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US Patent Application 20090010962 – Genetically Engineered Swine Influenza Virus and Uses Thereof

Posted by truthpills on 2009/11/05

Application Filed on June 1, 2005
Application Published on January 8, 2009

Abstract text

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.
Claims

1. An attenuated swine influenza virus comprising a mutation in a swine influenza NS1 gene that diminishes the ability of the NS1 gene product to antagonize the cellular interferon response, and permits the attenuated virus, at a multiplicity of infection of 0.001, to grow to titers between approximately 3 fold to approximately 7 fold lower than wild-type swine influenza in pig cells, as determined approximately 5 days post-infection when propagated under the same conditions.

2. The attenuated swine influenza virus of claim 1, wherein the attenuated virus is genetically engineered.
3. The attenuated swine influenza virus of claim 1, wherein the attenuated virus is a mutagenized virus or reassortant.
4. The attenuated swine influenza virus of claim 2, wherein the attenuated virus is a chimeric virus that expresses a heterologous sequence.
5. The attenuated swine influenza virus of claim 2, wherein the attenuated virus is a chimeric virus that expresses a tumor antigen.
6. The attenuated swine influenza virus of claim 2, wherein the attenuated virus is a chimeric virus that expresses an epitope of a foreign pathogen.
7. The attenuated swine influenza virus of claim 1, wherein pig cells are PK(D1) cells, PK(15) cells, PK13 cells, NSK cells, LLC-PK1 cells, LLC-PK1A cells, ESK-4 cells, ST cells, PT-K75 cells, PK-2a/CL 13 cells or SJPL cells.
8. (canceled)
9. The attenuated swine influenza virus of claim 2, wherein the mutation is a deletion at the carboxy terminus of the NS1 gene.
10. (canceled)
11. An attenuated swine influenza virus comprising a modified NS1 gene, wherein the attenuated swine influenza virus is TX/98/del 126, TX/98/del 99 or TX/98/del 73.
12. An attenuated swine influenza virus having an altered interferon antagonist phenotype, wherein said virus comprises a mutation in the NS1 gene resulting in a deletion of between the 105 carboxy terminal amino acid residues and the 160 carboxy terminal amino acid residues of NS1.
13. The attenuated swine influenza virus of claim 12, wherein the virus is attenuated by a mutation in the NS1 gene resulting in a deletion of all of the amino acid residues of NS1 except amino acid residues 1-95, amino acid residues 1-90, amino acid residues 1-85, amino acid residues 1-80, amino acid residues 1-75, amino acid residues 1-73, amino acid residues 1-70, amino acid residues 1-65, or amino acid residues 1-60, and wherein the amino terminal amino acid is number 1 and the mutation in the NS1 gene confers an altered interferon antagonist phenotype.
14. (canceled)
15. A immunogenic formulation comprising the attenuated swine influenza virus of claim 1, and a physiologically acceptable excipient.
16. A immunogenic formulation comprising the attenuated swine influenza virus of claim 11 or 12, and a physiologically acceptable excipient.
17. A pharmaceutical formulation comprising the attenuated swine influenza virus of claim 1, and a physiologically acceptable excipient.
18. A pharmaceutical formulation comprising the attenuated swine influenza virus of claim 11 or 12, and a physiologically acceptable excipient.
19-20. (canceled)
21. The immunogenic formulation of claim 15, wherein the attenuated swine influenza virus is genetically engineered.
22. The immunogenic formulation of claim 21, wherein the attenuated swine influenza virus is a chimeric virus that expresses a heterologous sequence.
23-26. (canceled)
27. The pharmaceutical formulation of claim 17, wherein the attenuated swine influenza virus is genetically engineered.
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