BACKGROUND: Rhinovirus and fungi are common environmental factors able to induce airway inflammation. They are associated with the production of chemical mediators by direct activation of epithelial cells. OBJECTIVE: To evaluate the effect of fungal stimulation of rhinovirus-infected nasal polyp epithelial cells (NPECs) on the activation and migration of eosinophils. METHODS: Rhinovirus-infected NPECs were stimulated with Alternaria and Aspergillus for 48 hours. Then, epithelial cells were co-cultured with freshly isolated eosinophils. An eosinophil migration study was performed with epithelial cell-conditioned media. Interleukin 6, interleukin 8, granulocyte-macrophage colony-stimulating factor, and tumor necrosis factor levels were measured to determine the activation of cells. RESULTS: Airborne fungi enhanced the production of cytokines from rhinovirus 16-infected NPECs compared with fungi stimulation or rhinovirus 16 infection alone. Rhinovirus 16-infected NPECs were co-cultured with eosinophils, and cytokine production was not significantly increased except tumor necrosis factor-alpha production by Aspergillus. Epithelial cell-conditioned media, which were stimulated with fungi, enhanced the migration of eosinophils. CONCLUSIONS: There was some synergism between rhinovirus 16 infection and airborne fungal exposure, enhancing the inflammatory response of airway epithelial cells.