Tags

Type your tag names separated by a space and hit enter

Immune recognition of novel isoforms and domains of the mugwort pollen major allergen Art v 1.
Mol Immunol. 2009 Jan; 46(3):416-21.MI

Abstract

Allergen isoforms can differ in their IgE and T cell recognition patterns, and thus might have an impact on the selection of candidates for molecule-based diagnostic and therapeutic approaches. The present study aimed at the identification and characterization of isoforms of Art v 1, the mugwort pollen major allergen. In addition, single Art v 1 domains were physicochemically and immunologically characterized. For this purpose, the Art v 1 cDNA was radiolabeled and used to screen a mugwort pollen cDNA library. Positive clones were sequenced and used for the production of recombinant proteins in Escherichia coli using the pHIS-Parallel2 vector. Protein purification was performed by affinity- and ion exchange chromatography. Antibody binding to the recombinant proteins was determined by immunoblot, ELISA, cross-inhibition experiments, and mediator release assays. We could identify 7 Art v 1 isoforms differing in 1-6 amino acid residues. Interestingly, all amino acid variations were restricted to the proline domain carrying the molecule's post-translational modifications. No significant difference in IgG or IgE reactivity could be observed between Art v 1 isoforms and the defensin domain produced in E. coli. When expressed in E. coli, the proline domain was not recognized by Art v 1-specific antibodies. Our results demonstrated that the relevant IgE epitopes of Art v 1 are located on the defensin domain and suggest the involvement of carbohydrates in the allergenicity of natural Art v 1. Plant-based expression systems could help to reveal possibly different glycosylation patterns and IgE binding properties of Art v 1 isoforms. These findings have direct implications on the development of novel tools for mugwort pollen allergy diagnosis and therapy.

Authors+Show Affiliations

Christian Doppler Laboratory for Allergy Diagnosis and Therapy, Department of Molecular Biology, University of Salzburg, Salzburg, Austria.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

19054564

Citation

Dedic, Azra, et al. "Immune Recognition of Novel Isoforms and Domains of the Mugwort Pollen Major Allergen Art V 1." Molecular Immunology, vol. 46, no. 3, 2009, pp. 416-21.
Dedic A, Gadermaier G, Vogel L, et al. Immune recognition of novel isoforms and domains of the mugwort pollen major allergen Art v 1. Mol Immunol. 2009;46(3):416-21.
Dedic, A., Gadermaier, G., Vogel, L., Ebner, C., Vieths, S., Ferreira, F., & Egger, M. (2009). Immune recognition of novel isoforms and domains of the mugwort pollen major allergen Art v 1. Molecular Immunology, 46(3), 416-21. https://doi.org/10.1016/j.molimm.2008.10.012
Dedic A, et al. Immune Recognition of Novel Isoforms and Domains of the Mugwort Pollen Major Allergen Art V 1. Mol Immunol. 2009;46(3):416-21. PubMed PMID: 19054564.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Immune recognition of novel isoforms and domains of the mugwort pollen major allergen Art v 1. AU - Dedic,Azra, AU - Gadermaier,Gabriele, AU - Vogel,Lothar, AU - Ebner,Christof, AU - Vieths,Stefan, AU - Ferreira,Fátima, AU - Egger,Matthias, Y1 - 2008/12/02/ PY - 2008/09/11/received PY - 2008/10/14/accepted PY - 2008/12/5/pubmed PY - 2009/3/12/medline PY - 2008/12/5/entrez SP - 416 EP - 21 JF - Molecular immunology JO - Mol. Immunol. VL - 46 IS - 3 N2 - Allergen isoforms can differ in their IgE and T cell recognition patterns, and thus might have an impact on the selection of candidates for molecule-based diagnostic and therapeutic approaches. The present study aimed at the identification and characterization of isoforms of Art v 1, the mugwort pollen major allergen. In addition, single Art v 1 domains were physicochemically and immunologically characterized. For this purpose, the Art v 1 cDNA was radiolabeled and used to screen a mugwort pollen cDNA library. Positive clones were sequenced and used for the production of recombinant proteins in Escherichia coli using the pHIS-Parallel2 vector. Protein purification was performed by affinity- and ion exchange chromatography. Antibody binding to the recombinant proteins was determined by immunoblot, ELISA, cross-inhibition experiments, and mediator release assays. We could identify 7 Art v 1 isoforms differing in 1-6 amino acid residues. Interestingly, all amino acid variations were restricted to the proline domain carrying the molecule's post-translational modifications. No significant difference in IgG or IgE reactivity could be observed between Art v 1 isoforms and the defensin domain produced in E. coli. When expressed in E. coli, the proline domain was not recognized by Art v 1-specific antibodies. Our results demonstrated that the relevant IgE epitopes of Art v 1 are located on the defensin domain and suggest the involvement of carbohydrates in the allergenicity of natural Art v 1. Plant-based expression systems could help to reveal possibly different glycosylation patterns and IgE binding properties of Art v 1 isoforms. These findings have direct implications on the development of novel tools for mugwort pollen allergy diagnosis and therapy. SN - 0161-5890 UR - https://www.unboundmedicine.com/medline/citation/19054564/Immune_recognition_of_novel_isoforms_and_domains_of_the_mugwort_pollen_major_allergen_Art_v_1_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0161-5890(08)00729-3 DB - PRIME DP - Unbound Medicine ER -