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Forschungsdatenbank PMU-SQQUID

Generation of an allergy vaccine by disruption of the three-dimensional structure of the cross-reactive calcium-binding allergen, Phl p 7.
Westritschnig, K; Focke, M; Verdino, P; Goessler, W; Keller, W; Twardosz, A; Mari, A; Horak, F; Wiedermann, U; Hartl, A; Thalhamer, J; Sperr, WR; Valent, P; Valenta, R;
J Immunol. 2004; 172(9):5684-5692
Originalarbeiten (Zeitschrift)

PMU-Autor/inn/en

Hartl Arnulf Josef

Abstract

The grass pollen allergen, Phl p 7, belongs to a family of highly cross-reactive calcium-binding pollen allergens. Because Phl p 7 contains most of the disease-eliciting epitopes of pollen-derived calcium-binding allergens, hypoallergenic variants were engineered according to the x-ray crystal structure of Phi p 7 for allergy vaccination. In three recombinant variants, amino acids essential for calcium binding were mutated, and two peptides comprising the N- and C-terminal half were obtained by synthetic peptide chemistry. As determined by circular dichroism analysis and size exclusion chromatography coupled to mass spectrometry, recombinant mutants showed altered structural fold and lacked calcium-binding capacity, whereas the two synthetic peptides had completely lost-their structural fold. Allergic patientsxxx IgE Ab binding was strongest reduced to the variant containing two mutations in each of the two calcium-binding sites and to the peptides. Basophil histamine release and skin test experiments in allergic patients identified the peptides as the vaccine candidates with lowest allergenic activity. Immunization of rabbits with the peptides induced IgG Abs that blocked allergic patientsxxx IgE binding to Phl p 7 and inhibited allergen-induced basophil degranulation. Our results indicate that disruption of an allergenxxxs three-dimensional structure represents a general strategy for the generation of hypoallergenic allergy vaccines, and demonstrate the importance of allergen-specific IgG Abs for the inhibition of immediate allergic symptoms.


Useful keywords (using NLM MeSH Indexing)

Allergens/chemistry

Allergens/genetics*

Allergens/immunology

Allergens/metabolism

Animals

Anti-Allergic Agents/administration*

dosage

Anti-Allergic Agents/chemical synthesis

Anti-Allergic Agents/immunology

Antigens, Plant

Basophils/immunology

Basophils/secretion

Binding Sites, Antibody/genetics

Binding, Competitive/immunology

Calcium-Binding Proteins/chemistry

Calcium-Binding Proteins/genetics*

Calcium-Binding Proteins/immunology

Calcium-Binding Proteins/metabolism

Cell Degranulation

Cell Line, Tumor

Cross Reactions

Desensitization, Immunologic/methods*

Dose-Response Relationship, Immunologic

Histamine Release/immunology

Humans

Immunoglobulin E/metabolism

Immunoglobulin G/biosynthesis

Immunoglobulin G/metabolism

Mice

Mutagenesis, Site-Directed

Peptide Fragments/administration*

dosage

Peptide Fragments/genetics

Peptide Fragments/immunology

Peptide Fragments/metabolism

Phleum/chemistry

Phleum/genetics

Phleum/immunology*

Plant Proteins/chemistry

Plant Proteins/genetics*

Plant Proteins/immunology

Plant Proteins/metabolism

Pollen/genetics

Pollen/immunology

Rabbits

Rats

Skin Tests

Structure-Activity Relationship

Vaccines/administration*

dosage

Vaccines/chemical synthesis*

Vaccines/genetics*

Vaccines/immunology