Chimeric influenza vaccines
The present disclosure relates to a chimeric influenza virus hemagglutinin (HA) polypeptide, comprising one or more stem domain sequence, each having at least 60% homology with a stem domain consensus sequence of H1 subtype HA (H1 HA) and/or H5 subtype HA (H5 HA), fused with one or more globular head domain sequence, each having at least 60% homology with a globular head domain consensus sequence of H1 subtype HA (H1 HA) or H5 subtype HA (H5 HA).
1. An immunogenic composition comprising a monoglycosylated chimeric influenza virus hemagglutinin (HA) polypeptide and an adjuvant;
wherein the monoglycosylated chimeric influenza virus HA polypeptide comprising an amino acid sequence of SEQ ID NO: 4;
wherein the monoglycosylated chimeric influenza virus HA polypeptide has only N-Acetylglucosamine (GlcNAc) at N28, N40, N182, N292, and N497; and
wherein the adjuvant is selected from the group consisting of Al(OH) 3 , AlPO 4 , an analog of α-galactosylceramide (α-GalCer), squalene and QS21.
2. The immunogenic composition of claim 1 , wherein the monoglycosylated chimeric influenza virus HA polypeptide further comprises only N-Acetylglucosamine (GlcNAc) at N171 and N303.
3. The immunogenic composition of claim 1 , wherein the adjuvant is Al(OH) 3 .
4. The immunogenic composition of claim 1 , wherein the analog of a-galactosylceramide is C34.
5. The immunogenic composition of claim 1 , wherein the monoglycosylated chimeric influenza virus HA polypeptide is existed as a trimer.
6. A method of immunizing a subject against influenza virus or preventing an influenza virus disease comprising administering an effective amount of the immunogenic composition of claim 1 to the subject.
7. The method of claim 6 , wherein the method elicits CD4 + and CD8 + T-cell immune responses.
8. The method of claim 6 , wherein the method induces stem-specific antibodies, with higher antibody-dependent cellular cytotoxicity (ADCC), better neutralizing and stronger cross-protection activities against H1, H3, H5 and H7 strains and subtypes.
9. The method of claim 6 , wherein the method enhances the vaccine efficacy with more IFN-γ, IL-4 and CD8+ memory T cells produced.
10. A method for preparing a chimeric influenza virus hemagglutinin (HA) polypeptide Swap H1/5, having one or more stem domain sequence fused with one or more globular head domain sequence, comprising: composing H1 as HA1 (amino acids 1-327 of SEQ ID NO: 8) And H5 as HA2 (amino acids 328503 of SEQ ID NO: 8) giving H1 as globular head and [H1+H5 (HA2)] as stem.
11. The method for preparing the chimeric influenza virus hemagglutinin (HA) polypeptide Swap H1/5 of claim 10 , wherein one or more glycosites on the chimeric influenza virus hemagglutinin (HA) polypeptide Swap H1/5 are monoglycosylated.
12. The method for preparing the chimeric influenza virus hemagglutinin (HA) polypeptide Swap H1/5 of claim 11 , wherein the monoglycosylated HA polypeptide Swap H1/5 has only N-Acetylglucosamine (GlcNAc) on each glycosite.
13. The method for preparing the chimeric influenza virus hemagglutinin (HA) polypeptide Swap H1/5 of claim 10 , wherein the chimeric influenza virus HA polypeptide Swap H1/5 comprises the amino acid sequence of SEQ ID NO: 8.
14. A method for preparing a chimeric influenza virus hemagglutinin (HA) polypeptide Swap H5/1, having one or more stem domain sequence fused with one or more globular head domain sequence, comprising: composing H5 as HA1 (amino acid 1-330 of SEQ ID NO: 12) And H1 as HA2 (amino acid 331-506 of SEQ ID NO: 12) giving H5 as globular head and [H5+H1 (HA2)] as stem.
15. The method for preparing the chimeric influenza virus hemagglutinin (HA) polypeptide Swap H5/1 of claim 14 , wherein one or more glycosites on the chimeric influenza virus hemagglutinin (HA) polypeptide Swap H5/1 are monoglycosylated.
16. The method for preparing the chimeric influenza virus hemagglutinin (HA) polypeptide Swap H5/1 of claim 15 , wherein the monoglycosylated HA polypeptide Swap H5/1 has only N-Acetylglucosamine (GlcNAc) on each glycosite.
17. The method for preparing the chimeric influenza virus hemagglutinin (HA) polypeptide Swap H5/1 of claim 14 , wherein the chimeric influenza virus HA polypeptide Swap H5/1 comprises the amino acid sequence of SEQ ID NO: 12.
18. A method for preparing a chimeric influenza virus hemagglutinin (HA) polypeptide chimeric H5/1, having one or more stem domain sequence fused with one or more globular head domain sequence, comprising:
(a) constructing the globular head domain with the amino acid sequence between residues C42 and C274 of SEQ ID NO: 4 (H3 numbering); and
(b) constructing the stem domain with portions of HA1 and HA2 subunits (amino acids 1˜41 of SEQ ID NO: 4 and 275-511 of SEQ ID NO: 4).
19. The method for preparing the chimeric influenza virus hemagglutinin (HA) polypeptide chimeric H5/1 of claim 18 , wherein one or more glycosites on the chimeric influenza virus hemagglutinin (HA) polypeptide chimeric H5/1 are monoglycosylated.
20. The method for preparing the chimeric influenza virus hemagglutinin (HA) polypeptide chimeric H5/1 of claim 19 , wherein the monoglycosylated HA polypeptide chimeric H5/1 has only N-Acetylglucosamine (GlcNAc) on each glycosite.
21. The method for preparing the chimeric influenza virus hemagglutinin (HA) polypeptide chimeric H5/1 of claim 18 , wherein the chimeric influenza virus HA polypeptide chimeric H5/1 comprises the amino acid sequence of SEQ ID NO: 4.