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Forschungsdatenbank PMU-SQQUID

Genetic engineering of the major timothy grass pollen allergen, Phl p 6, to reduce allergenic activity and preserve immunogenicity.
Vrtala, S; Focke, M; Kopec, J; Verdino, P; Hartl, A; Sperr, WR; Fedorov, AA; Ball, T; Almo, S; Valent, P; Thalhamer, J; Keller, W; Valenta, R;
J Immunol. 2007; 179(3):1730-1739
Originalarbeiten (Zeitschrift)

PMU-Autor/inn/en

Hartl Arnulf Josef

Abstract

On the basis of IgE epitope mapping data, we have produced three allergen fragments comprising aa 1-33, 1-57, and 31-110 of the major timothy grass pollen allergen Phi p 6 aa 1-110 by expression in Escherichia coli and chemical synthesis. Circular dichroism analysis showed that the purified fragments lack the typical a-helical fold of the complete allergen. Superposition of the sequences of the fragments onto the three-dimensional allergen structure indicated that the removal of only one of the four helices had led to the destabilization of the alpha helical structure of Phi p 6. The lack of structural fold was accompanied by a strong reduction of IgE reactivity and allergenic activity of the three fragments as determined by basophil histamine release in allergic patients. Each of the three Phi p 6 fragments adsorbed to CFA induced Phi p 6-specific IgG Abs in rabbits. However, immunization of mice with fragments adsorbed to an adjuvant allowed for human use (AluGel-S) showed that only the Phi p 6 aa 31-110 induced Phi p 6-specific IgG Abs. Anti-Phi p 6 IgG Abs induced by vaccination with Phi p 6 aa 31-110 inhibited patientsxxx IgE reactivity to the wild-type allergen as well as Phi p 6-induced basophil degranulation. Our results are of importance for the design of hypoallergenic allergy vaccines. They show that it has to be demonstrated that the hypoallergenic derivative induces a robust IgG response in a formulation that can be used in allergic patients.


Useful keywords (using NLM MeSH Indexing)

Allergens/administration*

dosage

Allergens/biosynthesis*

Allergens/genetics*

Allergens/immunology

Animals

Down-Regulation/genetics

Down-Regulation/immunology*

Female

Gene Expression Regulation/immunology

Humans

Immune Sera/biosynthesis

Immunoglobulin E/metabolism

Immunoglobulin G/biosynthesis

Immunoglobulin G/metabolism

Mice

Mice, Inbred BALB C

Peptide Fragments/administration*

dosage

Peptide Fragments/chemical synthesis

Peptide Fragments/genetics

Peptide Fragments/immunology

Phleum/genetics

Phleum/immunology

Plant Proteins/administration*

dosage

Plant Proteins/chemical synthesis*

Plant Proteins/genetics*

Plant Proteins/immunology

Pollen/genetics

Pollen/immunology

Protein Engineering/methods

Protein Folding

Protein Structure, Secondary

Rabbits

Recombinant Fusion Proteins/administration*

dosage

Recombinant Fusion Proteins/chemical synthesis*

Recombinant Fusion Proteins/genetics*

Recombinant Fusion Proteins/immunology

Vaccines/administration*

dosage

Vaccines/chemical synthesis

Vaccines/genetics*

Vaccines/immunology