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Ascorbate-dependent decrease of the mucosal immune inflammatory response to gliadin in coeliac disease patients

Bernardo, D.; Martínez-Abad, B.; Vallejo-Diez, S.; Montalvillo, E.; Benito, V.; Anta, B.; Fernández-Salazar, L.; Blanco-Quirós, A.; Garrote, J.A.; Arranz, E.

Published in Allergol Immunopathol (Madr). 2012;40:3-8. - vol.40 núm 01

English: [ Abstract | Full text | PDF ]

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 (100μg/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.

Key words: Ascorbate. Coeliac disease. Inhibition. IL-15. Therapy.

Introduction

Introduction Coeliac disease (CD) is a common gastro-intestinal disorder caused by a hypersensitivity reaction to wheat gliadin and similar proteins from rye and barley, affecting genetically predisposed individuals (HLA-DQ2/DQ8). The current treatment is a life-long strict gluten-free diet (GFD).¹^,² The most accepted model of the CD immunopathogenesis is the two-signal model, which establishes that gliadin has a dual effect on the CD duodenum, triggering the development of an innate immune response in the epithelium, and activating an adaptive immune response controlled by gluten-reactive T cells with a Th1 cytokine profile.³^,⁴ Innate immunity, and specifically interleukin (IL)-15,⁵^,⁶ plays a key role in the development of CD through a DQ2-independent mechanism.⁷ The induction of IL-15 seems to be involved in the initial stages of the disease leading to epithelial stress, increase tight-junction permeability, enterocyte apoptosis and dendritic cell (DC) activation,⁵^,⁶^,⁸^,⁹^,¹⁰^,¹¹^,¹² facilitating the development of the secondary adaptive response.³ Moreover, the gliadin amplifies the production of inflammatory cytokines through the nuclear factor (NF)-κB¹³ with a positive feedback by IL-15, which is also a potent NF-κB activator.¹⁴ Moreover, DCs are important players in the connection between the innate and the subsequent adaptive immune response,¹⁵ and require NF-κB for their development, survival, function and cytokine production.¹⁶^,¹⁷^,¹⁸ Thus, the IL-15/NF-κB axis is revealed to have an important role in the pathogenesis of CD and may represent a molecular target for strategies of immunomodulation.¹⁹ NF-κB is a heterogeneous collection of dimeric proteins subjected to a complex regulatory mechanism,²⁰^,²¹...

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Bernardo, D.^a; Martínez-Abad, B.^a; Vallejo-Diez, S.^a; Montalvillo, E.^a; Benito, V.^a; Anta, B.^a; Fernández-Salazar, L.^b; Blanco-Quirós, A.^a; Garrote, J.A.^a,c; Arranz, E.^a