Influenza vaccine compositions and methods of using same
AssigneeChildren's Hospital Medical Center
InventorsMing TAN, Xi Jason JIANG
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Abstract
The instant disclosure relates to pseudovirus nanoparticles (PVNPs) and compositions comprising PVNPs. The disclosed PVNPs may be comprised of fusion proteins that form an icosahedral structure and a nanoparticle shell. The disclosed fusion proteins may comprise a modified norovirus (NoV) S domain protein; a hemagglutinin I (HA1) antigen of the influenza hemagglutinin I (HA1) of influenza vims; and a peptide linker connecting the C-terminus of the NoV S domain to the HA1 antigen. The modified NoV S domain proteins form the interior nanoparticle shell of said PVNP composition and display the 60 HA1 antigens on the surface of the nanoparticle shell. Methods of making and using the PVNPs and compositions containing PVNPs are also disclosed.
Claims37
A pseudovirus nanoparticle (PVNP) composition or consisting of an aggregate of fusion proteins forming an icosahedral structure and a nanoparticle shell, said fusion protein comprising
- a) a modified norovirus (NoV) S domain protein;
- b) a hemagglutinin I (HA1) antigen of the influenza hemagglutinin I (HA1) of influenza virus;
- and
- c) a peptide linker, said peptide linker connecting the C-terminus of said NoV S domain to said HA1 antigen;
- wherein said modified NoV S domain protein forms the interior nanoparticle shell of said PVNP composition;
- wherein said inner nanoparticle shell displays 60 exposed C-termini of said S domain in a triangular pattern;
- wherein 60 HA1 antigens are displayed on the surface of said nanoparticle shell;
- wherein said 60 HA1 antigens form 20 HA1 trimers.
The PVNP composition of any preceding claim, wherein said NoV S domain sequence is wildtype at one or more of positions V57, Q58, or S136, and M140.
The PVNP composition of any preceding claim, wherein said NoV S domain sequence is wildtype at all positions V57, Q58, or 5136, and M140.
The PVNP of any preceding claim, wherein said protein sequence encoding a norovirus (NoV) S domain is wild-type at positions 57, 58, 136, and 140.
The PVNP composition of any preceding claim, wherein said HA1 antigen is of an H7N9 influenza virus subtype.
The PVNP composition of any preceding claim, wherein said HA1 antigen is of an H7N9 influenza virus subtype and has at least 90% sequence homology to SEQ ID NO:
- 2.
The PVNP composition of any preceding claim, wherein said HA1 antigen is of an H1N1 influenza virus subtype and has at least 90% sequence homology to SEQ ID NO:
- 3.
The PVNP composition of any preceding claim, wherein said HA1 antigen comprises or consists of a region covering the receptor binding site (RBS).
The PVNP composition of any preceding claim wherein said HA1 antigen is glycosylated.
The PVNP composition of any preceding claim, said linker comprises or consists of a sequence selected from HHHH (SEQ ID NO:
- 6), GGGG (SEQ ID NO:7), and GSGS (SEQ ID NO:8).
The PVNP composition of any preceding claim, wherein said fusion protein is tag-free.
The PVNP composition of any preceding claim, wherein said PVNP has T=1 icosahedral symmetry.
The PVNP composition of any preceding claim, wherein said PVNP induces substantial immunity to influenza virus infection or at least one symptom thereof when administered to a subject.
A method of immunizing an individual against one or both of an H7 IAV or H3N2 or H7N9 avian IAV, comprising administering the composition of any preceding claim.
A method of eliciting an immune response in a subject against an influenza virus in an individual in need thereof against an influenza virus, comprising administering a composition of any preceding claim, in an amount effective to produce an antigen-specific immune response in said individual.
A method of making a pseudovirus nanoparticle (PVNP), comprising
- a. providing a plasmid comprising gene sequences corresponding to a modified NoV S domain, a linker, and an HA1 antigen;
- b. expressing a gene product of said gene sequences in an E. coli or a eukaryotic cell system;
- c. lysing the cells of said cell system expressing the S-HA1 protein;
- and
- d. precipitating said gene product from said lysed cell system with (NH4)2SO4;
- wherein said sequence is tag-free;
- and
- wherein said concentration of (NH4)2SO4 is between about 0.7 and about 1.2 M.
A pseudovirus nanoparticle (PVNP), said PVNP comprising a plurality of fusion proteins, said fusion protein comprising a modified norovirus (NoV) S domain protein, a hemagglutinin I (HA1) antigen of the influenza hemagglutinin I (HA1) of an influenza virus, said S domain protein and HA1 antigen being connected via a peptide linker;
- wherein said plurality of said fusion proteins forms a nanoparticle having an icosahedral structure,
- wherein said nanoparticle has 60 HA1 antigens presented at the surface of said nanoparticle, said 60 HA1 antigens arranged to form 20 HA trimers;
- wherein said HA1 antigen has a sequence length of from about 225 to about 240 amino acids, and from about 95% to 100% sequence homology to a sequence selected from SEQ NO:
- 2, SEQ NO:
- 3, SEQ NO:
- 4 or SEQ NO:
- 5.
Dependencies are inferred from the claim text.