Non-transmissible pseudorabies virus gp50 mutants: a new generation of safe live vaccines

Abstract

Envelope glycoprotein gp50 of pseudorabies virus (PRV) is essential for virus entry, but is not required for subsequent steps in the viral replication cycle. Phenotypically-complemented gp50 null mutants can infect cells and can spread, both in vitro and in vivo, by direct cell-to-cell transmission. However, progeny virions released by the infected cells are non-infectious because they lack gp50. Therefore, these viruses cannot be transmitted from infected animals to contact animals. These properties could make PRV gp50 null mutants attractive candidates as safe non-transmissible live vaccines. To establish whether phenotypically-complemented PRV gp50 null mutants and gp50+gp63 double mutants could be used as live vaccines against Aujeszky's disease, the virulence and immunogenicity of these mutants were tested in pigs. Our results show that a gp50 null mutant has a greatly reduced virulence for pigs, and that pigs immunized with such a mutant were protected from clinical signs of Aujeszky's disease after a challenge inoculation with the virulent wild-type PRV strain NIA-3. PRV gp50+gp63 deletion mutants proved to be non-virulent for pigs and were somewhat less immunogenic, since immunized animals showed some fever and growth retardation after challenge inoculation. Replication of wild-type challenge virus was significantly reduced, but could not completely be prevented, in pigs immunized with a gp50 null mutant, and was reduced less in pigs immunized with a gp50+gp63 deletion mutant. Furthermore, infectious virus could not be recovered from oropharyngeal fluid or tissues from pigs inoculated with a gp50 null mutant or a gp50+gp63 deletion mutant. These results indicate that PRV gp50 null mutants and gp50+gp63 deletion mutants may be used as safe non-transmissible live vaccines against Aujeszky's disease, or as safe carrier vaccines for the delivery of heterologous gene products.