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 method of inducing an immune response against influenza B virus in a subject, comprising administering to the subject an immunogenic composition comprising a chimeric hemagglutinin (HA) polypeptide or a nucleic acid encoding the chimeric HA polypeptide, wherein:
the chimeric HA polypeptide comprises a hemagglutinin ectodomain from influenza B virus B/Yamagata/16/88, and wherein:
a. amino acid residues TIP and NIRLSTHNVINAERAPGGPYRL (SEQ ID NO: 1) in the 120 loop of the globular head domain of the influenza B virus B/Yamagata/16/88 hemagglutinin ectodomain are substituted with amino acid residues FIP and KIQLSTKNVINAEHAPGGPYRL (SEQ ID NO: 2), respectively;
b. amino acid residues PNVTSRNG (SEQ ID NO: 18) in the 150 loop of the globular head domain of the influenza B virus B/Yamagata/16/88 hemagglutinin ectodomain are substituted with amino acid residues PYQGKSS (SEQ ID NO:19);
c. amino acid residues RDNKTA (SEQ ID NO: 5) in the 160 loop of the globular head domain of the influenza B virus B/Yamagata/16/88 hemagglutinin ectodomain are substituted with amino acid residues KKNSTY (SEQ ID NO: 6); and
d. amino acid residues NKNQMKN (SEQ ID NO: 7) in the 190 helix of the globular head domain of the influenza B virus B/Yamagata/16/88 hemagglutinin ectodomain are substituted with amino acid residues NDAAMQT (SEQ ID NO: 8); or
II. the chimeric HA polypeptide comprises a hemagglutinin ectodomain from influenza B virus B/Yamagata/16/88, and wherein:
a. amino acid residues TIP and NIRLSTHNVINAERAPGGPYRL (SEQ ID NO: 1) in the 120 loop of the globular head domain of the influenza B virus B/Yamagata/16/88 hemagglutinin ectodomain are substituted with amino acid residues HIP and RIRLSTYNVINAETAPGGPYRL (SEQ ID NO: 51), respectively;
b. amino acid residues PNVTSRNG (SEQ ID NO: 18) in the 150 loop of the globular head domain of the influenza B virus B/Yamagata/16/88 hemagglutinin ectodomain are substituted with amino acid residues NASTGGQS (SEQ ID NO: 52);
c. amino acid residues RDNKTA (SEQ ID NO: 5) in the 160 loop of the globular head domain of the influenza B virus B/Yamagata/16/88 hemagglutinin ectodomain are substituted with amino acid residues KKKADTY (SEQ ID NO: 53); and
d. amino acid residues NKNQMKN (SEQ ID NO: 7) in the 190 helix of the globular head domain of the influenza B virus B/Yamagata/16/88 hemagglutinin ectodomain are substituted with amino acid residues ADAKMQT (SEQ ID NO: 54); or
III. the chimeric HA polypeptide comprises a hemagglutinin ectodomain from influenza B virus B/Yamagata/16/88, and wherein:
a. amino acid residues TIP and NIRLSTHNVINAERAPGGPYRL (SEQ ID NO: 1) in the 120 loop of the globular head domain of the influenza B virus B/Yamagata/16/88 hemagglutinin ectodomain are substituted with amino acid residues LIP and KIELSTSNVINAEVAPGGPYRL (SEQ ID NO: 55), respectively;
b. amino acid residues PNVTSRNG (SEQ ID NO: 18) in the 150 loop of the globular head domain of the influenza B virus B/Yamagata/16/88 hemagglutinin ectodomain are substituted with amino acid residues PFGSSNS (SEQ ID NO:56);
c. amino acid residues RDNKTA (SEQ ID NO: 5) in the 160 loop of the globular head domain of influenza B virus B/Yamagata/16/88 the hemagglutinin ectodomain are substituted with amino acid residues HQSGTY (SEQ ID NO: 57); and
d. amino acid residues NKNQMKN (SEQ ID NO: 7) in the 190 helix of the globular head domain of the influenza B virus B/Yamagata/16/88 hemagglutinin ectodomain are substituted with amino acid residues TTLKMHQ (SEQ ID NO: 58) or ATLKMHQ (SEQ ID NO: 70); or
IV. the chimeric HA polypeptide comprises a hemagglutinin ectodomain from influenza B virus B/Yamagata/16/88, and wherein:
a. amino acid residues TIP and NIRLSTHNVINAERAPGGPYRL (SEQ ID NO: 1) in the 120 loop of the globular head domain of the influenza B virus B/Yamagata/16/88 hemagglutinin ectodomain are substituted with amino acid residues NIP and RIELSTHNVINAEVAPGGPYRL (SEQ ID NO: 59), respectively;
