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Toll-like receptor expression in crypt epithelial cells, putative stem cells and intestinal myofibroblasts isolated from controls and patients with inflammatory bowel disease.

Brown M, Hughes KR, Moossavi S, Robins A, Mahida YR - Clin. Exp. Immunol. (2014)

Bottom Line: In conclusion, enhanced TLR-2 and TLR-4 expression by crypt epithelial cells in active inflammatory bowel disease likely reflects greater ability to respond to microbial products.Results from our studies using mucosal samples from patients with distal ulcerative colitis suggest that the enhanced expression of these TLRs could be constitutive.TLR-2, TLR-4 and TLR-5 expression by stem cells imply ability to respond to distinct bacterial products.

View Article: PubMed Central - PubMed

Affiliation: Institute of Infection, Immunity and Inflammation, University of Nottingham, UK; Nottingham Digestive Diseases Centre, University of Nottingham, UK.

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Quantitative surface Toll-like receptor (TLR)-2 and TLR-4 protein expression by colonic crypt epithelial cells. Isolated and disaggregated crypt epithelial cells were obtained from mucosal samples affected by active Crohn’s colitis (n = 8), active ulcerative colitis (n = 4) or from histologically normal control colonic tissue (n = 7). The cells were labelled with anti-BerEP4-fluorescein isothiocyanate (FITC) antibody and either anti-TLR2-allophycocyanin (APC), anti-TLR-4-APC or isotype control monoclonal antibodies and analysed by flow cytometry. Surface TLR-2 (a) and TLR-4 (b) protein expression was assessed in BerEP4-positive (gated) epithelial cells. Each data point represents the difference in median fluorescent intensity (MFI) between the primary and isotype control antibodies. Horizontal bars represent median values.
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fig04: Quantitative surface Toll-like receptor (TLR)-2 and TLR-4 protein expression by colonic crypt epithelial cells. Isolated and disaggregated crypt epithelial cells were obtained from mucosal samples affected by active Crohn’s colitis (n = 8), active ulcerative colitis (n = 4) or from histologically normal control colonic tissue (n = 7). The cells were labelled with anti-BerEP4-fluorescein isothiocyanate (FITC) antibody and either anti-TLR2-allophycocyanin (APC), anti-TLR-4-APC or isotype control monoclonal antibodies and analysed by flow cytometry. Surface TLR-2 (a) and TLR-4 (b) protein expression was assessed in BerEP4-positive (gated) epithelial cells. Each data point represents the difference in median fluorescent intensity (MFI) between the primary and isotype control antibodies. Horizontal bars represent median values.

Mentions: Toll-like receptor (TLR)-2 and TLR4 protein expression on the surface of isolated and disaggregated colonic crypt epithelial cells. The cells were incubated with anti-BerEP4-fluorescein isothiocyanate (FITC) antibody, followed by anti-TLR-2-allophycocyanin (APC), anti-TLR-4-APC or isotype control monoclonal antibodies. The cells were subsequently fixed and analysed by flow cytometry. TLR-2 (upper middle and right panels) and TLR-4 (lower middle and right panels) expression was determined in BerEP4-positive (gated) cells, which represent intestinal epithelial cells. The histograms show median fluorescent intensity of crypt epithelial cells labelled with anti-TLR-2 (red line, upper right panel) and anti-TLR-4 (blue line, lower right panel) monoclonal antibodies, or isotype control antibody (green line). The figure is representative of the experimental data summarized in Fig. 4, using crypt epithelial cells isolated from histologically normal control and inflamed (ulcerative colitis and Crohn’s colitis) colonic mucosal samples.


Toll-like receptor expression in crypt epithelial cells, putative stem cells and intestinal myofibroblasts isolated from controls and patients with inflammatory bowel disease.

Brown M, Hughes KR, Moossavi S, Robins A, Mahida YR - Clin. Exp. Immunol. (2014)

Quantitative surface Toll-like receptor (TLR)-2 and TLR-4 protein expression by colonic crypt epithelial cells. Isolated and disaggregated crypt epithelial cells were obtained from mucosal samples affected by active Crohn’s colitis (n = 8), active ulcerative colitis (n = 4) or from histologically normal control colonic tissue (n = 7). The cells were labelled with anti-BerEP4-fluorescein isothiocyanate (FITC) antibody and either anti-TLR2-allophycocyanin (APC), anti-TLR-4-APC or isotype control monoclonal antibodies and analysed by flow cytometry. Surface TLR-2 (a) and TLR-4 (b) protein expression was assessed in BerEP4-positive (gated) epithelial cells. Each data point represents the difference in median fluorescent intensity (MFI) between the primary and isotype control antibodies. Horizontal bars represent median values.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4231243&req=5

fig04: Quantitative surface Toll-like receptor (TLR)-2 and TLR-4 protein expression by colonic crypt epithelial cells. Isolated and disaggregated crypt epithelial cells were obtained from mucosal samples affected by active Crohn’s colitis (n = 8), active ulcerative colitis (n = 4) or from histologically normal control colonic tissue (n = 7). The cells were labelled with anti-BerEP4-fluorescein isothiocyanate (FITC) antibody and either anti-TLR2-allophycocyanin (APC), anti-TLR-4-APC or isotype control monoclonal antibodies and analysed by flow cytometry. Surface TLR-2 (a) and TLR-4 (b) protein expression was assessed in BerEP4-positive (gated) epithelial cells. Each data point represents the difference in median fluorescent intensity (MFI) between the primary and isotype control antibodies. Horizontal bars represent median values.
Mentions: Toll-like receptor (TLR)-2 and TLR4 protein expression on the surface of isolated and disaggregated colonic crypt epithelial cells. The cells were incubated with anti-BerEP4-fluorescein isothiocyanate (FITC) antibody, followed by anti-TLR-2-allophycocyanin (APC), anti-TLR-4-APC or isotype control monoclonal antibodies. The cells were subsequently fixed and analysed by flow cytometry. TLR-2 (upper middle and right panels) and TLR-4 (lower middle and right panels) expression was determined in BerEP4-positive (gated) cells, which represent intestinal epithelial cells. The histograms show median fluorescent intensity of crypt epithelial cells labelled with anti-TLR-2 (red line, upper right panel) and anti-TLR-4 (blue line, lower right panel) monoclonal antibodies, or isotype control antibody (green line). The figure is representative of the experimental data summarized in Fig. 4, using crypt epithelial cells isolated from histologically normal control and inflamed (ulcerative colitis and Crohn’s colitis) colonic mucosal samples.

Bottom Line: In conclusion, enhanced TLR-2 and TLR-4 expression by crypt epithelial cells in active inflammatory bowel disease likely reflects greater ability to respond to microbial products.Results from our studies using mucosal samples from patients with distal ulcerative colitis suggest that the enhanced expression of these TLRs could be constitutive.TLR-2, TLR-4 and TLR-5 expression by stem cells imply ability to respond to distinct bacterial products.

View Article: PubMed Central - PubMed

Affiliation: Institute of Infection, Immunity and Inflammation, University of Nottingham, UK; Nottingham Digestive Diseases Centre, University of Nottingham, UK.

Show MeSH
Related in: MedlinePlus