title: Recombinant phospholipase A1 (Ves v 1) from yellow jacket venom for improved diagnosis of hymenoptera venom hypersensitivity.
abstract: BACKGROUND
  Hymenoptera venoms are known to cause life-threatening IgE-mediated anaphylactic reactions in allergic individuals. Proper diagnosis of hymenoptera venom allergy using venom extracts is severely affected by molecular cross-reactivities. Although non-glycosylated marker allergens would facilitate the identification of the culprit venom, the major allergen phospholipase A1 (Ves v 1) from yellow jacket venom (YJV) remained unavailable so far.
  
  
  METHODS
  Expression of Ves v 1 as wild type and enzymatically inactivated mutant and Ves v 5 in insect cells yielded soluble proteins that were purified via affinity chromatography. Functionality of the recombinant allergens was assessed by enzymatic and biophysical analyses as well as basophil activation tests. Diagnostic relevance was addressed by ELISA-based analyses of sera of YJV-sensitized patients.
  
  
  RESULTS
  Both major allergens Ves v 1 and Ves v 5 could be produced in insect cells in secreted soluble form. The recombinant proteins exhibited their particular biochemical and functional characteristics and were capable for activation of human basophils. Assessment of IgE reactivity of sera of YJV-sensitized and double-sensitized patients emphasised the relevance of Ves v 1 in hymenoptera venom allergy. In contrast to the use of singular molecules the combined use of both molecules enabled a reliable assignment of sensitisation to YJV for more than 90% of double-sensitised patients.
  
  
  CONCLUSIONS
  The recombinant availability of Ves v 1 from yellow jacket venom will contribute to a more detailed understanding of the molecular and allergological mechanisms of insect venoms and may provide a valuable tool for diagnostic and therapeutic approaches in hymenoptera venom allergy.
authors.affiliation: Institute of Biochemistry and Molecular Biology, Department of Chemistry, University of Hamburg, Germany. ollert@lrz.tu-muenchen.de.
authors.forename: Henning
authors.initials: H
authors.lastname: Seismann
authors.forename: Simon
authors.initials: S
authors.lastname: Blank
authors.forename: Liliana
authors.initials: L
authors.lastname: Cifuentes
authors.forename: Ingke
authors.initials: I
authors.lastname: Braren
authors.forename: Reinhard
authors.initials: R
authors.lastname: Bredehorst
authors.forename: Thomas
authors.initials: T
authors.lastname: Grunwald
authors.forename: Markus
authors.initials: M
authors.lastname: Ollert
authors.forename: Edzard
authors.initials: E
authors.lastname: Spillner