title: Molecular evidence for Mycobacterium avium subspecies paratuberculosis (MAP) in Crohn's disease correlates with enhanced TNF-alpha secretion.
abstract: BACKGROUND
  Support for a role of Mycobacterium avium subspecies paratuberculosis in Crohn's disease is largely based on epidemiological evidence, as no data on mechanisms linking the presence of M. avium subspecies paratuberculosis with gut damage is available.
  
  
  OBJECTIVE
  To determine whether the presence of M. avium subspecies paratuberculosis contributes to the pathogenesis of Crohn's disease by promoting cytokine secretion within gut mucosa.
  
  
  METHODS
  A total of 235 subjects were recruited: 63 with Crohn's disease, 53 with ulcerative colitis, 45 with irritable bowel syndrome and 74 normal controls. M. avium subspecies paratuberculosis status was defined by nested PCR using IS900 sequence. Gut mucosal organ cultures were established to detect cytokine secretion patterns.
  
  
  RESULTS
  Significantly higher tumour necrosis factor-alpha concentrations were found in culture supernatants for Crohn's disease compared to ulcerative colitis (p<0.05), irritable bowel syndrome (p<0.01) and controls (p<0.0001). When tumour necrosis factor-alpha levels were correlated with the presence of M. avium subspecies paratuberculosis, significantly greater concentrations were only found in M. avium subspecies paratuberculosis-positive Crohn's disease patients (p<0.05). Tumour necrosis factor-alpha levels in M. avium subspecies paratuberculosis-positive Crohn's disease were significantly higher than in M. avium subspecies paratuberculosis-positive ulcerative colitis (p<0.01), M. avium subspecies paratuberculosis-positive irritable bowel syndrome (p<0.05) and M. avium subspecies paratuberculosis-positive controls (p<0.01) and all M. avium subspecies paratuberculosis-negative specimens.
  
  
  CONCLUSIONS
  The data link M. avium subspecies paratuberculosis with a pathogenic mechanism in Crohn's disease and is consistent with abnormal macrophage handling of M. avium subspecies paratuberculosis.
authors.affiliation: Hunter Immunology Unit, Hunter New England Area Health Service, John Hunter Hospital, Newcastle, NSW, Australia. robert.clancy@newcastle.edu.au
authors.forename: R
authors.initials: R
authors.lastname: Clancy
authors.forename: Z
authors.initials: Z
authors.lastname: Ren
authors.forename: J
authors.initials: J
authors.lastname: Turton
authors.forename: G
authors.initials: G
authors.lastname: Pang
authors.forename: A
authors.initials: A
authors.lastname: Wettstein
mesh_terms: D016022:Case-Control Studies; D003093:Colitis, Ulcerative / Q000503:physiopathology; D003424:Crohn Disease / Q000382:microbiology / Q000503:physiopathology; D005260:Female; D006801:Humans; D043183:Irritable Bowel Syndrome / Q000503:physiopathology; D008297:Male; D009162:Mycobacterium avium / Q000472:pathogenicity*; D016133:Polymerase Chain Reaction; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism*
chemical_list: D014409:Tumor Necrosis Factor-alpha