Identification of the immunodominant antigens from the venom of the social wasps Agelaia pallipes pallipes and Polybia paulista (Hymenoptera, Vespidae)

Abstract

Some patients allergic to vespid venoms present negative specific IgE tests when using the available extracts (imported from the producers countriesin the North Hemisphere) and can not be properly cared about their allergy to tropical wasps venom. Thus, it is important develop standadized venom extracts with the venoms of the local species (neotropical social wasps). In aditoin to this, it is necessary to investigate the lack of cross-reactivity betweem the venoms of wasps species endemic from temperate and tropical climates. This study intends to determine the major allergenic proteins of the venoms from Agelaia pallipes venom and Polybia paulista(the most common species in souhteast Brazil and therefore those which cause the most of stinfging accidents) using a proteomic approach to identify the immunodominant proteins of these venoms. Methods: The venoms of Agelaia pallipes pallipes and Polybia paulista will be submitted to bidimensional electrophoresis using immobilized pH gradient strips (3.0 -10.0) for the first dimension and 15% PAGE gel for the second dimension. Gels will made in duplicate. One of them will be transferred to a PVDF membrane and incubated with the serum from a sensitized patient diluted 1:5. Immunoblot detection was performed with biotinilated human anti-IgE and ECL (Enhanced chemiluminescence). On the other gel, after Coomassie Blue staining, the spots corresponding to the proteins recognized by specific IgE in the immunoblotting will be processed and analyzed using a MALDI-ToF and ESI-MS/MS mass spectrometry. Identification wil be performed with the use of proteomic bioinformatic tools. (AU)