IgE-Binding Epitope Mapping and Tissue Localization of the Major American Cockroach Allergen Per a 2
Allergy, Asthma & Immunology Research
; : 376-383, 2015.
Article
em En
| WPRIM
| ID: wpr-89603
Biblioteca responsável:
WPRO
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.Palavras-chave
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Assunto principal:
Peptídeos
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Periplaneta
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Taiwan
/
Sinais Direcionadores de Proteínas
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Imunoglobulina E
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Ensaio de Imunoadsorção Enzimática
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Imuno-Histoquímica
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Immunoblotting
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Reação em Cadeia da Polimerase
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Sequência de Aminoácidos
Limite:
Humans
País/Região como assunto:
Asia
Idioma:
En
Revista:
Allergy, Asthma & Immunology Research
Ano de publicação:
2015
Tipo de documento:
Article