title: Killing of Escherichia coli by Crohn's Disease Monocyte-derived Macrophages and Its Enhancement by Hydroxychloroquine and Vitamin D.
abstract: BACKGROUND
  Crohn's disease (CD) is associated with defective innate immunity, including impaired neutrophil chemotaxis, and mucosal invasion by bacteria, particularly adherent and invasive Escherichia coli that replicate inside macrophage phagolysosomes. We compared CD and healthy control (HC) macrophages for their abilities to kill E. coli and generate neutrophil chemoattractants and also assessed the effects of hydroxychloroquine (HCQ) and vitamin D on killing of phagocytosed E. coli.
  
  
  METHODS
  Peripheral blood monocyte-derived macrophages from CD and HC were compared for bacterial killing and generation of neutrophil chemoattractants in response to CD-derived E. coli. Escherichia coli replication was also assessed in the presence and absence of HCQ, alone and with antibiotics, and vitamin D.
  
  
  RESULTS
  Monocyte-derived macrophages from patients with CD were similar to HC in allowing replication of phagocytosed CD-derived E. coli: HM605 {CD: N = 10, mean fold replication in 3 hr = 1.08 (95% confidence interval [CI], 0.39-1.78); HC: N = 9, 1.50 (95% CI, 1.02-1.97); P = 0.15} and also in generation of neutrophil chemoattractants in response to E. coli (mean fold chemotaxis relative to control: CD = 2.55 [95% CI, 2.31-2.80]; HC = 2.65 [95% CI, 2.46-2.85], P = 0.42). HCQ and 1,25 OH2-vitamin D3 both caused dose-dependent inhibition of intramacrophage E. coli replication 3-hour postinfection; HCQ: 73.9% inhibition (P < 0.001) at 1 μg/mL, accompanied by raised intraphagosomal pH, and 1,25 OH2-vitamin D3: 80.7% inhibition (P < 0.05) at 80 nM. HCQ had synergistic effects with doxycycline and ciprofloxacin.
  
  
  CONCLUSIONS
  CD and HC macrophages perform similarly in allowing replication of phagocytosed E. coli and generating neutrophil chemoattractants. Replication of phagocytosed E. coli was substantially decreased by HCQ and vitamin D. These warrant further therapeutic trials in CD in combination with relevant antibiotics.
authors.affiliation: *Department of Gastroenterology, Institute of Translational Medicine, University of Liverpool, Liverpool, United Kingdom; †Department of Biochemistry, Institute of Integrative Biology, University of Liverpool, Liverpool, United Kingdom; ‡Immunology Unit and Center of Excellence in Immunology and Immune-mediated Disease, Department of Microbiology, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand; §Gastrointestinal Unit, Molecular Medicine Centre, University of Edinburgh, Edinburgh, Scotland; and ‖Department of Gastroenterology, Royal Liverpool University Hospital, Liverpool, United Kingdom.
authors.forename: Paul K
authors.initials: PK
authors.lastname: Flanagan
authors.forename: Direkrit
authors.initials: D
authors.lastname: Chiewchengchol
authors.forename: Helen L
authors.initials: HL
authors.lastname: Wright
authors.forename: Steven W
authors.initials: SW
authors.lastname: Edwards
authors.forename: Abdullah
authors.initials: A
authors.lastname: Alswied
authors.forename: Jack
authors.initials: J
authors.lastname: Satsangi
authors.forename: Sreedhar
authors.initials: S
authors.lastname: Subramanian
authors.forename: Jonathan M
authors.initials: JM
authors.lastname: Rhodes
authors.forename: Barry J
authors.initials: BJ
authors.lastname: Campbell
mesh_terms: D000328:Adult; D000818:Animals; D002478:Cells, Cultured; D003424:Crohn Disease / Q000276:immunology* / Q000473:pathology / Q000628:therapy; D016207:Cytokines / Q000378:metabolism; D004359:Drug Therapy, Combination; D004791:Enzyme Inhibitors / Q000627:therapeutic use; D004926:Escherichia coli / Q000302:isolation & purification*; D004927:Escherichia coli Infections / Q000276:immunology* / Q000473:pathology / Q000628:therapy; D005260:Female; D006801:Humans; D006886:Hydroxychloroquine / Q000627:therapeutic use*; D007113:Immunity, Innate; D007413:Intestinal Mucosa / Q000276:immunology / Q000473:pathology; D008264:Macrophages / Q000276:immunology; D008297:Male; D051379:Mice; D008875:Middle Aged; D010587:Phagocytosis; D014807:Vitamin D / Q000627:therapeutic use*; D014815:Vitamins / Q000627:therapeutic use
chemical_list: D016207:Cytokines; D004791:Enzyme Inhibitors; D014815:Vitamins; D014807:Vitamin D; D006886:Hydroxychloroquine
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