TY - JOUR
T1 - Tracheobronchial air-liquid interface cell culture: A model for innate mucosal defense of the upper airways?
AU - Kesimer, Mehmet
AU - Kirkham, Sara
AU - Pickles, Raymond J.
AU - Henderson, Ashley G.
AU - Alexis, Neil E.
AU - Demaria, Genevieve
AU - Knight, David
AU - Thornton, David J.
AU - Sheehan, John K.
N1 - HL-066973, NHLBI NIH HHS, United StatesHL-080098, NHLBI NIH HHS, United StatesHL-084934-02, NHLBI NIH HHS, United States, Wellcome Trust, United Kingdom
PY - 2009/1
Y1 - 2009/1
N2 - 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 tracheobron- chial 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. Copyright © 2009 the American Physiological Society.
AB - 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 tracheobron- chial 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. Copyright © 2009 the American Physiological Society.
KW - Human tracheobronchial epithelial cell culture
KW - Innate immunity
KW - Mucin
KW - Mucus
KW - Proteomics
U2 - 10.1152/ajplung.90388.2008
DO - 10.1152/ajplung.90388.2008
M3 - Article
C2 - 18931053
SN - 1522-1504
VL - 296
SP - L92-L100
JO - AJP: Lung Cellular and Molecular Physiology
JF - AJP: Lung Cellular and Molecular Physiology
IS - 1
ER -