Protein composition of apical surface fluid from the human airway cell line Calu-3: effect of ion transport mediators
Protein composition of apical surface fluid from the human airway cell line Calu-3: effect of ion transport mediators
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DOI：
10.1016/j.cccn.2004.04.006
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发表时间：
2004-09-01
影响因子：
5
通讯作者：
Shaffer, TH
中科院分区：
文献类型：
--
作者：
Babu, PBR;Chidekel, A;Shaffer, TH
背景：分析气道分泌物的蛋白质成分是检测和表征与气道疾病相关的生化改变的潜在手段。方法：我们使用在气液界面生长的气道上皮细胞系Calu-3来评估气道蛋白分泌物。为了观察细胞顶端分泌蛋白的变化，我们分析了离子转运介质处理过的Calu-3单层细胞顶端表面液体（ASF）洗涤液的蛋白含量。结果：抗菌和炎症蛋白的免疫分析筛选表明，Calu-3 ASF中存在可测量水平的溶菌酶和IL-8。RT-PCR和免疫分析表明，Calu-3细胞不产生clara细胞10 kDa蛋白（cc10）。缓激肽对Calu-3总蛋白分泌没有影响，而氨米洛利和腺苷显著增加Calu-3蛋白分泌。缓激肽不影响溶菌酶的分泌，但腺苷和阿米洛利可显著降低溶菌酶的分泌。缓激素和腺苷均未改变IL-8的分泌，但阿米洛利可显著降低IL-8的分泌。结论：抑菌蛋白溶菌酶和促炎细胞因子IL-8在Calu-3型ASF中存在，缓激肽、氨酰脲、腺苷等离子转运介质影响Calu-3型ASF蛋白的分泌。(C) 2004 Elsevier B.V.版权所有
Background: Analysis of the protein components of airway secretions is a potential means of detecting and characterizing biochemical alterations associated with airway diseases. Methods: We evaluated airway protein secretions using the airway epithelial cell line Calu-3 grown at an air-liquid interface. To observe changes in apically secreted proteins, we analyzed the protein content of apical surface fluid (ASF) washings of Calu-3 monolayers treated with ion transport mediators. Results: Immunoassay screening for antibacterial and inflammatory proteins indicated the presence of measurable levels of lysozyme and IL-8 in Calu-3 ASF. RT-PCR and immunoassay studies indicated that Calu-3 cells do not produce clara cell 10 kDa protein (CC 10). The total protein secretion of Calu-3 was not altered by bradykinin, but amiloride and adenosine significantly increased Calu-3 protein secretion. Lysozyme secretion was not altered by bradykinin, but amiloride and adenosine significantly reduced lysozyme secretion. IL-8 secretion was not altered by bradykinin or adenosine, but amiloride significantly decreased IL-8 secretion. Conclusion: Our results demonstrate the presence of antibacterial protein lysozyme and the pro-inflammatory cytokine IL-8 in Calu-3 ASF and that ion transport mediators such as bradykinin, amiloride and adenosine influence the secretion of Calu-3 ASF proteins. (C) 2004 Elsevier B.V. All rights reserved.