Macrolides have been shown to be effective in treating diffuse panbronchiolitis (DPB), although the precise modes of action remain unclear. At sites of airway inflammation, respiratory epithelium is considered an active participant in regulating neutrophil survival. We therefore examined the effect of erythromycin, clarithromycin, azithromycin, and josamycin on both neutrophil survival and on epithelial-derived factors, which influence neutrophil longevity. Media conditioned with transiently tumor necrosis factor (TNF)-alpha-stimulated A549 human airway epithelial cells prolonged neutrophil survival compared with control media. The presence of dexamethasone during neutrophil culture led to further prolongation of neutrophil survival. In contrast, none of the tested macrolides modulated neutrophil survival, suggesting a lack of direct effect of these drugs. On the other hand, pretreatment of TNF-alpha-stimulated A549 cells by erythromycin, clarithromycin, azithromycin, or dexamethasone, but not josamycin, decreased the neutrophil survival-enhancing effects in a dose-dependent manner. Neutralizing antibodies to granulocyte macrophage colony-stimulating factor (GM-CSF) dampened the prolonged neutrophil survival observed in TNF-alpha-stimulated A549 conditioned media. Erythromycin, clarithromycin, azithromycin, and dexamethasone inhibited TNF-alpha-induced GM-CSF expression in A549 cells at both the protein and messenger RNA levels. These results suggest that macrolides inhibit epithelial cell-mediated neutrophil survival by modulating GM-CSF release, which may, at least in part, explain the effectiveness of this family of drugs on DPB.