Influenza virus vaccines and uses thereof
Provided are influenza hemagglutinin stem domain polypeptides comprising (a) an influenza hemagglutinin HA1 domain that comprises an HA1 N-terminal stem segment comprising the amino acids from position 1 to position x, preferably from position p to position x, of the HA1 domain, covalently linked by a linking sequence of 0-50 amino acid residues to an HA1 C-terminal stem segment, comprising the amino acids from position y to and including the C-terminal amino acid of the HA1 domain; and (b) an influenza hemagglutinin HA2 domain, wherein the hemagglutinin stem domain polypeptide is resistant to protease cleavage at the junction between HA1 and HA2, and wherein one or more amino acid of the amino acids at positions 337, 340, 352, 353, 402, 406, 409, 413 and/or 416 have been mutated, as compared to the corresponding positions in wild-type influenza HA.
1. An influenza hemagglutinin (HA) stem domain polypeptide comprising the amino acid sequence of SEQ ID NO: 53, 54, 70, or 71:
(SEQ ID NO: 53)
DTICIGYHANNSTDTVDTVLEKNVTVTHSVNLLENGGGGKYVCSAKLRMV
TGLRN X 1 PS X 2 QSQGLFGAIAG X 3 X 4 EGGWTGMVDGWYGYHHQNEQGSGY
AADQKSTQNAINGITNKVNSVIEK X 5 NTQ X 6 TA X 7 GKE X 8 NK X 9 ERMKQI
EDKIEEIESKQIWCYNAELLVLLENERTLDFHDSNVKNLYEKVKSQLKNN
AKEIGNGCFEFYHKCNDECMESVKNGTYDYPKYSEESKLNREKIDGVSGR
DYKDDDDKLVPRGSPGSGYIPEAPRDGQAYVRKDGEWVLLSTFLGHHHHH
H,
(SEQ ID NO: 54)
DTICIGYHANNSTDTVDTVLEKNVTVTHSVNLLENGGGGKYVCSAKLRMV
TGLRN X 1 PS X 2 QSQGLFGAIAG X 3 X 4 EGGWTGMVDGWYGYHHQNEQGSGY
AADQKSTQNAINGITNKVNSVIEK X 5 NTQ X 6 TA X 7 GKE X 8 NK X 9 ERMKQI
EDKIEEIESKQIWCYNAELLVLLENERTLDFHDSNVKNLYEKVKSQLKNN
AKEIGNGCFEFYHKCNDECMESVKNGTYDYPKYSEESKLNREKIDG,
(SEQ ID NO: 70)
DTICIGYHANNSTDTVDTVLEKNVTVTHSVNLLENGGGGKYVCSAKLRMV
TGLRN X 1 PS X 2 QSQGLFGAIAG X 3 X 4 EGGWTGMVDGWYGYHHQNEQGSGY
AADQKSTQNAINGITNKVNSVIEK X 5 NTQ X 6 TA X 7 GKE X 8 NK X 9 ERMKQI
EDKIEEIESKQIWCYNAELLVLLENERTLDFHDSNVKNLYEKVKSQLKNN
AKEIGNGCFEFYHKCNDECMESVKNGTYDYPKYSEESKLNREKIDGVKLE
SMGVYQIEG,
(SEQ ID NO: 71)
DTICIGYHANNSTDTVDTVLEKNVTVTHSVNLLENGGGGKYVCSAKLRMV
TGLRN X 1 PS X 2 QSQGLFGAIAG X 3 X 4 EGGWTGMVDGWYGYHHQNEQGSGY
AADQKSTQNAINGITNKVNSVIEK X 5 NTQ X 6 TA X 7 GKE X 8 NK X 9 ERMKQI
EDKIEEIESKQIWCYNAELLVLLENERTLDFHDSNVKNLYEKVKSQLKNN
AKEIGNGCFEFYHKCNDECMESVKNGTYDYPKYSEESKLNREKIDGVKLE
SMGVYQILAIYSTVASSLVLLVSLGAISFWMCSNGSLQCRICI,
wherein X 1 is an amino acid selected from the group consisting of E, I, K, V, A, and T;
X 2 is an amino acid selected from the group consisting of I, K, R, T, F, N, S and Y;
X 3 is an amino acid selected from the group consisting of D, F, V, Y, A, I, N, S, and T;
X 4 is an amino acid selected from the group consisting of I, K, R, T, E, G and V;
X 5 is an amino acid selected from the group consisting of E, K, M, V, R, and T;
X 6 is an amino acid selected from the group consisting of F, I, N, S, T, Y, H, and L;
X 7 is an amino acid selected from the group consisting of A, G, I, R, T, V, F, and S;
X 8 is an amino acid selected from the group consisting of F, I, N, S, T, Y, G, E, K, M, and V; and
X 9 is an amino acid selected from the group consisting of H, I, L, N, R, and S.
2. The influenza HA stem domain polypeptide of claim 1 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 54:
(SEQ ID NO: 54)
DTICIGYHANNSTDTVDTVLEKNVTVTHSVNLLENGGGGKYVCSAKLRMV
TGLRN X 1 PS X 2 QSQGLFGAIAG X 3 X 4 EGGWTGMVDGWYGYHHQNEQGSGY
AADQKSTQNAINGITNKVNSVIEK X 5 NTQ X 6 TA X 7 GKE X 8 NK X 9 ERMKQI
EDKIEEIESKQIWCYNAELLVLLENERTLDFHDSNVKNLYEKVKSQLKNN
AKEIGNGCFEFYHKCNDECMESVKNGTYDYPKYSEESKLNREKIDG.
