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Batard, T; Didierlaurent, A; Chabre, H; Mothes, N; Bussières, L; Bohle, B; Couret, MN; Ball, T; Lemoine, P; Focks Tejkl, M; Chenal, A; Clément, G; Dupont, F; Valent, P; Krauth, MT; André, C; Valenta, R; Moingeon, P.
Characterization of wild-type recombinant Bet v 1a as a candidate vaccine against birch pollen allergy.
Int Arch Allergy Immunol. 2005; 136(3):239-249 Doi: 10.1159/000083950
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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.
Find related publications in this database (using NLM MeSH Indexing)
Allergens - chemistry Allergens - genetics Allergens - immunology Allergens - metabolism
Amino Acid Sequence -
Antibodies, Monoclonal -
Basophils - immunology
Betula - immunology
Escherichia coli - genetics Escherichia coli - metabolism
Histamine Release -
Humans -
Lymphocyte Activation -
Molecular Sequence Data -
Plant Proteins - chemistry Plant Proteins - genetics Plant Proteins - immunology Plant Proteins - isolation and purification Plant Proteins - metabolism
Pollen - immunology
Recombinant Proteins - biosynthesis Recombinant Proteins - chemistry Recombinant Proteins - immunology
Respiratory Hypersensitivity - immunology Respiratory Hypersensitivity - prevention and control
T-Lymphocytes - immunology
Vaccines, Synthetic - immunology

Find related publications in this database (Keywords)
Bet v 1
birch allergy
specific immunotherapy
vaccine
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