Identification of a new allergen from amaranthus retroflexus pollen, ama r 2

Tehrani, M. and Sankian, M. and Assarehzadegan, M. A. and Falak, R. and Noorbakhsh, R. and Moghadam, M. and Jabbari, F. and Varasteh, A. (2011) Identification of a new allergen from amaranthus retroflexus pollen, ama r 2. Allergology International, 60 (3). pp. 309-316.

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Abstract

Background: Pollinosis from Amaranthus retroflexus pollen is a common cause of respiratory allergy in Iran with a high positive rate (68.8) among Iranian allergic patients. The aim of the present study was to evaluate the allergenicity of the A. retroflexus pollen profilin. Methods: Using sera from twelve patients allergic to A. retroflexus pollen, IgE-binding proteins from the A. ret-roflexus pollen extract was identified by immunoblotting. The cDNA of A. retroflexus pollen profilin was amplified, then cloned into the pET-21b (+) vector, expressed in Escherichia coli, and finally purified by metal affinity chromatography. The IgE-binding capacity of the recombinant protein was then analyzed by the ELISA, im-munoblotting, and inhibition assays, as well as by the skin prick test (SPT). Results: Immunoblotting results indicated a 14.6 kDa protein with IgE-reactivity to 33 (4/12) among A. retroflexus pollen-allergic patients. Nucleotide sequencing of the cDNA revealed an open reading frame of 399 bp encoding for 133 amino acid residues which was belonged to the profilin family and designated as Ama r 2. A recombinant Ama r 2 (rAma r 2) was then produced in E. coli as a soluble protein which showed a strong IgE-reactivity via ELISA confirmed by the SPT. Inhibition experiments revealed high IgE cross-reactivities with the profilins from other plants. Conclusions: The profilin from the A. retroflexus pollen, Ama r 2, was firstly identified as an allergen. Moreover, rAma r 2 was produced in E. coli as a soluble immunoreactive protein with an IgE-reactivity similar to that of its natural counterpart. © 2011 Japanese Society of Allergology.

Item Type: Article
Additional Information: Cited By :17 Export Date: 16 February 2020 CODEN: ALINF Correspondence Address: Varasteh, A.; Immunology Research Center, Bu-Ali Research Institute, Mashhad University of Medical Sciences, Mashhad 9196773117, Iran; email: varasteha@mums.ac.ir
Uncontrolled Keywords: Allergen characterization Ama r 2 Amaranthus retroflexus Cloning Profilin Ama r2 pollen amino acid complementary DNA immunoglobulin E pollen antigen unclassified drug adult allergenicity Amaranthus retroflexus pollen article binding affinity chromatography clinical article cloning vector controlled study cross reaction DNA sequence enzyme linked immunosorbent assay Escherichia coli female human immunoblotting immunoreactivity male open reading frame pollen prick test priority journal
Subjects: QW Microbiology and Immunology
Divisions: Mashhad University of Medical Sciences
Depositing User: mr lib7 lib7
Date Deposited: 29 Apr 2020 05:30
Last Modified: 29 Apr 2020 05:30
URI: http://eprints.mums.ac.ir/id/eprint/17137

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