Non-spreading pestivirus
View Patent ↗The invention relates to vaccines used in the eradication or control of pestivirus infections, particularly used in pigs or ruminants. The invention provides nucleic acid, pestivirus-like particles and a pestivirus vaccine, comprising the nucleic acid or particles, which is capable of eliciting a proper immune response without having the ability to spread throughout the vaccinated animal, thereby avoiding the negative consequences of viral spread. Preferably, the immunological response allows for serological discrimination between vaccinated animals and wild-type pestivirus infected animals.
1. A recombinant nucleic acid that, upon inoculation of an animal, can elicit an immune response directed against immunodominant parts of structural proteins of a pestivirus without having the ability to spread through the inoculated animal, said recombinant nucleic acid comprising a genome of a pestivirus encoding at least one structural protein or an immunodominant part thereof, said genome having a functional deletion in a sequence encoding at least one protein related to viral spread.
2. A cell permissive for pestivirus infection comprising:
a nucleic acid construct encoding at least one pestiviral protein or substantial part thereof related to viral spread; and
a pestiviral genome containing a defect complemented by said construct;
wherein said cell allows for packaging of said genome and pestiviral protein or substantial part thereof in a pestivirus-like particle.
3. The cell of claim 2 , wherein said at least one pestiviral protein or substantial part thereof related to viral spread is stably expressed.
4. A process of making pestivirus-like particles comprising:
transfecting a cell with a recombinant nucleic acid comprising a pestiviral genome having a functional deletion in a sequence encoding at least one pestiviral protein related to viral spread, said genome being capable of RNA replication in a suitable cell and encoding at least one structural protein of pestivirus or at least one immunodominant part thereof,
wherein said cell comprises a second nucleic acid construct encoding at least one pestiviral protein or a substantial part thereof related to viral spread and allowing packaging of said recombinant pestiviral genome in a pestivirus-like particle;
allowing said pestiviral genome to replicate in said cell;
allowing the replicated recombinant pestiviral genome to package into a pestivirus-like particle comprising the protein encoded by the second nucleic acid; and
harvesting said pestivirus-like particles.
5. The process according to claim 4 wherein the second nucleic acid construct encoding the at least one pestiviral protein or substantial part thereof related to viral spread is stably expressed.
6. A process of making a vaccine comprising:
transfecting a cell with a recombinant nucleic acid comprising a pestiviral genome having a functional deletion in a sequence encoding at least one pestiviral protein related to viral spread, said genome being capable of RNA replication in a suitable cell and encoding at least one structural protein of pestivirus or at least one immunodominant part thereof;
wherein said cell comprises a second nucleic acid construct encoding at least one pestiviral protein or a substantial part thereof related to viral spread and allowing packaging of said recombinant pestiviral genome in a pestivirus-like particle;
allowing said pestiviral genome to replicate in said cell;
allowing the replicated recombinant pestiviral genome to package into a pestivirus-like particle comprising the protein encoded by the second nucleic acid;
harvesting said pestivirus-like particles; and
preparing a suspension of said pestivirus-like particles in a suitable diluent.
7. The process according to claim 6 comprising combining said suspension with an adjuvant.
8. The process according to claim 6 , wherein said functional deletion in the sequences encoding at least one structural protein or pestivirus or at least one immunodominant part thereof comprises a functional deletion in E 2 or E ms .