Molecular determinants of allergen-induced effector cell degranulation

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Presentation transcript:

Molecular determinants of allergen-induced effector cell degranulation Anna Gieras, MSc, Margarete Focke-Tejkl, PhD, Tanja Ball, PhD, Petra Verdino, PhD, Arnulf Hartl, PhD, Josef Thalhamer, PhD, Rudolf Valenta, MD Journal of Allergy and Clinical Immunology Volume 119, Issue 2, Pages 384-390 (February 2007) DOI: 10.1016/j.jaci.2006.09.034 Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 Localization of the P1 peptide in the context of the 3-dimensional structure of Phl p 1 visualized by means of surface (A-D) and backbone (E-H) representations. The 2 Phl p 1 monomers are shown in light gray and dark gray, and N- and C-termini are indicated. The P1 peptide comprising amino acids 86-116 of Phl p 1 has been colored, and the amino acids have been numbered. Journal of Allergy and Clinical Immunology 2007 119, 384-390DOI: (10.1016/j.jaci.2006.09.034) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 A, Levels of binding (mean OD values, y-axis) of the Phl p 1–specific monoclonal IgE to Phl p 1 and Phl p 1–derived peptides (x-axis). B and C, SDS-PAGE containing allergens (rPhl p 1 and rBet v 1) and peptide preparations (P1 monomer and P1 dimer) under reducing (Fig 2, B) and nonreducing (Fig 2, C) conditions. Molecular weights are displayed in kilodaltons at the margin. Journal of Allergy and Clinical Immunology 2007 119, 384-390DOI: (10.1016/j.jaci.2006.09.034) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 A, Induction of β-hexosaminidase release is displayed on RBL cells with rPhl p 1 and nPhl p 1. RBL cells were sensitized with mouse monoclonal IgE against the Phl p 1–derived peptide P1 (amino acids 86-116) and stimulated with increasing concentrations of allergens (x-axis). β-Hexosaminidase release is displayed on the y-axis. B, Allergenic activity of P1 monomer, P1 dimer, and P1 polymers. Sensitized RBL cells were stimulated with increasing concentrations of the peptides (x-axis), and the percentage of β-hexosaminidase release is displayed on the y-axis. Journal of Allergy and Clinical Immunology 2007 119, 384-390DOI: (10.1016/j.jaci.2006.09.034) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 Factors affecting magnitude of mediator release and cellular sensitivity. The magnitude of β-hexosaminidase release (y-axis) depends on the titers of IgE used for loading the cells. RBL cells loaded with different concentrations of IgE (1:50, 1:500, and 1:2000) were exposed to different concentrations of P1 dimer (0.01-1 μg/mL in A and 0.001-100 μg/mL in B, x-axis). Journal of Allergy and Clinical Immunology 2007 119, 384-390DOI: (10.1016/j.jaci.2006.09.034) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 5 Preincubation of the monoclonal P1-specific IgE with the P1 monomer inhibits binding to rPhl p 1. The P1-specific IgE was preincubated with increasing concentrations (x-axis) of the P1 monomer, control peptide P5, or buffer alone. OD values (y-axis) correspond to the amount of IgE bound to immobilized rPhl p 1. Journal of Allergy and Clinical Immunology 2007 119, 384-390DOI: (10.1016/j.jaci.2006.09.034) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 6 P1 monomer inhibits P1 polymer–induced β-hexosaminidase release. Sensitized RBL cells were incubated with increasing doses of P1 monomer (x-axis) and subsequently exposed to P1 polymer, and the percentage of released β-hexosaminidase was recorded (y-axis). Journal of Allergy and Clinical Immunology 2007 119, 384-390DOI: (10.1016/j.jaci.2006.09.034) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions