The role of IkappaB kinase 2, but not activation of NF-kappaB, in the release of CXCR3 ligands from IFN-gamma-stimulated human bronchial epithelial cells

J Immunol. 2007 Nov 1;179(9):6237-45. doi: 10.4049/jimmunol.179.9.6237.

Abstract

The severity of chronic obstructive pulmonary disease correlates with increased numbers of cytotoxic CD8(+) T lymphocytes in the lung parenchyma. CD8(+) T lymphocytes release IFN-gamma which stimulates airway epithelial cells to produce CXCR3 chemokines leading to further recruitment of CD8(+) T lymphocytes. To evaluate the signaling pathways involved in regulation of CXCR3 ligands, the human bronchial epithelial cell line BEAS-2B was stimulated with IFN-gamma and the release of the CXCR3 ligands was measured by ELISA. The release of CXCL9, CXCL10, and CXCL11 was inhibited by an IkappaB kinase 2 (IKK2) selective inhibitor 2-[(Aminocarbonyl)amino]-5-[4-fluorophenyl]-3-thiophenecarboxamide (TPCA-1) (EC(50) values were 0.50 +/- 0.03, 0.17 +/- 0.06, and 0.45 +/- 0.10 microM, respectively (n = 6)) and an IKK1/2 selective inhibitor 2-amino-6-(2'cyclopropylemethoxy-6'-hydroxy-phenyl)-4-piperidin-3-yl-pyridine-3-carbonitrile (EC(50) values 0.74 +/- 0.40, 0.27 +/- 0.06, and 0.88 +/- 0.29 microM, respectively (n = 6)). The glucocorticosteroid dexamethasone had no effect on CXCR3 ligand release. The release of CXCL10 was most sensitive to inhibition by IKK2 and a role for IKK2 in CXCL10 release was confirmed by overexpression of dominant-negative adenoviral constructs to IKK2 (68.2 +/- 8.3% n = 5), but not of IKK1. Neither phosphorylation of IkappaBalpha, translocation of p65 to the nucleus, or activation of a NF-kappaB-dependent reporter (Ad-NF-kappaB-luc) were detected following stimulation of BEAS-2B cells with IFN-gamma. These data suggest that IKK2 is also involved in the IFN-gamma-stimulated release of the CXCR3 ligands through a novel mechanism that is independent NF-kappaB.

MeSH terms

  • Amides / pharmacology
  • Animals
  • Bronchi / drug effects*
  • Bronchi / metabolism*
  • Cells, Cultured
  • Chemokines / genetics
  • Chemokines / metabolism
  • Dexamethasone / pharmacology
  • Epithelial Cells / drug effects*
  • Epithelial Cells / metabolism*
  • Gene Expression Regulation
  • Humans
  • I-kappa B Kinase / antagonists & inhibitors
  • I-kappa B Kinase / genetics
  • I-kappa B Kinase / metabolism*
  • Interferon-gamma / pharmacology*
  • Ligands
  • NF-kappa B / metabolism
  • Phosphorylation
  • Protein Kinase Inhibitors / pharmacology
  • Protein Transport
  • RNA, Messenger / genetics
  • Receptors, CXCR3 / metabolism*
  • STAT1 Transcription Factor / metabolism
  • Signal Transduction
  • Thiophenes / pharmacology

Substances

  • Amides
  • CXCR3 protein, human
  • Chemokines
  • Ligands
  • NF-kappa B
  • Protein Kinase Inhibitors
  • RNA, Messenger
  • Receptors, CXCR3
  • STAT1 Transcription Factor
  • Thiophenes
  • Dexamethasone
  • Interferon-gamma
  • 2-((aminocarbonyl)amino)-5-(4-fluorophenyl)-3-thiophenecarboxamide
  • I-kappa B Kinase