Abstract
BackgroundThe diagnosis of coeliac disease (CD) in individuals that have started a gluten-free diet (GFD) without an adequate previous diagnostic work-out is a challenge. Several immunological assays such as IFN-γ ELISPOT have been developed to avoid the need of prolonged gluten challenge to induce the intestinal damage. We aimed to evaluate the diagnostic accuracy of activated gut-homing CD8+ and TCRγδ+ T cells in blood after a 3-day gluten challenge and to compare it with the performance of IFN-γ ELISPOT in a HLA-DQ2.5 subsample.MethodsA total of 22 CD patients and 48 non-CD subjects, all of them following a GFD, underwent a 3-day 10-g gluten challenge. The percentage of two T cell subsets (CD8+ CD103+ β7hi CD38+/total CD8+ and TCRγδ+ CD103+ β7hi CD38+/total TCRγδ+) in fresh peripheral blood drawn baseline and 6 days after the challenge was determined by flow cytometry. IFN-γ ELISPOT assays were also performed in HLA-DQ2.5 participants. ROC curve analysis was used to assess the diagnostic performance of the CD8+ T cell response and IFN-γ ELISPOT.ResultsSignificant differences between the percentage of the two studied subsets of CD8+ and TCRγδ+ cells at days 0 and 6 were found only when considering CD patients (p < 10−3 vs. non-CD subjects). Measuring activated CD8+ T cells provided accurate CD diagnosis with 95% specificity and 97% sensitivity, offering similar results than IFN-γ ELISPOT.ConclusionsThe results provide a highly accurate blood test for CD diagnosis in patients on a GFD of easy implementation in daily clinical practice.
Highlights
The diagnosis of coeliac disease (CD) in individuals that have started a gluten-free diet (GFD) without an adequate previous diagnostic work-out is a challenge
We previously demonstrated that CD patients could be diagnosed in individuals on a GFD after a 3-day gluten challenge by analyzing only four markers in peripheral blood by flow cytometry [3]
The observed percentages of the studied TCRγδ+ T cells at day 6 were obtained with values of 5 cells or even less in the numerator (TCRγδ+ CD103+ β7hi CD38+ T cells/ total TCRγδ+ T cells) in some CD patients
Summary
The diagnosis of coeliac disease (CD) in individuals that have started a gluten-free diet (GFD) without an adequate previous diagnostic work-out is a challenge. Several immunological assays such as IFN-γ ELISPOT have been developed to avoid the need of prolonged gluten challenge to induce the intestinal damage. We aimed to evaluate the diagnostic accuracy of activated gut-homing CD8+ and TCRγδ+ T cells in blood after a 3-day gluten challenge and to compare it with the performance of IFN-γ ELISPOT in a HLA-DQ2.5 subsample. We previously demonstrated that CD patients could be diagnosed in individuals on a GFD after a 3-day gluten challenge by analyzing only four markers in peripheral blood by flow cytometry [3]. We performed a pilot study to compare this test with the production of IFN-γ measured by ELISPOT in a subgroup of HLA-DQ2.5 patients
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