Comparison of salt-soluble wheat allergens in a mouse model vs. known human wheat allergens: evidence from durum and ancient wheats

Abstract

Abstract Wheat allergies are among the major types of food allergies that are potentially life-threatening because of anaphylaxis. Monitoring changes to wheat allergens in novel wheat lines/genotypes is critical to prevent inadvertent introduction of potentially hyper-allergenic varieties from genetic modification. Nonetheless, validated methods for this purpose are unavailable at present. We have previously described a mouse model of wheat allergy using salt-soluble protein extract (SSPE) from durum wheat. As a proof-of-concept study, here we tested the hypothesis that wheat allergens in this mouse model will be identical to those reported for human wheat allergy. We created a mini plasma bank using hyper-IgE immune plasma obtained from Balb/cJ female mice (n=20) that had been sensitized with durum-SSPE along with alum adjuvant followed by repeated booster intraperitoneal injections with SSPE alone. Using hyper-IgE plasma we optimized an IgE-Western Blot (IgEWB) method to identify allergens present in SSPEs from durum wheat and an ancient tauschii wheat. The IgE-binding allergenic bands present in raw and boiled/reduced SSPEs from durum and tauschii wheats were sequenced using LC-MS/MS method. The most abundant allergens were identified and compared with a human wheat allergen database. There were 13 allergens present in durum-SSPE, of which 6 were present only in raw extract, 2 were present only in boiled/reduced extract, and 5 were present in both. There were 10 allergens present in tauschii-SSPE, of which 7 were present only in raw extract, and 3 were present in both raw and boiled/reduced extract. Between durum and tauschii wheats, 14 allergens were present in this mouse model, of which 10 are human wheat allergens. Supported by USDA/NIFA, MSU