Abstract
Epithelial cells at all mucosal surfaces are potentially apposed to bacteria, particularly in the intestine. It is established that intestinal epithelial cells (IECs) represent an important barrier between lamina propria cells and the potentially harmful lumenal contents. In addition, IECs are important immunoeffector cells with the capacity to release cytokines, chemokines, and other molecules involved in antigen presentation and immune defense. The interaction of IECs with intestinal bacteria can result in a decrease in barrier function and the development of inflammation, which is known to be an important factor in the development of intestinal pathology. The potential role of such crosstalk between bacteria and other intestinal cell types in normal physiology and/or pathophysiology is therefore a topic of intense investigation. In this chapter, we provide protocols for the identification of bacteria that are associated with the epithelium and mucosa in addition to functional assays examining the interactions of neutrophils with epithelial cells and epithelial cell-mediated killing of bacteria.
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References
Berg, R. D. (1996) The indigenous gastrointestinal microflora. Trends Microbiol. 4, 430–435.
Sansonetti, P. J. (2004) War and peace at mucosal surfaces. Nat. Rev. Immunol. 4, 953–964.
Ouellette, A. J. (2004) Defensin-mediated innate immunity in the small intestine. Best Pract. Res. Clin. Gastroenterol. 18, 405–419.
Swidsinski, A., Ladhoff, A., Pernthaler, S., et al. (2002) Mucosal flora in inflammatory bowel disease. Gastroenterology 122, 44–54.
Fiocchi, C. (1998) Inflammatory bowel disease; etiology and pathogenesis. Gastroenterology 115, 182–205.
Turnbull, P. C. and Richmond, J. E. (1978) A model of salmonella enteritis: the behaviour of, Salmonella enteritidis in chick intestine studies by light and electron microscopy. Br. J. Exp. Pathol. 59, 64–75.
Wallis, T. S., Hawker, R. J., Candy, D. C., et al. (1989) Quantification of the leucocyte influx into rabbit ileal loops induced by strains of, Salmonella typhimurium of different virulence. J. Med. Microbiol. 30, 149–156.
MacDermott, R. P. (1999) Chemokines in the inflammatory bowel diseases. J. Clin. Immunol. 19, 266–272.
Ajuebor, M. N. and Swain, M. G. (2002) Role of chemokines and chemokine receptors in the gastrointestinal tract. Immunology 105, 137–143.
Pavlick, K. P., Laroux, F. S., Fuseler, J., et al. (2002) Role of reactive metabolites of oxygen and nitrogen in inflammatory bowel disease (1,2). Free Radic. Biol. Med. 33, 311–322.
Nash, S., Stafford, J., and Madara, J. L. (1987) Effects of polymorphonuclear leukocyte transmigration on the barrier function of cultured intestinal epithelial monolayers. J. Clin. Invest. 80, 1104–1113.
Lee, C. A., Silva, M., Siber, A. M., Kelly, A. J., Galyov, E., and McCormick, B. A. (2000) A secreted Salmonella protein induces a proinflammatory response in epithelial cells, which promotes neutrophil migration. Proc. Natl. Acad. Sci. USA 97, 12,283–12,288.
Mrsny, R. J., Gewirtz, A. T., Siccardi, D., et al. (2004) Identification of hepoxilin, A3 in inflammatory events: a required role in neutrophil migration across intestinal epithelia. Proc. Natl. Acad. Sci. USA 101, 7421–7426.
Zasloff, M. (1992) Antibiotic peptides as mediators of innate immunity. Curr. Opin. Immunol. 4, 3–7.
Nissen-Meyer, J. and Nes, I. F. (1997) Ribosomally synthesized antimicrobial peptides: their function, structure, biogenesis, and mechanism of action. Arch. Microbiol. 167, 67–77.
Canny, G., Levy, O., Furuta, G. T., et al. (2002) Lipid mediator induced expression of bactericidal/permeability-increasing protein (BPI) in human mucosal epithelia. Proc. Natl. Acad. Sci. USA 99, 3902–3907.
