Overview -- Long-Term Goals & Project ObjectivesWe have developed a groundbreaking adjuvant-free Transdermal Sensitization/Oral Elicitation (TS/OE) mouse model of food allergy (Reviewed in Gonipeta et al 2015a). This model is based upon our discovery that food ingestion is not the only way to create a food allergy: We have shown that food allergies can develop upon dermal exposure to food allergens (see Preliminary Data). TS/OE mice simulate many key aspects of human food allergies. EPA regulators view our model as highly promising for evaluating allergenic potential of GM foods (Reviewed in Selgrade et al 2009).This project is about validating the TS/OE mouse model for wheat allergens. Once validated, the TS/OE model can be used to predict whether new genetically modified (GM) wheat varieties are likely to cause allergies in humans. With wheat eaten worldwide and added to all kinds of food products, keeping the world safe from GM wheat varieties that cause allergies, and even worse, life-threatening anaphylaxis, is a serious concern.The TS/OE model should also be of signal value in tackling an even more general issue: Are GM foods more allergenic than conventional non-GM foods? At present this question cannot be addressed effectively because no validated method exists to assess the allergenic potential of GM foods vs. non-GM foods (Domingo 2016, Ladics et al 2104, Selgrade et al 2009, Hollingworth et al 2003).We will use the TS/OE mouse model to determine the natural genetic variation in allergenicity in the world's 4 wheat genotypes -- AA, AABB, DD, and AABBDD. Based on strong preliminary data, our central hypothesis is that these wheat genotypes will show significant natural variation in allergenicity. Validating our model will create a new reference standard to quantitatively assess the allergenic potential of GM wheat in development. To test our central hypothesis we have two specific Aims.Aim 1. Validate the TS/OE model for transdermal sensitization: Determine the natural genetic variation in the intrinsic sensitization potencies from transdermal application of wheat genotypes AA, AABB, DD, and AABBDD.Hypothesis: The four distinct wheat genotypes differ in intrinsic sensitization potencies. We will test this using the TS/OE mouse model and identify specific IgE-binding allergens and immune mechanisms associated with different sensitization potencies due to transdermal application.im 2. Validate the TS/OE model for oral challenge: Determine the natural genetic variation in the oral allergic disease elicitation potencies of wheat genotypes AA, AABB, DD, and AABBDD.Hypothesis: The four wheat genotypes will differ in ability to elicit oral allergic disease in the TS/OE model. We will quantify disease-eliciting potencies using established disease assays and then identify the immune mechanisms associated with different disease elicitation potencies.