MAPK regulation of IL-4/IL-13 receptors contributes to the synergistic increase in CCL11/eotaxin-1 in response to TGF-β1 and IL-13 in human airway fibroblasts

J Immunol. 2012 Jun 15;188(12):6046-54. doi: 10.4049/jimmunol.1102760. Epub 2012 May 9.

Abstract

CCL11/eotaxin-1 is a potent eosinophilic CC chemokine expressed by primary human fibroblasts. The combination of TGF-β1 and IL-13 synergistically increases CCL11 expression, but the mechanisms behind the synergy are unclear. To address this, human airway fibroblast cultures from normal and asthmatic subjects were exposed to IL-13 alone or TGF-β1 plus IL-13. Transcriptional (nuclear run-on) and posttranscriptional (mRNA stability) assays confirmed that transcriptional regulation is critical for synergistic expression of CCL11. TGF-β1 plus IL-13 synergistically increased STAT-6 phosphorylation, nuclear translocation, and binding to the CCL11 promoter as compared with IL-13 alone. STAT-6 small interfering RNA significantly knocked down both STAT-6 mRNA expression and phosphorylation and inhibited CCL11 mRNA and protein expression. Regulation of the IL-4Rα complex by TGF-β1 augmented IL-13 signaling by dampening IL-13Rα2 expression, overcoming IL-13's autoregulation of its pathway and enhancing the expression of CCL11. Our data suggest that TGF-β1 induced activation of the MEK/ERK pathway reduces IL-13Rα2 expression induced by IL-13. Thus, TGF-β1, a pleiotropic cytokine upregulated in asthmatic airways, can augment eosinophilic inflammation by interfering with IL-13's negative feedback autoregulatory loop under MEK/ERK-dependent conditions.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Asthma / immunology
  • Asthma / metabolism*
  • Blotting, Western
  • Chemokine CCL11 / immunology
  • Chemokine CCL11 / metabolism
  • Chromatin Immunoprecipitation
  • Extracellular Signal-Regulated MAP Kinases / immunology*
  • Extracellular Signal-Regulated MAP Kinases / metabolism
  • Fibroblasts / immunology
  • Fibroblasts / metabolism*
  • Fluorescent Antibody Technique
  • Gene Expression Profiling
  • Gene Expression Regulation / immunology*
  • Humans
  • Inflammation / immunology
  • Inflammation / metabolism
  • Interleukin-13 / immunology
  • Interleukin-13 / metabolism
  • Lung / immunology
  • Lung / metabolism*
  • Receptors, Interleukin-13 / immunology
  • Receptors, Interleukin-13 / metabolism
  • Receptors, Interleukin-4 / immunology
  • Receptors, Interleukin-4 / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • STAT6 Transcription Factor / immunology
  • STAT6 Transcription Factor / metabolism
  • Signal Transduction / immunology
  • Transforming Growth Factor beta1 / immunology
  • Transforming Growth Factor beta1 / metabolism*

Substances

  • CCL11 protein, human
  • Chemokine CCL11
  • Interleukin-13
  • Receptors, Interleukin-13
  • Receptors, Interleukin-4
  • STAT6 Transcription Factor
  • STAT6 protein, human
  • Transforming Growth Factor beta1
  • Extracellular Signal-Regulated MAP Kinases