Journal article
Api m 6: a new bee venom allergen.
- Kettner A Institute of Biochemistry, University of Lausanne, Lausanne, Switzerland.
- Hughes GJ
- Frutiger S
- Astori M
- Roggero M
- Spertini F
- Corradin G
- 2001-05-10
Published in:
- The Journal of allergy and clinical immunology. - 2001
Allergens
Amino Acid Motifs
Amino Acid Sequence
Animals
Antibody Specificity
Antigens, Plant
Bee Venoms
Chromatography, Gel
Chromatography, High Pressure Liquid
Humans
Hypersensitivity, Immediate
Immunoglobulin E
Insect Proteins
Lymphocyte Activation
Molecular Sequence Data
Protein Denaturation
T-Lymphocytes
English
BACKGROUND
Characterization of the primary structure of allergens is a prerequisite for the design of new diagnostic and therapeutic tools for allergic diseases.
OBJECTIVE
The purpose of this study was the identification and characterization of a low-molecular-weight, IgE-binding, bee venom (BV) allergen.
METHODS
BV proteins were separated by using size exclusion chromatography and HPLC. IgE antibody binding to purified proteins was analyzed by means of immunoblotting, and T-cell response was analyzed by means of proliferation assay. Amino acid sequence was determined with 2 approaches, namely Edman degradation and carboxy terminal analysis with mass spectrometry.
RESULTS
Api m 6, which migrated as an 8-kd band in SDS-PAGE, was frequently (42%) recognized by IgE from BV-hypersensitive patients. In addition, PBMCs from BV-hypersensitive patients, as well as from a normal control subject, proliferated in response to this allergen. Api m 6 exists as 4 isoforms of 7190, 7400, 7598, and 7808 d, respectively. Amino acid sequences obtained from HPLC-purified preparations revealed that the isoforms were constituted of a common central core of 67 residues, only differing in the amino- and carboxy-terminal ends. Api m 6 showed no significant sequence homology with known proteins.
CONCLUSIONS
We have identified and sequenced a new BV allergen that elicits a strong IgE and T-cell response in a large number of BV-hypersensitive patients. Api m 6 should be considered in the diagnostic and therapeutic approach of BV immunotherapy on the basis of peptides or recombinant proteins.
Characterization of the primary structure of allergens is a prerequisite for the design of new diagnostic and therapeutic tools for allergic diseases.
OBJECTIVE
The purpose of this study was the identification and characterization of a low-molecular-weight, IgE-binding, bee venom (BV) allergen.
METHODS
BV proteins were separated by using size exclusion chromatography and HPLC. IgE antibody binding to purified proteins was analyzed by means of immunoblotting, and T-cell response was analyzed by means of proliferation assay. Amino acid sequence was determined with 2 approaches, namely Edman degradation and carboxy terminal analysis with mass spectrometry.
RESULTS
Api m 6, which migrated as an 8-kd band in SDS-PAGE, was frequently (42%) recognized by IgE from BV-hypersensitive patients. In addition, PBMCs from BV-hypersensitive patients, as well as from a normal control subject, proliferated in response to this allergen. Api m 6 exists as 4 isoforms of 7190, 7400, 7598, and 7808 d, respectively. Amino acid sequences obtained from HPLC-purified preparations revealed that the isoforms were constituted of a common central core of 67 residues, only differing in the amino- and carboxy-terminal ends. Api m 6 showed no significant sequence homology with known proteins.
CONCLUSIONS
We have identified and sequenced a new BV allergen that elicits a strong IgE and T-cell response in a large number of BV-hypersensitive patients. Api m 6 should be considered in the diagnostic and therapeutic approach of BV immunotherapy on the basis of peptides or recombinant proteins.
- Language
-
- English
- Open access status
- closed
- Identifiers
-
- DOI 10.1067/mai.2001.113867
- PMID 11344362
- Persistent URL
- https://sonar.ch/global/documents/93549
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