b. amino acid residues PNVTSRNG (SEQ ID NO: 18) in the 150 loop of the globular head domain of the influenza B virus B/Yamagata/16/88 hemagglutinin ectodomain are substituted with amino acid residues PDKGASS (SEQ ID NO:60);
c. amino acid residues RDNKTA (SEQ ID NO: 5) in the 160 loop of the globular head domain of the influenza B virus B/Yamagata/16/88 hemagglutinin ectodomain are substituted with amino acid residues KRGNQY (SEQ ID NO: 61); and
d. amino acid residues NKNQMKN (SEQ ID NO: 7) in the 190 helix of the globular head domain of the influenza B virus B/Yamagata/16/88 hemagglutinin ectodomain are substituted with amino acid residues VSTNMAK (SEQ ID NO: 62); or
V. the chimeric HA polypeptide comprises a hemagglutinin ectodomain from an influenza B virus, and wherein:
a. there are 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 hemagglutinin ectodomain, 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 hemagglutinin ectodomain with amino acid residues found in a corresponding region of the globular domain of an influenza A virus hemagglutinin;
b. there are 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 hemagglutinin ectodomain, 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 hemagglutinin ectodomain with amino acid residues found in a corresponding region of the globular domain of an influenza A virus hemagglutinin;
c. there are 2, 3, 4, 5 or more amino acid substitutions within the 160 loop of the globular head domain of the influenza B virus hemagglutinin ectodomain, 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 hemagglutinin ectodomain with amino acid residues found in a corresponding region of the globular domain of an influenza A virus hemagglutinin; and
d. there are 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 hemagglutinin ectodomain, 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 hemagglutinin ectodomain with amino acid residues found in a corresponding region of the globular domain of an influenza A virus hemagglutinin; or
VI. the nucleic acid comprises:
a. the nucleotide sequence set forth in SEQ ID NO: 20, or a complement thereof;
b. the nucleotide sequence set forth in SEQ ID NO: 22 or SEQ ID NO: 24, or a complement thereof; or
c. the nucleotide sequence set forth in SEQ ID NO: 43, SEQ ID NO: 45, SEQ ID NO: 47, or SEQ ID NO: 49, or a complement thereof; or
VII. the chimeric HA polypeptide comprises:
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 hemagglutinin ectodomain set forth in SEQ ID NO: 21;
d. the amino acid sequence of the influenza virus hemagglutinin 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 hemagglutinin ectodomain set forth in SEQ ID NO: 44, SEQ ID NO: 46, SEQ ID NO: 48, or SEQ ID NO: 50.
2. A method of immunizing a subject against influenza virus disease, comprising administering to the subject an immunogenic composition comprising a chimeric hemagglutinin (HA) polypeptide or a nucleic acid encoding the chimeric HA polypeptide, wherein:
I. the chimeric HA polypeptide comprises a hemagglutinin ectodomain from influenza B virus B/Yamagata/16/88, and wherein:
a. amino acid residues TIP and NIRLSTHNVINAERAPGGPYRL (SEQ ID NO: 1) in the 120 loop of the globular head domain of the influenza B virus B/Yamagata/16/88 hemagglutinin ectodomain are substituted with amino acid residues FIP and KIQLSTKNVINAEHAPGGPYRL (SEQ ID NO: 2), respectively;
b. amino acid residues PNVTSRNG (SEQ ID NO: 18) in the 150 loop of the globular head domain of the influenza B virus B/Yamagata/16/88 hemagglutinin ectodomain are substituted with amino acid residues PYQGKSS (SEQ ID NO:19);
c. amino acid residues RDNKTA (SEQ ID NO: 5) in the 160 loop of the globular head domain of the influenza B virus B/Yamagata/16/88 hemagglutinin ectodomain are substituted with amino acid residues KKNSTY (SEQ ID NO: 6); and
d. amino acid residues NKNQMKN (SEQ ID NO: 7) in the 190 helix of the globular head domain of the influenza B virus B/Yamagata/16/88 hemagglutinin ectodomain are substituted with amino acid residues NDAAMQT (SEQ ID NO: 8); or
