Chimeric viruses presenting non-native surface proteins and uses thereof
The present invention provides chimeric negative-stand RNA viruses that allow a subject, e.g., an avian, to be immunized against two infectious agents by using a single chimeric virus of the invention. In particular, the present invention provides chimeric influenza viruses engineered to express and incorporate into their virions a fusion protein comprising an ectodomain of a protein of an infectious agent and the transmembrane and cytoplasmic domain of an influenza virus protein. Such chimeric viruses induce an immune response against influenza virus and the infectious agent. The present invention also provides chimeric Newcastle Disease viruses (NDV) engineered to express and incorporate into their virions a fusion protein comprising the ectodomain of a protein of an infectious agent and the transmembrane and cytoplasmic domain of an NDV protein. Such chimeric viruses induce an immune response against NDV and the infectious agent.
1. A chimeric Newcastle Disease Virus (NDV), comprising a packaged genome comprising a nucleotide sequence encoding an F-fusion protein, wherein the F-fusion protein comprises the transmembrane and cytoplasmic domains of an NDV F protein and at least one epitope of an ectodomain of a protective antigen of an infectious agent other than NDV or an antigen associated with a disease that is anchored by the C-terminus of the antigen, so that the F-fusion protein is expressed and incorporated into the chimeric NDV, and wherein the antigen is not a paramyxovirus antigen.
2. A chimeric Newcastle Disease Virus (NDV), comprising a packaged genome comprising a nucleotide sequence encoding an HN-fusion protein, wherein the HN-fusion protein comprises the transmembrane and cytoplasmic domains of an NDV HN protein and at least one epitope of an ectodomain of a protective antigen of an infectious agent other than NDV or an antigen associated with a disease that is anchored by the N-terminus of the antigen, so that the HN protein-fusion protein is expressed and incorporated into the chimeric NDV, and wherein the antigen is not a paramyxovirus antigen.
3. The chimeric NDV of claim 1 in which the genome comprises a nucleotide sequence encoding an F-fusion protein, wherein the F-fusion protein comprises the transmembrane and cytoplasmic domains of an NDV F protein and an ectodomain of a protective antigen of an infectious agent other than NDV or an antigen associated with a disease that is anchored by the C-terminus of the antigen, so that the F-fusion protein is expressed and incorporated into the chimeric NDV, and wherein the antigen is not a paramyxovirus antigen.
4. The chimeric NDV of claim 1 in which the genome comprises a nucleotide sequence encoding an HN-fusion protein, wherein the HN-fusion protein comprises the transmembrane and cytoplasmic domains of an NDV HN protein and an ectodomain of a protective antigen of an infectious agent other than NDV or an antigen associated with a disease that is anchored by the C-terminus of the antigen, so that the F-fusion protein is expressed and incorporated into the chimeric NDV, and wherein the antigen is not a paramyxovirus antigen.
5. The chimeric NDV of claim 1 in which the genome comprises a nucleotide sequence encoding an F protein, so that the F protein is expressed and incorporated into the chimeric NDV.
6. The chimeric NDV of claim 1 in which the nucleotide sequence encoding the F-fusion protein replaces the nucleotide sequence encoding the NDV F protein and the F-fusion protein supplies the function of the F protein, wherein the at least one epitope of an ectodomain of a protective antigen of an infectious agent other than NDV or an antigen associated with a disease is selected to be incorporated into the F-fusion protein such that it supplies the function of the ectodomain of the F protein.
7. The chimeric NDV of claim 2 in which the genome comprises a nucleotide sequence encoding an HN protein, so that the HN protein is expressed and incorporated into the chimeric NDV.
8. The chimeric NDV of claim 2 in which the nucleotide sequence encoding the HN-fusion protein replaces the nucleotide sequence encoding the NDV HN protein and the HN-fusion protein supplies the function of the HN protein, wherein the at least one epitope of an ectodomain of a protective antigen of an infectious agent other than NDV or an antigen associated with a disease is selected to be incorporated into the HN-fusion protein such that it supplies the function of the ectodomain of the F protein.
9. A chimeric Newcastle Disease Virus (NDV), comprising a packaged genome comprising a nucleotide sequence encoding an NDV F-fusion protein, wherein the F-fusion protein comprises the transmembrane and cytoplasmic domains of an NDV F protein and an ectodomain of a hemagglutinin antigen of an influenza virus that is anchored by the C-terminus of the antigen, so that the F-fusion protein is expressed and incorporated into the chimeric NDV.
10. A chimeric Newcastle Disease Virus (NDV), comprising a packaged genome comprising a nucleotide sequence encoding an HN-fusion protein, wherein the HN-fusion protein comprises the transmembrane and cytoplasmic domains of an NDV HN protein and an ectodomain of a neuraminidase antigen of an influenza virus that is anchored by the N-terminus, so that the HN-fusion protein is expressed and incorporated into the chimeric NDV.
