IP Library › Granted Patent US 10,654,898
Granted Patent B2
US 10,654,898 · App. 15/545,218 · Granted May 19, 2020

Recombinant human/bovine parainfluenza virus 3 (B/HPIV3) expressing a chimeric RSV/BPIV3 F protein and uses thereof

Inventors: Peter Collins (Silver Spring, MD); Bo Liang (Boyds, MD); Shirin Munir (Bethesda, MD); Anne Schaap Nutt (Austin, TX); Ursula Buchholz (Silver Spring, MD); Natalie Mackow (East Setauket, NY); Peter Kwong (Washington, DC); Barney Graham (Rockville, MD); Jason McLellan (Norwich, VT)
Assignee: The United States of America, as represented by the Secretary, Department of Health and Human Serices
C07K14/005A61K39/12A61K39/155A61P31/14A61K2039/5254A61K2039/5256A61K2039/575A61K2039/70C07K2319/03C07K2319/033C12N2760/00022C12N2760/18534C12N2760/18571C12N2760/18622C12N2760/18634C12N2760/18641C12N2760/18643C12N2760/18671C12N2760/18722C12N2760/18734C12N2760/18741C12N2760/18743C12N2760/18771
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 10,654,898
App. No.
15/545,218
Granted
May 19, 2020
Kind
B2
Abstract

Recombinant paramyxoviruses including a viral genome encoding a heterologous gene are provided. In several embodiments, the recombinant paramyxovirus is a recombinant parainfluenza virus, such as a recombinant PIV3 including a viral genome encoding a heterologous respiratory syncytial virus F ectodomain linked to the transmembrane domain and the cytoplasmic tail of the F protein from the PIV3. Nucleic acid molecules including the genome of a recombinant paramyxoviruses are also provided. The recombinant viruses may advantageously be used in vaccine formulations, such as for vaccines against parainfluenza virus and respiratory syncytial virus.

Claims (70)

1. A recombinant paramyxovirus, comprising:

a viral genome comprising genes encoding parainfluenza virus N, P, M, F, HN, and L proteins, and further comprising a heterologous gene encoding a type I membrane protein comprising a recombinant respiratory syncytial virus (RSV) F ectodomain linked to a transmembrane domain (TM) and cytoplasmic tail (CT) of a parainfluenza virus F protein, wherein the RSV F ectodomain linked to the TM and CT of the parainfluenza virus F protein is encoded by a nucleotide sequence set forth as SEQ ID NO: 11, 22, or 23; and

wherein the recombinant paramyxovirus is a recombinant human/bovine parainfluenza virus 3 (B/HPIV3), a recombinant human parainfluenza virus 3 (HPIV3), or a recombinant bovine parainfluenza virus 3 (BPIV3).

2. The recombinant paramyxovirus of claim 1 , comprising:

the recombinant HPIV3, wherein the viral genome comprises genes encoding HPIV3 N, P, M, F, HN and L proteins, and wherein the RSV F ectodomain linked to the TM and CT of the parainfluenza virus F protein is encoded by the nucleotide sequence set forth as SEQ ID NO: 11;

the recombinant HPIV3, wherein the viral genome comprises genes encoding HPIV3 N, P, M, F, HN and L proteins, and wherein the RSV F ectodomain linked to the TM and CT of the parainfluenza virus F protein is encoded by the nucleotide sequence set forth as SEQ ID NO: 22 or 23;

the recombinant BPIV3, wherein the viral genome comprises genes encoding BPIV3 N, P, V, M, F, FIN, and L proteins, and wherein the RSV F ectodomain linked to the TM and CT of the parainfluenza virus F protein is encoded by the nucleotide sequence set forth as SEQ ID NO: 11;

the recombinant BPIV3, wherein the viral genome comprises genes encoding BPIV3 N, P, V, M, F, FIN, and L proteins, and wherein the RSV F ectodomain linked to the TM and CT of the parainfluenza virus F protein is encoded by the nucleotide sequence set forth as SEQ ID NO: 22 or 23;

the recombinant B/HPIV3, wherein the viral genome comprises genes encoding HPIV3 F and HN proteins and BPIV3 N, P, V, M, and L proteins, and wherein the RSV F ectodomain linked to the TM and CT of the parainfluenza virus F protein is encoded by the nucleotide sequence set forth as SEQ ID NO: 11; or

the recombinant B/HPIV3, wherein the viral genome comprises genes encoding HPIV3 F and HN proteins and BPIV3 N, P, V, M, and L proteins, and wherein the RSV F ectodomain linked to the TM and CT of the parainfluenza virus F protein is encoded by the nucleotide sequence set forth as SEQ ID NO: 22 or 23.

3. The recombinant paramyxovirus of claim 1 , wherein the heterologous gene encoding the recombinant RSV F ectodomain is the first or second gene downstream of a genomic promoter of the viral genome.

