title: Reduction of the in vivo allergenicity of Der p 2, the major house-dust mite allergen, by genetic engineering.
abstract: The major allergen of the house-dust mite Dermatophagoides pteronyssinus, Der p 2, is recognized by approximately 90% of mite-allergic patients. We have produced two recombinant fragments of Der p 2 comprising aa 1-53 and aa 54-129 and a hybrid molecule (aa 54-129+1-53), combining the two fragments in inverse order, by genetic engineering. The recombinant Der p 2 derivatives were expressed in E. coli and purified to homogeneity. rDer p 2 derivatives (fragments and hybrid) showed a considerably reduced beta sheet structure and IgE reactivity compared to the Der p 2 wild-type allergen. The allergenic activity of the Der p 2 derivatives was reduced more than tenfold as evaluated in vitro in basophil activation assays and in vivo by skin prick testing of mite-allergic patients. Immunization of mice and rabbits with rDer p 2 derivatives induced Der p 2-specific IgG antibodies, which inhibited the binding of allergic patients' IgE to Der p 2. Immunization of mice with rDer p 2 derivatives induced less allergenic IgE responses than immunization with rDer p 2. Thus the rDer p 2 derivatives exhibited less in vivo allergenic activity and allergenicity than the Der p 2 allergen but preserved immunogenicity and may hence represent candidates for specific immunotherapy of house-dust mite allergy.
authors.affiliation: Department of Pathophysiology, Center for Physiology and Pathophysiology, Medical University of Vienna, Waehringer Guertel 18-20, 1090 Vienna, Austria.
authors.forename: Kuan-Wei
authors.initials: KW
authors.lastname: Chen
authors.forename: Gudrun
authors.initials: G
authors.lastname: Fuchs
authors.forename: Karoline
authors.initials: K
authors.lastname: Sonneck
authors.forename: Anna
authors.initials: A
authors.lastname: Gieras
authors.forename: Ines
authors.initials: I
authors.lastname: Swoboda
authors.forename: Nikolas
authors.initials: N
authors.lastname: Douladiris
authors.forename: Birgit
authors.initials: B
authors.lastname: Linhart
authors.forename: Marija
authors.initials: M
authors.lastname: Jankovic
authors.forename: Tea
authors.initials: T
authors.lastname: Pavkov
authors.forename: Walter
authors.initials: W
authors.lastname: Keller
authors.forename: Nikolaos G
authors.initials: NG
authors.lastname: Papadopoulos
authors.forename: Peter
authors.initials: P
authors.lastname: Valent
authors.forename: Rudolf
authors.initials: R
authors.lastname: Valenta
authors.forename: Susanne
authors.initials: S
authors.lastname: Vrtala
mesh_terms: D000485:Allergens / Q000276:immunology* / Q000378:metabolism; D000818:Animals; D039741:Antigens, Dermatophagoides / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism; D060829:Arthropod Proteins; D001491:Basophils / Q000276:immunology / Q000378:metabolism; D003001:Cloning, Molecular; D040002:Dermatophagoides pteronyssinus / Q000276:immunology*; D005818:Genetic Engineering; D006801:Humans; D006969:Hypersensitivity, Immediate / Q000276:immunology*; D007073:Immunoglobulin E / Q000276:immunology*; D007074:Immunoglobulin G / Q000096:biosynthesis / Q000276:immunology; D051379:Mice; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D012882:Skin Tests; D001619:beta-N-Acetylhexosaminidases / Q000378:metabolism
chemical_list: D000485:Allergens; D039741:Antigens, Dermatophagoides; D060829:Arthropod Proteins; C451163:Dermatophagoides pteronyssinus antigen p 2; D007074:Immunoglobulin G; D011993:Recombinant Fusion Proteins; D007073:Immunoglobulin E; D001619:beta-N-Acetylhexosaminidases