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.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsSpringer Nature is developing a new tool to find and evaluate Protocols. Learn more
References
Tsukita, S., Furuse, M., and Itoh, M. (2001) Multifunctional strands in tight junctions, Nat Rev Mol Cell Biol 2, 285–293.
Capaldo, C. T., and Nusrat, A. (2009) Cytokine regulation of tight junctions, Biochim. Biophys. Acta 1788, 864–871.
Al-Sadi, R., Boivin, M., and Ma, T. (2009) Mechanism of cytokine modulation of epithelial tight junction barrier, Front Biosci 14, 2765–2778.
Schulzke, J. D., Ploeger, S., Amasheh, M., Fromm, A., Zeissig, S., Troeger, H., Richter, J., Bojarski, C., Schumann, M., and Fromm, M. (2009) Epithelial tight junctions in intestinal inflammation, Ann N Y Acad Sci 1165, 294–300.
Clayburgh, D. R., Shen, L., and Turner, J. R. (2004) A porous defense: the leaky epithelial barrier in intestinal disease, Lab Invest 84, 282–291.
Marchiando, A. M., Graham, W. V., and Turner, J. R. (2010) Epithelial barriers in homeostasis and disease, Annu Rev Pathol 5, 119–144.
Deli, M. A. (2009) Potential use of tight junction modulators to reversibly open membranous barriers and improve drug delivery, Biochim. Biophys. Acta 1788, 892–910.
Dharmsathaphorn, K., McRoberts, J. A., Mandel, K. G., Tisdale, L. D., and Masui, H. (1984) A human colonic tumor cell line that maintains vectorial electrolyte transport, Am J Physiol 246, G204–208.
Dharmsathaphorn, K., and Madara, J. L. (1990) Established intestinal cell lines as model systems for electrolyte transport studies, Methods Enzymol 192, 354–389.
Hillgren, K. M., Kato, A., and Borchardt, R. T. (1995) In vitro systems for studying intestinal drug absorption, Med Res Rev 15, 83–109.
Harhaj, N. S., and Antonetti, D. A. (2004) Regulation of tight junctions and loss of barrier function in pathophysiology, Int. J. Biochem. Cell Biol. 36, 1206–1237.
Schroder, K., Hertzog, P. J., Ravasi, T., and Hume, D. A. (2004) Interferon-gamma: an overview of signals, mechanisms and functions, J Leukoc Biol 75, 163–189.
Adams, R. B., Planchon, S. M., and Roche, J. K. (1993) IFN-gamma modulation of epithelial barrier function. Time course, reversibility, and site of cytokine binding, J Immunol 150, 2356–2363.
Colgan, S. P., Parkos, C. A., Matthews, J. B., D’Andrea, L., Awtrey, C. S., Lichtman, A. H., Delp-Archer, C., and Madara, J. L. (1994) Interferon-gamma induces a cell surface phenotype switch on T84 intestinal epithelial cells, Am J Physiol 267, C402–410.
Bruewer, M., Luegering, A., Kucharzik, T., Parkos, C. A., Madara, J. L., Hopkins, A. M., and Nusrat, A. (2003) Proinflammatory cytokines disrupt epithelial barrier function by apoptosis-independent mechanisms, J Immunol 171, 6164–6172.
Youakim, A., and Ahdieh, M. (1999) Interferon-gamma decreases barrier function in T84 cells by reducing ZO-1 levels and disrupting apical actin, Am J Physiol 276, G1279–1288.
Donato, R., Wood, S. A., Saunders, I., Gundsambuu, B., Yan Mak, K., Abbott, C. A., and Powell, B. C. (2009) Regulation of epithelial apical junctions and barrier function by Galpha13, Biochim. Biophys. Acta 1793, 1228–1235.
Drexler, H. G., and Uphoff, C. C. (2002) Mycoplasma contamination of cell cultures: Incidence, sources, effects, detection, elimination, prevention, Cytotechnology 39, 75–90.
Sanders, S. E., Madara, J. L., McGuirk, D. K., Gelman, D. S., and Colgan, S. P. (1995) Assessment of inflammatory events in epithelial permeability: a rapid screening method using fluorescein dextrans, Epithelial Cell Biol 4, 25–34.
Sander, G. R., Cummins, A. G., Henshall, T., and Powell, B. C. (2005) Rapid disruption of intestinal barrier function by gliadin involves altered expression of apical junctional proteins, FEBS Lett 579, 4851–4855.
Acknowledgements
This study was supported in part by Dairy Australia and the Women’s and Children’s Hospital Foundation. R. Donato was supported by a Flinders University Faculty of Science and Engineering Research award.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2011 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Donato, R.P. et al. (2011). Studying Permeability in a Commonly Used Epithelial Cell Line: T84 Intestinal Epithelial Cells. In: Turksen, K. (eds) Permeability Barrier. Methods in Molecular Biology, vol 763. Humana Press. https://doi.org/10.1007/978-1-61779-191-8_8
Download citation
DOI: https://doi.org/10.1007/978-1-61779-191-8_8
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-61779-190-1
Online ISBN: 978-1-61779-191-8
eBook Packages: Springer Protocols