Title
A novel and sensitive method for the detection of T cell stimulatory epitopes of α/β- and γ-gliadin
Author
Spaenij-Dekking, E.H.A.
Kooy-Winkelaar, E.M.C.
Nieuwenhuizen, W.F.
Drijfhout, J.W.
Koning, F.
TNO Voeding
Publication year
2004
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.
Subject
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
To reference this document use:
http://resolver.tudelft.nl/uuid:51b4bd37-5867-4d16-a65e-030e12dd799e
DOI
https://doi.org/10.1136/gut.2003.037952
TNO identifier
237982
ISSN
0017-5749
Source
Gut, 53 (9), 1267-1273
Document type
article