Recombinant phospholipase A1 (Ves v 1) from yellow jacket venom for improved diagnosis of hymenoptera venom hypersensitivity

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.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.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.Hymenoptera stings may cause life-threatening and sometimes fatal IgE-mediated anaphylactic reactions with the major threat emanating from the yellow jacket V. vulgaris and the honeybee A. mellifera. Although venom immunotherapy is highly effective, an adequate diagnosis and identification of the culprit venom is hampered by the use of crude venoms for measurement of specific IgE levels. Thereby, the main problem arises from serologic double-positivity for A. mellifera and V. vulgaris venom of up to 50% of patients that have IgE against hymenoptera venoms [1]. Apart from true double-sensitisation this phenomenon is largely attributed to molecular cross-reactivity, either based on the presence of cross-reactive epitopes in homolo