predominant. The epithelial mucins MUC1, MUC4, and MUC16 and the gel-forming mucins MUC5B and MUC5AC were identified in both secretions. Refractometry showed that the gel-forming mucins were the major contributors by mass to both secretions. When the composition of the IS was corrected for proteins that were most likely derived from saliva, serum, and migratory cells, there was considerable similarity between the two secretions, in particular, in the category of host defense proteins, which includes the mucins. This shows that the primary cell culture system is an important model for study of aspects of innate defense of the upper airways related specifically to mucus consisting solely of airway cell products.
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DOI：
10.1165/ajrcmb.22.6.3980
发表时间：
2000-06-01
影响因子：
6.4
作者：
Gerson, C;Sabater, J;Conner, GE
通讯作者：
Conner, GE
影响因子：
3.4
作者：
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DOI：
10.1152/ajplung.00108.2004
发表时间：
2004-10-01
影响因子：
4.9
作者：
Holmén, JM;Karlsson, NG;Davis, CW
通讯作者：
Davis, CW
影响因子：
4.8
作者：
Joo, NS;Lee, DJ;Wine, JJ
通讯作者：
Wine, JJ
影响因子：
6.1
作者：
Marano, F;Boland, S;Baeza-Squiban, A
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Baeza-Squiban, A