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Am J Physiol Lung Cell Mol Physiol 279: L1058-L1065, 2000;
1040-0605/00 $5.00
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Vol. 279, Issue 6, L1058-L1065, December 2000

IL-1beta induces eotaxin gene transcription in A549 airway epithelial cells through NF-kappa B

Sean Jedrzkiewicz1, Hidetoshi Nakamura1, Eric S. Silverman1, Andrew D. Luster2, Naresh Mansharamani1, Kwang Ho In1, Gen Tamura1, and Craig M. Lilly1

1 Combined Program in Pulmonary and Critical Care Medicine, Department of Medicine, Brigham and Women's Hospital and Harvard Medical School, Boston 02115; and 2 Infectious Disease Unit, Massachusetts General Hospital, Charlestown, Massachusetts 02129

Eotaxin is an asthma-related C-C chemokine that is produced in response to interleukin-1beta (IL-1beta ). We detected an increase in newly transcribed eotaxin mRNA in IL-1beta -stimulated airway epithelial cells. Transient transfection assays using promoter-reporter constructs identified a region as essential for IL-1beta -induced increases in eotaxin transcription. Using site-directed mutagenesis, we found that a nuclear factor-kappa B (NF-kappa B) site located 46 bp upstream from the transcriptional start site was both necessary and sufficient for IL-1beta induction of reporter construct activity. Electrophoretic mobility shift assay demonstrated that IL-1beta -stimulated airway epithelial cells produced p50 and p65 protein that bound this site in a sequence-specific manner. The functional importance of the NF-kappa B site was demonstrated by coexpression experiments in which increasing doses of p65 expression vector were directly associated with reporter activity exclusively in constructs with an intact NF-kappa B site (r2 = 0.97, P = 0.002). Moreover, IL-1beta -induced increases in eotaxin mRNA expression are inhibited by inhibitors of NF-kappa B. Our findings implicate NF-kappa B and its binding sequence in IL-1beta -induced transcriptional activation of the eotaxin gene.

cytokine; chemokine; eosinophil; asthma


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