Ex vivo whole blood secretion of interferon-gamma (IFN-gamma) and IFN-gamma-inducible protein-10 (IP-10) measured by ELISA are as sensitive as IFN-gamma ELISpot for the detection of gluten-reactive T cells in HLA-DQ2.5+ associated celiac disease
Details
Publication Year 2013-11-06,Volume 175,Issue #2,Page 305-15
Journal Title
Clinical and experimental immunology
Publication Type
Journal Article
Abstract
T-cell cytokine release assays are used to diagnose infectious diseases, but not autoimmune or allergic disease. Celiac disease (CD) is a common T-cell mediated disease diagnosed by the presence of gluten-dependent intestinal inflammation and serology. Many patients cannot be diagnosed with CD because they reduce dietary gluten before medical workup. Oral gluten challenge in CD patients treated with gluten-free diet (GFD) mobilizes gluten-reactive T cells measurable by IFN-gamma ELISpot or MHC Class II tetramers. Immuno-dominant peptides are quite consistent in the 90% of patients who possess HLA-DQ2.5. We aimed to develop whole blood assays to detect gluten-specific T-cells. Blood was collected before and after gluten challenge from GFD donors confirmed to have CD (n=27, all HLA-DQ2.5+), GFD donors confirmed not to have CD (n=6 HLA-DQ2.5+, 11 HLA-DQ2.5-), and donors with CD not following GFD (n=4, all HLA-DQ2.5+). Plasma IFN-gamma and IFN-gamma inducible protein-10 (IP-10) were measured by ELISA after whole blood incubation with peptides or gliadin, and correlated with IFN-gamma ELISpot. No T cell assay could distinguish between CD patients and controls prior to gluten challenge, but after gluten challenge the whole blood IFN-gamma ELISA and the ELISpot were both 85% sensitive and 100% specific for HLA-DQ2.5+ CD patients; the whole blood IP-10 ELISA was 94% sensitive and 100% specific. We conclude that whole blood cytokine release assays are sensitive and specific for detection of gluten-reactive T cells in CD; further clinical studies addressing the utility of these tests in patients with an uncertain diagnosis of CD is warranted.
Publisher
WILEY
Keywords
coeliac disease; diagnostics; gluten challenge
Research Division(s)
Immunology
NHMRC Grants
NHMRC/406656NHMRC/361646
Terms of Use/Rights Notice
© 2013 British Society for Immunology


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