Flavivirus vaccine vector against influenza virus
This invention relates to chimeric flavivirus vectors that can be used, for example, in the prevention and treatment of influenza virus infection, compositions including such viral vectors, and methods employing the vectors.
1. A flavivirus comprising an insertion of sequences encoding a heterologous peptide or protein between nucleotides encoding amino acids corresponding to amino acids 277 and 278 of the envelope protein of Japanese encephalitis virus (SEQ ID NO: 60).
2. The flavivirus of claim 1 , wherein the flavivirus is a chimeric flavivirus comprising structural sequences of a first flavivirus and non-structural sequences of a second flavivirus.
3. The flavivirus of claim 2 , comprising pre-membrane and envelope sequences of said first flavivirus and capsid and non-structural sequences of said second flavivirus.
4. The flavivirus of claim 2 , wherein said second flavivirus is a yellow fever virus.
5. The flavivirus of claim 2 , wherein said first flavivirus is selected from the group consisting of: Japanese encephalitis, Dengue-1, Dengue-2, Dengue-3, Dengue-4, Murray Valley encephalitis, St. Louis encephalitis, West Nile, Kunjin, Rocio encephalitis, Ilheus, Tick-borne encephalitis, Central European encephalitis, Siberian encephalitis, Russian Spring-Summer encephalitis, Kyasanur Forest Disease, Omsk Hemorrhagic fever, Louping ill, Powassan, Negishi, Absettarov, Hansalova, Apoi, and Hypr viruses.
6. The flavivirus of claim 1 , wherein the heterologous peptide or protein is an antigen.
7. The flavivirus of claim 6 , wherein the antigen is derived from an infectious agent.
8. The flavivirus of claim 7 , wherein the infectious agent is an influenza virus.
9. The flavivirus of claim 8 , wherein the antigen is selected from the group consisting of hemagglutinin, neuraminidase, or M2, or an immunogenic fragment thereof.
10. The flavivirus of claim 9 , wherein the M2 antigen comprises the M2e region of the M2 protein or a fragment thereof.
11. The flavivirus of claim 10 , wherein the antigen comprises a peptide of the sequence MSLLTEVETPIR (SEQ ID NO:1) or MSLLTEVETPIRNEWGSRSNDSSD (SEQ ID NO:2).
12. The flavivirus of claim 11 , wherein the antigen comprises amino and/or carboxy terminal glycine linker sequences.
13. The flavivirus of claim 1 , wherein the heterologous peptide or protein comprises a sequence of SEQ ID NO: 14.
14. The flavivirus of claim 1 , comprising a deletion in 3′-untranslated region or the NS1 sequences.
15. A nucleic acid molecule encoding the flavivirus of claim 1 , or the complement thereof.
16. A pharmaceutical composition comprising a flavivirus of claim 2 and a pharmaceutically acceptable carrier or diluent.
17. A method of producing a flavivirus, comprising culturing Vero cells into which RNA corresponding to the virus has been introduced at a temperature below 37° C., wherein the flavivirus comprises an insertion of sequences encoding a heterologous peptide or protein between nucleotides encoding amino acids corresponding to amino acids 277 and 278 of the envelope protein of Japanese encephalitis virus (SEQ ID NO: 60).
18. A method of propagating a flavivirus, comprising incubating Vero cells infected with said flavivirus at a temperature below 37° C., wherein the flavivirus comprises an insertion of sequences encoding a heterologous peptide or protein between nucleotides encoding amino acids corresponding to amino acids 277 and 278 of the envelope protein of Japanese encephalitis virus (SEQ ID NO: 60).
19. A flavivirus replicon comprising an insertion of sequences encoding a heterologous peptide or protein between nucleotides encoding amino acids corresponding to amino acids 277 and 278 of the envelope protein of Japanese encephalitis virus (SEQ ID NO: 60).