IP Library › Granted Patent US 11,865,173
Granted Patent B2
US 11,865,173 · App. 17/583,088 · Granted Jan 9, 2024

Influenza virus hemagglutinin proteins and uses thereof

Inventors: Peter Palese (New York, NY); Adolfo Garcia-Sastre (New York, NY); Megan Ermler (New York, NY); Florian Krammer (New York, NY)
Assignee: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
A61K39/145C07K14/005A61K2039/525A61K2039/53A61K2039/545A61K2039/70C12N2760/16122C12N2760/16134C12N2760/16171C12N2760/16222C12N2760/16234C12N2760/16271
View Patent ↗
Loading inventors, assignments & file history…
Monitor This Case
Get email alerts when status or documents change.
Order Certified Copies
Most orders are placed with the USPTO same day — all within 24 business hours.
Order via The Patent Place →
Pre-filled with this patent's details
Quick Facts
Patent No.
US 11,865,173
App. No.
17/583,088
Granted
Jan 9, 2024
Kind
B2
Abstract

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.

Claims (72)

1. A nucleic acid sequence encoding a chimeric hemagglutinin (HA) polypeptide, wherein the chimeric HA polypeptide comprises a hemagglutinin ectodomain from influenza B virus B/Yamagata/16/88, and 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 nucleic acid sequence of claim 1 , wherein the chimeric HA polypeptide further comprises the transmembrane and cytoplasmic tail domains of the influenza B virus hemagglutinin.

3. The nucleic acid sequence 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).

4. The nucleic acid sequence 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).

5. The nucleic acid sequence 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 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).

6. The nucleic acid sequence 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).

7. A nucleic acid sequence encoding a chimeric hemagglutinin (HA) polypeptide, wherein the chimeric HA polypeptide comprises:

a. the amino acid sequence set forth in SEQ ID NO: 21, or the amino acid 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.

8. The chimeric HA polypeptide of claim 7 , wherein the chimeric HA polypeptide comprises the amino acid sequence set forth in SEQ ID NO: 44.

9. The nucleic acid sequence of claim 7 , wherein the chimeric HA polypeptide comprises the amino acid sequence set forth in SEQ ID NO: 44 without the signal peptide.

10. The nucleic acid sequence of claim 7 , wherein the chimeric HA polypeptide comprises the amino acid sequence of the influenza virus HA ectodomain set forth in SEQ ID NO: 44.

11. The nucleic acid sequence of claim 1 , which is isolated.

12. The nucleic acid sequence of claim 1 , wherein the nucleic acid sequence is RNA or cDNA.

13. An isolated nucleic acid sequence encoding a chimeric hemagglutinin (HA) polypeptide, wherein the chimeric HA polypeptide comprises 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.

14. The nucleic acid sequence of claim 12 , wherein the chimeric HA polypeptide further comprises the transmembrane and cytoplasmic tail domains of the influenza B virus hemagglutinin.

15. A nucleic acid sequence encoding a chimeric hemagglutinin (HA) polypeptide, wherein the nucleic acid sequence 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.

16. The nucleic acid sequence of claim 15 , further comprising a nucleotide sequence comprising the 5′ non-coding region of an influenza B virus and a nucleotide sequence comprising the 3′ non-coding region of an influenza B virus.

17. The nucleic acid sequence of claim 15 , wherein the nucleic acid sequence is RNA or cDNA.

18. The nucleic acid sequence of claim 15 , which is isolated.

19. An expression vector comprising the nucleic acid sequence of claim 1 .

20. The expression vector of claim 19 , wherein the expression vector is a plasmid or a viral vector.

21. An influenza B virus comprising an HA segment that comprises the nucleic acid sequence of claim 1 .

22. An isolated host cell comprising the expression vector of claim 19 .

23. A method for producing an influenza virus comprising the nucleic acid sequence of claim 1 , comprising propagating the virus in a substrate that allows the virus to grow.

24. The nucleic acid sequence of claim 13 , wherein the chimeric HA polypeptide further comprises the transmembrane and cytoplasmic tail domains of the influenza B virus hemagglutinin.

25. The nucleic acid sequence of claim 13 , 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.

26. The nucleic acid sequence 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.

27. An influenza B virus comprising the nucleic acid sequence of claim 13 .

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Feb 8, 2022
From: PALESE, PETER; GARCIA-SASTRE, ADOLFO; ERMLER, MEGAN; KRAMMER, FLORIAN
To: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
Reel/Frame 058933/0384 →
Continuity (4)
Division 16305845
Provisional Application 62355679 · Jun 28, 2016
Provisional Application 62350701 · Jun 15, 2016
Related Publication 20220257749A1 · Aug 18, 2022
Cited By (3)
US 12,233,123 US 12,364,746 US 12,545,718