II. the chimeric HA polypeptide comprises a hemagglutinin ectodomain from influenza B virus B/Yamagata/16/88, and wherein:
a. amino acid residues TIP and NIRLSTHNVINAERAPGGPYRL (SEQ ID NO: 1) in the 120 loop of the globular head domain of the influenza B virus B/Yamagata/16/88 hemagglutinin ectodomain are substituted with amino acid residues HIP and RIRLSTYNVINAETAPGGPYRL (SEQ ID NO: 51), respectively;
b. amino acid residues PNVTSRNG (SEQ ID NO: 18) in the 150 loop of the globular head domain of the influenza B virus B/Yamagata/16/88 hemagglutinin ectodomain are substituted with amino acid residues NASTGGQS (SEQ ID NO: 52);
c. amino acid residues RDNKTA (SEQ ID NO: 5) in the 160 loop of the globular head domain of the influenza B virus B/Yamagata/16/88 hemagglutinin ectodomain are substituted with amino acid residues KKKADTY (SEQ ID NO: 53); and
d. amino acid residues NKNQMKN (SEQ ID NO: 7) in the 190 helix of the globular head domain of the influenza B virus B/Yamagata/16/88 hemagglutinin ectodomain are substituted with amino acid residues ADAKMQT (SEQ ID NO: 54); or
III. the chimeric HA polypeptide comprises a hemagglutinin ectodomain from influenza B virus B/Yamagata/16/88, and wherein:
a. amino acid residues TIP and NIRLSTHNVINAERAPGGPYRL (SEQ ID NO: 1) in the 120 loop of the globular head domain of the influenza B virus B/Yamagata/16/88 hemagglutinin ectodomain are substituted with amino acid residues LIP and KIELSTSNVINAEVAPGGPYRL (SEQ ID NO: 55), respectively;
b. amino acid residues PNVTSRNG (SEQ ID NO: 18) in the 150 loop of the globular head domain of the influenza B virus B/Yamagata/16/88 hemagglutinin ectodomain are substituted with amino acid residues PFGSSNS (SEQ ID NO:56);
c. amino acid residues RDNKTA (SEQ ID NO: 5) in the 160 loop of the globular head domain of influenza B virus B/Yamagata/16/88 the hemagglutinin ectodomain are substituted with amino acid residues HOSGTY (SEQ ID NO: 57); and
d. amino acid residues NKNQMKN (SEQ ID NO: 7) in the 190 helix of the globular head domain of the influenza B virus B/Yamagata/16/88 hemagglutinin ectodomain are substituted with amino acid residues TTLKMHQ (SEQ ID NO: 58) or ATLKMHQ (SEQ ID NO: 70); or
IV. the chimeric HA polypeptide comprises a hemagglutinin ectodomain from influenza B virus B/Yamagata/16/88, and wherein:
a. amino acid residues TIP and NIRLSTHNVINAERAPGGPYRL (SEQ ID NO: 1) in the 120 loop of the globular head domain of the influenza B virus B/Yamagata/16/88 hemagglutinin ectodomain are substituted with amino acid residues NIP and RIELSTHNVINAEVAPGGPYRL (SEQ ID NO: 59), respectively;
b. amino acid residues PNVTSRNG (SEQ ID NO: 18) in the 150 loop of the globular head domain of the influenza B virus B/Yamagata/16/88 hemagglutinin ectodomain are substituted with amino acid residues PDKGASS (SEQ ID NO:60);
c. amino acid residues RDNKTA (SEQ ID NO: 5) in the 160 loop of the globular head domain of the influenza B virus B/Yamagata/16/88 hemagglutinin ectodomain are substituted with amino acid residues KRGNQY (SEQ ID NO: 61); and
d. amino acid residues NKNQMKN (SEQ ID NO: 7) in the 190 helix of the globular head domain of the influenza B virus B/Yamagata/16/88 hemagglutinin ectodomain are substituted with amino acid residues VSTNMAK (SEQ ID NO: 62); or
V. the chimeric HA polypeptide comprises a hemagglutinin ectodomain from an influenza B virus, and wherein:
a. there are 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 hemagglutinin ectodomain, 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 hemagglutinin ectodomain with amino acid residues found in a corresponding region of the globular domain of an influenza A virus hemagglutinin;
b. there are 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 hemagglutinin ectodomain, 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 hemagglutinin ectodomain with amino acid residues found in a corresponding region of the globular domain of an influenza A virus hemagglutinin;
