Development of a high throughput Pseudomonas aeruginosa epithelial cell adhesion assay.
Pseudomonas aeruginosa colonizes the lungs of cystic fibrosis and mechanically ventilated patients by binding to specific carbohydrate residues on the surface of lung epithelial cells. Studies have shown that blocking this interaction may have therapeutic effects in vivo. To test compounds that may have an effect on the binding of P. aeruginosa to epithelial cells, we have developed a pseudomonal adhesion assay that is compatible with high throughput technology. This assay utilizes a 96-well culture plate assay and P. aeruginosa strains that have been modified to bioluminesce. This method has proven to be a rapid, sensitive and reproducible system for screening agents that inhibit bacterial adhesion.
Swanson
Britta
B
Department of Anesthesia and Perioperative Care and Cardiovascular Research Institute, San Francisco Medical Center, University of California, 513 Parnassus Avenue, S-261 94127-0542, USA. swansonb@sfsu.edu
Savel
Richard
R
Szoka
Frank
F
Sawa
Teiji
T
Wiener-Kronish
Jeanine
J
D001422:Bacterial Adhesion / Q000502:physiology*; D001431:Bacteriological Techniques / Q000379:methods*; D001681:Biological Assay; D050260:Carbohydrate Metabolism; D002241:Carbohydrates / Q000145:classification; D002448:Cell Adhesion / Q000502:physiology; D002460:Cell Line; D004847:Epithelial Cells / Q000382:microbiology*; D004848:Epithelium / Q000382:microbiology; D006801:Humans; D011550:Pseudomonas aeruginosa / Q000472:pathogenicity*; D015203:Reproducibility of Results
D002241:Carbohydrates