TY - JOUR
T1 - Regulation of antiprotease and antimicrobial protein secretion by airway submucosal gland serous cells
AU - Joo, Nam Soo
AU - Lee, Dennis J.
AU - Winges, Kimberly M.
AU - Rustagi, Arfun
AU - Wine, Jeffrey J.
PY - 2004/9/10
Y1 - 2004/9/10
N2 - Airway submucosal gland serous cells express the cystic fibrosis transmembrane conductance regulator (CFTR) and secrete antimicrobial, anti-inflammatory, and antioxidant molecules. In cystic fibrosis, diminished gland secretion may impair innate airway host defenses. We used Calu-3 cells as a serous cell model to study the types of proteins released, the pathways that release them, and the possible involvement of CFTR activity in protein release. Many proteins were secreted constitutively into the apical fluid and showed increased release to agonists. We identified some of them by high pressure liquid chromatography-mass spectrometry and reverse transcriptase PCR, including lysozyme, siderocalin (the protein NGAL), which inhibits bacterial growth by binding iron-containing siderophores, HSC-71, which is thought to have anti-inflammatory properties, and the serine protease inhibitors α-1-antitrypsin and α-1-antichymotrypsin, which may function as antimicrobials as well as play a potential role in diminishing the activation of epithelial Na+ channels by serine proteases. We used an enzyme-linked immunosorbent assay to quantify lysozyme secretion by Calu-3 cells in response to various agonists and inhibitors. Forskolin increased the lysozyme secretion rate (Jlyz) from 32 to 77 ng/hr/cm2 (n = 36, p < 0.005). Thapsigargin increased Jlyz from 40 to 63 ng/h/cm2 (n = 16, p < 0.005), and forskolin plus tliapsigargin further increased the forskolin-stimulated Jlyz by 48% (n = 9, p < 0.05). 1-Ethyl-benzimidazolinone and carbachol were less effective. Glibenclamide inhibited basal and stimulated Jlyz, but clotrimazole was without effect. CFTRinh172 caused a small (15%) but significant inhibition of forskolin-stimulated Jlyz without affecting basal Jlyz. Thus, Calu-3 cells secrete diverse proteins that in aggregate would be expected to suppress microbial growth, protect the airways from damage, and limit the activation of epithelial Na+ channels via serine proteases.
AB - Airway submucosal gland serous cells express the cystic fibrosis transmembrane conductance regulator (CFTR) and secrete antimicrobial, anti-inflammatory, and antioxidant molecules. In cystic fibrosis, diminished gland secretion may impair innate airway host defenses. We used Calu-3 cells as a serous cell model to study the types of proteins released, the pathways that release them, and the possible involvement of CFTR activity in protein release. Many proteins were secreted constitutively into the apical fluid and showed increased release to agonists. We identified some of them by high pressure liquid chromatography-mass spectrometry and reverse transcriptase PCR, including lysozyme, siderocalin (the protein NGAL), which inhibits bacterial growth by binding iron-containing siderophores, HSC-71, which is thought to have anti-inflammatory properties, and the serine protease inhibitors α-1-antitrypsin and α-1-antichymotrypsin, which may function as antimicrobials as well as play a potential role in diminishing the activation of epithelial Na+ channels by serine proteases. We used an enzyme-linked immunosorbent assay to quantify lysozyme secretion by Calu-3 cells in response to various agonists and inhibitors. Forskolin increased the lysozyme secretion rate (Jlyz) from 32 to 77 ng/hr/cm2 (n = 36, p < 0.005). Thapsigargin increased Jlyz from 40 to 63 ng/h/cm2 (n = 16, p < 0.005), and forskolin plus tliapsigargin further increased the forskolin-stimulated Jlyz by 48% (n = 9, p < 0.05). 1-Ethyl-benzimidazolinone and carbachol were less effective. Glibenclamide inhibited basal and stimulated Jlyz, but clotrimazole was without effect. CFTRinh172 caused a small (15%) but significant inhibition of forskolin-stimulated Jlyz without affecting basal Jlyz. Thus, Calu-3 cells secrete diverse proteins that in aggregate would be expected to suppress microbial growth, protect the airways from damage, and limit the activation of epithelial Na+ channels via serine proteases.
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U2 - 10.1074/jbc.M407077200
DO - 10.1074/jbc.M407077200
M3 - Article
C2 - 15234967
AN - SCOPUS:4644303141
SN - 0021-9258
VL - 279
SP - 38854
EP - 38860
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 37
ER -