Τίτλος:
Reduction of the in vivo allergenicity of Der p 2, the major house-dust mite allergen, by genetic engineering
Γλώσσες Τεκμηρίου:
Αγγλικά
Περίληψη:
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. © 2008 Elsevier Ltd. All rights reserved.
Συγγραφείς:
Chen, K.-W.
Fuchs, G.
Sonneck, K.
Gieras, A.
Swoboda, I.
Douladiris, N.
Linhart, B.
Jankovic, M.
Pavkov, T.
Keller, W.
Papadopoulos, N.G.
Valent, P.
Valenta, R.
Vrtala, S.
Περιοδικό:
Cellular and Molecular Immunology
Λέξεις-κλειδιά:
house dust allergen; immunoglobulin G antibody, allergenicity; animal cell; animal experiment; animal model; antibody specificity; antigen expression; article; basophil; cell activation; cell assay; controlled study; drug activity; Escherichia coli; female; genetic engineering; house dust allergy; human; immunization; immunogenicity; immunoreactivity; immunotherapy; in vitro study; in vivo study; mouse; nonhuman; prevention and control; prick test; priority journal; rabbit; rat; skin test; wild type, Allergens; Animals; Antigens, Dermatophagoides; Basophils; beta-N-Acetylhexosaminidases; Cloning, Molecular; Dermatophagoides pteronyssinus; Genetic Engineering; Humans; Hypersensitivity, Immediate; Immunoglobulin E; Immunoglobulin G; Mice; Rabbits; Recombinant Fusion Proteins; Skin Tests, Acari; Dermatophagoides pteronyssinus; Mus; Oryctolagus cuniculus; Pyroglyphidae
DOI:
10.1016/j.molimm.2008.01.006
