title: Molecular evolution of hypoallergenic hybrid proteins for vaccination against grass pollen allergy.
abstract: More than 10% of the population in Europe and North America suffer from IgE-associated allergy to grass pollen. In this article, we describe the development of a vaccine for grass pollen allergen-specific immunotherapy based on two recombinant hypoallergenic mosaic molecules, designated P and Q, which were constructed out of elements derived from the four major timothy grass pollen allergens: Phl p 1, Phl p 2, Phl p 5, and Phl p 6. Seventeen recombinant mosaic molecules were expressed and purified in Escherichia coli using synthetic genes, characterized regarding biochemical properties, structural fold, and IgE reactivity. We found that depending on the arrangement of allergen fragments, mosaic molecules with strongly varying IgE reactivity were obtained. Based on an extensive screening with sera and basophils from allergic patients, two hypoallergenic mosaic molecules, P and Q, incorporating the primary sequence elements of the four grass pollen allergens were identified. As shown by lymphoproliferation experiments, they contained allergen-specific T cell epitopes required for tolerance induction, and upon immunization of animals induced higher allergen-specific IgG Abs than the wild-type allergens and a registered monophosphoryl lipid A-adjuvanted vaccine based on natural grass pollen allergen extract. Moreover, IgG Abs induced by immunization with P and Q inhibited the binding of patients' IgE to natural allergens from five grasses better than IgG induced with the wild-type allergens or an extract-based vaccine. Our results suggest that vaccines based on the hypoallergenic grass pollen mosaics can be used for immunotherapy of grass pollen allergy.
authors: Division of Immunopathology, Department of Pathophysiology and Allergy Research, Center of Pathophysiology, Infectiology, and Immunology, Medical University of Vienna, 1090 Vienna, Austria;
authors: Birgit
authors: B
authors: Linhart
authors: Christian Doppler Laboratory for Allergy Research, Medical University of Vienna, 1090 Vienna, Austria;
authors: Margarete
authors: M
authors: Focke-Tejkl
authors: Division of Immunopathology, Department of Pathophysiology and Allergy Research, Center of Pathophysiology, Infectiology, and Immunology, Medical University of Vienna, 1090 Vienna, Austria;
authors: Milena
authors: M
authors: Weber
authors: Division of Immunopathology, Department of Pathophysiology and Allergy Research, Center of Pathophysiology, Infectiology, and Immunology, Medical University of Vienna, 1090 Vienna, Austria;
authors: Meena
authors: M
authors: Narayanan
authors: Biomay AG, 1090 Vienna, Austria; and.
authors: Angela
authors: A
authors: Neubauer
authors: Division of Immunopathology, Department of Pathophysiology and Allergy Research, Center of Pathophysiology, Infectiology, and Immunology, Medical University of Vienna, 1090 Vienna, Austria;
authors: Hannes
authors: H
authors: Mayrhofer
authors: Division of Hematology and Hemostaseology, Department of Internal Medicine I, Medical University of Vienna, 1090 Vienna, Austria.
authors: Katharina
authors: K
authors: Blatt
authors: Division of Immunopathology, Department of Pathophysiology and Allergy Research, Center of Pathophysiology, Infectiology, and Immunology, Medical University of Vienna, 1090 Vienna, Austria;
authors: Christian
authors: C
authors: Lupinek
authors: Division of Hematology and Hemostaseology, Department of Internal Medicine I, Medical University of Vienna, 1090 Vienna, Austria.
authors: Peter
authors: P
authors: Valent
authors: Division of Immunopathology, Department of Pathophysiology and Allergy Research, Center of Pathophysiology, Infectiology, and Immunology, Medical University of Vienna, 1090 Vienna, Austria; Christian Doppler Laboratory for Allergy Research, Medical University of Vienna, 1090 Vienna, Austria; rudolf.valenta@meduniwien.ac.at.
authors: Rudolf
authors: R
authors: Valenta
mesh_terms: D000485:Allergens / Q000235:genetics / Q000276:immunology / Q000494:pharmacology; D000818:Animals; D019020:Directed Molecular Evolution; D018984:Epitopes, T-Lymphocyte / Q000235:genetics / Q000276:immunology / Q000494:pharmacology; D005260:Female; D006801:Humans; D007114:Immunization; D007073:Immunoglobulin E / Q000235:genetics / Q000276:immunology; D007074:Immunoglobulin G / Q000235:genetics / Q000276:immunology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D031784:Phleum / Q000235:genetics / Q000276:immunology; D010940:Plant Proteins / Q000235:genetics / Q000276:immunology / Q000494:pharmacology; D011058:Pollen / Q000235:genetics / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000494:pharmacology; D006255:Rhinitis, Allergic, Seasonal / Q000235:genetics / Q000276:immunology / Q000517:prevention & control*
chemical_list: D000485:Allergens; D018984:Epitopes, T-Lymphocyte; D007074:Immunoglobulin G; D010940:Plant Proteins; D011993:Recombinant Fusion Proteins; D007073:Immunoglobulin E