Airway inflammation, including neutrophil influx is commonly seen in human pulmonary diseases. We developed an in vitro system where the adherence of neutrophils to bronchial epithelial cells could be examined. Primary cultures of nonhuman primate brenchial epithelial cells or transformed BEAS human bronchial epithelial cells were grown to confluence on collagen-coated culture plates. Cells were cocultured for 30 min following the addition of human neutrophils and PMA. Cultures were then inverted, fixed with methanol, and adherent neutrophils labeled with 1B4 mouse monoclonal anti-human neutrophil antibody followed by fluoresceinlabeled sheep anti-mouse IgG. Slides were examined using fluorescence microscopy. The 1B4 antibody allowed rapid identification of neutrophils adherent to the epithelial cell monolayers, which were not labeled by this technique. PMA increased the adherence of neutrophils to bronchial epithelial cells. Pretreatment of the neutrophils with anti-CD11/CD18 antibodies prevented the increase in PMA-stimulated adherence. We conclude that PMA-stimulated adherence to airway epithelial cells is in part dependent on the neurrophil CD11/CD18 adherence complex.
ASJC Scopus subject areas
- Cell Biology