11. The chimeric NDV of claim 9 , wherein the influenza virus is an avian influenza virus.
12. The chimeric NDV of claim 10 , wherein the influenza virus is an avian influenza virus.
13. The chimeric NDV of claim 1 , wherein the chimeric NDV has an NDV strain LaSota backbone.
14. The chimeric NDV of claim 2 , wherein the chimeric NDV has an NDV strain LaSota backbone.
15. The chimeric NDV of claim 9 , wherein the chimeric NDV has an NDV strain LaSota backbone.
16. The chimeric NDV of claim 1 , wherein the transmembrane and cytoplasmic domains of the F-fusion protein are from NDV strain LaSota.
17. The chimeric NDV of claim 2 , wherein the transmembrane and cytoplasmic domains of the F-fusion protein are from NDV strain LaSota.
18. The chimeric NDV of claim 9 , wherein the transmembrane and cytoplasmic domains of the F-fusion protein are from NDV strain LaSota.
19. The chimeric NDV of claim 5 , wherein the F protein is genetically modified at the cleavage site, such that fusogenic activity is increased.
20. The chimeric NDV of claim 9 in which the genome comprises a nucleotide sequence encoding an F protein, so that the F protein is expressed and incorporated into the chimeric NDV.
21. The chimeric NDV of claim 20 , wherein the F protein is genetically modified at the cleavage site, such that fusogenic activity is increased.
22. The chimeric NDV of claim 21 , wherein the genetically modified cleavage site comprises a multi-basic cleavage site.
23. The chimeric NDV of claim 9 in which the nucleotide sequence encoding the F-fusion protein replaces the nucleotide sequence encoding the NDV F protein and the F-fusion protein supplies the function of the F protein, wherein the ectodomain of a hemagglutinin antigen of an influenza virus is selected to be incorporated into the F-fusion protein such that it supplies the function of the ectodomain of the F protein.
24. The chimeric NDV of claim 1 , wherein the F-fusion protein either contains no amino acid residues of the ectodomain of the F protein, or the F-fusion protein contains a fragment of the ectodomain of the F protein that does not retain the activity of the ectodomain of the F protein.
25. The chimeric NDV of claim 2 , wherein the HN-fusion protein contains either no amino acid residues of the ectodomain of the HN protein, or it contains a fragment of the ectodomain of the HN protein that does not retain the activity of the ectodomain of the HN protein.
26. The chimeric NDV of claim 9 , wherein the F-fusion protein contains no amino acid residues of the ectodomain of the F protein.
27. The chimeric NDV of claim 9 , wherein the F-fusion protein contains a fragment of the ectodomain of the F protein that does not retain the activity of the ectodomain of the F protein.
28. The chimeric NDV of claim 10 , wherein the HN-fusion protein contains either no amino acid residues of the ectodomain of the HN protein, or it contains a fragment of the ectodomain of the HN protein that does not retain the activity of the ectodomain of the HN protein.
29. A chimeric Newcastle Disease Virus (NDV), comprising a packaged genome comprising a nucleotide sequence encoding an NDV F-fusion protein, wherein the F-fusion protein comprises the transmembrane and cytoplasmic domains of an NDV F protein and an ectodomain of a hemagglutinin antigen of an influenza virus that is anchored by the C-terminus of the antigen, so that the F-fusion protein is expressed and incorporated into the chimeric NDV, wherein the F-fusion protein contains a fragment of the ectodomain of the F protein that does not retain the activity of the ectodomain of the F protein, and wherein the fragment of the ectodomain of the F protein is between 1 and 15 residues of the ectodomain of an F protein that are immediately adjacent to the transmembrane domain of the F protein.
30. The chimeric NDV of claim 1 , wherein the sequence that encodes the F-fusion protein is inserted between the P and M genes of the NDV genome.
31. The chimeric NDV of claim 2 , wherein the sequence that encodes the HN-fusion protein is inserted between the P and M genes of the NDV genome.
32. The chimeric NDV of claim 9 , wherein the sequence that encodes the F-fusion protein is inserted between the P and M genes of the NDV genome.
33. The chimeric NDV of claim 1 , wherein the chimeric NDV is attenuated.
34. The chimeric NDV of claim 2 , wherein the chimeric NDV is attenuated.
35. The chimeric NDV of claim 9 , wherein the chimeric NDV is attenuated.
36. The chimeric NDV of claim 10 , wherein the chimeric NDV is attenuated.
37. An immunogenic composition comprising the chimeric NDV of claim 33 .
38. An immunogenic composition comprising the chimeric NDV of claim 34 .
39. An immunogenic composition comprising the chimeric NDV of claim 35 .
40. An immunogenic composition comprising the chimeric NDV of claim 36 .
41. A method for inducing an immune response to two infectious agents in an avian subject or a human subject, the method comprising administering the chimeric NDV of claim 1 .
42. A method for inducing an immune response to two infectious agents in an avian subject or a human subject, the method comprising administering the chimeric NDV of claim 2 .
43. A method for inducing an immune response to two infectious agents in an avian subject or a human subject, the method comprising administering the chimeric NDV of claim 9 .