Objectives Lung disease in cystic fibrosis (CF) is often exacerbated following acute upper respiratory tract infections. The majority of these exacerbations are associated with the human rhinovirus (HRV). Pathophysiology of these exacerbations is presently unclear. Potential mechanisms may involve impaired innate antiviral response in CF airway epithelial cells, the main replication site of HRV, or exaggerated inflammatory response by these cells. Furthermore, responses to HRV by CF cells may be adversely affected by chronic exposure to diffusible virulence factors of the opportunistic microorganism Pseudomonas aeruginosa , which frequently colonizes CF airways. Methods Here we examined HRV16 RNA load, production of the antiviral cytokine interferon-β and inflammatory chemokine interleukin-8, and expression of the interferon-responsive antiviral gene OAS1 in primary healthy and CF bronchial epithelial cells following infection with HRV16. Parallel cells were exposed to diffusible virulence factors of P. aeruginosa prior to infection with HRV16. Results We observed that CF cells exhibited high HRV16 RNA load, while production of interferon-β and interleukin-8, and expression of OAS1 were comparable to those in healthy cells. Pre-exposure to diffusible virulence factors of P. aeruginosa only mildly potentiated interleukin-8 response to HRV16, without affecting virus RNA load or interferon-b response. Conclusion CF cells demonstrate impaired HRV clearance with unaltered interferon-b or interleukin-8 responses. High HRV load in CF cells appear to be due to defective antiviral mechanisms unrelated to interferon-b.