Interleukin-8 production by the human colon epithelial cell line HT-29: modulation by interleukin-13

Br J Pharmacol. 1996 Sep;119(2):351-9. doi: 10.1111/j.1476-5381.1996.tb15993.x.

Abstract

1. We have determined which cytokines induce and modulate the production of the chemokine interleukin-8 (IL-8) by the human colonic epithelial cell line HT-29. 2. Growth arrested cell cultures were stimulated with the human recombinant cytokines interleukin-1 alpha (IL-1 alpha), tumour necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN-gamma), interleukin-13 (IL-13), interleukin-10 (IL-10) or vehicle added alone or in combination. The production of IL-8 was determined by enzyme-linked immunosorbent assay (ELISA) and IL-8 messenger RNA expression by Northern blot analysis. 3. The production of IL-8 in unstimulated cells was undetectable by both ELISA and Northern blot analysis. 4. HT-29 cells produced IL-8 following stimulation with IL-1 alpha or TNF-alpha in a time- and a concentration-dependent manner, while IFN-gamma, IL-10 and IL-13 did not induce IL-8 production by HT-29 cells. 5. IL-13 was found to up-regulate significantly (P < 0.01) the IL-1 alpha but not the TNF-alpha-induced IL-8 generation by HT-29 cells. In contrast, IL-10 had no effect on either IL-1 alpha or TNF-alpha-induced IL-8 production. 6. Experiments using cycloheximide demonstrated that this synergistic effect of IL-13 and IL-1 alpha on IL-8 secretion was not through de novo protein synthesis. Using actinomycin-D, we demonstrated that the IL-13 up-regulation was at the level of transcription rather than messenger RNA stability. 7. These findings suggest that colonic epithelial cells have a functional IL-13 receptor, which is coupled to an up-regulation of IL-1 alpha, but not TNF-alpha induced IL-8 generation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Drug Synergism
  • HT29 Cells / drug effects*
  • HT29 Cells / metabolism*
  • Humans
  • Interleukin-1 / pharmacology*
  • Interleukin-10 / pharmacology
  • Interleukin-13 / pharmacology*
  • Interleukin-8 / biosynthesis*
  • Interleukin-8 / metabolism
  • RNA, Messenger / metabolism

Substances

  • Interleukin-1
  • Interleukin-13
  • Interleukin-8
  • RNA, Messenger
  • Interleukin-10