Apical TLR ligation of intestinal epithelial cells drives a Th1-polarized regulatory or inflammatory type effector response in vitro

Immunobiology. 2011 Apr;216(4):518-27. doi: 10.1016/j.imbio.2010.08.005. Epub 2010 Aug 21.

Abstract

Intestinal epithelial cells (IEC) separate the mucosal immune system from the external milieu. Under inflammatory conditions, Toll-like receptor (TLR) expression by IEC is increased. In a transwell co-culture model immune modulation by IEC upon TLR ligation was studied. Human IEC (HT-29 and T84) grown on filters were apically or basolaterally exposed to TLR4 or TLR9 ligands and co-cultured with CD3/CD28-activated healthy donor PBMC in the basolateral compartment. TLR4 ligation of IEC (HT-29) enhanced the production of TNF-α and IEC-derived MDC and decreased numbers of Foxp3(+) regulatory T cells. Neutralization of TSLP abrogated TLR4-induced TNF-α secretion. In contrast, apical TLR9 ligation of IEC (HT-29 and T84) enhanced IFN-γ and IL-10 secretion and increased the number of activated T(h)1 cells. The increase in IFN-γ secretion depended on the presence of IEC. Furthermore, CD14 expression on monocytes was reduced coinciding with enhanced intracellular IL-10 and decreased TNF-α production. However, basolateral TLR9 ligand exposure of HT-29 cells resulted in enhanced IFN-γ, IL-6 and TNF-α, while IL-10 secretion remained unaltered. TLR4 and TLR9 ligands reduced IL-13 secretion in presence and absence of apically exposed IEC and enhanced IL-12 secretion in presence of IEC. These data suggest that TLR4 ligation of IEC drives an inflammatory, while apical TLR9 ligation drives a regulatory T(h)1 effector immune response in vitro in a polarized manner. IEC may be important modulators of the mucosal effector immune response.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line, Tumor
  • Cell Polarity / immunology*
  • Coculture Techniques
  • Cytokines / metabolism
  • Epithelial Cells / immunology*
  • HT29 Cells
  • Humans
  • Immunophenotyping
  • Inflammation / immunology
  • Intestinal Mucosa / cytology
  • Intestinal Mucosa / immunology
  • Intestines / cytology
  • Intestines / immunology*
  • Leukocytes, Mononuclear / immunology
  • Receptors, Cell Surface / immunology
  • Th1 Cells / immunology*
  • Toll-Like Receptors / immunology*

Substances

  • Cytokines
  • Receptors, Cell Surface
  • Toll-Like Receptors