Immunocytochemical localization of cyclo-oxygenase isoforms in cultured human airway structural cells

Clin Exp Allergy. 1999 Jul;29(7):965-72. doi: 10.1046/j.1365-2222.1999.00573.x.

Abstract

Background: Cyclo-oxygenase (COX) exists as two isoforms, COX-1, the constitutive isoform, and COX-2, which is inducible by cytokines or inflammatory stimuli and may participate in airway inflammation.

Objective: To determine the basal distribution of COX isoforms, and their regulation by interleukin-1 beta (IL-1beta), bradykinin (BK) and dexamethasone (Dex) in cultured airway structural cells.

Methods: We measured COX-1 and COX-2 in cultured human airway smooth muscle (HASM) cells, MRC5 fibroblasts and normal human epithelial cells (NHBE) using immunocytochemical analysis.

Results: The majority of all types of untreated cultured cells expressed COX-1 (75% of HASM, 75% of MRC5 fibroblasts and 72% of NHBE cells). Fibroblasts and smooth muscle cells showed low constitutive COX-2 expression (2 and 8%, respectively) but this was higher in NHBE cells (28%). IL-1beta (24 h incubation) or BK (4 h incubation) had no effect on COX-1 expression in any of the cells studied. In contrast, there was a two- and 1.5-fold rise in the percentage of NHBE cells expressing COX-2; a 7.5- and sixfold rise in the percentage of HASM cells expressing COX-2 and a 33.5- and 20.5-fold increase in the percentage of fibroblasts expressing COX-2 after IL-1beta or BK treatment, respectively. Pretreatment with dexamethasone abolished IL-1beta- and BK-stimulated COX-2 induction in all cells studied.

Conclusion: COX-1 is expressed constitutively in human airway fibroblasts, smooth muscle and epithelial cells but epithelial cells also show constitutive expression of COX-2. Both IL-1beta and BK induced COX-2 expression in all cells studied and this induction was blocked by dexamethasone. Immunocytochemical techniques can be successfully used to detect the distribution of COX isoforms in cell cultures.

MeSH terms

  • Blotting, Western
  • Bradykinin / pharmacology
  • Cells, Cultured
  • Cyclooxygenase 1
  • Cyclooxygenase 2
  • Dexamethasone / pharmacology
  • Epithelial Cells / cytology
  • Epithelial Cells / enzymology
  • Fibroblasts / cytology
  • Fibroblasts / enzymology
  • Humans
  • Immunoenzyme Techniques
  • Interleukin-1 / pharmacology
  • Isoenzymes / metabolism*
  • Lung / cytology
  • Lung / drug effects
  • Lung / enzymology*
  • Membrane Proteins
  • Muscle, Smooth / cytology
  • Muscle, Smooth / enzymology
  • Prostaglandin-Endoperoxide Synthases / metabolism*

Substances

  • Interleukin-1
  • Isoenzymes
  • Membrane Proteins
  • Dexamethasone
  • Cyclooxygenase 1
  • Cyclooxygenase 2
  • PTGS1 protein, human
  • PTGS2 protein, human
  • Prostaglandin-Endoperoxide Synthases
  • Bradykinin