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Published ahead of print on December 11, 2003, doi:10.1164/rccm.200207-765OC

Am. J. Respir. Crit. Care Med., Volume 169, Number 5, March 2004, 645-653

A more recent version of this article appeared on March 1, 2004
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Submitted on July 29, 2002
Accepted on December 9, 2003

Cytokine Secretion by Cystic Fibrosis Airway Epithelial Cells

Marie N Becker1, Mariam S Sauer1, Marianne S Muhlebach2, Andrew J Hirsh1, Qi Wu1, Margrith W Verghese1, and Scott H Randell3*

1 Medicine and Cystic Fibrosis/Pulmonary Research and Treatment Center, The University of North Carolina, Chapel Hill, NC, USA, 2 Pediatrics, The University of North Carolina, Chapel Hill, NC, USA, 3 Medicine and Cystic Fibrosis/Pulmonary Research and Treatment Center, The University of North Carolina, Chapel Hill, NC, USA; Cellular and Molecular Physiology, The University of North Carolina, Chapel Hill, NC, USA

* To whom correspondence should be addressed. E-mail: randell{at}med.unc.edu.

It is controversial whether mutations in CFTR intrinsically dysregulate inflammation. We characterized passage 2 human tracheobronchial epithelial cell cultures morphologically and physiologically and determined whether cytokine production or NF-{kappa}B activation was systematically altered in CF cells. Non-CF and CF cells originating from a total of 33 and 25 lungs, respectively, were available for culture on plastic or at an air-liquid interface until well-differentiated. Forskolin-stimulated short-circuit currents were present in representative polarized non-CF cultures and absent in CF cultures, whereas UTP stimulated currents were present in both. Constitutive or IL-1{beta}-induced IL-8 or IL-6 secretion or nuclear NF-{kappa}B activity was not significantly different between non-CF and CF cells, and RANTES and IL-10 were not detectable. Stimulation with TNF{alpha} or a synthetic Toll-like receptor 2 agonist, or variable doses and times of Staphylococcus aureus culture filtrate revealed a single dose- and time-dependent difference in IL-8 production by CF cells. Interestingly, while IL-8 secretion following stimulation with Pseudomonas aeruginosa filtrates was not greater in CF cells in the absence of human serum, it was variably greater in its presence. Thus, while exaggerated responses may develop under certain conditions, our results do not support an overall, intrinsically hyper-inflammatory phenotype in CF cells.
Key words: Bacterial Products, Electrophysiology, IL-8, Inflammation, NF-kB, Morphology




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