heat-labile toxin of e coli

Alexis Bretin, Ccily Lucas, Hang Thi Thu Nguyen, Alexander A. Crofts, Frdric M. Poly, M. Stephen Trent, Christopher J. Anderson, Christopher B. Medina, Kodi S. Ravichandran, Ashleigh P. Rogers, Steven J. Mileto & Dena Lyras, Jillian L. Pope, Ye Yang, Christian Jobin, Cristel Archambaud, Aurlie Derr-Bobillot, Pascale Serror, Steven X. Cho, Ina Rudloff, Marcel F. Nold, Nature Communications Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. Perturbed zinc homeostasis in rural 3-5-y-old Malawian children is associated with abnormalities in intestinal permeability attributed to tropical enteropathy. Enterotoxigenic E. coli (ETEC) produce heat-labile (LT) and/or heat-stable (ST) enterotoxins, and commonly cause diarrhea in resource-poor regions. Adaptation of the small intestine to microbial enteropathogens in Zambian children with stunting. J. Physiol. Escherichia coli Shiga-like, Heat-labile toxins and multidrug-resistant are important virulence factors. LT and mutant LT (E211K, mLT) were kindly provided by Dr. John D. Clements, Tulane University, New Orleans, Louisianna, USA. PubMed Central Mol. a Heatmap indicating key SLC genes modulated by heat-labile toxin (LT) compared to enzymatically inactive E112K LT mutant (mLT), or untreated () Caco-2 cells (left) and human small intestinal (ileal) enteroids (Hu235D, right). Physicians India 59, 420422 (2011). Importantly, ETEC infections have been linked to non-diarrheal sequelae, including environmental enteric dysfunction (EED), a condition characterized by impaired nutrient absorption, impaired growth7,8, and malnutrition9,10, adding significantly to the morbidity as well as deaths from diarrhea and other infections11. Acta Pathol. Cell Dev. Real-time qRT-PCR confirming LT-mediated modulation of genes in ileal (Hu235D) enteroids encoding b the major thiamine transporter SLC19A3 and c the SP1 cis-regulatory element. Article WebThe heat-stabile toxin of enterotoxigenic E. coli exerts its pathogenic effect in the host by which of the following mechanisms? Lancet Glob. El-Kamary, S. S., Cohen, M. B., Bourgeois, A. L., Van De Verg, L., Bauers, N., Reymann, M., Chen, W. H. (2013). ETEC secretes a heat-labile enterotoxin (LT), which resembles cholera toxin, but the actual mechanism of LT HNF4 activates the transcription of SMAD4, and in turn, SMAD4 activates the transcription of HNF470. At the end of treatment cell culture supernatants media were transferred to black-walled microplates (Greiner 655086) and processed as directed (Secrete-Pair Luminescence, GeneCopoeia, Inc. Rockville, MD), followed by luminescence detection (Synergy H1, BioTek). These assays demonstrated that HNF4-mediated transcription was dramatically reduced in cells treated with LT (Fig. Therefore, repeated infections in this model appear to recapitulate impacts observed following repeated ETEC infection in children, and our data suggest that these features are at least in part driven by LT. To address whether enteropathic changes to the small intestine can be prevented by vaccination, and to further define the role of LT, we vaccinated mouse dams with heat-labile toxin and examined the brush border ultrastructure in suckling mice. Immun. ELISA plates were coated with 2 g/mL of CTB, CTX or the Escherichia coli heat-labile enterotoxin B subunit (LTB). Decreasing cAMP Importantly, the long-term morbidity associated with ETEC infections does not appear to correlate with the severity of diarrhea as both mild illness4, and perhaps asymptomatic colonization may lead to growth faltering. Nucleic Acids Res. El-Kamary, S. S., Cohen, M. B., Bourgeois, A. L., Van De Verg, L., Bauers, N., Reymann, M., Chen, W. H. (2013). HMS is the recipient of a Senior Research Career Scientist award # IK6BX006189. Harris, J. Gastroenterology 68, 1732 (1975). WebChemicals and Drugs 104. Rev. E. coli heat-labile toxin (LT): LT is a heat-labile protein. g Length of microvilli (dashed horizontal lines represent geometric means). Slider with three articles shown per slide. Three biological replicates per treatment (untreated and LT treated) were submitted to the Genome Technology Access Center (GTAC) at Washington University in St. Louis School of Medicine for RNA-seq library preparation. a Kinetic ELISA data of from triplicate samples of breast milk anti-LT (IgA, and IgG in n=2 immunized dams and 1 un-immunized control). Gastroenterol. Soc. The specificity of the SLC19A3 antibodies was validated in our laboratory previously using different approaches that include overexpression of tagged protein or gene silencing85. 6, 445454 (2021). LT(K63), a non-toxic mutant and LT(R72), a low toxic mutant of E. coli heat-labile enterotoxin are frequently used mucosal adjuvants. Infect. Genome-wide analysis of cAMP-response element binding protein occupancy, phosphorylation, and target gene activation in human tissues. Notably, cholera toxin (CT), LT, and dibutyryl-cyclic AMP all induce hypersecretion and impact the architecture of gastrointestinal epithelia in rodent small intestine24, while small intestinal biopsies of patients with acute cholera exhibit marked changes in the ultrastructure of the intestinal brush border, the major absorptive surface in the small intestine25,26, including shortening and disruption of the microvilli. 