Studying permeability in a commonly used epithelial cell line: T84 intestinal epithelial cells

Methods Mol Biol. 2011:763:115-37. doi: 10.1007/978-1-61779-191-8_8.

Abstract

The integrity, or barrier function, of the intestinal epithelium is of paramount importance in -maintaining good health. This is largely imparted by a single layer of epithelial cells linked by the transmembrane tight junction protein complex near their apical surface. Disruption of epithelial permeability via the tight junctions can contribute to disease progression. The cytokine IFNγ is involved in many inflammatory processes and has been shown to dramatically increase permeability via changes at the tight junction in experimental models. One of its key effectors is the transcription factor, -IRF-1. In our studies of the role of IRF-1 in barrier function using the human T84 intestinal epithelial cell monolayer model, we have found that induction of IRF-1 alone is insufficient to change permeability and that if IRF-1 is involved in mediating the permeability effects of IFNγ, then other factors must also be required.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blotting, Western
  • Cell Line
  • Dextrans / analysis
  • Diffusion Chambers, Culture
  • Electrophoresis, Polyacrylamide Gel
  • Epithelial Cells / cytology
  • Epithelial Cells / metabolism*
  • Fluorescein-5-isothiocyanate / analogs & derivatives
  • Fluorescein-5-isothiocyanate / analysis
  • Fluorescent Antibody Technique
  • Humans
  • Immunohistochemistry
  • Inflammation / immunology
  • Inflammation / metabolism
  • Interferon Regulatory Factor-1 / genetics
  • Interferon Regulatory Factor-1 / immunology
  • Interferon Regulatory Factor-1 / metabolism*
  • Interferon-gamma / genetics
  • Interferon-gamma / immunology
  • Interferon-gamma / metabolism*
  • Intestinal Mucosa / cytology
  • Intestinal Mucosa / metabolism*
  • Intestines / cytology
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism*
  • Occludin
  • Permeability
  • Plasmids
  • Tight Junctions / genetics
  • Tight Junctions / immunology
  • Tight Junctions / metabolism*
  • Transfection

Substances

  • Dextrans
  • Interferon Regulatory Factor-1
  • Membrane Proteins
  • OCLN protein, human
  • Occludin
  • fluorescein isothiocyanate dextran
  • Interferon-gamma
  • Fluorescein-5-isothiocyanate