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. 2019 May:43:325-332.
doi: 10.1016/j.ebiom.2019.03.071. Epub 2019 Apr 26.

Inflammation associated ethanolamine facilitates infection by Crohn's disease-linked adherent-invasive Escherichia coli

Michael J Ormsby  1 , Michael Logan  2 , Síle A Johnson  1 , Anne McIntosh  1 , Ghaith Fallata  1 , Rodanthi Papadopoulou  3 , Eleftheria Papachristou  3 , Georgina L Hold  4 , Richard Hansen  5 , Umer Z Ijaz  2 , Richard K Russell  5 , Konstantinos Gerasimidis  3 , Daniel M Wall  6
Affiliations

"VSports" Inflammation associated ethanolamine facilitates infection by Crohn's disease-linked adherent-invasive Escherichia coli

Michael J Ormsby et al. EBioMedicine. 2019 May.

Abstract (VSports手机版)

Background: The predominance of specific bacteria such as adherent-invasive Escherichia coli (AIEC) within the Crohn's disease (CD) intestine remains poorly understood with little evidence uncovered to support a selective pressure underlying their presence VSports手机版. Intestinal ethanolamine is however readily accessible during periods of intestinal inflammation, and enables pathogens to outcompete the host microbiota under such circumstances. .

Methods: Quantitative RT-PCR (qRT-PCR) to determine expression of genes central to ethanolamine metabolism; transmission electron microscopy to detect presence of bacterial microcompartments (MCPs); in vitro infections of both murine and human macrophage cell lines examining intracellular replication of the AIEC-type strain LF82 and clinical E. coli isolates in the presence of ethanolamine; determination of E. coli ethanolamine utilization (eut) operon transcription in faecal samples from healthy patients, patients with active CD and the same patients in remission following treatment V体育安卓版. .

Results: Growth on the intestinal short chain fatty acid propionic acid (PA) stimulates significantly increased transcription of the eut operon (fold change relative to glucose: >16. 9; p-value <. 01). Additionally ethanolamine was accessible to intra-macrophage AIEC and stimulated significant increases in growth intracellularly when it was added extracellularly at concentrations comparable to those in the human intestine V体育ios版. Finally, qRT-PCR indicated that expression of the E. coli eut operon was increased in children with active CD compared to healthy controls (fold change increase: >4. 72; P < . 02). After clinical remission post-exclusive enteral nutrition treatment, the same CD patients exhibited significantly reduced eut expression (Pre vs Post fold change decrease: >15. 64; P < . 01). .

Interpretation: Our data indicates a role for ethanolamine metabolism in selecting for AIEC that are consistently overrepresented in the CD intestine VSports最新版本. The increased E. coli metabolism of ethanolamine seen in the intestine during active CD, and its decrease during remission, indicates ethanolamine use may be a key factor in shaping the intestinal microbiome in CD patients, particularly during times of inflammation. FUND: This work was funded by Biotechnology and Biological Sciences Research Council (BBSRC) grants BB/K008005/1 & BB/P003281/1 to DMW; by a Tenovus Scotland grant to MJO; by Glasgow Children's Hospital Charity, Nestle Health Sciences, Engineering and Physical Sciences Research Council (EPSRC) and Catherine McEwan Foundation grants awarded to KG; and by a Natural Environment Research Council (NERC) fellowship (NE/L011956/1) to UZI. The IBD team at the Royal Hospital for Children, Glasgow are supported by the Catherine McEwan Foundation and Yorkhill IBD fund. RKR and RH are supported by NHS Research Scotland Senior fellowship awards. .

Keywords: Adherent-invasive Escherichia coli; Biomarker; Crohn's disease; Ethanolamine V体育平台登录. .

