Ex-vivo whole blood secretion of interferon (IFN)- gamma and IFN- gamma -inducible protein-10 measured by enzyme-linked immunosorbent assay are as sensitive as IFN- gamma enzyme-linked immunospot for the detection of gluten-reactive T cells in human leucocyte antigen (HLA)-DQ2.5+-associated coeliac disease

• Published in: Clinical and experimental immunology Vol. 175; no. 2; pp. 305 - 315
• Main Authors: Ontiveros, N, Tye-Din, JA, Hardy, MY, Anderson, R P
• Format: Journal Article
• Language: English
• Published: 01-02-2014
• ISSN: 0009-9104; 1365-2249
• DOI: 10.1111/cei.12232

T cell cytokine release assays are used to diagnose infectious diseases, but not autoimmune or allergic disease. Coeliac 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 interferon (IFN)- gamma enzyme-linked immunospot (ELISPOT) or major histocompatibility complex (MHC) class II tetramers. Immunodominant 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 enzyme-linked immunosorbent assay (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.