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

• Published in: Toxicon (Oxford) Vol. 82; pp. 104 - 111
• Main Authors: Justo Jacomini, Débora Laís, Gomes Moreira, Susana Margarida, Campos Pereira, Franco Dani, Zollner, Ricardo de Lima, Brochetto Braga, Márcia Regina
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01-05-2014
• Subjects: Allergen; Allergens - genetics; Allergens - immunology; Animals; Antibody Specificity; Cloning, Molecular; Cross Reactions; Cross-immunoreactivity; Humans; Hyaluronoglucosaminidase - immunology; Hypersensitivity - immunology; Immunoglobulin E (IgE); Immunoglobulin E - immunology; Inclusion Bodies - immunology; Polybia paulista; Recombinant hyaluronidase; Recombinant Proteins - biosynthesis; Recombinant Proteins - immunology; Venom; Wasp Venoms - enzymology; Wasp Venoms - genetics; Wasp Venoms - immunology; Wasps - immunology; Recombinant hyaluronidase; Polybia paulista; Cross-immunoreactivity; Immunoglobulin E (IgE); Venom; Allergen
• ISSN: 0041-0101; 1879-3150
• DOI: 10.1016/j.toxicon.2014.02.016

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 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).
•First time that a Brazilian social wasp had its venom allergen Pp-Hyal heterologously expressed.•Pp-Hyal-rec was expressed in Escherichia coli DE3 (BL21) cells in insoluble form.•Specific IgEs from allergic patients recognized Pp-Hyal rec at a higher intensity than the native allergen.•Pp-Hyal-rec allergen is a potential candidate to allergy diagnosis and immunotherapy.