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1 Cardiovascular Research Institute, University of California at San Francisco, San Francisco, CA, USA
2 Department of Medicine /Physiology, University of California at San Francisco, San Francisco, CA, USA
3 Lab. di Genetica Molecolare, Istituto Giannina Gaslini, Genova, Italy
* To whom correspondence should be addressed. E-mail: xhfang{at}itsa.ucsf.edu.
Previous studies in intact lung suggest that CFTR may play a role in cAMP regulated fluid transport from the distal airspaces 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 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 0.89 ± 0.11 µl/cm2/h and increased to 1.35 ± 0.11 µl/cm2/h (mean ± SE, n=18, P<0.05) by stimulation with cAMP agonists. The CFTR inhibitor, CFTRinh-172, inhibited cAMP stimulated but not basal fluid transport. In short-circuit current studies with an apical-to-basolateral transepithelial Cl- gradient, apical application of CFTRinh-172 reversed the forskolin-induced decrease in Isc. 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 vectorial fluid transport.
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