title: Altered pattern of tumor necrosis factor-alpha production in peripheral blood monocytes from Crohn's disease.
abstract: OBJECTIVE
  To evaluate the inflammatory state in Crohn's disease (CD) patients and correlate it with genetic background and microbial spreading.
  
  
  METHODS
  By means of flow cytometry, production of tumor necrosis factor-alpha (TNF-α) was measured in peripheral blood monocytes from patients suffering from CD, ulcerative colitis (UC) and in healthy subjects after stimulation of the NOD2 and TLR pathways. CD patients were genotyped for the three most commonNOD2variants (R702W, G908R and L1007Pfs*2) and basal production of TNF-α was correlated toNOD2genotype. Also, production of TNF-α was correlated to plasmatic levels of LPS Binding Protein (LBP), soluble (s) CD14 and to the activity state of the disease.
  
  
  RESULTS
  The patients with CD were characterized by a significantly higher monocyte basal expression of TNF-α compared with healthy subjects and UC patients, and after stimulation with Pam3CSK4(ligand of TLR2/1) and MDP-L18 (ligand of NOD2) this difference was maintained, while other microbial stimuli (LPS, ligand of TLR4 and PolyI:C, ligand of TLR3) induced massive activation in CD monocytes as well as in UC and in healthy control cells. There was no significant difference in the production of TNF-α between patients who carried CD-associated heterozygous or homozygous variants inNOD2and patients with wild typeNOD2genotype. Although serum LBP levels have been shown to correlate positively with the state of activity of the disease, TNF-α production did not show a clear correlation with either LBP or sCD14 levels in plasma. Moreover, no clear correlation was seen between TNF-α production and activity indices in either CD or UC.
  
  
  CONCLUSIONS
  Peripheral monocytes from CD express higher basal and stimulated TNF-α than controls, regardless ofNOD2genotype and without a clear correlation with disease activity.
authors.affiliation: Claudia Loganes, Samuele Naviglio, Elisa Piscianz, Department of Medicine, Surgery and Health Sciences, University of Trieste, 34128 Trieste, Italy.
authors.forename: Claudia
authors.initials: C
authors.lastname: Loganes
authors.affiliation: Claudia Loganes, Samuele Naviglio, Elisa Piscianz, Department of Medicine, Surgery and Health Sciences, University of Trieste, 34128 Trieste, Italy.
authors.forename: Alessia
authors.initials: A
authors.lastname: Pin
authors.affiliation: Claudia Loganes, Samuele Naviglio, Elisa Piscianz, Department of Medicine, Surgery and Health Sciences, University of Trieste, 34128 Trieste, Italy.
authors.forename: Samuele
authors.initials: S
authors.lastname: Naviglio
authors.affiliation: Claudia Loganes, Samuele Naviglio, Elisa Piscianz, Department of Medicine, Surgery and Health Sciences, University of Trieste, 34128 Trieste, Italy.
authors.forename: Martina
authors.initials: M
authors.lastname: Girardelli
authors.affiliation: Claudia Loganes, Samuele Naviglio, Elisa Piscianz, Department of Medicine, Surgery and Health Sciences, University of Trieste, 34128 Trieste, Italy.
authors.forename: Anna Monica
authors.initials: AM
authors.lastname: Bianco
authors.affiliation: Claudia Loganes, Samuele Naviglio, Elisa Piscianz, Department of Medicine, Surgery and Health Sciences, University of Trieste, 34128 Trieste, Italy.
authors.forename: Stefano
authors.initials: S
authors.lastname: Martelossi
authors.affiliation: Claudia Loganes, Samuele Naviglio, Elisa Piscianz, Department of Medicine, Surgery and Health Sciences, University of Trieste, 34128 Trieste, Italy.
authors.forename: Alberto
authors.initials: A
authors.lastname: Tommasini
authors.affiliation: Claudia Loganes, Samuele Naviglio, Elisa Piscianz, Department of Medicine, Surgery and Health Sciences, University of Trieste, 34128 Trieste, Italy.
authors.forename: Elisa
authors.initials: E
authors.lastname: Piscianz
mesh_terms: D000209:Acute-Phase Proteins; D000293:Adolescent; D015415:Biomarkers / Q000097:blood; D002352:Carrier Proteins / Q000097:blood; D016022:Case-Control Studies; D002478:Cells, Cultured; D002648:Child; D003093:Colitis, Ulcerative / Q000097:blood / Q000175:diagnosis / Q000276:immunology; D003424:Crohn Disease / Q000097:blood* / Q000175:diagnosis / Q000235:genetics / Q000276:immunology; D005260:Female; D020022:Genetic Predisposition to Disease; D014644:Genetic Variation; D006801:Humans; D007113:Immunity, Innate; D018836:Inflammation Mediators / Q000097:blood*; D007963:Leukocytes, Mononuclear / Q000276:immunology / Q000378:metabolism*; D018950:Lipopolysaccharide Receptors / Q000097:blood; D008297:Male; D008562:Membrane Glycoproteins / Q000097:blood; D053473:Nod2 Signaling Adaptor Protein / Q000235:genetics; D010641:Phenotype; D012720:Severity of Illness Index; D014409:Tumor Necrosis Factor-alpha / Q000097:blood*; D015854:Up-Regulation; D000099231:Lipopolysaccharide-Binding Protein
chemical_list: D000209:Acute-Phase Proteins; D015415:Biomarkers; D002352:Carrier Proteins; D018836:Inflammation Mediators; D018950:Lipopolysaccharide Receptors; D008562:Membrane Glycoproteins; D053473:Nod2 Signaling Adaptor Protein; D014409:Tumor Necrosis Factor-alpha; D000099231:Lipopolysaccharide-Binding Protein; C429018:NOD2 protein, human
keywords: Activity index; Crohn’s disease; Dysbiosis; LPS-binding protein; NOD2 variants; Toll like receptors; Tumor necrosis factor-α; Ulcerative colitis