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Vol. 136, No. 3, 2005
Issue release date: March 2005
Section title: Original Paper
Int Arch Allergy Immunol 2005;136:239–249
(DOI:10.1159/000083950)

Characterization of Wild-Type Recombinant Bet v 1a as a Candidate Vaccine against Birch Pollen Allergy

Batard T.a · Didierlaurent A.a · Chabre H.a · Mothes N.b · Bussières L.a · Bohle B.b · Couret M.-N.a · Ball T.b · Lemoine P.a · Focks Tejkl M.b · Chenal A.d · Clément G.e · Dupont F.f · Valent P.c · Krauth M.-T.c · André C.a · Valenta R.b · Moingeon P.a
aStallergènes, R&D, Antony, France; bDepartment of Pathophysiology and cDepartment of Internal Medicine, Medical University, Vienna, Austria; dDépartement d’Ingénierie et d’Etude des Protéines (DIEP) and eLaboratoire d’Immunoallergie alimentaire, CEA/Saclay, Gif-sur-Yvette, France; fHenogen, Charleroi, Belgium

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Article / Publication Details

First-Page Preview
Abstract of Original Paper

Received: 7/29/2004
Accepted: 10/18/2004
Published online: 3/16/2005
Issue release date: March 2005

Number of Print Pages: 11
Number of Figures: 8
Number of Tables: 1

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

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

Abstract

Background: We describe the production in Escherichia coli as a recombinant protein of clinical grade wild-type Bet v 1a (rBet v 1a), to be used as a candidate vaccine against birch pollen allergy. Methods: This recombinant protein was purified by hydrophobic interaction and ion exchange chromatography and characterized by SDS-PAGE, immunoprint and circular dichroism in parallel with natural Bet v 1 (nBet v 1) purified from a birch pollen extract. We also compared rBet v 1 and nBet v 1 for their capacity to induce histamine release from basophils and to stimulate T lymphocyte proliferation. Results: rBet v 1a appears in SDS-PAGE as an 18-kDa monomeric protein, whereas purified nBet v 1 comprises a mixture of isoforms (resolving as three distinct bands and six spots after 1-dimensional and 2-dimensional electrophoresis, respectively). Both recombinant and natural purified Bet v 1 molecules are recognized by IgE from birch pollen-allergic patients as well as anti-Bet v 1 murine monoclonal antibodies, suggesting that the recombinant protein is correctly folded in a native configuration. Circular dichroism analysis confirmed that the two Bet v 1 molecules exhibit similar 3-dimensional structures, even if rBet v 1a appears more compact and stable in thermodenaturation/renaturation experiments. Both rBet v 1 and nBet v 1 induce the degranulation of sensitized basophils and proliferation of Bet v 1-specific T lymphocytes in a similar manner. Conclusions: On the basis of these structural and biological properties, rBet v 1a is a valid candidate vaccine against birch pollen allergy, currently evaluated in humans.

© 2005 S. Karger AG, Basel


  

Author Contacts

Correspondence to: Dr. P. Moingeon
Research and Development, Stallergènes
6, rue Alexis de Tocqueville
FR–92160 Antony (France)
Tel. +33 1 55 59 25 35, Fax +33 1 55 59 21 02, E-Mail pmoingeon@stallergenes.fr

  

Article Information

Received: July 29, 2004
Accepted after revision: October 19, 2004
Published online: February 16, 2005
Number of Print Pages : 11
Number of Figures : 8, Number of Tables : 1, Number of References : 42

  

Publication Details

International Archives of Allergy and Immunology

Vol. 136, No. 3, Year 2005 (Cover Date: March 2005)

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

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


Article / Publication Details

First-Page Preview
Abstract of Original Paper

Received: 7/29/2004
Accepted: 10/18/2004
Published online: 3/16/2005
Issue release date: March 2005

Number of Print Pages: 11
Number of Figures: 8
Number of Tables: 1

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

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


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Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in goverment regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
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