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Vol. 145, No. 3, 2008
Issue release date: February 2008
Section title: Original Paper
Int Arch Allergy Immunol 2008;145:193–206
(DOI:10.1159/000109288)

Characterization of a Hypoallergenic Recombinant Bet v 1 Variant as a Candidate for Allergen-Specific Immunotherapy

Kahlert H. · Suck R. · Weber B. · Nandy A. · Wald M. · Keller W. · Cromwell O. · Fiebig H.
aAllergopharma Joachim Ganzer KG, Reinbek, Germany; bInstitute of Chemistry, Structural Biology, Graz, Austria

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Article / Publication Details

First-Page Preview
Abstract of Original Paper

Received: 11/20/2006
Accepted: 6/18/2007
Published online: 10/2/2007

Number of Print Pages: 14
Number of Figures: 9
Number of Tables: 3

ISSN: 1018-2438 (Print)
eISSN: 1423-0097 (Online)

For additional information: http://www.karger.com/IAA

Abstract

Background: Recombinant allergens and especially their hypoallergenic variants are promising candidates for a more effective and safer specific immunotherapy. Methods: Physicochemical and immunological characteristics of a folding variant of recombinant Bet v 1 (rBet v 1-FV) were investigated in comparison to natural Bet v 1 (nBet v 1) and the correctly folded recombinant Bet v 1 (rBet v 1-WT) by SDS-PAGE, size exclusion chromatography, multi-angle light scattering, circular dichroism, immunoblotting and enzyme allergosorbent test inhibition assay for detection of IgE reactivity and ELISA with Bet v 1-specific monoclonal antibodies. The functional IgE reactivity of the different Bet v 1 proteins was investigated using basophil activation in terms of CD203c expression and histamine release. T cell reactivity was investigated using T cell lines raised from birch pollen-allergic subjects against nBet v 1. Immunogenicity was investigated in mice. Results: Physicochemical characterization revealed purity, homogeneity and monomeric properties of rBet v 1-FV. Unlike nBet v 1 and rBet v 1-WT, rBet v 1-FV showed almost no IgE binding in immunoblots. The reduction of allergenicity was further proved by IgE-binding inhibition assays, basophil activation and histamine release. T cell reactivity was completely conserved, as demonstrated by proliferation of Bet v 1-specific T cell lines with multiple epitope specificities. rBet v 1-FV showed strong immunogenicity in mice. Conclusions: Due to its reduced IgE reactivity and decreased capacity to activate basophils, but retained T cell reactivity and strong immunogenicity, rBet v 1-FV proved to be a very promising candidate for specific immunotherapy in birch pollen-allergic subjects.


  

Author Contacts

Correspondence to: Dr. Helga Kahlert
Allergopharma, Joachim Ganzer KG
Hermann-Körner-Strasse 52
DE–21465 Reinbek (Germany)
Tel. +49 40 727 65 109, Fax +49 40 722 7713, E-Mail helga.kahlert@allergopharma.de

  

Article Information

Received: November 20, 2006
Accepted after revision: June 18, 2007
Published online: October 2, 2007
Number of Print Pages : 14
Number of Figures : 9, Number of Tables : 3, Number of References : 51

  

Publication Details

International Archives of Allergy and Immunology

Vol. 145, No. 3, Year 2008 (Cover Date: February 2008)

Journal Editor: Valenta, R. (Vienna)
ISSN: 1018–2438 (Print), eISSN: 1423–0097 (Online)

For additional information: http://www.karger.com/IAA


Article / Publication Details

First-Page Preview
Abstract of Original Paper

Received: 11/20/2006
Accepted: 6/18/2007
Published online: 10/2/2007

Number of Print Pages: 14
Number of Figures: 9
Number of Tables: 3

ISSN: 1018-2438 (Print)
eISSN: 1423-0097 (Online)

For additional information: http://www.karger.com/IAA


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