title: Distinctive colonic mucosal cytokine signature in new-onset, untreated pediatric Crohn disease.
abstract: OBJECTIVE
  The aim of the study was to compare the colonic mucosal immune response in children with new, untreated Crohn disease (CD-New), CD in remission (CD-Remission), and unaffected children (CTRL [controls]).
  
  
  METHODS
  We performed flow cytometry of mitogen-stimulated colonic lamina propria mononuclear cells isolated from colonic biopsies and 72-hour biopsy explant cultures, and analyzed the supernatant by an unbiased multiplex cytokine array of 45 analytes.
  
  
  RESULTS
  Thirty-six children were studied (mean age 14 ± 3 years, 14 girls): 12 CD-New, 11 CD-Remission, and 13 CTRL. We found that stimulation of lamina propria mononuclear cells isolated from colonic biopsies induced comparable intracellular cytokine levels of interferon (IFN-γ), interleukin (IL)-17, and tumor necrosis factor (TNF)-α in T cells from CD-New, CD-Remission, and CTRL, suggesting that mucosal innate inflammation plays a larger role than activated T cells in CD-New. To measure factors released during the ongoing inflammatory response in CD-New, we cultured colonic biopsy explants and uncovered 13/45 factors that were significantly higher in CD-New versus CD-Remission, whereas 10 were increased in CD-New over CTRL. Ingenuity Pathway Analysis software revealed the anticipated interconnectivity of TNF-α, IL-6, and CSF-2 in CD-New of the colon. A novel subnetwork of chemokines was, however, evident, whereas IL-17a appeared as a peripheral factor. Principal component analysis and hierarchal clustering showed that CD-New and CD-Remission separated into distinct subgroups based on the 13 factors.
  
  
  CONCLUSIONS
  At diagnosis of inflammatory bowel disease, the colonic cytokine response contains a predominance of innate immune factors, with chemoattractants and vascular adhesion molecules playing a central role.
authors.affiliation: *Department of Pediatrics, Connecticut Children's Medical Center, Hartford †Department of Immunology, University of Connecticut Health Center, Farmington.
authors.forename: Francisco A
authors.initials: FA
authors.lastname: Sylvester
authors.forename: Andrew
authors.initials: A
authors.lastname: Draghi
authors.forename: Antoine
authors.initials: A
authors.lastname: Menoret
authors.forename: Marina L
authors.initials: ML
authors.lastname: Fernandez
authors.forename: Zhu
authors.initials: Z
authors.lastname: Wang
authors.forename: Anthony T
authors.initials: AT
authors.lastname: Vella
mesh_terms: D000293:Adolescent; D015815:Cell Adhesion Molecules / Q000378:metabolism; D018925:Chemokines / Q000378:metabolism; D002648:Child; D003106:Colon / Q000378:metabolism*; D003424:Crohn Disease / Q000276:immunology* / Q000378:metabolism; D016207:Cytokines / Q000378:metabolism*; D018450:Disease Progression; D005260:Female; D006801:Humans; D007113:Immunity, Innate; D007249:Inflammation / Q000378:metabolism*; D007371:Interferon-gamma / Q000378:metabolism; D020381:Interleukin-17 / Q000378:metabolism; D015850:Interleukin-6 / Q000378:metabolism; D007413:Intestinal Mucosa / Q000276:immunology* / Q000378:metabolism; D008297:Male; D025341:Principal Component Analysis; D013601:T-Lymphocytes / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
chemical_list: D015815:Cell Adhesion Molecules; D018925:Chemokines; D016207:Cytokines; D020381:Interleukin-17; D015850:Interleukin-6; D014409:Tumor Necrosis Factor-alpha; D007371:Interferon-gamma