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Direct cloning and tetramer staining to measure the frequency of intestinal gluten‐reactive T cells in celiac disease
Author(s) -
Bodd Michael,
Ráki Melinda,
Bergseng Elin,
Jahnsen Jørgen,
Lundin Knut E. A.,
Sollid Ludvig M.
Publication year - 2013
Publication title -
european journal of immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.272
H-Index - 201
eISSN - 1521-4141
pISSN - 0014-2980
DOI - 10.1002/eji.201343382
Subject(s) - gluten , biology , immunology , t cell , tissue transglutaminase , microbiology and biotechnology , immune system , biochemistry , enzyme
Knowledge of the frequency of disease‐driving CD4 + T cells in lesions of chronic human inflammatory diseases is limited. In celiac disease (CD), intestinal gluten‐reactive CD4 + T cells, which recognize gluten peptides only in the context of the disease‐associated HLA‐DQ molecules, are key pathogenic players. By cloning CD4 + T cells directly from intestinal biopsies of CD patients, we found that 0.5–1.8% of CD4 + T cells were gluten reactive. About half of the gluten‐reactive T cells were specific for either the immuno‐dominant DQ2.5‐glia‐α1a or DQ2.5‐glia‐α2 epitopes, suggesting that direct visualization of gluten‐specific T cells could be possible. Assessed by flow cytometry, tetramer‐positive T cells were present in 10/10 untreated CD patients with a frequency of 0.1–1.2% of CD4 + T cells. Gluten‐specific T cells were also detectable in most treated CD patients (7/10). Moreover, the frequency of gluten‐specific T cells correlated with the degree of histological damage in the gut mucosa as scored by Marsh‐grading, and also with serum IgA anti‐transglutaminase 2 antibody levels. Tetramer staining of gluten‐reactive T cells in biopsy material is a useful tool for future studies of such cells in CD and could also potentially serve as a diagnostic supplement in selected cases.