c. there are 2, 3, 4, 5 or more amino acid substitutions within the 160 loop of the globular head domain of the influenza B virus hemagglutinin ectodomain, 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 hemagglutinin ectodomain with amino acid residues found in a corresponding region of the globular domain of an influenza A virus hemagglutinin; and
d. there are 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 hemagglutinin ectodomain, 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 hemagglutinin ectodomain with amino acid residues found in a corresponding region of the globular domain of an influenza A virus hemagglutinin; or
VI. the nucleic acid comprises:
a. the nucleotide sequence set forth in SEQ ID NO: 20, or a complement thereof;
b. the nucleotide sequence set forth in SEQ ID NO: 22 or SEQ ID NO: 24, or a complement thereof; or
c. the nucleotide sequence set forth in SEQ ID NO: 43, SEQ ID NO: 45, SEQ ID NO: 47, or SEQ ID NO: 49, or a complement thereof; or
VII. the chimeric HA polypeptide comprises:
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 hemagglutinin ectodomain set forth in SEQ ID NO: 21;
d. the amino acid sequence of the influenza virus hemagglutinin 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 hemagglutinin ectodomain set forth in SEQ ID NO: 44, SEQ ID NO: 46, SEQ ID NO: 48, or SEQ ID NO: 50.
3. A method of preventing an influenza virus disease in a subject, comprising administering to the subject an immunogenic composition comprising a chimeric hemagglutinin (HA) polypeptide or a nucleic acid encoding the chimeric HA polypeptide, wherein:
I. the chimeric HA polypeptide comprises a hemagglutinin ectodomain from influenza B virus B/Yamagata/16/88, and wherein:
a. amino acid residues TIP and NIRLSTHNVINAERAPGGPYRL (SEQ ID NO: 1) in the 120 loop of the globular head domain of the influenza B virus B/Yamagata/16/88 hemagglutinin ectodomain are substituted with amino acid residues FIP and KIQLSTKNVINAEHAPGGPYRL (SEQ ID NO: 2), respectively;
b. amino acid residues PNVTSRNG (SEQ ID NO: 18) in the 150 loop of the globular head domain of the influenza B virus B/Yamagata/16/88 hemagglutinin ectodomain are substituted with amino acid residues PYQGKSS (SEQ ID NO:19);
c. amino acid residues RDNKTA (SEQ ID NO: 5) in the 160 loop of the globular head domain of the influenza B virus B/Yamagata/16/88 hemagglutinin ectodomain are substituted with amino acid residues KKNSTY (SEQ ID NO: 6); and
d. amino acid residues NKNQMKN (SEQ ID NO: 7) in the 190 helix of the globular head domain of the influenza B virus B/Yamagata/16/88 hemagglutinin ectodomain are substituted with amino acid residues NDAAMQT (SEQ ID NO: 8); or
II. the chimeric HA polypeptide comprises a hemagglutinin ectodomain from influenza B virus B/Yamagata/16/88, and wherein:
a. amino acid residues TIP and NIRLSTHNVINAERAPGGPYRL (SEQ ID NO: 1) in the 120 loop of the globular head domain of the influenza B virus B/Yamagata/16/88 hemagglutinin ectodomain are substituted with amino acid residues HIP and RIRLSTYNVINAETAPGGPYRL (SEQ ID NO: 51), respectively;
b. amino acid residues PNVTSRNG (SEQ ID NO: 18) in the 150 loop of the globular head domain of the influenza B virus B/Yamagata/16/88 hemagglutinin ectodomain are substituted with amino acid residues NASTGGQS (SEQ ID NO: 52);
c. amino acid residues RDNKTA (SEQ ID NO: 5) in the 160 loop of the globular head domain of the influenza B virus B/Yamagata/16/88 hemagglutinin ectodomain are substituted with amino acid residues KKKADTY (SEQ ID NO: 53); and
d. amino acid residues NKNQMKN (SEQ ID NO: 7) in the 190 helix of the globular head domain of the influenza B virus B/Yamagata/16/88 hemagglutinin ectodomain are substituted with amino acid residues ADAKMQT (SEQ ID NO: 54); or
III. the chimeric HA polypeptide comprises a hemagglutinin ectodomain from influenza B virus B/Yamagata/16/88, and wherein:
