Strong allergenicity of Pru av 3, the lipid transfer protein from cherry, is related to high stability against thermal processing and digestion Stephan.

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Strong allergenicity of Pru av 3, the lipid transfer protein from cherry, is related to high stability against thermal processing and digestion Stephan Scheurer, PhD, Iris Lauer, PhD, Kay Foetisch, PhD, Mar San Miguel Moncin, MD, Mechthild Retzek, Christina Hartz, MSc, Ernesto Enrique, MD, Jonas Lidholm, PhD, Anna Cistero-Bahima, MD, Stefan Vieths, PhD Journal of Allergy and Clinical Immunology Volume 114, Issue 4, Pages 900-907 (October 2004) DOI: 10.1016/j.jaci.2004.06.017 Copyright © 2004 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 1 Cherry extract (50 μg total protein) was digested with pepsin, separated by SDS-PAGE, silver stained (A), and tested for IgE reactivity with a pool of cherry allergic patients' sera (B). Two hours without pepsin (1); 30 seconds (2); 1 minute (3); 15 minutes (4); 30 minutes (5); 1 hour (6); 2 hours (7); BSA without pepsin (8); BSA, 2 hours (9). ∗Cherry allergens. Journal of Allergy and Clinical Immunology 2004 114, 900-907DOI: (10.1016/j.jaci.2004.06.017) Copyright © 2004 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 2 Purified recombinant cherry allergens (Pru av 1–HisTag, Pru av 3–HisTag, Pru av 4; 2 μg each) were digested with pepsin (20 μL; as long as 2 hours). Analysis of IgE reactivity was performed with a pool of cherry allergic patients' sera (1:6 dilution) sensitized to Pru av 1, Pru av 3, or Pru av 4, respectively. Buffer control (C1, H2O; C2, HCl). Journal of Allergy and Clinical Immunology 2004 114, 900-907DOI: (10.1016/j.jaci.2004.06.017) Copyright © 2004 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 3 CD spectra of rPru av 1 (A) and Pru av 3 (B) were recorded at different temperatures: 20°C, 40°C, 70°C, 100°C, and after cooling down to 20°C. The spectral range was 185 to 255 nm, and the scanning rate was 50 nm/min. Ten scans were accumulated. MRD, Magnetic relaxation dispersion. Journal of Allergy and Clinical Immunology 2004 114, 900-907DOI: (10.1016/j.jaci.2004.06.017) Copyright © 2004 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 4 Analysis of IgE reactivity of heat-processed foods containing cherry by using sera from 2 patients allergic to cherry with sensitization to Pru av 1 (patient 2) and additional sensitization to Pru av 3 (patient 5). Fresh sweet cherries from the market (1); cherry fruit juice, 35% fruit content (Rapp's, Karben, Germany) (2); preserved cherries, sweetened (Kaiserkirschen mit Stein, Erlenhof, REWE-Erlenhof, Germany) (3); cherry jam, 45% fruit content (Erlenhof) (4). Journal of Allergy and Clinical Immunology 2004 114, 900-907DOI: (10.1016/j.jaci.2004.06.017) Copyright © 2004 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 5 SDS-PAGE and silver staining (left) and immunoblotting (right) of 2 commercial cherry SPT extracts (A and B). Right, Detection of Pru av 1 and Pru av 3 by specific IgG (mouse anti–Pru av 1, rabbit anti–Pru p 3) and specific IgE from patients allergic to cherry (patient [pt] 3 sensitized to Pru av 1; pt 13 sensitized to Pru av 3). Journal of Allergy and Clinical Immunology 2004 114, 900-907DOI: (10.1016/j.jaci.2004.06.017) Copyright © 2004 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 6 Histamine release from passively sensitized basophils from a cherry LTP–sensitized patient (patient 12) (A) and a Pru av 1–sensitized patient (patient 9) (B), induced by cherry extract (CE) prepared from fresh fruits, CE digested by pepsin, and CE prepared from 3 processed food products containing cherries (1, cherry fruit juice; 2, preserved cherries; 3, cherry jam). Journal of Allergy and Clinical Immunology 2004 114, 900-907DOI: (10.1016/j.jaci.2004.06.017) Copyright © 2004 American Academy of Allergy, Asthma and Immunology Terms and Conditions