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10.1053/j.gastro.2014.06.024 http://scihub22266oqcxt.onion/10.1053/j.gastro.2014.06.024 C4170054!4170054 !24960189     free     free     free Warning: file_get_contents(https://eutils.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&id=24960189 &cmd=llinks): Failed to open stream: HTTP request failed! 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Activation of pattern recognition receptors up-regulates metallothioneins, thereby increasing intracellular accumulation of zinc, autophagy, and bacterial clearance by macrophages |pdf=|usr=}}{{24960189 }} gab.com Text Activation of pattern recognition receptors up-regulates metallothioneins, thereby increasing intracellular accumulation of zinc, autophagy, and bacterial clearance by macrophages JO: Gastroenterology http://www.kidney.de/pget.php?&tw=1&pmid=24960189 #FOAvip BACKGROUND & AIMS: Continuous stimulation of pattern recognition receptors (PRRs), including nucleotide-binding oligomerization domain-2 (NOD2) (variants in NOD2 have been associated with Crohn's disease), alters the phenotype of myeloid-derived cells, reducing production of inflammatory cytokines and increasing microbe clearance. We investigated the mechanisms by which microbial clearance increases in macrophages under these conditions. METHODS: Monocytes were purified from human peripheral blood mononuclear cells and differentiated to monocyte-derived macrophages (MDMs). We also isolated human intestinal macrophages. Bacterial clearance by MDMs was assessed in gentamicin protection assays. Effects of intracellular zinc and autophagy were measured by flow cytometry, immunoblot, reverse-transcription polymerase chain reaction, and microscopy experiments. Small interfering RNAs were used to knock down specific proteins in MDMs. NOD2-/- and C57BL/6J mice, maintained in a specific pathogen-free facility, were given antibiotics, muramyl dipeptide (to stimulate NOD2), or dextran sodium sulfate; intestinal lamina propria cells were collected and analyzed. RESULTS: Chronic stimulation of human MDMs through NOD2 up-regulated the expression of multiple genes encoding metallothioneins, which bind and regulate levels of intracellular zinc. Intestinal myeloid-derived cells are stimulated continually through PRRs; metallothionein expression was up-regulated in human and mouse intestinal myeloid-derived cells. Continuous stimulation of NOD2 increased the levels of intracellular zinc, thereby increasing autophagy and bacterial clearance. The metal-regulatory transcription factor-1 (MTF-1) was required for regulation of metallothionein genes in human MDMs. Knockdown of MTF-1 did not affect baseline clearance of bacteria by MDMs. However, the increase in intracellular zinc, autophagy, and bacterial clearance observed with continuous NOD2 stimulation was impaired in MDMs upon MTF-1 knockdown. The addition of zinc or induction of autophagy restored bacterial clearance to MDMs after metallothionein knockdown. NOD2 synergized with the PRRs Toll-like receptors 5 and 9 increase the effects of metallothioneins in MDMs. In mice, the intestinal microbiota contributed to the regulation in expression of metallothioneins, levels of zinc, autophagy, and bacterial clearance by intestinal macrophages. CONCLUSIONS: In studies of human MDMs and in mice, continuous stimulation of PRRs induces expression of metallothioneins. This leads to increased levels of intracellular zinc and enhanced clearance of bacteria via autophagy in macrophages. Twit Text FOAVip Activation of pattern recognition receptors up-regulates metallothioneins, thereby increasing intracellular accumulation of zinc, autophagy, and bacterial clearance by macrophages ????Gastroenterology http://www.kidney.de/pget.php?&tw=1&pmid=24960189 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=24960189 #FOAvip English Wikipedia <ref>{{Cite pmid|24960189 }}</ref> Activation of pattern recognition receptors up-regulates metallothioneins, thereby increasing intracellular accumulation of zinc, autophagy, and bacterial clearance by macrophages #MMPMID24960189 Lahiri A ; Abraham C Gastroenterology 2014[Oct]; 147 (4 ): 835-46 PMID24960189 show ga BACKGROUND & AIMS: Continuous stimulation of pattern recognition receptors (PRRs), including nucleotide-binding oligomerization domain-2 (NOD2) (variants in NOD2 have been associated with Crohn's disease), alters the phenotype of myeloid-derived cells, reducing production of inflammatory cytokines and increasing microbe clearance. We investigated the mechanisms by which microbial clearance increases in macrophages under these conditions. METHODS: Monocytes were purified from human peripheral blood mononuclear cells and differentiated to monocyte-derived macrophages (MDMs). We also isolated human intestinal macrophages. Bacterial clearance by MDMs was assessed in gentamicin protection assays. Effects of intracellular zinc and autophagy were measured by flow cytometry, immunoblot, reverse-transcription polymerase chain reaction, and microscopy experiments. Small interfering RNAs were used to knock down specific proteins in MDMs. NOD2-/- and C57BL/6J mice, maintained in a specific pathogen-free facility, were given antibiotics, muramyl dipeptide (to stimulate NOD2), or dextran sodium sulfate; intestinal lamina propria cells were collected and analyzed. RESULTS: Chronic stimulation of human MDMs through NOD2 up-regulated the expression of multiple genes encoding metallothioneins, which bind and regulate levels of intracellular zinc. Intestinal myeloid-derived cells are stimulated continually through PRRs; metallothionein expression was up-regulated in human and mouse intestinal myeloid-derived cells. Continuous stimulation of NOD2 increased the levels of intracellular zinc, thereby increasing autophagy and bacterial clearance. The metal-regulatory transcription factor-1 (MTF-1) was required for regulation of metallothionein genes in human MDMs. Knockdown of MTF-1 did not affect baseline clearance of bacteria by MDMs. However, the increase in intracellular zinc, autophagy, and bacterial clearance observed with continuous NOD2 stimulation was impaired in MDMs upon MTF-1 knockdown. The addition of zinc or induction of autophagy restored bacterial clearance to MDMs after metallothionein knockdown. NOD2 synergized with the PRRs Toll-like receptors 5 and 9 increase the effects of metallothioneins in MDMs. In mice, the intestinal microbiota contributed to the regulation in expression of metallothioneins, levels of zinc, autophagy, and bacterial clearance by intestinal macrophages. CONCLUSIONS: In studies of human MDMs and in mice, continuous stimulation of PRRs induces expression of metallothioneins. This leads to increased levels of intracellular zinc and enhanced clearance of bacteria via autophagy in macrophages. |*Autophagy/drug effects [MESH] |Animals [MESH] |Anti-Bacterial Agents/pharmacology [MESH] |Caspase 1/metabolism [MESH] |Cells, Cultured [MESH] |DNA-Binding Proteins/genetics/metabolism [MESH] |Gene Expression Regulation [MESH] |Humans [MESH] |Intestinal Mucosa/*metabolism [MESH] |Intestines/drug effects/microbiology/pathology [MESH] |Macrophages/drug effects/*metabolism/microbiology/pathology [MESH] |Metallothionein/genetics/*metabolism [MESH] |Mice [MESH] |Mice, Inbred C57BL [MESH] |Mice, Knockout [MESH] |NF-kappa B/metabolism [MESH] |Nod2 Signaling Adaptor Protein/agonists/deficiency/genetics/*metabolism [MESH] |RNA Interference [MESH] |Time Factors [MESH] |Toll-Like Receptors/metabolism [MESH] |Transcription Factor MTF-1 [MESH] |Transcription Factors/genetics/metabolism [MESH] |Transfection [MESH]Activation of pattern recognition receptors up-regulates metallothione(epmc).pdf DeepDyve Pubget Overpricing