a. amino acid residues TIP and NIRLSTHNVINAERAPGGPYRL (SEQ ID NO: 1) in the 120 loop of the globular head domain of the influenza B virus B/Yamagata/16/88 hemagglutinin ectodomain are substituted with amino acid residues LIP and KIELSTSNVINAEVAPGGPYRL (SEQ ID NO: 55), respectively;
b. amino acid residues PNVTSRNG (SEQ ID NO: 18) in the 150 loop of the globular head domain of the influenza B virus B/Yamagata/16/88 hemagglutinin ectodomain are substituted with amino acid residues PFGSSNS (SEQ ID NO:56);
c. amino acid residues RDNKTA (SEQ ID NO: 5) in the 160 loop of the globular head domain of influenza B virus B/Yamagata/16/88 the hemagglutinin ectodomain are substituted with amino acid residues HQSGTY (SEQ ID NO: 57); and
d. amino acid residues NKNQMKN (SEQ ID NO: 7) in the 190 helix of the globular head domain of the influenza B virus B/Yamagata/16/88 hemagglutinin ectodomain are substituted with amino acid residues TTLKMHQ (SEQ ID NO: 58) or ATLKMHQ (SEQ ID NO: 70); or
IV the chimeric HA polypeptide comprises a hemagglutinin ectodomain from influenza B virus B/Yamagata/16/88, and wherein:
a. amino acid residues TIP and NIRLSTHNVINAERAPGGPYRL (SEQ ID NO: 1) in the 120 loop of the globular head domain of the influenza B virus B/Yamagata/16/88 hemagglutinin ectodomain are substituted with amino acid residues NIP and RIELSTHNVINAEVAPGGPYRL (SEQ ID NO: 59), respectively;
b. amino acid residues PNVTSRNG (SEQ ID NO: 18) in the 150 loop of the globular head domain of the influenza B virus B/Yamagata/16/88 hemagglutinin ectodomain are substituted with amino acid residues PDKGASS (SEQ ID NO:60);
c. amino acid residues RDNKTA (SEQ ID NO: 5) in the 160 loop of the globular head domain of the influenza B virus B/Yamagata/16/88 hemagglutinin ectodomain are substituted with amino acid residues KRGNQY (SEQ ID NO: 61); and
d. amino acid residues NKNQMKN (SEQ ID NO: 7) in the 190 helix of the globular head domain of the influenza B virus B/Yamagata/16/88 hemagglutinin ectodomain are substituted with amino acid residues VSTNMAK (SEQ ID NO: 62); or
V. the chimeric HA polypeptide comprises a hemagglutinin ectodomain from an influenza B virus, and wherein:
a. there are 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 hemagglutinin ectodomain, 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 hemagglutinin ectodomain with amino acid residues found in a corresponding region of the globular domain of an influenza A virus hemagglutinin;
b. there are 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 hemagglutinin ectodomain, 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 hemagglutinin ectodomain with amino acid residues found in a corresponding region of the globular domain of an influenza A virus hemagglutinin;
c. there are 2, 3, 4, 5 or more amino acid substitutions within the 160 loop of the globular head domain of the influenza B virus hemagglutinin ectodomain, 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 hemagglutinin ectodomain with amino acid residues found in a corresponding region of the globular domain of an influenza A virus hemagglutinin; and
d. there are 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 hemagglutinin ectodomain, 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 hemagglutinin ectodomain with amino acid residues found in a corresponding region of the globular domain of an influenza A virus hemagglutinin; or
VI. the nucleic acid comprises:
a. the nucleotide sequence set forth in SEQ ID NO: 20, or a complement thereof;
b. the nucleotide sequence set forth in SEQ ID NO: 22 or SEQ ID NO: 24, or a complement thereof; or
c. the nucleotide sequence set forth in SEQ ID NO: 43, SEQ ID NO: 45, SEQ ID NO: 47, or SEQ ID NO: 49, or a complement thereof; or
VII. the chimeric HA polypeptide comprises:
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 hemagglutinin ectodomain set forth in SEQ ID NO: 21;
d. the amino acid sequence of the influenza virus hemagglutinin 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 hemagglutinin ectodomain set forth in SEQ ID NO: 44, SEQ ID NO: 46, SEQ ID NO: 48, or SEQ ID NO: 50.
