title: Ascorbate-dependent decrease of the mucosal immune inflammatory response to gliadin in coeliac disease patients.
abstract: BACKGROUND
  The IL-15/NF-κB axis has an important role in coeliac disease (CD) and may represent a molecular target for immunomodulation. Ascorbate (vitamin C) is known to show inhibitory effects on NF-κB. Therefore, we studied if ascorbate supplementation to gliadin gliadin-stimulated biopsy culture could down-regulate the mucosal immune response to gliadin in CD.
  
  
  METHODS
  Duodenal biopsy explants from treated CD patients were gliadin challenged in vitro (100 μg/ml) with and without 20mM ascorbate. An extra tissue explant in basal culture was used as internal control. Secretion levels of nitrites (3h), and IFNγ, TNFα, IFNα, IL-17, IL-13, and IL-6 (24h) were measured on the supernatants. IL-15 was assayed by western-blot on whole protein duodenal explants.
  
  
  RESULTS
  The addition of ascorbate to in vitro culture gliadin-challenged biopsies blocked the secretion of nitrites (p=0.013), IFNγ (p=0.0207), TNFα (p=0.0099), IFNα (p=0.0375), and IL-6 (p=0.0036) compared to samples from non-ascorbate supplemented culture. Cytokine secretion was downregulated by ascorbate even to lower values than those observed in basal cultures (IFNγ: p=0.0312; TNFα: p=0.0312; IFNα: p=0.0312; and IL-6: p=0.0078). Gliadin-challenge induced IL-15 production in biopsies from treated CD patients, while the addition of ascorbate to culture medium completely inhibited IL-15 production. Moreover, the inhibition of IL-15 by ascorbate took place even in the only treated CD-patient who had basal IL-15 production.
  
  
  CONCLUSIONS
  Ascorbate decreases the mucosal inflammatory response to gluten in an intestinal biopsy culture model, so it might have a role in future supplementary therapy in CD.
authors.affiliation: Mucosal Immunology Lab, Department of Paediatrics & Immunology, Universidad de Valladolid-CSIC, Spain.
authors.forename: D
authors.initials: D
authors.lastname: Bernardo
authors.forename: B
authors.initials: B
authors.lastname: Martínez-Abad
authors.forename: S
authors.initials: S
authors.lastname: Vallejo-Diez
authors.forename: E
authors.initials: E
authors.lastname: Montalvillo
authors.forename: V
authors.initials: V
authors.lastname: Benito
authors.forename: B
authors.initials: B
authors.lastname: Anta
authors.forename: L
authors.initials: L
authors.lastname: Fernández-Salazar
authors.forename: A
authors.initials: A
authors.lastname: Blanco-Quirós
authors.forename: J A
authors.initials: JA
authors.lastname: Garrote
authors.forename: E
authors.initials: E
authors.lastname: Arranz
mesh_terms: D000328:Adult; D000368:Aged; D001205:Ascorbic Acid / Q000494:pharmacology*; D001706:Biopsy; D002446:Celiac Disease / Q000188:drug therapy* / Q000276:immunology; D016207:Cytokines / Q000096:biosynthesis; D005260:Female; D005903:Gliadin / Q000276:immunology*; D006801:Humans; D018928:Immunity, Mucosal / Q000187:drug effects; D007249:Inflammation / Q000517:prevention & control*; D019409:Interleukin-15 / Q000037:antagonists & inhibitors / Q000502:physiology; D008297:Male; D008875:Middle Aged
chemical_list: D016207:Cytokines; D019409:Interleukin-15; D005903:Gliadin; D001205:Ascorbic Acid