Vaccines comprising mutant attenuated influenza viruses
The invention provides a vaccine comprising an effective amount of an isolated recombinant influenza virus comprising a mutant M gene segment that is mutated so that upon viral replication the mutant M gene expresses a functional M1 protein and a mutant M2 protein with a deletion of the cytoplasmic tail and either lacking a transmembrane domain or having a mutated transmembrane domain.
1. A vaccine comprising an amount of a live, attenuated isolated recombinant influenza virus comprising a mutant M gene segment that is mutated so that upon viral infection of a cell the mutant M gene expresses a functional M1 protein and a mutant M2 protein with a M2 extracellular domain or a portion thereof having at least 9 residues and a deletion of entire cytoplasmic tail and a deletion of at least 10 residues of the transmembrane domain, wherein the amount per said influenza virus isolate in the vaccine is 0.1 to 200 micrograms influenza virus hemagglutinin (HA),
wherein a) the mutant M2 protein in the attenuated isolated recombinant virus further comprises a heterologous protein at the C-terminus of the extracellular domain, b) the mutant M2 protein in the attenuated isolated recombinant virus further comprises a heterologous immunogenic protein of a pathogen, c) the HA in the attenuated isolated recombinant virus is modified at the HA cleavage site, or d) the attenuated isolated recombinant virus further comprises another attenuating mutation.
2. The vaccine of claim 1 wherein the mutant M2 protein in the attenuated isolated recombinant virus further comprises a heterologous protein at the C-terminus of the extracellular domain.
3. The vaccine of claim 1 wherein the mutant M2 protein in the attenuated isolate recombinant virus further comprises a heterologous immunogenic protein of a pathogen.
4. The vaccine of claim 1 wherein the HA in the attenuated isolated recombinant virus is modified at the HA cleavage site.
5. The vaccine of claim 1 wherein the attenuated isolated recombinant virus further comprises another attenuating mutation.
6. A vaccine comprising an amount of a live, attenuated isolated recombinant influenza virus comprising a mutant M gene segment that is mutated so that upon viral infection of a cell the mutant M gene expresses a functional M1 protein and a mutant M2 protein with a M2 extracellular domain or a portion thereof having at least 9 residues and a deletion of the entire cytoplasmic tail and a deletion of at least 10 residues of the transmembrane domain, wherein the amount per said influenza virus isolate in the vaccine is 0.1 to 200 micrograms influenza virus hemagglutinin (HA), wherein the recombinant virus comprises influenza A HA H2 or H3.
7. The vaccine of claim 6 , wherein the amount is 1 to 50 micrograms influenza virus hemagglutinin (HA).
8. The vaccine of claim 6 wherein the attenuated isolated recombinant influenza virus is a reassortant virus.
9. The vaccine of claim 8 , wherein the reassortant comprises at least six gene segments from a master vaccine strain.
10. The vaccine of claim 6 , wherein the mutant M2 protein lacks the transmembrane domain of M2.
11. A vaccine comprising an amount of a live, attenuated isolated recombinant influenza virus comprising a mutant M gene segment that is mutated so that upon viral infection of a cell the mutant M gene expresses a functional M1 protein and a mutant M2 protein with a M2 extracellular domain or a portion thereof having at least 9 residues and a deletion of the entire cytoplasmic tail and a deletion of at least 10 residues of the transmembrane domain, wherein the amount per said influenza virus isolate in the vaccine is 0.1 to 200 micrograms influenza virus hemagglutinin (HA), wherein the vaccine further comprises a different influenza virus.
12. The vaccine of claim 11 , wherein the HA in the attenuated isolated recombinant influenza virus comprises H5 HA.
13. The vaccine of claim 11 , further comprising two influenza viruses that are different from the attenuated isolated recombinant influenza virus.
14. The vaccine of claim 11 wherein the amount of the attenuated isolated recombinant influenza virus in the vaccine is 1 to 50 micrograms influenza virus hemagglutinin (HA).
15. The vaccine of claim 11 , where the amount of the attenuated isolated recombinant influenza virus is up to 1 microgram influenza virus hemagglutinin (HA).
16. The vaccine of claim 11 wherein the attenuated isolated recombinant influenza virus comprises influenza A HA.
17. The vaccine of claim 16 , wherein the HA in the attenuated isolated recombinant influenza virus is not H1 and not H3.
18. The vaccine of claim 11 wherein the attenuated isolated recombinant influenza virus is a reassortant virus.
19. The vaccine of claim 18 wherein the reassortant comprises at least six gene segments from a master vaccine strain.
20. The vaccine of claim 11 , wherein the mutant M2 protein lacks the transmembrane domain of M2.
21. A method to immunize a human or an avian, comprising:
administering to the human or avian an effective amount of a vaccine comprising a live, attenuated isolated recombinant influenza virus comprising a mutant M gene segment that is mutated so that upon viral infection of a cell the mutant M gene expresses a functional M1 protein and a mutant M2 protein with a M2 extracellular domain or a portion thereof having at least 9 residues and a deletion of the entire cytoplasmic tail and a deletion of at least 10 residues of the transmembrane domain, wherein the amount per said influenza virus isolate in the vaccine is 0.1 to 200 micrograms of influenza virus hemagglutinin (HA).
22. The method of claim 21 , wherein the vaccine is administered to an avian.
23. The method of claim 21 , wherein the vaccine is administered to a human.
24. A method to immunize a vertebrate, comprising:
administering to the vertebrate a vaccine comprising an effective amount of a live, attenuated isolated recombinant influenza virus comprising a mutant M gene segment that is mutated so that upon viral infection of a cell the mutant M gene expresses a functional M1 protein and a mutant M2 protein with a M2 extracellular domain or a portion thereof having at least 9 residues and a deletion of the entire cytoplasmic tail and a deletion of at least 10 residues of the transmembrane domain, wherein the amount per said influenza virus isolate in the vaccine is 10 3 to 10 7 PFU/kg.
25. The method of claim 24 , wherein the mutant M2 protein consists of the extracellular domain.
26. The method of claim 24 , wherein the attenuated isolated recombinant virus comprises influenza A HA.
27. The method of claim 24 , wherein the attenuated isolated recombinant influenza virus is a reassortant virus.
28. The method of claim 27 , wherein the reassortant comprises at least six gene segments from a master vaccine strain.
29. The method of claim 24 , wherein the HA is H5 HA.
30. The method of claim 24 , wherein the HA is H1 HA.
31. The method of claim 24 , wherein the mutant M2 protein lacks the transmembrane domain of M2.