3. The influenza HA stem domain polypeptide of claim 1 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 70:
(SEQ ID NO: 70)
DTICIGYHANNSTDTVDTVLEKNVTVTHSVNLLENGGGGKYVCSAKLRMV
TGLRN X 1 PS X 2 QSQGLFGAIAG X 3 X 4 EGGWTGMVDGWYGYHHQNEQGSGY
AADQKSTQNAINGITNKVNSVIEK X 5 NTQ X 6 TA X 7 GKE X 8 NK X 9 ERMKQI
EDKIEEIESKQIWCYNAELLVLLENERTLDFHDSNVKNLYEKVKSQLKNN
AKEIGNGCFEFYHKCNDECMESVKNGTYDYPKYSEESKLNREKIDGVKLE
SMGVYQIEG.
4. The influenza HA stem domain polypeptide of claim 1 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 71:
(SEQ ID NO: 71)
DTICIGYHANNSTDTVDTVLEKNVTVTHSVNLLENGGGGKYVCSAKLRMV
TGLRN X 1 PS X 2 QSQGLFGAIAG X 3 X 4 EGGWTGMVDGWYGYHHQNEQGSGY
AADQKSTQNAINGITNKVNSVIEK X 5 NTQ X 6 TA X 7 GKE X 8 NK X 9 ERMKQI
EDKIEEIESKQIWCYNAELLVLLENERTLDFHDSNVKNLYEKVKSQLKNN
AKEIGNGCFEFYHKCNDECMESVKNGTYDYPKYSEESKLNREKIDGVKLE
SMGVYQILAIYSTVASSLVLLVSLGAISFWMCSNGSLQCRICI.
5. An influenza hemagglutinin (HA) stem domain polypeptide, wherein the polypeptide comprises a peptide having the amino acid sequence selected from the group consisting of SEQ ID NO: 55 to SEQ ID NO:76.
6. An influenza hemagglutinin (HA) stem domain polypeptide, wherein the polypeptide comprises a peptide having the amino acid sequence selected from the group consisting of SEQ ID NO: 55, SEQ ID NO: 66 and SEQ ID NO: 73.
7. A nucleic acid molecule encoding the influenza HA stem domain polypeptide of claim 1 .
8. A vector comprising the nucleic acid molecule of claim 7 .
9. A composition comprising the influenza HA stem domain polypeptide of claim 1 .
10. A vaccine comprising an effective amount of the influenza HA stem domain polypeptide of claim 1 .
11. A method of inducing an immune response against influenza HA protein in a subject, the method comprising:
administering an effective amount of the influenza HA stem domain polypeptide of claim 1 to the subject to induce an immune response against influenza HA protein.
12. The influenza HA stem domain polypeptide of claim 6 , wherein the polypeptide comprises SEQ ID NO: 55.
13. The influenza HA stem domain polypeptide of claim 6 , wherein the polypeptide comprises SEQ ID NO: 66.
14. The influenza HA stem domain polypeptide of claim 6 , wherein the polypeptide comprises SEQ ID NO: 73.
15. The influenza HA stem domain polypeptide of claim 1 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 53:
(SEQ ID NO: 53)
DTICIGYHANNSTDTVDTVLEKNVTVTHSVNLLENGGGGKYVCSAKLRMV
TGLRN X 1 PS X 2 QSQGLFGAIAG X 3 X 4 EGGWTGMVDGWYGYHHQNEQGSGY
AADQKSTQNAINGITNKVNSVIEK X 5 NTQ X 6 TA X 7 GKE X 8 NK X 9 ERMKQI
EDKIEEIESKQIWCYNAELLVLLENERTLDFHDSNVKNLYEKVKSQLKNN
AKEIGNGCFEFYHKCNDECMESVKNGTYDYPKYSEESKLNREKIDGVSGR
DYKDDDDKLVPRGSPGSGYIPEAPRDGQAYVRKDGEWVLLSTFLGHHHHH
H.
16. A composition comprising the influenza HA stem domain polypeptide of claim 5 .
17. A composition comprising the influenza HA stem domain polypeptide of claim 6 .
18. A method of inducing an immune response against influenza HA protein in a subject, the method comprising:
administering an effective amount of the influenza HA stem domain polypeptide of claim 5 to the subject to induce an immune response against influenza HA protein.
19. A method of inducing an immune response against influenza HA protein in a subject, the method comprising:
administering an effective amount of the influenza HA stem domain polypeptide of claim 6 to the subject to induce an immune response against influenza HA protein.
20. The influenza HA stem domain polypeptide of claim 1 , wherein the HA stem domain polypeptide comprises an influenza HA1 domain and an influenza HA2 domain, and the HA stem domain polypeptide is resistant to protease cleavage at the junction between the HA1 domain and the HA2 domain.
21. A nucleic acid molecule encoding the influenza HA stem domain polypeptide of claim 5 .
22. A vector comprising the nucleic acid molecule of claim 21 .
23. A nucleic acid molecule encoding the influenza HA stem domain polypeptide of claim 6 .
24. A vector comprising the nucleic acid molecule of claim 23 .
25. A vaccine comprising an effective amount of the influenza HA stem domain polypeptide of claim 5 .
26. A vaccine comprising an effective amount of the influenza HA stem domain polypeptide of claim 6 .