TY - JOUR
TI - Reduction of the in vivo allergenicity of Der p 2, the major house-dust mite allergen, by genetic engineering
AU - Chen, K.-W.
AU - Fuchs, G.
AU - Sonneck, K.
AU - Gieras, A.
AU - Swoboda, I.
AU - Douladiris, N.
AU - Linhart, B.
AU - Jankovic, M.
AU - Pavkov, T.
AU - Keller, W.
AU - Papadopoulos, N.G.
AU - Valent, P.
AU - Valenta, R.
AU - Vrtala, S.
JO - Cellular and Molecular Immunology
PY - 2008
VL - 45
TODO - 9
SP - 2486-2498
PB - 
SN - null
TODO - 10.1016/j.molimm.2008.01.006
TODO - 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
TODO - 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.
ER -