Fibronectin and alpha5beta1 integrin mediate binding of Pseudomonas aeruginosa to repairing airway epithelium.
Initial infection of the airway by Pseudomonas aeruginosa may occur through a variety of bacterial strategies including binding to epithelial receptors present at the surface of the respiratory epithelium. In order to characterize the adherence sites for P. aeruginosa in damaged and repairing bronchial tissue, an ex vivo model of airway epithelial injury and repair was developed using primary cell cultures of nasal cells from 14 subjects with polyposis. P. aeruginosa strongly adhered to flattened dedifferentiated (FD) bronchial and nasal cytokeratin 13-positive epithelial cells in the process of migration for repair. In in vitro experiments, competitive binding inhibition assays demonstrated that alpha5beta1 integrins and cellular fibronectin, in particular the RGD sequence, are receptors involved in P. aeruginosa adherence to FD nasal epithelial cells. Fluorescent cell sorting analysis and immunofluorescence techniques revealed that the alpha5beta1 integrins are overexpressed and apically exposed in FD nasal epithelial cells. One 50 kDa outer membrane protein was identified in piliated and nonpiliated strains of P. aeruginosa that was involved in binding to cellular fibronectin and alpha5beta1 epithelial integrins. These results demonstrate that Pseudomonas aeruginosa adherence is related to the dedifferentiation of airway epithelium during the repair process which unmasks and upregulates the alpha5beta1 integrin expression and induces active synthesis of cellular fibronectin. These epithelial receptors are then used by a Pseudomonas aeruginosa 50 kDa outer membrane protein as sites of bacterial adherence.
Roger
P
P
INSERM U514, CHR Maison Blanche, Reims, France.
Puchelle
E
E
Bajolet-Laudinat
O
O
Tournier
J M
JM
Debordeaux
C
C
Plotkowski
M C
MC
Cohen
J H
JH
Sheppard
D
D
de Bentzmann
S
S
D001422:Bacterial Adhesion / Q000502:physiology*; D001425:Bacterial Outer Membrane Proteins / Q000502:physiology; D001980:Bronchi / Q000378:metabolism / Q000382:microbiology* / Q000473:pathology; D002454:Cell Differentiation; D002478:Cells, Cultured; D004848:Epithelium / Q000378:metabolism / Q000382:microbiology / Q000473:pathology; D005353:Fibronectins / Q000378:metabolism / Q000502:physiology*; D005434:Flow Cytometry; D006801:Humans; D007150:Immunohistochemistry; D009297:Nasal Mucosa / Q000378:metabolism / Q000382:microbiology* / Q000473:pathology; D011552:Pseudomonas Infections / Q000382:microbiology / Q000473:pathology; D011550:Pseudomonas aeruginosa / Q000502:physiology*; D017469:Receptors, Fibronectin / Q000378:metabolism / Q000502:physiology*; D012141:Respiratory Tract Infections / Q000382:microbiology / Q000473:pathology
D001425:Bacterial Outer Membrane Proteins; D005353:Fibronectins; D017469:Receptors, Fibronectin