Influenza virus hemagglutinin proteins and uses thereof
Provided herein are chimeric hemagglutinin (HA) polypeptides and uses thereof for inducing an immune response (e.g., an antibody response) against influenza virus. Also provided herein are methods of generating antibodies to the chimeric HA polypeptides in a subject.
1. A chimeric hemagglutinin (HA) polypeptide comprising a hemagglutinin ectodomain from influenza B virus B/Yamagata/16/88, wherein:
I. a. the following amino acid residues in the 120 loop of the globular head domain of influenza B virus B/Yamagata/16/88 hemagglutinin TIP and NIRLSTHNVINAERAPGGPYRL (SEQ ID NO: 1) are substituted with amino acid residues FIP and KIQLSTKNVINAEHAPGGPYRL (SEQ ID NO: 2);
b. the following amino acid residues in the 150 loop of the globular head domain of influenza B virus B/Yamagata/16/88 hemagglutinin PNVTSRNG (SEQ ID NO: 18) are substituted with amino acid residues PYQGKSS (SEQ ID NO:19);
c. the following amino acid residues in the 160 loop of the globular head domain of influenza B virus B/Yamagata/16/88 hemagglutinin RDNKTA (SEQ ID NO: 5) are substituted with amino acid residues KKNSTY (SEQ ID NO: 6); and
d. the following amino acid residues in the 190 helix of the globular head domain of influenza B virus B/Yamagata/16/88 hemagglutinin NKNQMKN (SEQ ID NO: 7) are substituted with amino acid residues NDAAMQT (SEQ ID NO: 8);
II. a. the following amino acid residues in the 120 loop of the globular head domain of influenza B virus B/Yamagata/16/88 hemagglutinin TIP and NIRLSTHNVINAERAPGGPYRL (SEQ ID NO: 1) are substituted with amino acid residues HIP and RIRLSTYNVINAETAPGGPYRL (SEQ ID NO: 51);
b. the following amino acid residues in the 150 loop of the globular head domain of influenza B virus B/Yamagata/16/88 hemagglutinin PNVTSRNG (SEQ ID NO: 18) are substituted with amino acid residues NASTGGQS (SEQ ID NO: 52);
c. the following amino acid residues in the 160 loop of the globular head domain of influenza B virus B/Yamagata/16/88 hemagglutinin RDNKTA (SEQ ID NO: 5) are substituted with amino acid residues KKKADTY (SEQ ID NO: 53); and
d. the following amino acid residues in the 190 helix of the globular head domain of influenza B virus B/Yamagata/16/88 hemagglutinin NKNQMKN (SEQ ID NO: 7) are substituted with amino acid residues ADAKMQT (SEQ ID NO: 54);
III. a. the following amino acid residues in the 120 loop of the globular head domain of influenza B virus B/Yamagata/16/88 hemagglutinin TIP and NIRLSTHNVINAERAPGGPYRL (SEQ ID NO: 1) are substituted with amino acid residues LIP and KIELSTSNVINAEVAPGGPYRL (SEQ ID NO: 55);
b. the following amino acid residues in the 150 loop of the globular head domain of influenza B virus B/Yamagata/16/88 hemagglutinin PNVTSRNG (SEQ ID NO: 18) are substituted with amino acid residues PFGSSNS (SEQ ID NO:56);
c. the following amino acid residues in the 160 loop of the globular head domain of influenza B virus B/Yamagata/16/88 hemagglutinin RDNKTA (SEQ ID NO: 5) are substituted with amino acid residues HQSGTY (SEQ ID NO: 57); and
d. the following amino acid residues in the 190 helix of the globular head domain of influenza B virus B/Yamagata/16/88 hemagglutinin NKNQMKN (SEQ ID NO: 7) are substituted with amino acid residues TTLKMHQ (SEQ ID NO: 58) or ATLKMHQ (SEQ ID NO: 70); or
IV. a. the following amino acid residues in the 120 loop of the globular head domain of influenza B virus B/Yamagata/16/88 hemagglutinin TIP and NIRLSTHNVINAERAPGGPYRL (SEQ ID NO: 1) are substituted with amino acid residues NIP and RIELSTHNVINAEVAPGGPYRL (SEQ ID NO: 59);
b. the following amino acid residues in the 150 loop of the globular head domain of influenza B virus B/Yamagata/16/88 hemagglutinin PNVTSRNG (SEQ ID NO: 18) are substituted with amino acid residues PDKGASS (SEQ ID NO:60);
c. the following amino acid residues in the 160 loop of the globular head domain of influenza B virus B/Yamagata/16/88 hemagglutinin RDNKTA (SEQ ID NO: 5) are substituted with amino acid residues KRGNQY (SEQ ID NO: 61); and
d. the following amino acid residues in the 190 helix of the globular head domain of influenza B virus B/Yamagata/16/88 hemagglutinin NKNQMKN (SEQ ID NO: 7) are substituted with amino acid residues VSTNMAK (SEQ ID NO: 62).