6CE). Chem. The nuclear receptor HNF4 drives a brush border gene program conserved across murine intestine, kidney, and embryonic yolk sac. PubMed Cell Biol. 3b). Genet. Through further analyzes, it is found that LT(K63/R72) exhibits characteristics more closely related to LT (K63) than LT(R72), which is a low toxic mutant of E. coli heat-labile enterotoxin. To further examine the impact of LT on HNF4-mediated transcription, we introduced a transcriptional reporter plasmid containing six tandem copies of the HNF4 transcriptional response element (5-CAAAGGTCA-3) linked to a human codon-optimized Gaussia princeps luciferase into Caco-2 cells. In a cyclic adenosine monophosphate assay using Caco2 human colon epithelial cells, the 7A12B3 mAb was found to be a potent inhibitor of CTX, J. Med. PKA largely regulates transcription by phosphorylation of transcription factors, including the cyclic AMP response element binding protein (CREB) and the cAMP-response element modulator (CREM), which bind cAMP-response elements (CRE) in the promoter regions of target genes21,22,23. Data represent geometric mean length from n=5 mice per group in three independent experiments. Enterotoxigenic Escherichia coli degrades the host MUC2 mucin barrier to facilitate critical pathogen-enterocyte interactions in human small intestine. Det verkar p samma stt som koleratoxinet genom att hja cAMP-niver genom ADP-ribosylering av alfa-subenheten fr ett Gs-protein som leder till den konstitutiva For GO terms with an adjusted statistical significance of FDR 0.05, heatmaps were automatically generated for each respective term to show how genes co-vary or co-express across the term in relation to a given biological process or molecular function. Enterotoxigenic Escherichia coli isolated from patients with severe cholera-like disease. J. Biol. Acta 1803, 10131019 (2010). in 1956 in E. coli isolated from cases of adult diarrhoea in adults in Kolkata. Klipstein, F. A., Short, H. B., Engert, R. F., Jean, L. & Weaver, G. A. Pellets were then washed once in ice-cold sterile PBS, then resuspended to a density of ~1105 colony forming units (CFU) per 20l inoculum. We found that transcription of many SLC genes was altered in Caco-2 cells as well as small intestinal enteroids (Fig. WebThis product Mouse Anti-Escherichia Coli Labile toxin A chain with code 4330-1088 is available. As one of the crucial enterotoxins secreted by enterotoxigenic Escherichia coli (ETEC), heat-labile enterotoxin (LT) enhances bacterial adherence both in vivo and in vitro; however, the underlying mechanism remains unclear. Controlling the false discovery rate: a practical and powerful approach to multiple testing. Roy, S. K. et al. LPS consists of three components: (1) the genus-specific somatic O polysaccharide, (2) a core polysaccharide common to all Enterobacteriaceae (common antigen) and (3) lipid A. Fast SYBR Green master mix (Applied Biosystems/Thermo Fisher) was used for qPCR analysis. PubMed Central Cells were then resuspended in 1:1 CM and primary medium with Y-27632 and SB431541 as described above. d Uptake of [3H]-thiamine by Hu235D cells is impaired following LT treatment. MathSciNet Lancet Infect. Rohlff, C., Ahmad, S., Borellini, F., Lei, J. Google Scholar. Different in cDNA was generated using SuperscriptTM ViloTM cDNA synthesis kit (Invitrogen by Thermo Fisher) after normalizing RNA concentrations. CREB sequence matches were identified in the upstream sequences of 1,302 genes. Primary source data and original images are available on Figshare as detailed in Supplementary Table6 of the supplementary information. Diarrheal illness contributes to malnutrition, stunted growth, impaired cognitive development, and high morbidity rates in children worldwide. 18, 18031808 (2011). Vaccine 39, 42664277 (2021). Postema, M. M., Grega-Larson, N. E., Neininger, A. C. & Tyska, M. J. IRTKS (BAIAP2L1) elongates epithelial microvilli using EPS8-dependent and independent mechanisms. Google Scholar. He, L., Vasiliou, K. & Nebert, D. W. Analysis and update of the human solute carrier (SLC) gene superfamily. Studies in mice were performed under protocol 20-0438 approved by the Washington University in Saint Louis Institutional Animal Care and Use Committee. Web(heat labile) for prevention of carry-over contamination. WebChemicals and Drugs 104. 6A, B), leading us to speculate that LT-mediated phosphorylation of HNF4 by PKA, interrupts this critical transcription module (Supplementary Fig. Cell lines used in these are routinely validated for response to the heat-labile toxin by cAMP assays (Arbor Assays, Ann Arbor, MI). Published: 07 January 2022 Holotoxin disassembly by protein disulfide isomerase is less efficient for Escherichia coli heat-labile enterotoxin than cholera EBioMedicine 70, 103509 (2021). Immunofluorescence intensity of HNF4 signal in sections from n=5 control mice, and n=6 ETEC-challenged mice. & Hochberg, Y. Am. J. Biol. CFU in each inoculum were determined by plating serial dilutions LB-agar plates. Nakamura, T., Datta, R., Sherman, M. L. & Kufe, D. Regulation of c-jun gene expression by cAMP in HL-60 myeloid leukemia cells. close . Many of the SLC proteins are enriched in the small intestinal brush border, where they transport critical nutrients, including amino acids, oligopeptides, sugars, and vitamins. 105, 11321138 (2017). Media (EMEM, ATCC) were exchanged 24h after transfection, and after an additional 24h, baseline samples of media were removed from each well and stored at 80C. Burden of enterotoxigenic Escherichia coli and shigella non-fatal diarrhoeal infections in 79 low-income and lower middle-income countries: a modelling analysis. As well as small intestinal enteroids ( Fig harris, J. Gastroenterology 68, 1732 ( )... 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