Functional influenza virus-like particles (VLPs)
The present invention discloses and claims virus like particles (VLPs) that express and/or contains seasonal influenza virus proteins, avian influenza virus proteins and/or influenza virus proteins from viruses with pandemic potential. The invention includes vector constructs comprising said proteins, cells comprising said constructs, formulations and vaccines comprising VLPs of the inventions. The invention also includes methods of making and administrating VLPs to vertebrates, including methods of inducing substantial immunity to either seasonal and avian influenza, or at least one symptom thereof.
1. A vaccine comprising an influenza virus-like particle (VLP) and a pharmaceutically acceptable carrier or excipient, wherein the VLP comprises influenza M1, HA and NA proteins, wherein the vaccine induces substantial immunity to influenza virus infection in an animal susceptible to influenza,
wherein the M1 protein is derived from the influenza strain A/Indonesia/5/05,
wherein the HA protein is an H7 protein, and
wherein the NA protein is an N9 protein.
2. The vaccine of claim 1 , wherein the HA protein exhibits hemagglutinin activity.
3. The vaccine of claim 1 , wherein the NA protein exhibits neuraminidase activity.
4. The vaccine of claim 1 , wherein the animal is a human.
5. The vaccine of claim 1 , wherein the influenza VLP further comprises an adjuvant.
6. The vaccine of claim 5 , wherein the adjuvant comprises paucilamellar nonphospholipid vesicles.
7. The vaccine of claim 1 , wherein the vaccine further comprises a second VLP, wherein the second VLP comprises a non-H7 HA protein, a non-N9 NA protein, and an M1 protein derived from the influenza strain A/Indonesia/5/05.
8. The vaccine of claim 1 , wherein the VLP proteins consist essentially of influenza M1, HA and NA proteins.
9. The vaccine of claim 1 , wherein the VLP proteins consist of influenza M1, HA and NA proteins.
10. The vaccine of claim 1 wherein the M1 protein comprises SEQ ID NO: 49.
11. A method of formulating a vaccine that induces substantial immunity to influenza virus infection to an animal susceptible to influenza, comprising adding an effective dose of an influenza virus-like particle (VLP) to a pharmaceutically acceptable carrier or excipient, wherein the VLP comprises influenza M1, HA and NA proteins, wherein the vaccine induces substantial immunity to influenza virus infection to the animal
wherein the M1 protein is derived from the influenza strain A/Indonesia/5/05,
wherein the HA protein is an H7 protein, and
wherein the NA protein is an N9 protein.
12. The method of claim 11 , wherein the VLP proteins consist essentially of influenza M1, HA and NA proteins.
13. The method of claim 11 , wherein the VLP proteins consist of influenza M1, HA and NA proteins.
14. The method of claim 11 , wherein the influenza VLP has been treated to inactivate baculovirus.
15. The method of claim 14 , wherein the inactivation treatment comprises incubating a sample comprising VLPs in about 0.2% of β-propyl lactone (BPL) for about 3 hours at about 25° C.
16. The method of claim 11 wherein the M1 protein comprises SEQ ID NO: 49.
17. A virus-like particle (VLP) comprising an influenza virus M1 protein, an influenza virus H7 hemagglutinin protein, and an influenza virus N9 neuraminidase protein, wherein the M1 protein is derived from the influenza strain A/Indonesia/5/05.
18. The VLP of claim 17 , wherein the VLP proteins consist essentially of influenza M1, HA and NA proteins.
19. The VLP of claim 17 , wherein the VLP proteins consist of influenza M1, HA and NA proteins.
20. The VLP of claim 17 , wherein the VLP is expressed from a eukaryotic cell comprising one or more nucleic acids encoding the H7, N9 and M1 proteins under conditions that permit the formation of VLPs.
21. The VLP of claim 20 , wherein the eukaryotic cell is selected from the group consisting of yeast, insect, amphibian, avian and mammalian cells.
22. The VLP of claim 20 , wherein the eukaryotic cell is an insect cell.
23. The VLP of claim 22 , wherein the insect cell is Sf9.
24. The VLP of claim 17 , wherein the VLP elicits neutralizing antibodies in a human or animal that are protective against influenza infection when administered to the human or animal.
25. The VLP of claim 17 , wherein the M1 protein comprises SEQ ID NO: 49.