HLA-DQ:gluten tetramer test in blood gives better detection of coeliac patients than biopsy after 14-day gluten challenge

Abstract

NCT02464150.

HLA-DQ–Gluten Tetramer Blood Test Accurately Identifies Patients With and Without Celiac Disease in Absence of Gluten Consumption

“…The CD4 þ T-cell reactivity in relation to celiac disease has been assessed by either HLA-DQ-gluten tetramer analysis or by T-cell cytokine release assays (enzyme-linked immunospot [ELISPOT]/enzyme-linked immunosorbent assay [ELISA]). 13,[15][16][17]25,[27][28][29] The HLA-DQ-gluten tetramer approach uses fluorescent complexes of gluten peptides tethered to disease-specific HLA molecules for direct detection of gluten-specific cells in PBMCs by flow cytometry. 20 In the ELISPOT test, PBMCs are incubated with gluten-epitopes overnight, followed by an interferon-g ELISPOT assay.…”

“…However, 1 index test-positive GS subject had inadequate prior exclusion of celiac disease, and 1 index test-negative TCD subject, diagnosed with celiac disease in the early 1970s, did not respond with duodenal changes after a 2-week gluten challenge in another related study. 25 Both subjects may thus have contributed to a lower than actual estimate of sensitivity and specificity. By comparison, the highest reported estimates of diagnostic sensitivity of 2-to 12-week gluten challenge followed by duodenal biopsy are approximately 70%.…”

“…One of them, diagnosed as a child in the early 1970s, participated also in a 2-week gluten challenge study and remained both histology and antibody negative. 25 In Controls With High Frequency of HLA-DQ-Gluten Tetramer þ Cells, These Cells Were Specific to Gluten on Stimulation In Vitro Four of 5 index test-positive controls, who were found to be seropositive for either anti-TG2 IgA or anti-DGP IgG, accepted invitation for diagnostic workup of celiac disease in addition to 2 index test-negative and -seronegative controls with a frequency of gluten-specific T cells on the same level as UCD subjects. All 6 control subjects underwent duodenal biopsies and celiac disease-specific serology, 6 to 12 months after the index test.…”

Recent Progress and Recommendations on Celiac Disease From the Working Group on Prolamin Analysis and Toxicity

“…The procedure can be used for diagnostic purposes after gluten challenge (80) and may perform better than a 2-week gluten challenge followed by upper endoscopy with biopsy (81). Furthermore, employing an improved methodology, such HLA-DQ:gluten tetramer + cells can be detected without gluten challenge and clearly distinguish CD patients and healthy individuals (82,83). When the HLA tetramer technology was coupled with the CyTof technology, it was found that these cells carry a surprisingly rare phenotype with a profile suggesting that they may help plasma cells.…”

Gluten Immunogenic Peptides as Standard for the Evaluation of Potential Harmful Prolamin Content in Food and Human Specimen

“…A systematic study of hundreds of gluten epitopes by ELISPOT revealed that three highly immunogenic peptides, derived from α-gliadin (ELQPFPQPELPYPQPQ), ω-gliadin/C-hordein (EQPFPQPEQPFPWQP), and β-hordein (EPEQPIPEQPQPYPQQ), account for 90% of the celiac-specific immuno-response elicited by the full proteome of wheat, barley and rye proteins [27,39,43,56]. Recently, a HLA-DQ gluten tetramer test comprising only five gluten T cells epitope sequences (Table 1) could serve to identify celiac patients with 100% sensitivity and 90% specificity vs. control with gluten containing diet [57]. Those results also suggested that a limited number of peptides represent more than 90% of the celiac specific T-cell response.…”