rAed a 4: A New 67-kDa Aedes aegypti Mosquito Salivary Allergen for the Diagnosis of Mosquito Allergy
| Autor: | Peng, Zhikang, Caihe, Li, Beckett, Andrew N, Guan, Qingdong, James, Anthony A, Simons, F Estelle R |
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| Rok vydání: | 2016 |
| Předmět: |
Allergy
Immunology Food Allergies Mosquito allergy Enzyme-Linked Immunosorbent Assay Aedes aegypti Aedes Clinical Research parasitic diseases Hypersensitivity Animals Humans Salivary Proteins and Peptides Skin Tests Inflammatory and immune system fungi Insect Bites and Stings Immunoglobulin E Allergens Recombinant Proteins Vector-Borne Diseases Infectious Diseases Immunoglobulin G Insect Proteins Salivary allergen/antigen Aed a 4 |
| Zdroj: | International archives of allergy and immunology, vol 170, iss 3 |
| Popis: | BackgroundAccurate diagnosis of mosquito allergy has been hampered by the laborious task of obtaining mosquito salivary allergens. We have previously studied 3 recombinant (r) Aedes aegypti mosquito salivary allergens: rAed a 1, rAed a 2 and rAed a 3. Here, we report the expression, purification, identification and evaluation of rAed a 4, a 67-kDa α-glucosidase.MethodsrAed a 4 was expressed using a baculovirus/insect cell system, purified by a combination of anion- and cation-exchange chromatography, and identified by immunoblotting. A. aegypti saliva extract was prepared in our laboratory. An indirect enzyme-linked immunosorbent assay (ELISA) was developed to measure rAed a 4-specific immunoglobulin E (IgE) and IgG antibodies in sera from 13 individuals with a positive mosquito-bite test from a laboratory-reared mosquito. Sera from 18 individuals with a negative bite test served as controls.ResultsPurified rAed a 4 bound to the IgE in mosquito-allergic sera, as detected by ELISA and immunoblotting. The binding of rAed a 4 to IgE could be inhibited in a dose-dependent manner by the addition of an A. aegypti extract. Mosquito-allergic individuals had significantly higher mean levels of rAed a 4-specific IgE and IgG than controls. Using the mean of the controls ± 2 SD as a cut-off level, 46% of the 13 allergic individuals had a positive IgE, while none of the controls was positive (p < 0.001).ConclusionsAed a 4 is a major allergen in mosquito saliva. Its recombinant form has the hydrolase function and can be used for the diagnosis of mosquito allergy. |
| Databáze: | OpenAIRE |
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