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"VSports在线直播" Figures

Fig. 1
Fig. 1
PA stimulates AIEC degradation of ethanolamine. Anaerobic growth (OD600nm) of LF82, LF82-PA and their corresponding eutR knock out mutants in minimal media (NCE) supplemented with ethanolamine (20 mM), MgSO4 (1 mM), trace metals and sodium thiosulphate (40 mM). Data were analysed using a two-way ANOVA with Tukey; p < .001 ***. Only significant differences between LF82 and LF82-PA are shown. There were no significant differences between mutant strains at any time point.
Fig. 2
Fig. 2
Growth on PA stimulates the production of bacterial MCPs for the utilization of ethanolamine as a carbon source. For TEM, cultures of LF82 were grown in NCE media supplemented with cobalamin (200 nM) and either (a) glucose or (b) PA, at a final concentration of 20 mM. (c) A close up of a MCP-containing outer membrane vesicle from a PA supplemented culture is shown. qRT-PCR was conducted on LF82 grown in NCE media supplemented with cobalamin (200 nM) and either glucose (G), propionic acid (PA) 1,2-propanediol (1,2-PD) or ethanolamine (E) at a final concentration of 20 mM. Relative fold change of (d) prpB, (e) pduC and (f) eutS were measured relative to their expression in the presence of glucose, using 16S rRNA as an internal control. Four independent biological replicates were performed. Data are expressed as relative fold change ± SD and were analysed using a one-way ANOVA with Tukey; p < .05 *; p < .001 ***.
Fig. 3
Fig. 3
Extracellular ethanolamine increases intracellular replication of LF82-PA. Intra-macrophage (RAW264.7) survival and replication of wild type, PA-adapted, and LF82ΔeutR at 24 h post-infection with or without ethanolamine supplementation. For all values, the mean ± SD of three independent biological replicates are shown. Statistical analyses were preformed using GraphPad Prism, with data analysed by two-way ANOVA (p < .05 *; p < .01 **; p < .001 ***).
Fig. 4
Fig. 4
qRT-PCR of eutS in healthy patients, active Crohn's disease patients and Crohn's disease patients following EEN. Ethanolamine utilization was determined by abundance of eutS transcripts. eutS was amplified using primers designed against LF82. Transcript levels were normalized to 16S rRNA transcripts. Healthy samples (n = 9); Crohn's disease samples pre-treatment (CD pre; n = 10); Crohn's disease samples post-treatment (CD post; n = 10). Crohn's disease samples pre- and post-treatment were paired. Patients were marked as responders (squares) and non-responders (triangles) based on their drop in calprotectin levels (Supplementary Table S1). Statistical analyses were preformed using GraphPad Prism, with data analysed by one-way ANOVA followed by Dunns multiple comparisons post-test (p < .05 *; p < .01 **).
Supplementary Fig. S1
Supplementary Fig. S1
AIEC are unable to utilize butyrate or acetate as sole carbon sources for effective growth. The growth of commensal (F-18) and adherent and invasive (LF82) E. coli strains in minimal media (M9) supplemented with 20 mM butyrate (a) or acetate (b) was monitored over time.
Supplementary Fig. S2
Supplementary Fig. S2
Extracellular ethanolamine increases intracellular replication of LF82-PA in THP-1 cells. Intra-macrophage (THP-1) survival and replication of wild type, PA adapted, and ΔeutR mutants of each at 4 h post-infection with or without ethanolamine supplementation. For all values, the mean ± SD of three independent biological replicates are shown. Statistical analyses were preformed using GraphPad Prism, with data analysed by two-way ANOVA (p < .05 *; p < .01 **; p < .001 ***).
Supplementary Fig. S3
Supplementary Fig. S3
Extracellular ethanolamine increases intracellular replication of clinical E. coli isolates. Intra-macrophage (RAW 264.7) survival and replication of wild type and PA adapted LF82, clinical isolates B94, B115, B122 and B125 and commensal E. coli, F-18 24 h post-infection with or without ethanolamine supplementation. For all values, the mean ± SD of three independent biological replicates are shown. Statistical analyses were preformed using GraphPad Prism, with data analysed by two-way ANOVA (p < .05 *; p < .01 **; p < .001 ***).
Supplementary Fig. S4
Supplementary Fig. S4
Total E. coli was determined by abundance of 16S transcripts amplified using primers designed against E. coli. Healthy samples (n = 9); Crohn's disease samples pre-treatment (CD pre; n = 10); Crohn's disease samples post-treatment (CD post; n = 10). Crohn's disease samples pre- and post-treatment were paired. Statistical analyses were preformed using GraphPad Prism, with data analysed by one-way ANOVA followed by Dunns multiple comparisons post test (p < .05 *; p < .01 **).
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References

    1. Baumgart M., Dogan B., Rishniw M., Weitzman G., Bosworth B., Yantiss R. Culture independent analysis of ileal mucosa reveals a selective increase in invasive Escherichia coli of novel phylogeny relative to depletion of Clostridiales in Crohn's disease involving the ileum. ISME J. 2007;1:403–418. - PubMed
    1. Darfeuille-Michaud A., Neut C., Barnich N., Lederman E., Di Martino P., Desereumaux P. Presence of adherent Escherichia coli strains in ileal mucosa of patients with Crohn's disease. Gastroenterology. 1998;115(6):1405–1413. - PubMed (VSports最新版本)
    1. Martin H.M., Campbell B.J., Hart C.A., Mpofu C., Nayar M., Singh R. Enhanced Escherichia coli adherence and invasion in Crohn's disease and colon cancer. Gastroenterology. 2004;127(1):80–93. - PubMed
    1. Martinez-Medina M., Aldeguer X., Lopez-Siles M., González-Huix F., López-Oliu C., Dahbi G. Molecular diversity of Escherichia coli in the human gut: new ecological evidence supporting the role of adherent-invasive E. coli (AIEC) in Crohn's disease. Inflamm Bowel Dis. 2009;15(6):872–882. - PubMed
    1. Agus A., Denizot J., Thévenot J., Martinez-Medina M., Massier S., Sauvanet P. Western diet induces a shift in microbiota composition enhancing susceptibility to Adherent-Invasive E. coli infection and intestinal inflammation. Sci Rep. 2016:19032. http://www.ncbi.nlm.nih.gov/pubmed/26742586 6(July 2015). Available from. - PMC - PubMed

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