Christian Schwager1, Skadi Kull1, Jochen Behrends2, Niels Röckendorf3, Frauke Schocker1, Andreas Frey3, Arne Homann1, Wolf-Meinhard Becker1, Uta Jappe4. 1. Division of Clinical and Molecular Allergology, Research Center Borstel, Priority Research Area Asthma and Allergy, Airway Research Center North (ARCN), German Center for Lung Research (DZL), Borstel, Germany. 2. Core Facility Fluorescence Cytometry, Research Center Borstel, Borstel, Germany. 3. Division of Mucosal Immunology and Diagnostics, Research Center Borstel, Priority Research Area Asthma and Allergy, Airway Research Center North (ARCN), German Center for Lung Research (DZL), Borstel, Germany. 4. Division of Clinical and Molecular Allergology, Research Center Borstel, Priority Research Area Asthma and Allergy, Airway Research Center North (ARCN), German Center for Lung Research (DZL), Borstel, Germany; Interdisciplinary Allergy Outpatient Clinic, Department of Internal Medicine, University of Luebeck, Luebeck, Germany. Electronic address: ujappe@fz-borstel.de.
Abstract
BACKGROUND: Peanut allergy is one of the most common and most severe food allergies in Western countries and its accurate diagnosis to prevent potential life-threatening allergic reactions is crucial. However, aqueous extracts used for routine diagnostic measurements are devoid of lipophilic allergens such as oleosins. We have recently succeeded in the isolation and purification of these unique proteins, and the present study evaluates their allergenic potential and clinical relevance. OBJECTIVE: We sought to assess allergenicity and sensitization prevalence of oleosins obtained from both raw and in-shell roasted peanuts. In addition, we tested the utilization of natural and recombinant oleosins for allergy diagnostic purposes. METHODS: Oleosin sensitization, prevalence, and impact of thermal processing were analyzed by immunoblot with sera from 52 peanut-allergic individuals displaying different clinical phenotypes. The application of natural and recombinant oleosins for allergy diagnostics was investigated by basophil activation test (BAT). IgE-binding epitopes were identified by oligopeptide microarray. RESULTS: Sensitization to oleosins was observed exclusively in peanut-allergic subjects suffering from severe systemic reactions. IgE-binding capacity of oleosins derived from in-shell roasted peanuts was increased as shown by immunoblot analysis and BAT. Both natural and recombinant molecules can be used to identify oleosin-sensitized patients by BAT. A linear epitope of Ara h 15 was determined that displays high similarity to other seed-derived oleosins. CONCLUSIONS: Oleosins are clinically relevant peanut allergens and most likely associated with severe allergic symptoms. In-shell roasting increases their allergenicity, which is consistent with the observation that most allergic reactions are in connection with roasted peanuts.
BACKGROUND:Peanutallergy is one of the most common and most severe food allergies in Western countries and its accurate diagnosis to prevent potential life-threatening allergic reactions is crucial. However, aqueous extracts used for routine diagnostic measurements are devoid of lipophilic allergens such as oleosins. We have recently succeeded in the isolation and purification of these unique proteins, and the present study evaluates their allergenic potential and clinical relevance. OBJECTIVE: We sought to assess allergenicity and sensitization prevalence of oleosins obtained from both raw and in-shell roasted peanuts. In addition, we tested the utilization of natural and recombinant oleosins for allergy diagnostic purposes. METHODS:Oleosin sensitization, prevalence, and impact of thermal processing were analyzed by immunoblot with sera from 52 peanut-allergic individuals displaying different clinical phenotypes. The application of natural and recombinant oleosins for allergy diagnostics was investigated by basophil activation test (BAT). IgE-binding epitopes were identified by oligopeptide microarray. RESULTS: Sensitization to oleosins was observed exclusively in peanut-allergic subjects suffering from severe systemic reactions. IgE-binding capacity of oleosins derived from in-shell roasted peanuts was increased as shown by immunoblot analysis and BAT. Both natural and recombinant molecules can be used to identify oleosin-sensitized patients by BAT. A linear epitope of Ara h 15 was determined that displays high similarity to other seed-derived oleosins. CONCLUSIONS:Oleosins are clinically relevant peanut allergens and most likely associated with severe allergic symptoms. In-shell roasting increases their allergenicity, which is consistent with the observation that most allergic reactions are in connection with roasted peanuts.
Authors: Anna Pomés; Janet M Davies; Gabriele Gadermaier; Christiane Hilger; Thomas Holzhauser; Jonas Lidholm; Andreas L Lopata; Geoffrey A Mueller; Andreas Nandy; Christian Radauer; Sanny K Chan; Uta Jappe; Jörg Kleine-Tebbe; Wayne R Thomas; Martin D Chapman; Marianne van Hage; Ronald van Ree; Stefan Vieths; Monika Raulf; Richard E Goodman Journal: Mol Immunol Date: 2018-04-04 Impact factor: 4.407
Authors: Uta Jappe; Christian Schwager; Andra B Schromm; Nestor González Roldán; Karina Stein; Holger Heine; Katarzyna A Duda Journal: Front Immunol Date: 2019-02-14 Impact factor: 7.561