Overproduction of the CFTR R domain leads to increased levels of asialoGM1 and increased Pseudomonas aeruginosa binding by epithelial cells.
Mutations in cystic fibrosis transmembrane conductance regulator (CFTR), particularly the common DeltaF508 mutation, have been associated with alterations in glycolipid sialylation and the availability of receptors for Pseudomonas aeruginosa binding. The surface properties of 9HTEo- tracheal epithelial cell lines transfected with plasmids that overproduce the regulatory (R) domain of CFTR (pCEP-R) and lack cyclic adenosine monophosphate-stimulated Cl- conductance were compared with control cell lines with normal CFTR function. There was increased bacterial adherence to the mutant cell lines with abnormal CFTR activity. Cell lines with overexpression of the R domain had surface properties similar to cells expressing the common DeltaF508 mutation in CF. P. aeruginosa adherence correlated with the increased numbers of asialoGM1 residues available on the surface of the epithelial cells with altered CFTR function; and antibody to asialoGM1, a P. aeruginosa pilin receptor, was able to compete with piliated bacteria for epithelial binding sites. The pCEP-R cell lines with increased bacterial binding were also associated with increased production of interleukin-8 in response to adherent P. aeruginosa compared with cells transfected with the empty vector pCEP. P. aeruginosa pil mutants that lack the adhesin specific for the asialoGM1 receptor did not discriminate between epithelial cells with normal or deficient CFTR function. These results confirm a direct relationship between aberrant CFTR function and increased levels of apical asialoGM1, and support the role of these asialylated glycolipids as P. aeruginosa receptors that initiate an epithelial proinflammatory response in response to bacterial ligands.
Department of Pediatrics, College of Physicians and Surgeons, Columbia University, New York, USA.
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Bryan
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Kube
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Perez
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Davis
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Prince
D001422:Bacterial Adhesion; D002461:Cell Line, Transformed; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics / Q000378:metabolism*; D004847:Epithelial Cells / Q000382:microbiology; D010861:Fimbriae, Bacterial; D005677:G(M1) Ganglioside / Q000378:metabolism*; D006801:Humans; D010957:Plasmids; D011550:Pseudomonas aeruginosa / Q000502:physiology*; D014132:Trachea / Q000382:microbiology*
C505032:CFTR protein, human; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D005677:G(M1) Ganglioside; C018835:asialo GM1 ganglioside