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Am. J. Respir. Cell Mol. Biol., Volume 23, Number 3, September, 2000 396-403

Exaggerated Activation of Nuclear Factor-kappa B and Altered Ikappa B-beta Processing in Cystic Fibrosis Bronchial Epithelial Cells

Annapurna Venkatakrishnan, Arlene A. Stecenko, Gayle King, Thomas R. Blackwell, Kenneth L. Brigham, John W. Christman, and Timothy S. Blackwell

Department of Medicine, Division of Allergy, Pulmonary, and Critical Care Medicine, Vanderbilt University School of Medicine, and Department of Veterans Affairs Medical Center, Nashville, Tennessee

In cystic fibrosis (CF), inflammatory mediator production by airway epithelial cells is a critical determinant of chronic airway inflammation. To determine whether altered signal transduction through the nuclear factor (NF)-kappa B pathway occurs in CF epithelial cells and results in excessive generation of inflammatory cytokines, we evaluated tumor necrosis factor (TNF)-alpha -induced production of the NF-kappa B-dependent cytokine interleukin (IL)-8 and activation of NF-kappa B in three different human bronchial epithelial cell lines: (1) BEAS cells that express wild-type CF transmembrane conductance regulator (CFTR), (2) IB3 cells with mutant CFTR, and (3) C38 cells, which are "corrected" IB3 cells complemented with wild-type CFTR. Treatment of cells with TNF-alpha (30 ng/ml) resulted in markedly elevated NF-kappa B activation and production of IL-8 by IB3 cells compared with BEAS and C38 cells. Despite the differences in NF- kappa B activation, no differences in basal levels of Ikappa B-alpha or TNF-alpha - induced Ikappa B-alpha processing and degradation were detected among the cell lines. In contrast, the basal level of Ikappa B-beta was increased in the IB3 cells. Treatment with TNF-alpha resulted in increased formation of hypophosphorylated Ikappa B-beta and increased nuclear localization of Ikappa B-beta in IB3 cells compared with the other cell types. These findings provide additional evidence of a dysregulated inflammatory response in CF.




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