Xiaopeng Li, Xiao Xiao Tang, Luis G Vargas Buonfiglio, Alejandro P Comellas, Ian M Thornell, Shyam Ramachandran, Philip H Karp, Peter J Taft, Kelsey Sheets, Mahmoud H Abou Alaiwa, Michael J Welsh, David K Meyerholz, David A Stoltz, Joseph Zabner

While pathological and clinical data suggest that small airways are involved in early cystic fibrosis (CF) lung disease development, little is known about how the lack of cystic fibrosis transmembrane conductance regulator (CFTR) function contributes to disease pathogenesis in these small airways. Large and small airway epithelia are exposed to different airflow velocities, temperatures, humidity, and CO2 concentrations. The cellular composition of these two regions is different and small airways lack submucosal glands. In order to better understand the ion transport properties and impacts of lack of CFTR function on host defense function in small airways, we adapted a novel protocol to isolate small airway epithelial cells from CF and non-CF pigs and established an organotypic culture model. Compared to non-CF large airways, non-CF small airway epithelia cultures had higher Cl(-) and bicarbonate (HCO3 (-)) short-circuit currents and higher airway surface liquid (ASL) pH under 5% CO2 conditions. CF small airway epithelia were characterized by minimal Cl(-) and HCO3 (-) transport and decreased ASL pH, and had impaired bacterial killing compared to non-CF small airways. In addition, CF small airway epithelia had a higher ASL viscosity than non-CF small airways. Thus, the activity of CFTR is higher in the small airways, where it plays a role in alkalinization of ASL, enhancement of antimicrobial activity and lowering mucus viscosity. These data provide insight to explain why the small airways are a susceptible site for the bacterial colonization.