AAV mediated influenza vaccines
A non-replicating recombinant adeno-associated associated virus (rAAV) having an AAV capsid having packaged therein a vector genome which comprises AAV inverted terminal repeat sequences and at least one nucleic acid sequence encoding four different immunoglobulin regions (a), (b), (c) and (d) is provided. The rAAV-expressed immunoglobulins are useful for providing passive immunization against influenza A and influenza B. Also described herein are compositions containing the rAAV. Methods of vaccinating patients against influenza are provided.
1. A non-replicating recombinant adeno-associated virus (rAAV) comprising an AAV capsid and a vector genome packaged in the AAV capsid, wherein the vector genome comprises AAV inverted terminal repeat sequences and an expression cassette comprising a nucleic acid sequence encoding an anti-influenza fusion protein having the amino acid sequence of SEQ ID NO: 27.
2. The rAAV according to claim 1 , wherein the vector genome further comprises a 5′ UTR which comprises is-a fragment of human c-myc 5′ UTR.
3. The rAAV according to claim 1 , wherein the vector genome comprises a single-stranded AAV 5′ inverted terminal repeat and an AAV 3′ inverted terminal repeat.
4. The rAAV according to claim 1 , wherein the nucleic acid sequence encoding the anti-influenza fusion protein is of: SEQ ID NO: 21, or SEQ ID NO: 26.
5. The rAAV according to claim 1 , wherein the rAAV has an AAVhu68 capsid, wherein the AAVhu68 capsid comprises:
(a) a heterogenous population of AAVhu68 vp1 proteins selected from one or more of: vp1 proteins produced by expression from a nucleic acid sequence which encodes the predicted amino acid sequence of 1 to 736 of SEQ ID NO:16, vp1 proteins produced from SEQ ID NO: 18, or vp1 proteins produced from a nucleic acid sequence at least 70% identical to SEQ ID NO:18 which encodes the predicted amino acid sequence of 1 to 736 of SEQ ID NO:16;
(b) a heterogenous population of AAVhu68 vp2 proteins selected from one or more of: vp2 proteins produced by expression from a nucleic acid sequence which encodes the predicted amino acid sequence of at least about amino acids 138 to 736 of SEQ ID NO:16, vp2 proteins produced from a sequence comprising at least nucleotides 412 to 2211 of SEQ ID NO:18, or vp2 proteins produced from a nucleic acid sequence at least 70% identical to at least nucleotides 412 to 2211 of SEQ ID NO:18 which encodes the predicted amino acid sequence of at least about amino acids 138 to 736 of SEQ ID NO:16; and
(c) a heterogenous population of AAVhu68 vp3 proteins selected from one or more of: vp3 proteins produced by expression from a nucleic acid sequence which encodes the predicted amino acid sequence of at least about amino acids 203 to 736 of SEQ ID NO:16, vp3 proteins produced from a sequence comprising at least nucleotides 607 to 2211 of SEQ ID NO:18, or vp3 proteins produced from a nucleic acid sequence at least 70% identical to at least nucleotides 607 to 2211 of SEQ ID NO:18 which encodes the predicted amino acid sequence of at least about amino acids 203 to 736 of SEQ ID NO:16.
6. The rAAV according to claim 5 , wherein the nucleic acid sequence encoding the vp1 proteins, vp2 proteins, and vp3 proteins is SEQ ID NO: 18, or a sequence at least 80% to at least 99% identical to SEQ ID NO: 18 which encodes the amino acid sequence of SEQ ID NO:16.
7. The rAAV according to claim 6 , wherein the sequence is at least 80% to 97% identical to SEQ ID NO: 18.
8. The rAAV according to claim 1 , wherein the rAAV has an AAV9 capsid, wherein said AAV9 capsid comprises vp1 capsid proteins having an amino acid sequence of SEQ ID NO: 17 or an amino acid sequence encoded by SEQ ID NO: 36.
9. A composition comprising a carrier, diluent or excipient and a stock of at least the non-replicating rAAV according to claim 1 .
10. The composition according to claim 9 , wherein the composition is formulated for intranasal administration.
11. The composition according to claim 9 , wherein the composition is formulated for intramuscular or intravenous administration.
12. The A method for immunizing a human against influenza, wherein the method comprises intranasally administering to a human in need thereof about 10 9 genome copies (GC) to about 10 14 GC of the rAAV according to claim 1 .
13. A product which comprises a container comprising the rAAV according to claim 1 , optional diluent, and instructions for administration.
14. The rAAV according to claim 1 , wherein the expression cassette comprises a nucleic acid sequence of SEQ ID NO: 26.
15. The rAAV according to claim 1 , wherein the expression cassette further comprises a chicken beta actin promoter.
16. The rAAV according to claim 1 , wherein the expression cassette further comprises a rabbit beta globin polyA.
17. The rAAV according to claim 1 , wherein the expression cassette comprises the sequence of nucleotides 1992 to 4298 of SEQ ID NO: 5.
18. The composition according to claim 9 , wherein the composition is formulated for delivery to the lung.