Recombinant influenza virus vaccines for influenza
Disclosed are recombinant chimeric influenza virus vaccines and live attenuated influenza virus (LAIV) vaccines expressing foreign (RSV) neutralizing epitopes or conserved M2e epitopes that are capable of providing broader cross-protection against influenza virus and/or protecting against respiratory syncytial virus (RSV) without vaccine-enhanced RSV disease (ERD).
1. A recombinant influenza virus comprising a chimeric hemagglutinin (HA) fusion protein, wherein the HA fusion protein comprises an influenza A hemagglutinin (HA) protein or fragment thereof comprising at least the HA head domain, and
one or more repeats of three or more influenza virus matrix protein 2 extracellular (M2e) domains.
2. The recombinant influenza virus of claim 1 , wherein the chimeric HA fusion protein comprises one or more M2e domains from a human influenza A subtype, one or more M2e domains from a swine influenza A subtype, and one or more M2e domains from an avian influenza A subtype.
3. The recombinant influenza virus of claim 1 , wherein at least one M2e domain comprises a partial or full human, swine, or avian M2e domain comprising the amino acid sequence SEQ ID NO:3 or an amino acid sequence having at least 90% sequence identity to SEQ ID NO:3, and wherein at least one M2e domain comprises an avian M2e domain comprising the amino acid sequence SEQ ID NO:4 or an amino acid sequence having at least 90% sequence identity to SEQ ID NO:4.
4. The recombinant influenza virus of claim 1 , wherein the chimeric HA fusion protein further comprises a signal peptide at the N-terminus or in the middle HA head domain of the HA protein.
5. The recombinant influenza virus of claim 1 , wherein the HA protein is derived from a seasonal or pandemic influenza virus.
6. The recombinant influenza virus of claim 1 , wherein the chimeric HA protein comprises an amino acid sequence having a formula selected from the group consisting of:
X 1 -([hM2e] n -[sM2e] n -[aM2e] n ) n -X 2 ,
X 1 -([hM2e] n -[aM2e] n -[sM2e] n ) n -X 2 ,
X 1 -([sM2e] n -[hM2e] n -[aM2e] n ) n -X 2 ,
X 1 -([sM2e] n -[aM2e] n -[hM2e] n ) n -X 2 ,
X 1 -([aM2e] n -[sM2e] n -[hM2e] n ) n -X 2 , and
X 1 -([aM2e] n -[hM2e] n -[sM2e] n ) n -X 2 ;
wherein “X 1 ” consists of a signal peptide of HA protein,
wherein “X 2 ” consists of an HA protein domain other than the signal peptide,
wherein “hM2e” consists of a human M2e domain,
wherein “sM2e” consists of a swine M2e domain,
wherein “aM2e” consists of an avian M2e domain,
wherein each “n” is independently an integer from one to five, and
wherein “-” consists of a peptide linker or a peptide bond.
7. The recombinant influenza virus of claim 1 , wherein the chimeric HA fusion protein comprises an amino acid sequence having a formula selected from the group consisting of:
X 3 -[hM2e]-X 4 ,
X 3 -[sM2e]-X 4 , and
X 3 -[aM2e]-X 4 ,
wherein “X 3 ” consists of from aa1 to aa171 of an HA derived from A/PR8 virus or seasonal influenza vaccine strains,
wherein “hM2e” consists of a human M2e domain,
wherein “sM2e” consists of a swine M2e domain,
wherein “aM2e” consists of an avian M2e domain,
wherein “X 4 ” consists of from aa172 to the end of HA derived from A/PR8 virus or seasonal influenza vaccine strains,
wherein “-” consists of a peptide linker or a peptide bond.
8. The recombinant influenza virus of claim 1 , wherein the chimeric HA fusion protein comprises an amino acid sequence having a formula selected from the group consisting of:
X 1 -(hM2e-hM2e-sM2e-aM2e-aM2e) n -X 2 ,
X 3 -[hM2e]-X 4 ,
X 3 -[sM2e]-X 4 , and
X 3 -[aM2e]-X 4 ,
wherein “X 1 ” consists of a signal peptide of HA protein,
wherein “X 2 ” consists of an HA protein domain other than the signal peptide,
wherein “X 3 ” consists of from aa1 to aa171 of an HA derived from A/PR8 virus or seasonal influenza vaccine strains,
wherein “X 4 ” consists of from aa172 to the end of HA derived from A/PR8 virus or seasonal influenza vaccine strains,
wherein “hM2e” consists of a human M2e domain,
wherein “sM2e” consists of a swine M2e domain,
wherein “aM2e” consists of an avian M2e domain,
wherein each “n” is independently an integer from one to five, and
wherein “-” consists of a peptide linker or a peptide bond.
9. The recombinant influenza virus of claim 1 formulated as a whole inactivated virus, split viral vaccine, or live attenuated influenza vaccine.
10. The recombinant influenza virus of claim 1 , wherein the influenza virus is an A/Ann Arbor/6/60 (H2N2), A/PR/8/34 (H1N1), or A/Leningrad/134/17/57 (H2N2), influenza virus type A strain.
11. A cross-protective vaccine comprising the recombinant influenza virus of claim 1 .
12. The vaccine of claim 11 , further comprising an adjuvant.
13. The vaccine of claim 12 , wherein the adjuvant is selected from the group consisting of AS04 (alum plus monophosphoryl lipid A), MF59 (oil-in-water emulsion adjuvant), and toll-like receptor agonist adjuvants (monophosphoryl lipid A plus CpG).
14. The vaccine of claim 11 , further comprising a neuraminidase.
15. A method of vaccinating a subject for influenza A, comprising administering the vaccine of claim 11 to a subject in need thereof by intranasal, intramuscular, subcutaneous, microneedle skin, transdermal, or sublingual administration.