2. The chimeric HA polypeptide of claim 1 , wherein
a. the following amino acid residues in the 120 loop of the globular head domain of influenza B virus B/Yamagata/16/88 hemagglutinin TIP and NIRLSTHNVINAERAPGGPYRL (SEQ ID NO: 1) are substituted with amino acid residues HIP and RIRLSTYNVINAETAPGGPYRL (SEQ ID NO: 51);
b. the following amino acid residues in the 150 loop of the globular head domain of influenza B virus B/Yamagata/16/88 hemagglutinin PNVTSRNG (SEQ ID NO: 18) are substituted with amino acid residues NASTGGQS (SEQ ID NO: 52);
c. the following amino acid residues in the 160 loop of the globular head domain of influenza B virus B/Yamagata/16/88 hemagglutinin RDNKTA (SEQ ID NO: 5) are substituted with amino acid residues KKKADTY (SEQ ID NO: 53); and
d. the following amino acid residues in the 190 helix of the globular head domain of influenza B virus B/Yamagata/16/88 hemagglutinin NKNQMKN (SEQ ID NO: 7) are substituted with amino acid residues ADAKMQT (SEQ ID NO: 54).
3. The chimeric HA polypeptide of claim 1 , wherein
a. the following amino acid residues in the 120 loop of the globular head domain of influenza B virus B/Yamagata/16/88 hemagglutinin TIP and NIRLSTHNVINAERAPGGPYRL (SEQ ID NO: 1) are substituted with amino acid residues LIP and KIELSTSNVINAEVAPGGPYRL (SEQ ID NO: 55);
b. the following amino acid residues in the 150 loop of the globular head domain of influenza B virus B/Yamagata/16/88 hemaglutinin PNVTSRNG (SEQ ID NO: 18) are substituted with amino acid residues PFGSSNS (SEQ ID NO:56);
c. the following amino acid residues in the 160 loop of the globular head domain of influenza B virus B/Yamagata/16/88 hemagglutinin RDNKTA (SEQ ID NO: 5) are substituted with amino acid residues HQSGTY (SEQ ID NO: 57); and
d. the following amino acid residues in the 190 helix of the globular head domain of influenza B virus B/Yamagata/16/88 hemagglutinin NKNQMKN (SEQ ID NO: 7) are substituted with amino acid residues TTLKMHQ (SEQ ID NO: 58).
4. The chimeric HA polypeptide of claim 1 , wherein
a. the following amino acid residues in the 120 loop of the globular head domain of influenza B virus B/Yamagata/16/88 hemagglutinin TIP and NIRLSTHNVINAERAPGGPYRL (SEQ ID NO: 1) are substituted with amino acid residues NIP and RIELSTHNVINAEVAPGGPYRL (SEQ ID NO: 59);
b. the following amino acid residues in the 150 loop of the globular head domain of influenza B virus B/Yamagata/16/88 hemagglutinin PNVTSRNG (SEQ ID NO: 18) are substituted with amino acid residues PDKGASS (SEQ ID NO:60);
c. the following amino acid residues in the 160 loop of the globular head domain of influenza B virus B/Yamagata/16/88 hemagglutinin RDNKTA (SEQ ID NO: 5) are substituted with amino acid residues KRGNQY (SEQ ID NO: 61); and
d. the following amino acid residues in the 190 helix of the globular head domain of influenza B virus B/Yamagata/16/88 hemagglutinin NKNQMKN (SEQ ID NO: 7) are substituted with amino acid residues VSTNMAK (SEQ ID NO: 62).
