Mosquito salivary allergen Aed a 3: cloning, comprehensive molecular analysis, and clinical evaluation

Allergy. 2016 May;71(5):621-8. doi: 10.1111/all.12812. Epub 2016 Mar 6.

Abstract

Background: Allergic reactions to mosquito bites are an increasing clinical concern. Due to the lack of availability of mosquito salivary allergens, they are underdiagnosed. Here, we reported a newly cloned mosquito Aedes (Ae.) aegypti salivary allergen.

Methods: A cDNA encoding a 30-kDa Ae. aegypti salivary protein, designated Aed a 3, was isolated from an expression library. The full-length cDNA was cloned into a baculovirus expression vector, and recombinant Aed a 3 (rAed a 3) was expressed, purified, and characterized. Skin prick tests with purified rAed a 3 and Ae. aegypti bite tests were performed in 43 volunteers. Serum rAed a 3-specific IgE levels were measured in 28 volunteers.

Results: The primary nucleotide sequence, deduced amino acid sequence, and IgE-binding sites of Aed a 3 were identified. rAed a 3-selected antibodies recognized a 30-kDa Ae. aegypti saliva protein. rAed a 3 bound IgE in mosquito-allergic volunteers and the binding could be inhibited by the addition of natural mosquito extract dose dependently. Immediate skin test reactions to rAed a 3 correlated significantly with mosquito bite-induced reactions. Of the bite test-positive volunteers, 32% had a positive rAed a 3 skin test and 46% had specific IgE. No bite test-negative volunteers reacted to rAed a 3 in either the skin tests or the IgE assays, confirming the specificity of the assay.

Conclusions: Aed a 3 that corresponds to the Aegyptin protein is a major mosquito salivary allergen. Its recombinant form has biological activity and is suitable for use in skin tests and specific IgE assays in mosquito-allergic individuals.

Keywords: Aed a 3; Aedes aegypti; insect allergy; mosquito; salivary allergen/antigen.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aedes
  • Allergens / chemistry
  • Allergens / genetics*
  • Allergens / immunology*
  • Amino Acid Sequence
  • Animals
  • Cloning, Molecular
  • DNA, Complementary
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • Humans
  • Immunoglobulin E / immunology
  • Insect Bites and Stings / diagnosis
  • Insect Bites and Stings / immunology
  • Insect Proteins / chemistry
  • Insect Proteins / genetics*
  • Insect Proteins / immunology*
  • Models, Molecular
  • Protein Binding
  • Protein Conformation
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / immunology
  • Salivary Proteins and Peptides / chemistry
  • Salivary Proteins and Peptides / genetics*
  • Salivary Proteins and Peptides / immunology*
  • Sensitivity and Specificity
  • Sequence Analysis, DNA
  • Skin Tests

Substances

  • Allergens
  • DNA, Complementary
  • Insect Proteins
  • Recombinant Proteins
  • Salivary Proteins and Peptides
  • Immunoglobulin E