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Contribution of CFTR to apical-basolateral fluid transport in cultured human alveolar epithelial type II cells

¹Cardiovascular Research Institute and ²Department of Medicine/Physiology, University of California, San Francisco, California; and ³Laboratorio di Genetica Molecolare, Istituto Giannina Gaslini, Genoa, Italy

Submitted 20 April 2005 ; accepted in final form 29 August 2005

Previous studies in intact lung suggest that CFTR may play a role in cAMP-regulated fluid transport from the distal air spaces of the lung. However, the potential contribution of different epithelial cells (alveolar epithelial type I, type II, or bronchial epithelial cells) to CFTR-regulated fluid transport is unknown. In this study we determined whether the CFTR gene is expressed in human lung alveolar epithelial type II (AT II) cells and whether the CFTR chloride channel contributes to cAMP-regulated fluid transport in cultured human AT II cells. Human AT II cells were isolated and cultured on collagen I-coated Transwell membranes for 120–144 h with an air-liquid interface. The cultured cells retained typical AT II-like features based on morphologic studies. Net basal fluid transport was 0.9 ± 0.1 µl·cm^–2·h^–1 and increased to 1.35 ± 0.11 µl·cm^–2·h^–1 (mean ± SE, n = 18, P < 0.05) by stimulation with cAMP agonists. The CFTR inhibitor, CFTR_inh-172, inhibited cAMP stimulated but not basal fluid transport. In short-circuit current (I_sc) studies with an apical-to-basolateral transepithelial Cl^– gradient, apical application of CFTR_inh-172 reversed the forskolin-induced decrease in I_sc. Real time RT-PCR demonstrated CFTR transcript expression in human AT II cells at a level similar to that in airway epithelial cells. We conclude that CFTR is expressed in cultured human AT II cells and may contribute to cAMP-regulated apical-basolateral fluid transport.

lung epithelial cells; cystic fibrosis transmembrane conductance regulator; ion channel; cyclic adenosine monophosphate; pulmonary edema; alveolar fluid clearance

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