5. A chimeric hemagglutinin (HA) polypeptide comprising:
a. the amino acid sequence set forth in SEQ ID NO: 21, or the amino acid sequence set forth in SEQ ID NO: 21 without the signal peptide;
b. the amino acid sequence set forth in SEQ ID NO: 23 or SEQ ID NO: 25, or the amino acid sequence set forth in SEQ ID NO: 23 or SEQ ID NO: 25 without the signal peptide;
c. the amino acid sequence of the influenza virus HA ectodomain set forth in SEQ ID NO: 21;
d. the amino acid sequence of the influenza virus HA ectodomain set forth in SEQ ID NO: 23 or SEQ ID NO: 25;
e. the amino acid sequence set forth in SEQ ID NO: 44, SEQ ID NO: 46, SEQ ID NO: 48, or SEQ ID NO: 50, or the amino acid sequence set forth in SEQ ID NO: 44, SEQ ID NO: 46, SEQ ID NO: 48, or SEQ ID NO: 50 without the signal peptide; or
f. the amino acid sequence of the influenza virus HA ectodomain set forth in SEQ ID NO: 44, SEQ ID NO: 46, SEQ ID NO: 48, or SEQ ID NO: 50.
6. An influenza virus engineered to express a chimeric hemagglutinin (HA) polypeptide comprising a hemagglutinin ectodomain from an influenza B virus comprising the following:
a. 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more amino acid substitutions within the 120 loop of the globular head domain of the influenza B virus HA, wherein the amino acid substitutions substitute 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more amino acid residues in the 120 loop of the globular head of the influenza B virus HA with amino acid residues found in a corresponding region of the globular domain of an influenza A virus HA;
b. 2, 3, 4, 5, 6, 7, 8, 9 or more amino acid substitutions within the 150 loop of the globular head domain of the influenza B virus HA, wherein the amino acid substitutions substitute 2, 3, 4, 5, 6, 7, 8, 9 or more amino acid residues in the 150 loop of the globular head of the influenza B virus HA with amino acid residues found in a corresponding region of the globular domain of an influenza A virus HA;
c. 2, 3, 4, 5 or more amino acid substitutions within the 160 loop of the globular head domain of the influenza B virus HA, wherein the amino acid substitutions substitute 2, 3, 4, 5 or more amino acid residues in the 160 loop of the globular head of the influenza B virus HA with amino acid residues found in a corresponding region of the globular domain of an influenza A virus HA; and
d. 2, 3, 4, 5, 6, 7, 8 or more amino acid substitutions within the 190 helix of the globular head domain of the influenza B virus HA, wherein the amino acid substitutions substitute 2, 3, 4, 5, 6, 7, 8 or more amino acid residues in the 190 helix of the globular head of the influenza B virus HA with amino acid residues found in a corresponding region of the globular domain of an influenza A virus HA.
7. An influenza B virus comprising the chimeric HA polypeptide of claim 5 .
8. An immunogenic composition comprising the influenza virus of claim 6 .
9. The chimeric HA polypeptide of claim 5 , wherein the chimeric HA polypeptide comprises the amino acid sequence set forth in SEQ ID NO: 44.
10. The chimeric HA polypeptide of claim 5 , wherein the chimeric HA polypeptide comprises the amino acid sequence set forth in SEQ ID NO: 44 without the signal peptide.
11. The chimeric HA polypeptide of claim 5 , wherein the chimeric HA polypeptide comprises the amino acid sequence of the influenza virus HA ectodomain set forth in SEQ ID NO: 44.
