title: Monocyte-derived macrophages from Crohn's disease patients are impaired in the ability to control intracellular adherent-invasive Escherichia coli and exhibit disordered cytokine secretion profile.
abstract: BACKGROUND
  Ileal lesions of Crohn's disease [CD] patients are colonised by adherent-invasive Escherichia coli [AIEC] able to survive in macrophage cell lines. We analysed the ability of monocyte-derived macrophages [MDM] from CD patients to control AIEC intracellular replication and the pro-inflammatory cytokine response of the infected-MDM.
  
  
  METHODS
  Peripheral blood MDM were obtained from 24 CD genotyped for NOD2 and ATG16L1 mutations, 5 ulcerative colitis [UC] patients and 12 healthy controls [HC]. The numbers of intracellular bacteria were determined using gentamicin assay. Cytokine secretion was quantified by ELISA assay.
  
  
  RESULTS
  We observed that higher levels of bacteria were internalised within MDM from CD patients than MDM from HC or UC patients. MDM from CD patients were unable to restrict AIEC intracellular replication. Infection of MDM from CD patients with AIEC resulted in significantly increased secretion of IL-6 and tumour necrosis factor alpha [TNF α] than did infection with non-pathogenic E. coli. AIEC-infected MDM from CD patients exhibited a disordered cytokines secretion compared with MDM from UC patients and HC. AIEC-infected MDM from patients with quiescent CD released significantly higher amounts of IL-6 and TNF-alpha than those with active disease or those from HC. The level of secreted TNF-alpha was correlated to the number of intracellular AIEC in MDM from CD patients. Treatment of MDM with infliximab did not change the MDM behaviour.
  
  
  CONCLUSIONS
  MDM from CD patients are unable to restrict intracellular AIEC replication, leading to disordered inflammatory response influenced by disease activity.
authors: Clermont Université, UMR 1071 Inserm/Université d'Auvergne, Clermont-Ferrand, France Department of Hepato-Gastroenterology, University Hospital Estaing of Clermont-Ferrand, Université d'Auvergne, Clermont-Ferrand, France.
authors: Emilie
authors: E
authors: Vazeille
authors: Clermont Université, UMR 1071 Inserm/Université d'Auvergne, Clermont-Ferrand, France Department of Hepato-Gastroenterology, University Hospital Estaing of Clermont-Ferrand, Université d'Auvergne, Clermont-Ferrand, France.
authors: Anthony
authors: A
authors: Buisson
authors: Clermont Université, UMR 1071 Inserm/Université d'Auvergne, Clermont-Ferrand, France.
authors: Marie-Agnès
authors: MA
authors: Bringer
authors: Clermont Université, UMR 1071 Inserm/Université d'Auvergne, Clermont-Ferrand, France Department of Hepato-Gastroenterology, University Hospital Estaing of Clermont-Ferrand, Université d'Auvergne, Clermont-Ferrand, France.
authors: Marion
authors: M
authors: Goutte
authors: Université Clermont 1, CHU Clermont-Ferrand, Pôle Santé PubliqueCentre Nationale de la Recherche en Santé, Institut de l'image pour les Sciences Interventionnelles Techniques, UMR6284 Clermont-Ferrand France.
authors: Lemlih
authors: L
authors: Ouchchane
authors: UMR843, INSERM, Assistance Publique Hôpitaux de Paris et Université, Paris Diderot, France.
authors: Jean-Pierre
authors: JP
authors: Hugot
authors: Clermont Université, UMR 1071 Inserm/Université d'Auvergne, Clermont-Ferrand, France.
authors: Amélie
authors: A
authors: de Vallée
authors: Clermont Université, UMR 1071 Inserm/Université d'Auvergne, Clermont-Ferrand, France.
authors: Nicolas
authors: N
authors: Barnich
authors: Clermont Université, UMR 1071 Inserm/Université d'Auvergne, Clermont-Ferrand, France Department of Hepato-Gastroenterology, University Hospital Estaing of Clermont-Ferrand, Université d'Auvergne, Clermont-Ferrand, France.
authors: Gilles
authors: G
authors: Bommelaer
authors: Clermont Université, UMR 1071 Inserm/Université d'Auvergne, Clermont-Ferrand, France Department of Hepato-Gastroenterology, University Hospital Estaing of Clermont-Ferrand, Université d'Auvergne, Clermont-Ferrand, France.
authors: Arlette
authors: A
authors: Darfeuille-Michaud
mesh_terms: D000328:Adult; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology; D000071183:Autophagy-Related Proteins; D001422:Bacterial Adhesion / Q000276:immunology; D058491:Bacterial Load / Q000187:drug effects; D002352:Carrier Proteins / Q000235:genetics; D002478:Cells, Cultured; D003093:Colitis, Ulcerative / Q000276:immunology* / Q000382:microbiology; D003424:Crohn Disease / Q000235:genetics / Q000276:immunology* / Q000382:microbiology; D004926:Escherichia coli / Q000254:growth & development* / Q000302:isolation & purification; D004927:Escherichia coli Infections / Q000276:immunology*; D005260:Female; D005838:Genotype; D006801:Humans; D000069285:Infliximab / Q000494:pharmacology; D015850:Interleukin-6 / Q000378:metabolism; D016209:Interleukin-8 / Q000378:metabolism; D008264:Macrophages / Q000276:immunology* / Q000378:metabolism* / Q000382:microbiology; D008297:Male; D008875:Middle Aged; D053473:Nod2 Signaling Adaptor Protein / Q000235:genetics; D020641:Polymorphism, Single Nucleotide; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
chemical_list: C518959:ATG16L1 protein, human; D000894:Anti-Inflammatory Agents, Non-Steroidal; D000071183:Autophagy-Related Proteins; D002352:Carrier Proteins; D015850:Interleukin-6; D016209:Interleukin-8; C429018:NOD2 protein, human; D053473:Nod2 Signaling Adaptor Protein; D014409:Tumor Necrosis Factor-alpha; D000069285:Infliximab
keywords: Crohn’s disease; Macrophages; adherent-invasive E. coli; cytokines; infliximab