Tranquet, OlivierGaudin, Jean-CharlesPatil, SaritaSteinbrecher, JohannaMatsunaga, KayokoTeshima, ReikoSakai, ShinobuLarré, ColetteDenery-Papini, Sandra2017-12-062017Tranquet, Olivier, Jean-Charles Gaudin, Sarita Patil, Johanna Steinbrecher, Kayoko Matsunaga, Reiko Teshima, Shinobu Sakai, Colette Larré, and Sandra Denery-Papini. 2017. “A chimeric IgE that mimics IgE from patients allergic to acid-hydrolyzed wheat proteins is a novel tool for in vitro allergenicity assessment of functionalized glutens.” PLoS ONE 12 (11): e0187415. doi:10.1371/journal.pone.0187415. http://dx.doi.org/10.1371/journal.pone.0187415.http://nrs.harvard.edu/urn-3:HUL.InstRepos:34492943Background: Acid-hydrolyzed wheat proteins (acid-HWPs) have been shown to provoke severe allergic reactions in Europe and Japan that are distinct from classical wheat allergies. Acid-HWPs were shown to contain neo-epitopes induced by the deamidation of gluten proteins. However, products with variable rates of deamidation can be found. Objectives: In this work, we studied the effect of the extent of wheat proteins deamidation on its allergenicity. A recombinant chimeric IgE was produced and compared to patients’ IgE for its capacity to assess the IgE-mediated triggering potential of acid-HWPs. Methods: Sera from acid-HWP allergic patients were analyzed via ELISA and a functional basophil assay for their IgE reactivity to wheat proteins with different deamidation levels. A chimeric mouse/human IgE (chIgE-DG1) specific for the main neo-epitope, QPEEPFPE, involved in allergy to acid-HWPs was characterized with respect to its functionality and its reactivity compared to that of patients’ IgE. Results: Acid-HWPs with medium (30%) and high (50–60%) deamidation levels displayed a markedly stronger IgE binding and capacity to activate basophils than those of samples with weak (15%) deamidation levels. The monoclonal chIgE-DG1 allowed basophil degranulation in the presence of deamidated wheat proteins. ChIgE-DG1 was found to mimic patients’ IgE reactivity and displayed the same ability to rank acid-HWP products in a degranulation assay. Conclusion: Increasing the deamidation level of products from 15% to 60% resulted in an approximately 2-fold increase in their antigenicity and a 100-fold increase in their eliciting potential. The chimeric ChIgE-DG1 may be a useful tool to evaluate functionalized glutens for their allergenic potential. By mimicking patient sera reactivity, chIgE-DG1 also provided data on the patients' IgE repertoire and on the functionality of certain repeated epitopes in gluten proteins.en-USBiology and Life SciencesBiochemistryProteinsGlutenOrganismsEukaryotaPlantsGrassesWheatPost-Translational ModificationDeamidationCell BiologyCellular TypesAnimal CellsBlood CellsWhite Blood CellsBasophilsImmune CellsImmunologyMedicine and Health SciencesCell PhysiologyCell DegranulationClinical MedicineClinical ImmunologyAllergiesAllergic DiseasesFood AllergiesAllergensJournal Article2017-12-0610.1371/journal.pone.0187415