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A novel and sensitive method for the detection of T cell stimulatory epitopes of α/β- and γ-gliadin

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Author: Spaenij-Dekking, E.H.A. · Kooy-Winkelaar, E.M.C. · Nieuwenhuizen, W.F. · Drijfhout, J.W. · Koning, F.
Type:article
Date:2004
Institution: TNO Voeding
Source:Gut, 9, 53, 1267-1273
Identifier: 237982
doi: doi:10.1136/gut.2003.037952
Keywords: Nutrition · Food technology · alcohol · CD4 antigen · epitope · gliadin · gliadin antibody · gluten · pepsin A · trypsin · analytic method · animal experiment · antigen detection · article · barley · celiac disease · cell clone · cereal · controlled study · food · food analysis · human · human cell · mouse · nonhuman · priority journal · protein analysis · quantitative analysis · rye · sensitivity and specificity · T lymphocyte activation · T lymphocyte subpopulation · triticale · wheat · Animals · Antibodies, Monoclonal · Binding, Competitive · Cell Division · Cereals · Enzyme-Linked Immunosorbent Assay · Epitopes, T-Lymphocyte · Food · Food Analysis · Gliadin · Humans · Lymphocyte Activation · Mice · Mice, Inbred BALB C · Peptide Fragments · T-Lymphocytes

Abstract

Background: It is now generally accepted that coeliac disease (CD) is caused by inflammatory T cell responses to gluten peptides bound to HLA-DQ2 or -DQ8 molecules. There is overwhelming evidence that CD patients can mount T cell responses to peptides found in both α-gliadin and γ-gliadin molecules. Assays that would detect the presence or absence of such peptides in food would thus be accurate indicators of safely for consumption by CD patients. Aims: The development of a sensitive method to detect T cell stimulatory epitopes of α-gliadin and γ-gliadin molecules in food products. Methods: Monoclonal antibodies (mAb) were raised against peptides encoding the T cell stimulatory epitopes of α-gliadin (amino acids (aa) 59-71) and aa γ-gliadin (aa 142-153 and aa 147-159). These mAb competition assays were developed that quantitatively detect T cell stimulatory epitopes present on both intact proteins and peptides of sizes recognisable by CD4+ T cells. Results: With the mAb based competition assays, T cell epitopes were detected in pepsin/trypsin digests of wheat proteins and ethanol extracts of various food products, with detection levels lower than those reached with gluten specific T cells. Moreover, the presence of T cell stimulatory epitopes was also detected in preparations of barley, rye, and triticale, other cereals known to be toxic for CD patients. Conclusions: A new antibody based method has been developed, detecting the presence of T cell stimulatory gluten peptides. This can be used to further ensure the safety of food consumed by CD patients.