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

doi:10.4168/aair.2015.7.4.376

中山醫學大學機構典藏 CSMUIR > 醫學科技學院 > 醫學檢驗暨生物技術學系暨碩士班 > 期刊論文 > Item 310902500/15909

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题名:	IgE-Binding Epitope Mapping and Tissue Localization of the Major American Cockroach Allergen Per a 2
作者:	MF, Lee CW, Chang PP, Song GY, Hwang SJ, Lin YH, Chen
贡献者:	中山醫學大學
关键词:	Cockroach allergy;IgE-binding epitope;Per a 2;specific immunotherapy;synthetic peptide
日期:	2015
上传时间:	2016-09-05T08:20:27Z (UTC)
ISSN:	2092-7355
摘要:	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.
URI:	http://dx.doi.org/10.4168/aair.2015.7.4.376 https://ir.csmu.edu.tw:8080/ir/handle/310902500/15909
關聯:	Allergy Asthma Immunol Res. 2015 Jul;7(4):376-83.
显示于类别:	[醫學檢驗暨生物技術學系暨碩士班] 期刊論文

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