Activated gut-homing CD8⁺ T cells for coeliac disease diagnosis on a gluten-free diet

Background: 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.

Methods: A 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⁺ β7^hi CD38⁺/total CD8⁺ and TCRγδ⁺ CD103⁺ β7^hi 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.

Results: Significant 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.

Conclusions: The results provide a highly accurate blood test for CD diagnosis in patients on a GFD of easy implementation in daily clinical practice.