IgE-Binding Epitope Mapping and Tissue Localization of the Major American Cockroach Allergen Per a 2
Allergy, Asthma & Immunology Research
;
: 376-383, 2015.
Article
in English
| WPRIM
| ID: wpr-89603
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.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Peptides
/
Periplaneta
/
Taiwan
/
Protein Sorting Signals
/
Immunoglobulin E
/
Enzyme-Linked Immunosorbent Assay
/
Immunohistochemistry
/
Immunoblotting
/
Polymerase Chain Reaction
/
Amino Acid Sequence
Limits:
Humans
Country/Region as subject:
Asia
Language:
English
Journal:
Allergy, Asthma & Immunology Research
Year:
2015
Type:
Article
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