4. The method of claim 1 , wherein the subject is human.
5. The method of claim 2 , wherein the subject is human.
6. The method of claim 3 , wherein the subject is human.
7. The method of claim 1 , wherein the chimeric HA polypeptide further comprises the transmembrane and cytoplasmic tail domains of the influenza B virus hemagglutinin.
8. The method of claim 2 , wherein the chimeric HA polypeptide further comprises the transmembrane and cytoplasmic tail domains of the influenza B virus hemagglutinin.
9. The method of claim 3 , wherein the chimeric HA polypeptide further comprises the transmembrane and cytoplasmic tail domains of the influenza B virus hemagglutinin.
10. The method of claim 2 , wherein the chimeric HA polypeptide comprises a hemagglutinin ectodomain from an influenza B virus, and wherein:
a. there are 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 hemagglutinin ectodomain, 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 hemagglutinin ectodomain with amino acid residues found in a corresponding region of the globular domain of an influenza A virus hemagglutinin;
b. there are 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 hemagglutinin ectodomain, 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 hemagglutinin ectodomain with amino acid residues found in a corresponding region of the globular domain of an influenza A virus hemagglutinin;
c. there are 2, 3, 4, 5 or more amino acid substitutions within the 160 loop of the globular head domain of the influenza B virus hemagglutinin ectodomain, 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 hemagglutinin ectodomain with amino acid residues found in a corresponding region of the globular domain of an influenza A virus hemagglutinin; and
d. there are 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 hemagglutinin ectodomain, 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 hemagglutinin ectodomain with amino acid residues found in a corresponding region of the globular domain of an influenza A virus hemagglutinin.
11. The method of claim 1 , wherein the immunogenic composition comprises a chimeric HA polypeptide.
12. The method of claim 1 , wherein the immunogenic composition comprises a nucleic acid encoding a chimeric HA polypeptide.
13. The method of claim 2 , wherein the immunogenic composition comprises a chimeric HA polypeptide.
14. The method of claim 2 , wherein the immunogenic composition comprises a nucleic acid encoding a chimeric HA polypeptide.
15. The method of claim 3 , wherein the immunogenic composition comprises a chimeric HA polypeptide.
16. The method of claim 3 , wherein the immunogenic composition comprises a nucleic acid encoding a chimeric HA polypeptide.
17. The method of claim 2 , wherein the method further comprises a certain period of time after administration of the immunogenic composition, administrating to the subject a second immunogenic composition, wherein the second immunogenic composition comprises a second chimeric HA polypeptide or a nucleic acid encoding a second chimeric HA polypeptide, wherein the chimeric HA and the second chimeric HA are not the same, wherein the second chimeric HA polypeptide comprises a hemagglutinin ectodomain from an influenza B virus, and wherein:
a. there are 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 hemagglutinin ectodomain, 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 hemagglutinin ectodomain with amino acid residues found in a corresponding region of the globular domain of an influenza A virus hemagglutinin;
b. there are 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 hemagglutinin ectodomain, 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 hemagglutinin ectodomain with amino acid residues found in a corresponding region of the globular domain of an influenza A virus hemagglutinin;
c. there are 2, 3, 4, 5 or more amino acid substitutions within the 160 loop of the globular head domain of the influenza B virus hemagglutinin ectodomain, 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 hemagglutinin ectodomain with amino acid residues found in a corresponding region of the globular domain of an influenza A virus hemagglutinin; and
d. there are 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 hemagglutinin ectodomain, 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 hemagglutinin ectodomain with amino acid residues found in a corresponding region of the globular domain of an influenza A virus hemagglutinin.
18. The method of claim 1 , wherein the globular head domain of the hemagglutinin ectodomain further comprises 1, 2, 3, 4, or 5 amino acid substitutions outside of the 120 loop, 150 loop, 160 loop, and/or 190 helix.
19. The method of claim 2 , wherein the globular head domain of the hemagglutinin ectodomain further comprises 1, 2, 3, 4, or 5 amino acid substitutions outside of the 120 loop, 150 loop, 160 loop, and/or 190 helix.
20. The method of claim 3 , wherein the globular head domain of the hemagglutinin ectodomain further comprises 1, 2, 3, 4, or 5 amino acid substitutions outside of the 120 loop, 150 loop, 160 loop, and/or 190 helix.