Up-regulation of MUC2 and IL-1β expression in human colonic epithelial cells by Shigella and its interaction with mucins.

<h4>Background</h4>The entire gastrointestinal tract is protected by a mucous layer, which contains complex glycoproteins called mucins. MUC2 is one such mucin that protects the colonic mucosa from invading microbes. The initial interaction between microbes and mucins is an important ste...

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Autores principales:	Radhakrishnan Prakash, Subramaniya Bharathi Raja, Halagowder Devaraj, Sivasitambaram Niranjali Devaraj
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2011
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/644ab2a9ebdf41f78e94abe2d3495769
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spelling	oai:doaj.org-article:644ab2a9ebdf41f78e94abe2d34957692021-11-18T07:34:57ZUp-regulation of MUC2 and IL-1β expression in human colonic epithelial cells by Shigella and its interaction with mucins.1932-620310.1371/journal.pone.0027046https://doaj.org/article/644ab2a9ebdf41f78e94abe2d34957692011-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22073249/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203<h4>Background</h4>The entire gastrointestinal tract is protected by a mucous layer, which contains complex glycoproteins called mucins. MUC2 is one such mucin that protects the colonic mucosa from invading microbes. The initial interaction between microbes and mucins is an important step for microbial pathogenesis. Hence, it was of interest to investigate the relationship between host (mucin) and pathogen interaction, including Shigella induced expression of MUC2 and IL-1β during shigellosis.<h4>Methods</h4>The mucin-Shigella interaction was revealed by an in vitro mucin-binding assay. Invasion of Shigella dysenteriae into HT-29 cells was analyzed by Transmission electron microscopy. Shigella induced mucin and IL-1β expression were analyzed by RT-PCR and Immunofluorescence.<h4>Results</h4>The clinical isolates of Shigella were found to be virulent by a congo-red binding assay. The in vitro mucin-binding assay revealed both Shigella dysenteriae and Shigella flexneri have binding affinity in the increasing order of: guinea pig small intestinal mucin<guinea pig colonic mucin< Human colonic mucin. Invasion of Shigella dysenteriae into HT-29 cells occurs within 2 hours. Interestingly, in Shigella dysenteriae infected conditions, significant increases in mRNA expression of MUC2 and IL-1β were observed in a time dependent manner. Further, immunofluorescence analysis of MUC2 shows more positive cells in Shigella dysenteriae treated cells than untreated cells.<h4>Conclusions</h4>Our study concludes that the Shigella species specifically binds to guinea pig colonic mucin, but not to guinea pig small intestinal mucin. The guinea pig colonic mucin showed a greater binding parameter (R), and more saturable binding, suggesting the presence of a finite number of receptor binding sites in the colonic mucin of the host. In addition, modification of mucins with TFMS and sodium metaperiodate significantly reduced mucin-bacterial binding; suggesting that the mucin-Shigella interaction occurs through carbohydrate epitopes on the mucin backbones. Overproduction of MUC2 may alter adherence and invasion of Shigella dysenteriae into human colonic epithelial cells.Radhakrishnan PrakashSubramaniya Bharathi RajaHalagowder DevarajSivasitambaram Niranjali DevarajPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 6, Iss 11, p e27046 (2011)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Radhakrishnan Prakash Subramaniya Bharathi Raja Halagowder Devaraj Sivasitambaram Niranjali Devaraj Up-regulation of MUC2 and IL-1β expression in human colonic epithelial cells by Shigella and its interaction with mucins.
description	<h4>Background</h4>The entire gastrointestinal tract is protected by a mucous layer, which contains complex glycoproteins called mucins. MUC2 is one such mucin that protects the colonic mucosa from invading microbes. The initial interaction between microbes and mucins is an important step for microbial pathogenesis. Hence, it was of interest to investigate the relationship between host (mucin) and pathogen interaction, including Shigella induced expression of MUC2 and IL-1β during shigellosis.<h4>Methods</h4>The mucin-Shigella interaction was revealed by an in vitro mucin-binding assay. Invasion of Shigella dysenteriae into HT-29 cells was analyzed by Transmission electron microscopy. Shigella induced mucin and IL-1β expression were analyzed by RT-PCR and Immunofluorescence.<h4>Results</h4>The clinical isolates of Shigella were found to be virulent by a congo-red binding assay. The in vitro mucin-binding assay revealed both Shigella dysenteriae and Shigella flexneri have binding affinity in the increasing order of: guinea pig small intestinal mucin<guinea pig colonic mucin< Human colonic mucin. Invasion of Shigella dysenteriae into HT-29 cells occurs within 2 hours. Interestingly, in Shigella dysenteriae infected conditions, significant increases in mRNA expression of MUC2 and IL-1β were observed in a time dependent manner. Further, immunofluorescence analysis of MUC2 shows more positive cells in Shigella dysenteriae treated cells than untreated cells.<h4>Conclusions</h4>Our study concludes that the Shigella species specifically binds to guinea pig colonic mucin, but not to guinea pig small intestinal mucin. The guinea pig colonic mucin showed a greater binding parameter (R), and more saturable binding, suggesting the presence of a finite number of receptor binding sites in the colonic mucin of the host. In addition, modification of mucins with TFMS and sodium metaperiodate significantly reduced mucin-bacterial binding; suggesting that the mucin-Shigella interaction occurs through carbohydrate epitopes on the mucin backbones. Overproduction of MUC2 may alter adherence and invasion of Shigella dysenteriae into human colonic epithelial cells.
format	article
author	Radhakrishnan Prakash Subramaniya Bharathi Raja Halagowder Devaraj Sivasitambaram Niranjali Devaraj
author_facet	Radhakrishnan Prakash Subramaniya Bharathi Raja Halagowder Devaraj Sivasitambaram Niranjali Devaraj
author_sort	Radhakrishnan Prakash
title	Up-regulation of MUC2 and IL-1β expression in human colonic epithelial cells by Shigella and its interaction with mucins.
title_short	Up-regulation of MUC2 and IL-1β expression in human colonic epithelial cells by Shigella and its interaction with mucins.
title_full	Up-regulation of MUC2 and IL-1β expression in human colonic epithelial cells by Shigella and its interaction with mucins.
title_fullStr	Up-regulation of MUC2 and IL-1β expression in human colonic epithelial cells by Shigella and its interaction with mucins.
title_full_unstemmed	Up-regulation of MUC2 and IL-1β expression in human colonic epithelial cells by Shigella and its interaction with mucins.
title_sort	up-regulation of muc2 and il-1β expression in human colonic epithelial cells by shigella and its interaction with mucins.
publisher	Public Library of Science (PLoS)
publishDate	2011
url	https://doaj.org/article/644ab2a9ebdf41f78e94abe2d3495769
work_keys_str_mv	AT radhakrishnanprakash upregulationofmuc2andil1bexpressioninhumancolonicepithelialcellsbyshigellaanditsinteractionwithmucins AT subramaniyabharathiraja upregulationofmuc2andil1bexpressioninhumancolonicepithelialcellsbyshigellaanditsinteractionwithmucins AT halagowderdevaraj upregulationofmuc2andil1bexpressioninhumancolonicepithelialcellsbyshigellaanditsinteractionwithmucins AT sivasitambaramniranjalidevaraj upregulationofmuc2andil1bexpressioninhumancolonicepithelialcellsbyshigellaanditsinteractionwithmucins
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Up-regulation of MUC2 and IL-1β expression in human colonic epithelial cells by Shigella and its interaction with mucins.

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