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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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Side population cells (putative stem cells) express Toll-like receptor (TLR)-2, TLR-4 and TLR-5 mRNA transcripts. Side population cells in isolated and disaggregated colonic crypt epithelial cells were identified (as described in Fig. 5a) and sorted for extraction of RNA, for subsequent reverse transcription–polymerase chain reaction (RT–PCR) using primer pairs specific for TLR-2, TLR-4 and TLR-5. Lane M represents the 100 base pair size ladder. The figure is representative of crypt epithelial cells isolated from three operation resection specimens.
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fig06: Side population cells (putative stem cells) express Toll-like receptor (TLR)-2, TLR-4 and TLR-5 mRNA transcripts. Side population cells in isolated and disaggregated colonic crypt epithelial cells were identified (as described in Fig. 5a) and sorted for extraction of RNA, for subsequent reverse transcription–polymerase chain reaction (RT–PCR) using primer pairs specific for TLR-2, TLR-4 and TLR-5. Lane M represents the 100 base pair size ladder. The figure is representative of crypt epithelial cells isolated from three operation resection specimens.

Mentions: Side population cells present in isolated and disaggregated crypt cell preparations from normal control colon were characterized by flow cytometry (Fig. 5a), as described previously 24. Sorted side population cells were labelled by anti-BerEP4 (Fig. 5b), anti-TLR-2 and anti-TLR-4 (Fig. 5c) antibodies. When studied by RT–PCR, sorted side population cells also expressed transcripts for TLR-2, TLR-4 and TLR-5 (Fig. 6). In contrast to other disaggregated crypt epithelial cells, side population/putative stem cells adhere readily to monolayers of intestinal myofibroblasts 24. Such co-cultures were used to demonstrate immunoreactivity for not only BerEP4 (Fig. 7a), but also TLR-2 (Fig. 7b), TL-4 (Fig. 7c) and TLR-5 (Fig. 7d). In contrast to the epithelial cells, myofibroblast immunoreactivity for TLR-2, TLR-4 and TLR-5 in these co-cultures was weak.


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)

Side population cells (putative stem cells) express Toll-like receptor (TLR)-2, TLR-4 and TLR-5 mRNA transcripts. Side population cells in isolated and disaggregated colonic crypt epithelial cells were identified (as described in Fig. 5a) and sorted for extraction of RNA, for subsequent reverse transcription–polymerase chain reaction (RT–PCR) using primer pairs specific for TLR-2, TLR-4 and TLR-5. Lane M represents the 100 base pair size ladder. The figure is representative of crypt epithelial cells isolated from three operation resection specimens.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig06: Side population cells (putative stem cells) express Toll-like receptor (TLR)-2, TLR-4 and TLR-5 mRNA transcripts. Side population cells in isolated and disaggregated colonic crypt epithelial cells were identified (as described in Fig. 5a) and sorted for extraction of RNA, for subsequent reverse transcription–polymerase chain reaction (RT–PCR) using primer pairs specific for TLR-2, TLR-4 and TLR-5. Lane M represents the 100 base pair size ladder. The figure is representative of crypt epithelial cells isolated from three operation resection specimens.
Mentions: Side population cells present in isolated and disaggregated crypt cell preparations from normal control colon were characterized by flow cytometry (Fig. 5a), as described previously 24. Sorted side population cells were labelled by anti-BerEP4 (Fig. 5b), anti-TLR-2 and anti-TLR-4 (Fig. 5c) antibodies. When studied by RT–PCR, sorted side population cells also expressed transcripts for TLR-2, TLR-4 and TLR-5 (Fig. 6). In contrast to other disaggregated crypt epithelial cells, side population/putative stem cells adhere readily to monolayers of intestinal myofibroblasts 24. Such co-cultures were used to demonstrate immunoreactivity for not only BerEP4 (Fig. 7a), but also TLR-2 (Fig. 7b), TL-4 (Fig. 7c) and TLR-5 (Fig. 7d). In contrast to the epithelial cells, myofibroblast immunoreactivity for TLR-2, TLR-4 and TLR-5 in these co-cultures was weak.

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