Cystic fibrosis (CF) is characterized by intense neutrophil migration into the airways. Increasing evidence indicates that interaction between neutrophils and airway epithelial cells contributes to the modulation of the inflammatory response. Blood neutrophils were reported to express connexins and form gap junctions with endothelial cells, thereby establishing gap junctional communication. We tested whether altered communication between human neutrophils and airway epithelial cells may contribute to the exaggerated inflammatory response observed in CF patients. Microinjections did not reveal dye coupling between activated blood neutrophils. By contrast, diffusion of calcein between neutrophils and airway epithelial cells of CF or non-CF origin was observed in transmigration and adhesion assays. This diffusion was prevented with probenicid, an inhibitor of ATP-dependent organic anion pumps, but not with gap junction blockers. Finally, RT-PCR failed to detect mRNAs for six connexins in blood neutrophils. These results suggest that gap junctional communication does not contribute to neutrophil-airway epithelial cell interaction.