Platelet-activating factor induction of activator protein-1 signaling in bronchial epithelial cells

Trici D. Levan, John W. Bloom, Deanna G. Adams, Jennifer L. Hensel, Marilyn Halonen

Research output: Contribution to journalArticlepeer-review

11 Scopus citations

Abstract

Platelet-activating factor (PAF) has been implicated in the pathogenesis of allergic and inflammatory events in the airway. In the present study, we sought to determine if PAF receptors are present on human bronchial epithelial cells and whether PAF binding to these receptors leads to activation of activator protein-1 (AP-1)-mediated transcription. Radioligand binding studies demonstrated specific binding sites for the PAF antagonist [3H]WEB 2086 (3-[4-(2-chlorophenyl)-9-methyl-6H-thieno[3,2-f]- [1,2,4]triazolo[4,3- a][1,4]diazepine-2-yl]-1-(4-morpholinyl)-1-propanone) on primary bronchial epithelial cells with an equilibrium dissociation constant (K(d)) = 9.8 nM and maximal density of binding sites (B(max)) = 42.4 fmol/mg of protein. The expression of PAF receptors in these cells was further confirmed by reverse transcriptase-polymerase chain reaction, which revealed amplification products derived from PAF receptor mRNA corresponding to transcripts 1 and 2. In the bronchial epithelial cell line BEAS-2B transfected with an expression plasmid for the human PAF receptor, PAF stimulation increased AP-1 DNA binding activity as determined by electrophoretic mobility shift assays. The Fos and Jun family proteins were identified as components of the DNA-protein complexes by anti-peptide antibodies in gel supershift assays. Additionally, PAF significantly induced AP-1 mediated transcription which was dependent on the expression of PAF receptors. The PAF antagonist WEB 2086 blocked the PAF effect but not that induced by 12-O-tetradecanoyl phorbol-13-acetate, indicating the specificity of the PAF response. These results indicate that activation of airway epithelial cells through stimulation of PAF receptors includes up-regulation of the nuclear transcription factor AP-1 and AP-1 transcriptional activity.

Original languageEnglish (US)
Pages (from-to)135-140
Number of pages6
JournalMolecular pharmacology
Volume53
Issue number1
DOIs
StatePublished - Jan 1998

ASJC Scopus subject areas

  • Molecular Medicine
  • Pharmacology

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