Immunoprotective influenza antigen and its use in vaccination
View Patent ↗The present invention relates to an influenza antigen, comprising a fusion product of at least the extracellular part of a conserved influenza membrane protein or a functional fragment thereof and a presenting carrier, which may be a presenting (poly)peptide or a non-peptidic structure, such as glycans, peptide mimetics, synthetic polymers. The invention further relates to a vaccine against influenza, comprising at least an antigen of the invention, optionally in the presence of one or more excipients. The invention also relates to use of the antigen, a method for preparing the antigen and acceptor cells expressing the antigen.
1. A human influenza immunogenic composition comprising a fusion product, said fusion product comprising
(i) an antigen comprising an immunogenic extracellular part of an M2 membrane protein of a human influenza A virus, wherein said extracellular immunogenic part consists of SEQ ID NOs: 1, 2, or 3, or an immunogenic fragment thereof that induces antibodies to human influenza A virus, and
(ii) a presenting carrier.
2. The composition of claim 1 , wherein the presenting carrier is heterologous peptide or polypeptide.
3. The composition of claim 2 , wherein the heterologous peptide or polypeptide is selected from the group consisting of a hepatitis B core protein, C3d, a polypeptide comprising multiple copies of C3d, tetanus toxin fragment C and yeast Ty particles.
4. The composition of claim 1 , wherein the presenting carrier is a non-peptidic structure.
5. The composition of claim 4 , wherein the presenting non-peptidic structure is selected from the group consisting of glycans, polyethylene glycols, peptide mimetics, and synthetic polymers.
6. The composition of claim 1 , wherein the presenting carrier enhances the immunogenicity of the antigen.
7. The composition of claim 6 , wherein the presenting carrier comprises an epitope recognized by an influenza-specific T helper cell or cytotoxic T cell.
8. The composition of claim 1 , wherein the composition comprises Lactococci cells expressing said fusion product in or on their cell membrane, and said cells can release said fusion product.
9. The composition of claim 1 , wherein the fusion product is in an isolated form.
10. The composition of claim 1 , wherein the fusion product is anchored in the membrane of an acceptor cell expressing the fusion product.
11. The composition of claim 1 , wherein the fusion product is part of a lipid bilayer or cell wall.
12. The composition of claim 1 , wherein the composition comprises Lactococci cells expressing said fusion product in or on their cell wall.
13. The composition of claim 1 , further comprising an influenza antigen selected from the group consisting of hemagglutinin, neuraminidase, nucleoprotein and native M2.
14. A method of obtaining a human influenza immunogenic composition, comprising
providing a fusion product, said fusion product comprising
(i) an antigen comprising an immunogenic extracellular part of an M 2 membrane protein of a human influenza A virus, wherein said extracellular immunogenic part consists of SEQ ID NOs: 1, 2, or 3, or an immunogenic fragment thereof that induces antibodies to human influenza A virus, and
(ii) a presenting carrier; and mixing said fusion product with an excipient.
15. The method according to claim 14 , wherein the presenting carrier is heterologous peptide or polypeptide.
16. The method according to claim 14 , wherein the heterologous peptide or polypeptide is selected from the group consisting of a hepatitis B core protein, C3d, a polypeptide comprising multiple copies of C3d, tetanus toxin fragment C and yeast Ty particles.
17. The method according to claim 14 , wherein the presenting carrier is a non-peptidic structure.
18. The method according to claim 14 , wherein the presenting non-peptidic structure is selected from the group consisting of glycans, polyethylene glycols, peptide mimetics, and synthetic polymers.
19. A human influenza immunogenic composition obtained by the following steps:
providing a nucleic acid construct that encodes a fusion product, said fusion product comprising (i) an antigen comprising an immunogenic extracellular part of an M2 membrane protein of a human influenza A virus, wherein said extracellular immunogenic part consists of SEQ ID NOs: 1, 2, or 3, or an immunogenic fragment thereof that induces antibodies to human influenza A virus, and (ii) a heterologous peptide or polypeptide presenting carrier;
introducing the nucleic acid construct into an acceptor cell form a transformed acceptor cell;
culturing the transformed acceptor cell in a culture medium under conditions that permit expression of the fusion product; and
optionally isolating the fusion product from the acceptor cell or its culture medium; and
optionally admixing the fusion product with an excipient, thereby obtaining a human influenza immunogenic composition.
20. The influenza immunogenic composition of claim 19 , wherein the heterologous peptide or polypeptide is selected from the group consisting of a hepatitis B core protein, C3d, a polypeptide comprising multiple copies of C3d, tetanus toxin fragment C and yeast Ty particles.
21. The influenza immunogenic composition of claim 20 , wherein the influenza immunogenic composition comprises a cytokine.
22. The influenza immunogenic composition of claim 20 , wherein the influenza immunogenic composition comprises a vaccine adjuvant that is not Freund's adjuvant.
23. An influenza immunogenic composition for an animal species comprising a fusion product, said fusion product comprising
(i) an antigen comprising an immunogenic extracellular part of an M2 membrane protein of an influenza A virus, wherein said extracellular immunogenic part consists of SEQ ID NOs: 1, 2, or 3, or an immunogenic fragment thereof that induces antibodies to human influenza A virus; and
(ii) a presenting carrier.
24. The composition of claim 23 , wherein the presenting carrier is heterologous peptide or polypeptide.
25. The composition of claim 24 , wherein the heterologous peptide or polypeptide is selected from the group consisting of a hepatitis B core protein, C3d, a polypeptide comprising multiple copies of C3d, tetanus toxin fragment C and yeast Ty particles.
26. The composition of claim 24 , wherein the presenting carrier is a non-peptidic structure.
27. The composition of claim 25 , wherein the presenting non-peptidic structure is selected from the group consisting of glycans, polyethylene glycols, peptide mimetics, and synthetic polymers.
28. A human influenza immunogenic composition comprising a fusion polypeptide, said fusion polypeptide comprising
(i) an antigen comprising an immunogenic extracellular part of an M2 membrane protein of an influenza A virus, wherein said extracellular immunogenic part consists of SEQ ID NOs: 1, 2, or 3, or an immunogenic fragment thereof that induces antibodies to human influenza A virus; and
(ii) a heterologous peptide or polypeptide presenting carrier,
said fusion polypeptide being the expression product of a gene construct comprising a coding sequence for said antigen of (i) linked to a coding sequence for said presenting carrier of (ii).
29. The influenza immunogenic composition of claim 28 , wherein said heterologous peptide or polypeptide presenting carrier is selected from the group consisting of a hepatitis B core protein, C3d, a polypeptide comprising multiple copies of C3d, and tetanus toxin fragment C.
30. The influenza immunogenic composition of claim 20 , wherein said heterologous peptide or polypeptide presenting carrier is the hepatitis B core protein.