RESUMEN
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).
Asunto(s)
Alérgenos/genética , Alérgenos/inmunología , Hialuronoglucosaminidasa/inmunología , Inmunoglobulina E/inmunología , Venenos de Avispas/enzimología , Venenos de Avispas/inmunología , Avispas/inmunología , Animales , Especificidad de Anticuerpos , Clonación Molecular , Reacciones Cruzadas , Humanos , Hipersensibilidad/inmunología , Cuerpos de Inclusión/inmunología , Proteínas Recombinantes/biosíntesis , Proteínas Recombinantes/inmunología , Venenos de Avispas/genéticaRESUMEN
A urticária é uma condição frequente; consiste de lesões eritematopapulosas pruriginosas, isoladas ou agrupadas, fugazes, geralmente circulares, podendo variar em forma e tamanho. Convencionalmente, a urticária pode ser dividida, quanto a sua duração, em duas formas: aguda e crônica. Na forma crônica as lesões estão presentes diariamente ou quase diariamente, permanecendo menos de 24 horas na maior parte dos casos, durante um período superior a seis semanas, frequentemente tendo impacto na qualidade de vida. A anti-IgE é um anticorpo monoclonal humanizado aprovado para o uso em asma de difícil controle. Atualmente, várias linhas de evidência indicam que a anti-IgE pode ser benéfica no tratamento da urticária espontânea crônica e física. Neste artigo revisamos as principais e atuais publicações sobre o uso de anti-IgE para o tratamento da urticária crônica refratária aos tratamentos convencionais. Outros estudos ainda são necessários para melhor compreender os mecanismos envolvidos na resposta favorável a esta terapia.
Urticaria is a common condition, consisting of fleeting erythematopapulous pruritic lesions, appearing isolated or grouped, usually circular, but varying in shape and size. Conventionally, hives can be divided into acute and chronic, according to their duration. In the chronic form, lesions are present daily or almost daily, remaining present for less than 24 hours in most cases, over a period longer than 6 weeks, usually with an impact on quality of life. Anti-immunoglobulin E (anti-IgE) is a humanized monoclonal antibody approved for use in difficult-to-control asthma. Currently, several lines of evidence indicate that anti-IgE antibodies can be beneficial in the treatment of chronic spontaneous and physical urticaria. In this paper, we review major and current publications on the use of anti-IgE for the treatment of chronic urticaria refractory to conventional treatment. Further studies are needed to better understand the mechanisms involved in the favorable response of chronic urticaria to this therapy.