12. The chimeric HA polypeptide of claim 1 , which further comprises the transmembrane and cytoplasmic tail domain of the influenza B virus hemagglutinin.
13. An influenza virus engineered to express the chimeric HA polypeptide of claim 1 .
14. An influenza virus comprising the chimeric HA polypeptide of claim 1 .
15. An influenza B virus engineered to express the chimeric HA polypeptide of claim 9 .
16. An influenza B virus comprising the chimeric HA polypeptide of claim 9 .
17. An isolated chimeric hemagglutinin (HA) polypeptide comprising a hemagglutinin ectodomain from an influenza B virus comprising the following:
a. 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more amino acid substitutions within the 120 loop of the globular head domain of the influenza B virus HA, wherein the amino acid substitutions substitute 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more amino acid residues in the 120 loop of the globular head of the influenza B virus HA with amino acid residues found in a corresponding region of the globular domain of an influenza A virus HA;
b. 2, 3, 4, 5, 6, 7, 8, 9 or more amino acid substitutions within the 150 loop of the globular head domain of the influenza B virus HA, wherein the amino acid substitutions substitute 2, 3, 4, 5, 6, 7, 8, 9 or more amino acid residues in the 150 loop of the globular head of the influenza B virus HA with amino acid residues found in a corresponding region of the globular domain of an influenza A virus HA;
c. 2, 3, 4, 5 or more amino acid substitutions within the 160 loop of the globular head domain of the influenza B virus HA, wherein the amino acid substitutions substitute 2, 3, 4, 5 or more amino acid residues in the 160 loop of the globular head of the influenza B virus HA with amino acid residues found in a corresponding region of the globular domain of an influenza A virus HA; and
d. 2, 3, 4, 5, 6, 7, 8 or more amino acid substitutions within the 190 helix of the globular head domain of the influenza B virus HA, wherein the amino acid substitutions substitute 2, 3, 4, 5, 6, 7, 8 or more amino acid residues in the 190 helix of the globular head of the influenza B virus HA with amino acid residues found in a corresponding region of the globular domain of an influenza A virus HA.
18. The chimeric HA polypeptide of claim 1 which is isolated.
19. The chimeric HA polypeptide of claim 5 which is isolated.
20. An influenza B virus engineered to express the chimeric HA polypeptide of claim 1 .
21. An immunogenic composition comprising the chimeric HA polypeptide of claim 1 .
22. An immunogenic composition comprising the chimeric HA polypeptide of claim 5 .
23. An immunogenic composition comprising the influenza virus of claim 13 .
24. An immunogenic composition comprising the influenza virus of claim 14 .
25. The chimeric HA polypeptide of claim 1 , wherein:
a. the following amino acid residues in the 120 loop of the globular head domain of influenza B virus B/Yamagata/16/88 hemagglutinin TIP and NIRLSTHNVINAERAPGGPYRL (SEQ ID NO: 1) are substituted with amino acid residues FIP and KIQLSTKNVINAEHAPGGPYRL (SEQ ID NO: 2);
b. the following amino acid residues in the 150 loop of the globular head domain of influenza B virus B/Yamagata/16/88 hemagglutinin PNVTSRNG (SEQ ID NO: 18) are substituted with amino acid residues PYQGKSS (SEQ ID NO:19);
c. the following amino acid residues in the 160 loop of the globular head domain of influenza B virus B/Yamagata/16/88 hemagglutinin RDNKTA (SEQ ID NO: 5) are substituted with amino acid residues KKNSTY (SEQ ID NO: 6); and
d. the following amino acid residues in the 190 helix of the globular head domain of influenza B virus B/Yamagata/16/88 hemagglutinin NKNQMKN (SEQ ID NO: 7) are substituted with amino acid residues NDAAMQT (SEQ ID NO: 8).
26. An immunogenic composition comprising the isolated chimeric HA polypeptide of claim 17 .
27. The influenza virus of claim 6 , wherein the chimeric HA polypeptide further comprises the transmembrane and cytoplasmic tail domains of the influenza B virus hemagglutinin.