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Interleukin‐8 production by the human colon epithelial cell line HT‐29: modulation by interleukin‐13
Author(s) -
Kolios George,
Robertson Duncan A.F.,
Jordan Nicola J.,
Minty Adrian,
Caput Daniel,
Ferrara Pascal,
Westwick John
Publication year - 1996
Publication title -
british journal of pharmacology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.432
H-Index - 211
eISSN - 1476-5381
pISSN - 0007-1188
DOI - 10.1111/j.1476-5381.1996.tb15993.x
Subject(s) - cycloheximide , cytokine , microbiology and biotechnology , interleukin , northern blot , cell culture , biology , tumor necrosis factor alpha , interleukin 8 , messenger rna , endocrinology , immunology , protein biosynthesis , biochemistry , gene , genetics
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‐1α (IL‐1α), tumour necrosis factor‐α (TNF‐α), interferon‐γ (IFN‐γ), 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‐1α or TNF‐α in a time‐ and a concentration‐dependent manner, while IFN‐γ, 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‐1α but not the TNF‐α‐induced IL‐8 generation by HT‐29 cells. In contrast, IL‐10 had no effect on either IL‐1α or TNF‐α‐induced IL‐8 production. 6 Experiments using cycloheximide demonstrated that this synergistic effect of IL‐13 and IL‐1α 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‐1α, but not TNF‐α induced IL‐8 generation.