Interferon inducing genetically engineered attenuated viruses
View Patent ↗The present invention relates to genetically engineered attenuated viruses and methods for their production. In particular, the present invention relates to engineering live attenuated viruses which contain a modified NS gene segment. Recombinant DNA techniques can be utilized to engineer site specific mutations into one or more noncoding regions of the viral genome which result in the down-regulation of one or more viral genes. Alternatively, recombinant DNA techniques can be used to engineer a mutation, including but not limited to an insertion, deletion, or substitution of an amino acid residue(s) or an epitope(s) into a coding region of the viral genome so that altered or chimeric viral proteins are expressed by the engineered virus.
1. An attenuated genetically engineered influenza A virus with an interferon-inducing phenotype containing a knockout of the NS1 gene segment.
2. An attenuated genetically engineered influenza A virus containing a deletion of the entire NS1 gene segment which is infectious and replicates in an interferon (IFN) deficient cell line.
3. The attenuated virus of claim 1 or 2 which further encodes a heterologous sequence.
4. The attenuated virus of claim 3 in which the heterologous sequence encodes a viral antigenic peptide.
5. The attenuated virus of claim 3 in which the heterologous sequence encodes a tumor antigenic peptide.
6. A formulation comprising an attenuated genetically engineered influenza A virus containing a complete deletion in its NS1 gene segment in a suitable pharmaceutical formulation.
7. A formulation comprising a genetically engineered attenuated chimeric influenza A virus that expresses a heterologous sequence, said virus having an interferon-inducing phenotype and a viral genome wherein the complete NS1 coding sequences have been deleted.
8. The formulation of claim 7 , wherein the heterologous sequence encodes a viral antigen.
9. The formulation of claim 8 , wherein the viral antigen is human immunodeficiency virus gp120, hepatitis B virus surface antigen, a herpes virus glycoprotein, or VP1 of poliovirus.
10. The formulation of claim 7 , wherein the heterologous sequence encodes a tumor specific antigen.
11. The formulation of claim 7 , wherein the heterologous sequence encodes an immunoglobulin gene or a portion thereof.
12. The formulation of claim 7 , wherein the heterologous sequence is the malarial epitope (ME 1) of Plasmodium yoelii.
13. The formulation of claim 7 which is formulated for an intranasal administration.
14. A method of inducing an immune response comprising administering to a subject the formulation of claim 7 .
15. A method of inducing an immune response comprising administering to a subject the formulation of claim 13 .
16. The method of claim 14 , wherein the subject is an animal.
17. The method of claim 15 , wherein the subject is an animal.
18. The method of claim 14 , wherein the subject is a human.
19. The method of claim 15 , wherein the subject is a human.
20. The method of claim 14 , wherein the formulation is administered to the subject orally, intradermally, intramusclarly, intraperitoneally, intravenously, or subcutaneously.
21. The method of claim 15 , wherein the formulation is administered to the subject intranasally.
22. The formulation of claim 7 , wherein the heterologous sequence encodes a bacterial antigen.
23. The formulation of claim 7 , wherein the heterologous sequence encodes a parasitic antigen.