|
KMID : 1225720150070040376
|
|
Allergy, Asthma & Immunology Research : AAIR
2015 Volume.7 No. 4 p.376 ~ p.383
|
|
|
IgE-Binding Epitope Mapping and Tissue Localization of the Major American Cockroach Allergen Per a 2
|
|
Lee Mey-Fann
Chang Chia-Wei
Song Pei-Pong
Hwang Guang-Yuh
Lin Shyh-Jye
Chen Yi-Hsing
|
|
|
Abstract
|
|
|
Purpose: 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.
Methods: 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.
Results: 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 localized partly in the mouth and midgut of the cockroach, with the most intense staining observed in the hindgut, suggesting that the Per a 2 allergen might be excreted through the feces.
Conclusions: Information on the IgE-binding epitope of Per a 2 may be used for designing more specific diagnostic and therapeutic approaches to cockroach allergy.
|
|
|
KEYWORD
|
|
|
Cockroach allergy, IgE-binding epitope, synthetic peptide, specific immunotherapy, Per a 2
|
|
|
FullTexts / Linksout information
|
|
|
|
|
Listed journal information
|
|
   
|
|