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Isolation, expression and immunological characterization of a calcium-binding protein from Parietaria pollen.
- Source :
-
Molecular immunology [Mol Immunol] 2008 May; Vol. 45 (9), pp. 2465-73. Date of Electronic Publication: 2008 Mar 04. - Publication Year :
- 2008
-
Abstract
- The diagnosis and therapy of allergic disorders are usually performed with crude extracts which are a heterogeneous mixture of proteins with different allergenic potency. The knowledge of the allergenic composition is a key step for diagnostic and therapeutic options. Parietaria judaica pollen represents one of the main sources of allergens in the Mediterranean area and its major allergens have already been identified (Par j 1 and Par j 2). In addition, inhibition studies performed using a calcium-binding protein (CBP) from grass pollen (Phl p 7) showed the presence of a homologue of this cross-reactive allergen in the Parietaria extract. Screening of a cDNA library allowed us to isolate a 480bp cDNA containing the information for an 87 AA long protein with high level of homology to calcium-binding proteins from other allergenic sources. It was expressed as a recombinant allergen in Escherichia coli and purified by affinity chromatography. Its expression allowed us to study the prevalence of this allergen in a population of allergic patients in southern Europe. Immunoblotting and inhibition studies showed that this allergen shares a pattern of IgE epitopes in common with other 2-EF-hand calcium-binding proteins from botanically non-related species. The immunological properties of the Pj CBP were investigated by CD63 activation assay and CFDA-SE staining. In conclusion, DNA recombinant technology allowed the isolation, expression and immunological characterization of a cross-reactive calcium-binding protein allergen from Parietaria judaica pollen.
- Subjects :
- Allergens genetics
Allergens immunology
Allergens metabolism
Amino Acid Sequence
Antigens, Plant immunology
Antigens, Plant metabolism
Base Sequence
Basophils metabolism
Calcium-Binding Proteins genetics
Calcium-Binding Proteins metabolism
Cell Proliferation
Cloning, Molecular
Humans
Immunoglobulin E metabolism
Leukocytes, Mononuclear immunology
Leukocytes, Mononuclear metabolism
Molecular Sequence Data
Pollen chemistry
Sequence Alignment
Allergens isolation & purification
Basophils immunology
Calcium-Binding Proteins immunology
Calcium-Binding Proteins isolation & purification
Immunoglobulin E immunology
Parietaria immunology
Pollen immunology
Subjects
Details
- Language :
- English
- ISSN :
- 0161-5890
- Volume :
- 45
- Issue :
- 9
- Database :
- MEDLINE
- Journal :
- Molecular immunology
- Publication Type :
- Academic Journal
- Accession number :
- 18289680
- Full Text :
- https://doi.org/10.1016/j.molimm.2008.01.009