Development of orf virus as a bifunctional recombinant vaccine: Surface display of Echinococcus granulosus antigen EG95 by fusion to membrane structural proteins

The parapoxvirus, orf virus (ORFV) causes superficial skin lesions in infected sheep. Unattenuated ORFV is used globally to vaccinate against orf. Recombinant poxviruses are proven delivery systems and we investigated strategies to express the immunogenic Echinococcus granulosus peptide EG95 from ORFV with the aim of developing a recombinant bivalent vaccine. EG95 is an oncosphere protein of the cestode E. granulosus, a parasite responsible for causing cystic hydatid disease in a wide range of hosts including humans and grazing animals such as sheep. Recombinant viruses were produced in which EG95 was expressed by itself or fused to ORFV envelope-associated structural proteins 10 kDa and F1L. Infection studies in sheep showed that specific antibodies were produced against ORFV and EG95 and that the antibody levels against EG95 were comparable to that of animals immunized with purified EG95 in Quil A adjuvant, an immunization regime that is known to afford protection. A single exposure to the dual vaccine has potential for protecting lambs against orf and for priming against EG95 so as to respond strongly to a later injection of EG95 protein.

► An immunogenic peptide EG95 from the tapeworm Echinococcus granulosus protects sheep against cystic hydatid disease.
► The parapoxvirus orf virus was modified to display EG95 on the virion surface or to produce the EG95 ‘stand-alone’ peptide.
► Sheep infected with orf virus recombinants produced antibodies against EG95 and orf virus.
► A single exposure to the dual vaccine has the potential to protect lambs against orf and for priming against EG95.
► Orf virus has the potential to be developed as a recombinant bivalent vaccine.