Fellermann, K., Wehkamp, J., Herrlinger, K. R., and Stange, E. F. (2003) Crohn’s disease: a defensin deficiency syndrome? Eur. J. Gastroenterol. Hepatol. 15, 627–634.
Costerton, J. W., Veeh, R., Shirtliff, M., Pasmore, M., Post, C., and Ehrlich, G. (2003) The application of biofilm science to the study and control of chronic bacterial infections. J. Clin. Invest. 112, 1466–1477.
Wilson, M. (2001) Bacterial biofilms and human disease. Sci. Prog. 84, 235–254.
Amann, R., Krumholz, L., and Stahl, D. A. (1990) Fluorescent-oligonucleotide probing of whole cells for determinative, phylogenetic, and environmental studies in microbiology. J. Bacteriol. 172, 762–770.
Bohnert, J., Hübner, B., and Botzenhart, K. (2002) Rapid identification of Enterobacteriaceae using a novel 23S rR. N.A-targeted oligonucleotide probe. Int. J. Hyg. Environ. Health 203, 77–82.
Franks, A. H., Harmsen, H. J., Raangs, G. C., Jansen, G. J., Schut, F., and Welling, G. W. (1998) Variations of bacterial populations in human feces measured by fluorescent in situ hybridization with group-specific 16S rR. N.A-targeted oligonucleotide probes. Appl. Environ. Microbiol. 64, 3336–3345.
Harmsen, H. J., Raangs, G. C., He, T., Degener, J. E., and Welling, G. W. (2002) Extensive set of 16S rRNA-based probes for detection of bacteria in human feces. Appl. Environ. Microbiol. 68, 2982–2990.
Elsbach, P. and Weiss, J. (1992) Oxygen-independent antimicrobial systems of phagocytes, in Inflammation: Basic principles and Clinical Correlates, Second Edition (Gallin J. I., Goldstein I. M., and Snyderman R., eds.), Raven, New York, pp. 603–636.
Cereijido, M., Robbins, E. S., Dolan, W. J., Rotunno, C. A., and Sabitini, D. D. (1978) Polarized monolayers formed by epithelial cells on a permeable and translucent support. J. Cell Biol. 77, 853–880.
McCormick, B.A., Colgan, S. P., Delp-Archer, C., Miller, S. I., and Madara, J. L. (1993) Salmonella typhimurium attachment to human intestinal epithelial monolayers: transcellular signaling to subepithelial neutrophils. J. Cell Biol. 123, 895–907.
Henson, P. M. and Oades, Z. G. (1975) Stimulation of human neutrophils by soluble and insoluble immunoglobulin aggregates: secretion of granule constituents and increased oxidation of glucose. J. Clin. Invest. 56, 1053–1061.
Lee, C. A. and Falkow, S. (1990) The ability of Salmonella to enter mammalian cells is affected by bacterial growth state. Proc. Natl. Acad. Sci. USA 87, 4304–4308.
Lissner, C. R., Swanson, R. N., and O’Brien, A. D. (1983) Genetic control of the innate resistance of mice to Salmonella typhimurium expression of the Ity gene in peritoneal and splenic macrophages is located in vitro. J. Immunol. 131, 3006–3013.
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Canny, G., Swidsinski, A., McCormick, B.A. (2006). Interactions of Intestinal Epithelial Cells With Bacteria and Immune Cells: Methods to Characterize Microflora and Functional Consequences. In: Colgan, S.P. (eds) Cell-Cell Interactions. Methods in Molecular Biology™, vol 341. Humana Press. https://doi.org/10.1385/1-59745-113-4:17
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DOI: https://doi.org/10.1385/1-59745-113-4:17
Publisher Name: Humana Press
Print ISBN: 978-1-58829-523-1
Online ISBN: 978-1-59745-113-0
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