Differential activation of cytokine secretion in primary human colonic fibroblast/myofibroblast cultures

Scand J Gastroenterol. 2001 Apr;36(4):389-98. doi: 10.1080/003655201300051216.


Background: Fibroblasts and myofibroblasts are known to secrete a wide spectrum of cytokines, but the individual spectrum is tissue-specific. We investigated the effect of cell activation on cytokine secretion of isolated human colonic fibroblasts/myofibroblasts from control patients and patients with mucosal inflammation.

Methods: Primary cultures of human colonic submucosal fibroblasts/myofibroblasts were incubated with IL-1alpha (100 U/ml), IL-Ibeta (10 ng/ml), IL-10 (10 ng/ml), TNF (10 ng/ml), PMA (10 ng/ml), LPS (50 ng/ml), IL-4 (10 ng/ml), or a combination of IL-1 and TNF. Secreted cytokines were determined by ELISA. NF-kappaB activation was demonstrated by electrophoretic mobility-shift assays (EMSA).

Results: Incubation of colonic fibroblasts/myofibroblasts with IL-1, LPS, TNF and PMA induced secretion of IL-6, IL-8, M-CSF and GM-CSF. IL-8 and IL-6 secretion could be stimulated by IL-1alpha, IL-1beta, TNF, PMA and LPS within 6 h of incubation. IL-6 secretion was stimulated from 0.5 +/- 0.01 pg/h x microg fibroblast protein to 18.5 +/- 2.6 pg/h x microg fibroblast protein with IL-1beta (P < 0.01). IL-8 secretion was stimulated from 1.0 +/- 0.1 pg/h x microg fibroblast protein to 41.1 +/- 3.6 pg/h x microg (P < 0.005). IL-4 and IL-10 did not change cytokine secretion significantly. No significant differences between cultures from normal and inflamed mucosa were observed. TNF and IL-1 induced NF-kappaB activation. ALLN, a proteasome and NF-kappaB activation inhibitor, reduced TNF-mediated IL-8, GM-CSF and M-CSF induction significantly, whereas induction of IL-6 secretion remained unchanged.

Conclusion: Human colonic myofibroblasts can secrete large amounts of IL-6, IL-8, M-CSF and GM-CSF upon stimulation. The induction of IL-8, M-CSF and GM-CSF, but not of IL-6 secretion, is mediated mainly by NF-kappaB activation. The cytokine profile and the total amounts of cytokines released suggest that colonic myofibroblasts can play a role in leukocyte recruitment and during mucosal inflammation. They therefore have to be regarded as an important part of the mucosal immune system.

Publication types

  • Clinical Trial
  • Comparative Study
  • Controlled Clinical Trial

MeSH terms

  • Adult
  • Analysis of Variance
  • Biopsy, Needle
  • Cells, Cultured
  • Colitis, Ulcerative / metabolism*
  • Colitis, Ulcerative / pathology
  • Colon / metabolism*
  • Crohn Disease / pathology
  • Cytokines / analysis
  • Cytokines / drug effects
  • Cytokines / metabolism*
  • Electrophoresis
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • Fibroblasts / drug effects
  • Fibroblasts / metabolism*
  • Humans
  • Immunohistochemistry
  • Interleukin-1 / pharmacology
  • Interleukin-10 / pharmacology
  • Intestinal Mucosa / cytology
  • Intestinal Mucosa / drug effects
  • Intestinal Mucosa / metabolism*
  • Lipopolysaccharides / pharmacology
  • Male
  • Middle Aged
  • NF-kappa B / drug effects
  • NF-kappa B / metabolism
  • Probability
  • Reference Values
  • Sensitivity and Specificity
  • Tumor Necrosis Factor-alpha / pharmacology


  • Cytokines
  • Interleukin-1
  • Lipopolysaccharides
  • NF-kappa B
  • Tumor Necrosis Factor-alpha
  • Interleukin-10