Spatial clustering of the IgE epitopes on the major timothy grass pollen allergen Phl p 1: Importance for allergenic activity Sabine Flicker, PhD, Peter.

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Spatial clustering of the IgE epitopes on the major timothy grass pollen allergen Phl p 1: Importance for allergenic activity Sabine Flicker, PhD, Peter Steinberger, PhD, Tanja Ball, PhD, Maria-Theresa Krauth, MD, Petra Verdino, PhD, Peter Valent, MD, Steve Almo, PhD, Rudolf Valenta, MD Journal of Allergy and Clinical Immunology Volume 117, Issue 6, Pages 1336-1343 (June 2006) DOI: 10.1016/j.jaci.2006.02.012 Copyright © 2006 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 1 rPhl p 1–specific IgE Fabs cross-react with natural group 1 allergens from a variety of grass species (1, oat; 2, sweet vernal grass; 3, Bermuda grass; 4, rye grass; 5, common reed; 6, Kentucky blue grass; 7, rye; 8, wheat; 9, maize). The positions of the molecular weights are indicated in kilodaltons on the left margin. Journal of Allergy and Clinical Immunology 2006 117, 1336-1343DOI: (10.1016/j.jaci.2006.02.012) Copyright © 2006 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 2 A, Summary of recombinant and synthetic Phl p 1 fragments. Complete mature rPhl p 1 without the 24-amino-acid leader peptide comprises 240 amino acids (amino acids 1-240). The borders and designations of the Phl p 1 fragments and peptides are displayed. B, Antibody reactivity of nitrocellulose-bound rPhl p 1 fragments. Fragments 1 through 6 (Fig 2, A) were exposed to the IgE-derived monoclonal IgG1 (IgG1 αPhl p 1) and to serum IgE from 4 representative allergic patients (patients 1 through 4). Journal of Allergy and Clinical Immunology 2006 117, 1336-1343DOI: (10.1016/j.jaci.2006.02.012) Copyright © 2006 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 3 The C-terminal Phl p 1 fragment T147 induces histamine release from basophils of patients with Phl p 1 allergy comparable with complete rPhl p 1. Granulocytes from 2 patients with grass pollen allergy (A and B) were exposed to different concentrations (x-axis) of complete rPhl p 1, rT147, and GST. Histamine release is expressed as percentage of total histamine on the y-axis. Journal of Allergy and Clinical Immunology 2006 117, 1336-1343DOI: (10.1016/j.jaci.2006.02.012) Copyright © 2006 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 4 A, Frontal view onto the backbone representation of dimeric Phl p 1 (PDB entry 1N10). The N- and C-terminal portions (Na, Nb, Ca, and Cb) of the 2 monomers are indicated. B, The same representation as in Fig 4, A, rotated 90° along the vertical axis. The major IgE epitope-containing C-terminal regions are colored red, with the remainder of the backbone shown in blue. Journal of Allergy and Clinical Immunology 2006 117, 1336-1343DOI: (10.1016/j.jaci.2006.02.012) Copyright © 2006 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 5 A model for efficient FcεRI cross-linking by spatially clustered IgE epitopes on Phl p 1. The C-terminal IgE epitope–containing domain of Phl p 1 (C) focuses Phl p 1–specific IgE and induces efficient aggregation of FcεRI. Journal of Allergy and Clinical Immunology 2006 117, 1336-1343DOI: (10.1016/j.jaci.2006.02.012) Copyright © 2006 American Academy of Allergy, Asthma and Immunology Terms and Conditions