Genetically engineered swine influenza virus and uses thereof
The present invention relates, in general, to attenuated swine influenza viruses having an impaired ability to antagonize the cellular interferon (IFN) response, and the use of such attenuated viruses in vaccine and pharmaceutical formulations. In particular, the invention relates to attenuated swine influenza viruses having modifications to a swine NS1 gene that diminish or eliminate the ability of the NS1 gene product to antagonize the cellular IFN response. These viruses replicate in vivo, but demonstrate decreased replication, virulence and increased attenuation, and therefore are well suited for use in live virus vaccines, and pharmaceutical formulations.
1. A genetically engineered attenuated swine influenza virus comprising eight viral segments of swine influenza virus A/Swine/Texas/4199-2/98, wherein the viral segments comprise PB1, PB2, PA, hemagglutinin (HA), nucleoprotein (NP), neuraminidase (NA), M and NS, wherein the NS viral segment comprises a swine influenza virus A/Swine/Texas/4199-2/98 NS1 gene with a mutation resulting in an NS1 protein with a deletion of all of the amino acid residues except amino acid residues 1 to 126, and wherein the amino terminal residue of the NS1 protein is number 1.
2. An immunogenic formulation comprising the genetically engineered attenuated swine influenza virus of claim 1 .
3. A genetically engineered attenuated swine influenza virus, wherein the virus comprises a swine influenza virus NS1 gene with a mutation resulting in a swine influenza virus NS1 protein having a deletion of 90 to 95 amino acid residues from the carboxy-terminus of the swine influenza virus NS1 protein, and wherein the swine influenza virus NS1 gene is from A/Swine/Texas/4199-2/98.
4. The genetically engineered attenuated swine influenza virus of claim 3 , wherein the deletion is a deletion of 90, 93 or 95 amino acid residues from the carboxy-terminus of the swine influenza virus NS1 protein.
5. The genetically engineered attenuated swine influenza virus of claim 3 , wherein the deletion is a deletion of all of the amino acid residues of NS1 except amino acid residues 1 to 126, wherein the amino terminal amino acid is number 1.
6. The genetically engineered attenuated swine influenza virus of claim 3 , wherein the genetically engineered attenuated swine influenza virus is A/Swine/Texas/4199-2/98.
7. The genetically engineered attenuated swine influenza virus of claim 3 , wherein the swine influenza virus is H1N1, H1N2, H3N2, H3N1, H9N2, or H5N1.
8. The genetically engineered attenuated swine influenza virus of claim 3 , wherein the genetically engineered attenuated swine influenza virus is a reassortant.
9. The genetically engineered attenuated swine influenza virus of claim 3 , wherein the genetically engineered attenuated swine influenza virus is a chimeric virus comprising a heterologous sequence.
10. The genetically engineered attenuated swine influenza virus of claim 3 , wherein the genetically engineered attenuated swine influenza virus is engineered to express an epitope from a different virus or at least one segment derived from a different virus.
11. The genetically engineered attenuated swine influenza virus of claim 5 , wherein the swine influenza virus is H1N1, H1N2, H3N2, H3N1, H9N2, or H5N1.
12. The genetically engineered attenuated swine influenza virus of claim 5 , wherein the genetically engineered attenuated swine influenza virus is a reassortant.
13. The genetically engineered attenuated swine influenza virus of claim 5 , wherein the genetically engineered attenuated swine influenza virus is a chimeric virus comprising a heterologous sequence.
14. The genetically engineered attenuated swine influenza virus of claim 5 , wherein the genetically engineered attenuated swine influenza virus is engineered to express an epitope from a different virus or at least one segment derived from a different virus.
15. An immunogenic formulation comprising the genetically engineered attenuated swine influenza virus of claim 3 .
16. An immunogenic formulation comprising the genetically engineered attenuated swine influenza virus of claim 5 .
17. An immunogenic formulation comprising the genetically engineered attenuated swine influenza virus of claim 6 .
18. An immunogenic formulation comprising the genetically engineered attenuated swine influenza virus of claim 7 .
19. An immunogenic formulation comprising the genetically engineered attenuated swine influenza virus of claim 11 .
20. A method for preventing swine influenza virus disease in a pig, comprising administering to the pig an effective amount of an immunogenic formulation of claim 2 .
21. A method for preventing swine influenza virus disease in a pig, comprising administering to the pig an effective amount of an immunogenic formulation of claim 15 .
22. A method for preventing swine influenza virus disease in a pig, comprising administering to the pig an effective amount of an immunogenic formulation of claim 16 .
23. A method for preventing swine influenza virus disease in a pig, comprising administering to the pig an effective amount of an immunogenic formulation of claim 17 .
24. A method for preventing swine influenza virus disease in a pig, comprising administering to the pig an effective amount of an immunogenic formulation of claim 18 .
25. A method for preventing swine influenza virus disease in a pig, comprising administering to the pig an effective amount of an immunogenic formulation of claim 19 .
26. The method of claim 20 , wherein the immunogenic formulation is administered to the pig intranasally.
27. The method of claim 21 , wherein the immunogenic formulation is administered to the pig intranasally.
28. The method of claim 22 , wherein the immunogenic formulation is administered to the pig intranasally.