IgE-Binding Epitope Mapping and Tissue Localization of the Major American Cockroach Allergen Per a 2

Allergy, Asthma & Immunology Research
M F LeeYi-Hsing Chen

Abstract

Cockroaches are the second leading allergen in Taiwan. Sensitization to Per a 2, the major American cockroach allergen, correlates with clinical severity among patients with airway allergy, but there is limited information on IgE epitopes and tissue localization of Per a 2. This study aimed to identify Per a 2 linear IgE-binding epitopes and its distribution in the body of a cockroach. The cDNA of Per a 2 was used as a template and combined with oligonucleotide primers specific to the target areas with appropriate restriction enzyme sites. Eleven overlapping fragments of Per a 2 covering the whole allergen molecule, except 20 residues of signal peptide, were generated by PCR. Mature Per a 2 and overlapping deletion mutants were affinity-purified and assayed for IgE reactivity by immunoblotting. Three synthetic peptides comprising the B cell epitopes were evaluated by direct binding ELISA. Rabbit anti-Per a 2 antibody was used for immunohistochemistry. Human linear IgE-binding epitopes of Per a 2 were located at the amino acid sequences 57-86, 200-211, and 299-309. There was positive IgE binding to 10 tested Per a 2-allergic sera in 3 synthetic peptides, but none in the controls. Immunostaining revealed that Per a 2 was localize...Continue Reading

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Citations

Dec 27, 2015·Allergy·D C DoP Gao
Oct 30, 2016·Annals of Allergy, Asthma & Immunology : Official Publication of the American College of Allergy, Asthma, & Immunology·Mey-Fann LeePei-Pong Song
Apr 20, 2017·Current Allergy and Asthma Reports·Anna PomésL Karla Arruda
Nov 15, 2016·International Journal of Molecular Medicine·Xunliang TongJi-Fu Wei

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Datasets Mentioned

BETA
GM-CSF
GU188391

Methods Mentioned

BETA
ELISA
PCR
column chromatography
enzyme-linked immunosorbent assay

Software Mentioned

Oligo
exe structure

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