Reactivity of IgE to the allergen hyaluronidase from Polybia paulista (Hymenoptera, Vespidae) venom

Autor(es)

Justo Jacomini, Debora Lais ^[1] ; Gomes Moreira, Susana Margarida ^[2] ; Campos Pereira, Franco Dani ^[1] ; Zollner, Ricardo de Lima ^[3] ; Brochetto Braga, Marcia Regina ^{[1, 4]}

Afiliação do(s) autor(es)

^[1] Univ Estadual Paulista, Lab Biol Mol Artropodes LBMA IB RC UNESP, BR-13506900 Rio Claro, SP - Brazil

^[2] Univ Fed Rio Grande do Norte, Ctr Biociencias, BR-59072970 Natal, RN - Brazil

^[3] Univ Estadual Campinas, UNICAMP, Fac Ciencias Med, Lab Imunol & Alergia Expt LIAE, Dept Clin Med, FCM, BR-13083887 Campinas, SP - Brazil

^[4] Univ Estadual Paulista, Ctr Estudos Venenos & Anim Peconhentos CEVAP, BR-18610307 Botucatu, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: Toxicon; v. 82, p. 104-111, MAY 2014.
• Citações Web of Science: 12

Resumo

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 Sao 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 Ni2+ 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). (C) 2014 Elsevier Ltd. All rights reserved. (AU)

• Processo FAPESP: 09/51539-1 - Analise imunologica comparada da eficiencia e especificidade da hialuronidase recombinante de polybia paulista (hymenoptera: vespidae) expressa em bacteria e levedura
• Beneficiário: Debora Lais Justo Jacomini
• Modalidade de apoio: Bolsas no Brasil - Doutorado