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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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Toll-like receptor (TLR)-2 and TLR-4 protein expression in colonic crypt epithelial cells and myofibroblasts. Colonic crypt epithelial cells were isolated from histologically normal colonic (lanes 1 and 2) mucosal samples and those affected by active Crohn’s colitis (lanes 3 and 4) and active ulcerative colitis (lanes 5 and 6). Primary human colonic myofibroblasts were isolated from histologically normal colonic mucosal samples (lanes 7–10). Cell lysates were used for Western blot analysis. For crypt epithelial cell lysates, 20 μg of total protein was applied per lane. Although higher amounts of myofibroblast cell lysates (40 μg in each of lanes 7 and 8; 80 μg in each of lanes 9 and 10) were used, bands for TLR-2 and TLR-4 were stronger in lysates of crypt epithelial cells (lanes 1–6).
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fig08: Toll-like receptor (TLR)-2 and TLR-4 protein expression in colonic crypt epithelial cells and myofibroblasts. Colonic crypt epithelial cells were isolated from histologically normal colonic (lanes 1 and 2) mucosal samples and those affected by active Crohn’s colitis (lanes 3 and 4) and active ulcerative colitis (lanes 5 and 6). Primary human colonic myofibroblasts were isolated from histologically normal colonic mucosal samples (lanes 7–10). Cell lysates were used for Western blot analysis. For crypt epithelial cell lysates, 20 μg of total protein was applied per lane. Although higher amounts of myofibroblast cell lysates (40 μg in each of lanes 7 and 8; 80 μg in each of lanes 9 and 10) were used, bands for TLR-2 and TLR-4 were stronger in lysates of crypt epithelial cells (lanes 1–6).

Mentions: Using conventional RT–PCR, myofibroblasts isolated from normal control and active IBD mucosal samples showed PCR products specific for TLR-2 and TLR-4 (not shown). TLR-2 and TLR-4 protein expression was confirmed by Western blot analysis, but the level of expression was much lower than that for isolated crypt epithelial cells (Fig. 8).


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)

Toll-like receptor (TLR)-2 and TLR-4 protein expression in colonic crypt epithelial cells and myofibroblasts. Colonic crypt epithelial cells were isolated from histologically normal colonic (lanes 1 and 2) mucosal samples and those affected by active Crohn’s colitis (lanes 3 and 4) and active ulcerative colitis (lanes 5 and 6). Primary human colonic myofibroblasts were isolated from histologically normal colonic mucosal samples (lanes 7–10). Cell lysates were used for Western blot analysis. For crypt epithelial cell lysates, 20 μg of total protein was applied per lane. Although higher amounts of myofibroblast cell lysates (40 μg in each of lanes 7 and 8; 80 μg in each of lanes 9 and 10) were used, bands for TLR-2 and TLR-4 were stronger in lysates of crypt epithelial cells (lanes 1–6).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig08: Toll-like receptor (TLR)-2 and TLR-4 protein expression in colonic crypt epithelial cells and myofibroblasts. Colonic crypt epithelial cells were isolated from histologically normal colonic (lanes 1 and 2) mucosal samples and those affected by active Crohn’s colitis (lanes 3 and 4) and active ulcerative colitis (lanes 5 and 6). Primary human colonic myofibroblasts were isolated from histologically normal colonic mucosal samples (lanes 7–10). Cell lysates were used for Western blot analysis. For crypt epithelial cell lysates, 20 μg of total protein was applied per lane. Although higher amounts of myofibroblast cell lysates (40 μg in each of lanes 7 and 8; 80 μg in each of lanes 9 and 10) were used, bands for TLR-2 and TLR-4 were stronger in lysates of crypt epithelial cells (lanes 1–6).
Mentions: Using conventional RT–PCR, myofibroblasts isolated from normal control and active IBD mucosal samples showed PCR products specific for TLR-2 and TLR-4 (not shown). TLR-2 and TLR-4 protein expression was confirmed by Western blot analysis, but the level of expression was much lower than that for isolated crypt epithelial cells (Fig. 8).

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