Reduction of the in vivo allergenicity of Der p 2, the major house-dust mite allergen, by genetic engineering.
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.
Chen
Kuan-Wei
KW
Department of Pathophysiology, Center for Physiology and Pathophysiology, Medical University of Vienna, Waehringer Guertel 18-20, 1090 Vienna, Austria.
Fuchs
Gudrun
G
Sonneck
Karoline
K
Gieras
Anna
A
Swoboda
Ines
I
Douladiris
Nikolas
N
Linhart
Birgit
B
Jankovic
Marija
M
Pavkov
Tea
T
Keller
Walter
W
Papadopoulos
Nikolaos G
NG
Valent
Peter
P
Valenta
Rudolf
R
Vrtala
Susanne
S
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
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