Stabilized pre-fusion RSV F proteins
Stable pre-fusion respiratory syncitial virus (RSV) F proteins, immunogenic compositions including the proteins and uses thereof for the prevention and/or treatment of RSV infection are described.
1. A recombinant pre-fusion respiratory syncytial virus (RSV) Fusion (F) protein comprising a mutation of the amino acid residue at position 215, a mutation of the amino acid residue at position 357, and a mutation of the amino acid residue at position 371, wherein the amino acid positions correspond to the numbering of the amino acid sequence of SEQ ID NO: 13.
2. The pre-fusion RSV F protein according to claim 1 , comprising a mutation of the amino acid residue Ser at position 215 to Pro (S215P), a mutation of the amino acid residue Thr at position 357 to Lys (T357K) and a mutation of the amino acid residue Asn at position 371 to Tyr (N371Y).
3. The pre-fusion RSV F protein according to claim 1 , further comprising at least one mutation selected from the group consisting of:
a) a mutation of the amino acid residue at position 67; and
b) a mutation of the amino acid residue at position 486.
4. The pre-fusion RSV F protein according to claim 3 , wherein the at least one further mutation is selected from the group consisting of:
c) a mutation of the amino acid residue Asn/Thr at position 67 to Ile; and
d) a mutation of the amino acid residue Asp at position 486 to Asn.
5. The pre-fusion RSV F protein according to claim 1 , wherein the protein comprises at least one epitope that is specific to the pre-fusion conformation F protein, wherein the at least one epitope is recognized by a pre-fusion specific monoclonal antibody, comprising a heavy chain CDR1 region of SEQ ID NO: 1, a heavy chain CDR2 region of SEQ ID NO: 2, a heavy chain CDR3 region of SEQ ID NO: 3 and a light chain CDR1 region of SEQ ID NO: 4, a light chain CDR2 region of SEQ ID NO: 5, and a light chain CDR3 region of SEQ ID NO: 6 and/or a pre-fusion specific monoclonal antibody, comprising a heavy chain CDR1 region of SEQ ID NO: 7, a heavy chain CDR2 region of SEQ ID NO: 8, a heavy chain CDR3 region of SEQ ID NO: 9 and a light chain CDR1 region of SEQ ID NO: 10, a light chain CDR2 region of SEQ ID NO: 67, and a light chain CDR3 region of SEQ ID NO: 11.
6. The pre-fusion RSV F protein according to claim 1 , wherein the protein is trimeric.
7. The pre-fusion RSV F protein according to claim 1 , comprising a truncated F1 domain and a heterologous trimerization domain linked to the truncated F1 domain.
8. The pre-fusion RSV F protein according to claim 7 , wherein the heterologous trimerization domain comprises the amino acid sequence GYIPEAPRDGQAYVRKDGEWVLLSTFL (SEQ ID NO: 14).
9. The pre-fusion RSV F protein according to claim 7 , wherein the trimerization domain is linked to amino acid residue 513 of the RSV F protein.
10. The pre-fusion RSV F protein according to claim 1 , wherein the RSV F protein comprises the amino acid sequence of SEQ ID NO: 21.
11. A nucleic acid molecule encoding the pre-fusion RSV F protein according to claim 1 .
12. The nucleic acid molecule according to claim 11 , wherein the nucleic acid molecule has been codon-optimized for expression in mammalian cells.
13. A vector comprising the nucleic acid molecule according to claim 11 .
14. A composition comprising a pre-fusion RSV F protein according to claim 1 .
15. A method of inducing an immune response against RSV F protein in a subject in need thereof, the method comprising administering to the subject the pre-fusion RSV F protein according to claim 1 .
16. A vaccine comprising the pre-fusion RSV F protein according to claim 1 .
17. A method of prophylaxis and/or treatment of RSV infection in a subject in need thereof, the method comprising administering to the subject the pre-fusion RSV F protein according to claim 1 .
18. A host cell comprising the nucleic acid molecule according to claim 11 .
19. A method of preparing the pre-fusion RSV F protein according to claim 1 , comprising growing a host cell comprising a nucleic acid molecule encoding the pre-fusion RSV F protein under conditions for expression of the pre-fusion RSV F protein.
20. A nucleic acid molecule encoding the pre-fusion RSV F protein according to claim 10 .
21. A composition comprising a pre-fusion RSV F protein according to claim 10 .
22. A composition comprising the nucleic acid molecule according to claim 11 .
23. A composition comprising the nucleic acid molecule according to claim 20 .
24. A method of inducing an immune response against RSV F protein in a subject in need thereof, the method comprising administering to the subject the pre-fusion RSV F protein according to claim 10 .
25. A method of inducing an immune response against RSV F protein in a subject in need thereof, the method comprising administering to the subject the nucleic acid molecule according to claim 11 .
26. A method of inducing an immune response against RSV F protein in a subject in need thereof, the method comprising administering to the subject the nucleic acid molecule according to claim 20 .
27. A vaccine comprising the pre-fusion RSV F protein according to claim 10 .
28. A vaccine comprising the nucleic acid molecule according to claim 11 .
29. A vaccine comprising the nucleic acid molecule according to claim 20 .
30. A method of prophylaxis and/or treatment of RSV infection in a subject in need thereof, the method comprising administering to the subject the pre-fusion RSV F protein according to claim 10 .
31. A method of prophylaxis and/or treatment of RSV infection in a subject in need thereof, the method comprising administering to the subject the nucleic acid molecule according to claim 11 .
32. A method of prophylaxis and/or treatment of RSV infection in a subject in need thereof, the method comprising administering to the subject the nucleic acid molecule according to claim 20 .