Wheat-dependent exercise-induced anaphylaxis (WDEIA) is a severe IgE-mediated allergic reaction provoked by the combination of wheat-ingestion with intensive physical exercise over the next few hours. Among wheat proteins, -5 gliadin, which is one of the components of fast -gliadin, has been reported as a major allergen in the anaphylaxis. In this study, we detected IgE-binding epitopes within the primary sequence of -5 gliadin using arrays of overlapping peptides synthesized on derivatized cellulose membranes. Sera from four patients with WDEIA having specific IgE to the fast -gliadin were used to probe the membrane. Seven epitopes, QQIPQQQ, QQLPQQQ, QQFPQQQ, QQSPEQQ, QQSPQQQ, QQYPQQQ, and PYPP, were detected within the primary sequence of -5 gliadin. By using sera of 15 patients, 4 of them, QQIPQQQ, QQFPQQQ, QQSPEQQ, and QQSPQQQ, were found to be dominant epitopes. Mutational analysis of the QQIPQQQ and QQFPQQQ indicated that amino acids at positions Gln 1 , Pro 4 , Gln 5 , Gln 6 , and Gln 7 were critical for IgE binding. These results will provide a useful tool for developing safer wheat products in addition to diagnostic and immunotherapy techniques for WDEIA.Food-dependent exercise-induced anaphylaxis is a distinct form of food allergy induced by physical exercise (1). Food items such as shrimp (2), hazelnut (3), buckwheat (4), corn (5), and celery (6) are responsible for the development of food-dependent exercise-induced anaphylaxis. However, of all of the various kinds of food, wheat is reported to be the allergen with the highest frequency in Japan (7). Symptoms are typically generalized urticaria and severe allergic reactions such as shock or hypotension. Because of this serious reaction, it is important to determine the causative food to avoid the allergic reaction. A challenge test consisting of ingestion of the assumed food followed by intense physical exercise is the only reliable method to determine the causative food and to diagnose the disease. However, the challenge test is not always safe because in some cases the test induces an anaphylactic shock. In addition, the most reliable treatment for this disease is to avoid taking the causative food or, alternatively, to take a rest after meals. However, in the case of wheat allergy, elimination causes a decline in the quality of life for the patients. Thus, an in vitro diagnostic method as well as hypoallergenic wheat is necessary for patients with wheat-dependent exercise-induced anaphylaxis (WDEIA).
1Recent studies have revealed the IgE-binding epitopes of several food allergens including egg (8), milk (9, 10), soybean (11), and peanut (12), whereas the IgE-binding epitopes for wheat allergen are controversial. Wheat protein is composed of water/salt-soluble proteins and water/salt-insoluble proteins. Proteins in the water/salt-soluble fraction, such as ␣-amylase inhibitor, peroxidase, glycerinaldehyde-3-phosphate dehydrogenase, serpin, and triosephosphate isomerase, have been considered to be major allergens in patients with bakers' asthma (13-15...