Clonagem e expressão do gene da glicoproteína D do herpesvírus equídeo tipo 1 em Pichia pastoris e avaliação de sua imunogenicidade

Abstract

Equine herpesvirus 1 (EHV-1) has a worldwide distribution and causes serious damage to the equidoculture and the productive chain of this agribusiness, as an agent of outbreaks of respiratory, reproductive and neurological disease, in young and adult animals. EHV-1 glycoprotein D (gDEHV-1) is an enveloped glycoprotein essential for the agent penetration into the host cells and has already demonstrated the ability to induce protection against this agent in murine and equine models. The objectives of this work were: to clone, express and demonstrate the immunogenicity of gDEHV-1 expressed in Pichia pastoris, in a murine model. For this, the immune response induced by two vaccines was evaluated: one formulated with gDEHV-1 and another with gDEHV-1 associated with EHV-1; compared to three vaccines: one commercial, formulated with EHV-1 associated with EHV-4, another experimental, formulated with EHV-1, and the control, formulated with PBS. In all vaccines the adjuvant used was 10% aluminum hydroxide. The results showed that recombinant D-glycoprotein EHV-1, expressed in P. pastoris, demonstrated its antigenicity and immunogenicity. Also, it conserved epitopes that allowed its identification by commercial and experimental vaccine-induced anti-EHV-1 specific antibodies formulated with inactivated whole virus. The experimental vaccine containing inactivated whole virus (EHV-1) plus the recombinant D-glycoprotein of EHV-1 was significantly superior in the induction of specific antibodies against gDEHV-1 in the indirect ELISA when compared to the vaccines containing whole virus or subunits of the virus.