title: Mosquito salivary allergen Aed a 3: cloning, comprehensive molecular analysis, and clinical evaluation.
abstract: 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.
  
  
  
  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.
  
  
  
  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.
  
  
  
  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.
authors.affiliation: Department of Pediatrics and Child Health, University of Manitoba, Winnipeg, MB, Canada.|Department of Immunology, University of Manitoba, Winnipeg, MB, Canada.
authors.forename: Z
authors.initials: Z
authors.lastname: Peng
authors.affiliation: Department of Pediatrics and Child Health, University of Manitoba, Winnipeg, MB, Canada.|Department of Biochemistry and Medical Genetics, University of Manitoba, Winnipeg, MB, Canada.
authors.forename: W W
authors.initials: WW
authors.lastname: Xu
authors.affiliation: Center for Drug Design, Academic Health Center, University of Minnesota, Minneapolis, MN, USA.
authors.forename: Y
authors.initials: Y
authors.lastname: Sham
authors.affiliation: Department of Pediatrics and Child Health, University of Manitoba, Winnipeg, MB, Canada.
authors.forename: H
authors.initials: H
authors.lastname: Lam
authors.affiliation: Department of Pediatrics and Child Health, University of Manitoba, Winnipeg, MB, Canada.
authors.forename: D
authors.initials: D
authors.lastname: Sun
authors.affiliation: Department of Pediatrics and Child Health, University of Manitoba, Winnipeg, MB, Canada.
authors.forename: L
authors.initials: L
authors.lastname: Cheng
authors.affiliation: Department of Pediatrics and Child Health, University of Manitoba, Winnipeg, MB, Canada.
authors.forename: N F
authors.initials: NF
authors.lastname: Rasic
authors.affiliation: Department of Pediatrics and Child Health, University of Manitoba, Winnipeg, MB, Canada.|Department of Immunology, University of Manitoba, Winnipeg, MB, Canada.
authors.forename: Q
authors.initials: Q
authors.lastname: Guan
authors.affiliation: Departments of Microbiology & Molecular Genetics and Molecular Biology & Biochemistry, University of California, Irvine, CA, USA.
authors.forename: A A
authors.initials: AA
authors.lastname: James
authors.affiliation: Department of Pediatrics and Child Health, University of Manitoba, Winnipeg, MB, Canada.|Department of Immunology, University of Manitoba, Winnipeg, MB, Canada.
authors.forename: F E R
authors.initials: FE
authors.lastname: Simons
mesh_terms: D000328:Adult; D000330:Aedes; D000485:Allergens / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D018076:DNA, Complementary; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D007073:Immunoglobulin E / Q000276:immunology; D007299:Insect Bites and Stings / Q000175:diagnosis / Q000276:immunology; D019476:Insect Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D008958:Models, Molecular; D011485:Protein Binding; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D012471:Salivary Proteins and Peptides / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D012882:Skin Tests
chemical_list: D000485:Allergens; D018076:DNA, Complementary; D019476:Insect Proteins; D011994:Recombinant Proteins; D012471:Salivary Proteins and Peptides; D007073:Immunoglobulin E
keywords: Aed a 3; Aedes aegypti; insect allergy; mosquito; salivary allergen/antigen