title: Reactivity of IgE to the allergen hyaluronidase from Polybia paulista (Hymenoptera, Vespidae) venom.
abstract: To date, there are no allergenic extracts or components available in Brazil to diagnosis and treatment of patients with venom allergy from social wasp (Vespidae Family; Polistinae Subfamily) despite of the great number of existing species. We evaluated the immunogenic potential of the Hyal recombinant protein (Pp-Hyal-rec) which was expressed in an insoluble form in comparison with the allergenic native protein (Pp-Hyal-nat) for recognition of immunoglobulin E (IgE) in the serum of allergic patients to venom of the endemic social wasp Polybia paulista from São Paulo State, Brazil. Hyal cDNA from the venom of the social wasp P. paulista (Pp-Hyal) (GI: 302201582) was cloned into the expression vector pET-28a in Escherichia coli DE3 (BL21) cells. Solubilization and purification of Pp-Hyal-rec from inclusion bodies were performed using Ni(2+) affinity chromatography (Ni-NTA-Agarose) under denaturing conditions. Both the native (Pp-Hyal-nat) and the recombinant (Pp-Hyal-rec) purified allergens were used for Western blotting to assess the levels of Pp-Hyal-IgE specific in the serum of 10 patients exclusively reactive to the venom of the social wasp P. paulista. The immune sera specifically recognized the band corresponding to the Pp-Hyal-rec protein (40 kDa) at a higher intensity than the native allergen (39 kDa). The sera recognized other proteins in P. paulista crude venom extract to a lesser extent, likely corresponding to other venom allergens such as phospholipase (34 kDa), Antigen 5 (25 kDa), and proteases. The recognition pattern of the immune sera to the Pp-Hyal-rec allergen strongly suggests that this recombinant antigen could be used for developing a diagnostic allergy test as well as for specific immunotherapy (IT).
authors.affiliation: Laboratório de Biologia Molecular de Artrópodes-LBMA-IB-RC-UNESP (Univ Estadual Paulista), Av. 24-A, n° 1515, CEP 13506-900, Bela Vista, Rio Claro, SP, Brazil.
authors.forename: Débora Laís
authors.initials: DL
authors.lastname: Justo Jacomini
authors.affiliation: Centro de Biociências, Universidade Federal do Rio Grande do Norte, Av. Sen. Salgado Filho, 3000, CEP 59072-970, Candelária, Natal, RN, Brazil. Electronic address: ssmargarida@gmail.com.
authors.forename: Susana Margarida
authors.initials: SM
authors.lastname: Gomes Moreira
authors.affiliation: Laboratório de Biologia Molecular de Artrópodes-LBMA-IB-RC-UNESP (Univ Estadual Paulista), Av. 24-A, n° 1515, CEP 13506-900, Bela Vista, Rio Claro, SP, Brazil.
authors.forename: Franco Dani
authors.initials: FD
authors.lastname: Campos Pereira
authors.affiliation: Laboratório de Imunologia e Alergia Experimental-LIAE, Departamento de Clínica Médica, Faculdade de Ciências Médicas, FCM, Universidade Estadual de Campinas-UNICAMP, Rua Tessália Vieira de Camargo n° 126, Cidade Universitária "Zeferino Vaz", CEP 13083-887 Campinas, SP, Brazil. Electronic address: zollner@unicamp.br.
authors.forename: Ricardo de Lima
authors.initials: Rde L
authors.lastname: Zollner
authors.affiliation: Laboratório de Biologia Molecular de Artrópodes-LBMA-IB-RC-UNESP (Univ Estadual Paulista), Av. 24-A, n° 1515, CEP 13506-900, Bela Vista, Rio Claro, SP, Brazil; Centro de Estudos de Venenos e Animais Peçonhentos-CEVAP (Univ Estadual Paulista), Rua José Barbosa de Barros, 1780, Fazenda Experimental Lageado, CEP 18610-307 Botucatu, SP, Brazil. Electronic address: mrbbraga@rc.unesp.br.
authors.forename: Márcia Regina
authors.initials: MR
authors.lastname: Brochetto Braga
mesh_terms: D000485:Allergens / Q000235:genetics* / Q000276:immunology*; D000818:Animals; D000918:Antibody Specificity; D003001:Cloning, Molecular; D003429:Cross Reactions; D006801:Humans; D006821:Hyaluronoglucosaminidase / Q000276:immunology*; D006967:Hypersensitivity / Q000276:immunology; D007073:Immunoglobulin E / Q000276:immunology*; D002479:Inclusion Bodies / Q000276:immunology; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology; D014862:Wasp Venoms / Q000201:enzymology* / Q000235:genetics / Q000276:immunology*; D014863:Wasps / Q000276:immunology*
chemical_list: D000485:Allergens; D011994:Recombinant Proteins; D014862:Wasp Venoms; D007073:Immunoglobulin E; D006821:Hyaluronoglucosaminidase
keywords: Allergen; Cross-immunoreactivity; Immunoglobulin E (IgE); Polybia paulista; Recombinant hyaluronidase; Venom