4. The recombinant paramyxovirus of claim 2 , wherein the recombinant paramyxovirus comprises:

the recombinant HPIV3, wherein the viral genome comprises, from upstream to downstream, a HPIV3 genomic promoter followed by the genes encoding the HPIV3 N, P, M, F, HN, and L proteins, and wherein the heterologous gene encoding the recombinant RSV F ectodomain is located between the genomic promoter and the gene encoding the N protein or between the genes encoding the N and the P proteins;

the recombinant BPIV3, wherein the viral genome comprises, from upstream to downstream, a BPIV3 genomic promoter followed by the genes encoding the BPIV3 N, P, M, F, HN, and L proteins, and wherein the heterologous gene encoding the recombinant RSV F ectodomain is located between the genomic promoter and the gene encoding the N protein or between the genes encoding the N and the P proteins; or

the recombinant B/HPIV3, wherein the viral genome comprises, from upstream to downstream, a BPIV3 genomic promoter followed by the genes encoding the BPIV3 N, P, and M proteins, the HPIV3 F and HN proteins, and the BPIV3 L protein, and wherein the heterologous gene encoding the recombinant RSV F ectodomain is located between the genomic promoter and the gene encoding the N protein or between the genes encoding the N and the P proteins.

5. The recombinant paramyxovirus of claim 2 , comprising the recombinant B/HPIV3, wherein the HPIV3 F and HN proteins and BPIV3 N, P, M, and L proteins comprise amino acid sequences at least 90% identical to SEQ ID NOs: 43, 101, 47, 48, 49, 52, respectively.

6. The recombinant paramyxovirus of claim 1 , comprising the recombinant B/HPIV3 or the recombinant HPIV3, wherein the viral genome comprises a gene encoding the HPIV3 HN protein, and wherein the HPIV3 HN protein comprises a threonine and a proline at residues 263 and 370, respectively.

7. The recombinant paramyxovirus of claim 1 , comprising:

the recombinant HPIV3, wherein the heterologous gene comprises the nucleotide sequence set forth as SEQ ID NO: 11 (GS RSV F_HEK_DS-Cav1_H3TMCT);

the recombinant B/HPIV3, wherein the heterologous gene comprises the nucleotide sequence set forth as SEQ ID NO: 11 (GS RSV F_HEK_DS-Cav1_H3TMCT); or

the recombinant B/HPIV3, wherein the heterologous gene comprises the nucleotide sequence set forth as SEQ ID NO: 22 (GA RSV F_HEK_DS-Cav1_B3TMCT) or SEQ ID NO: 23 (GS RSV F HEK DS-Cav1_B3TMCT).

8. The recombinant paramyxovirus of claim 1 , comprising:

the recombinant HPIV3, and wherein the viral genome comprises, from upstream to downstream, a HPIV3 genomic promoter followed by HPIV3 N, P, M, F, HN, and L genes, and wherein the heterologous gene encoding the recombinant RSV F ectodomain is located between the genomic promoter and the gene encoding the N protein, and wherein the RSV F ectodomain linked to the TM and CT of the parainfluenza virus F protein is encoded by the nucleotide sequence set forth as SEQ ID NO: 11 ;

the recombinant HPIV3, and wherein the viral genome comprises, from upstream to downstream, a HPIV3 genomic promoter followed by HPIV3 N, P, M, F, HN, and L genes, and wherein the heterologous gene encoding the recombinant RSV F ectodomain is located between the genes encoding the N and P proteins, and wherein the RSV F ectodomain linked to the TM and CT of the parainfluenza virus F protein is encoded by the nucleotide sequence set forth as SEQ ID NO: 11;

the recombinant HPIV3, and wherein the viral genome comprises, from upstream to downstream, a HPIV3 genomic promoter followed by HPIV3 N, P, M, F, HN, and L genes, and wherein the heterologous gene encoding the recombinant RSV F ectodomain is located between the genomic promoter and the gene encoding the N protein, and wherein the RSV F ectodomain linked to the TM and CT of the parainfluenza virus F protein is encoded by the nucleotide sequence set forth as SEQ ID NO: 22 or 23;

the recombinant HPIV3, and wherein the viral genome comprises, from upstream to downstream, a HPIV3 genomic promoter followed by HPIV3 N, P, M, F, HN, and L genes, and wherein the heterologous gene encoding the recombinant RSV F ectodomain is located between the genes encoding the N and P proteins, and wherein the RSV F ectodomain linked to the TM and CT of the parainfluenza virus F protein is encoded by the nucleotide sequence set forth as SEQ ID NO: 22 or 23 ;

the recombinant BPIV3, and wherein the viral genome comprises, from upstream to downstream, a BPIV3 genomic promoter followed by BPIV3 N, P, M, F, HN, and L genes, and wherein the heterologous gene encoding the recombinant RSV F ectodomain is located between the genomic promoter and the gene encoding the N protein, and wherein the RSV F ectodomain-linked to the TM and CT of the parainfluenza virus F protein is encoded by the nucleotide sequence set forth as SEQ ID NO: 22 or 23;

the recombinant BPIV3, and wherein the viral genome comprises, from upstream to downstream, a BPIV3 genomic promoter followed by BPIV3 N, P, M, F, HN, and L genes, and wherein the heterologous gene encoding the recombinant RSV F ectodomain is located between the genes encoding the N and P proteins, and wherein the RSV F ectodomain linked to the TM and CT of the parainfluenza virus F protein is encoded by the nucleotide sequence set forth as SEQ ID NO: 22 or 23 ;

the recombinant B/HPIV3, and wherein the viral genome comprises, from upstream to downstream, a BPIV3 genomic promoter followed by BPIV3 N, P, and M genes, HPIV3 F and HN genes, and a BPIV3 L gene, and wherein the heterologous gene encoding the recombinant RSV F ectodomain is located between the genomic promoter and the gene encoding the N protein, and wherein the RSV F ectodomain linked to the TM and CT of the parainfluenza virus F protein is encoded by the nucleotide sequence set forth as SEQ ID NO: 22 or 23 ;

the recombinant B/HPIV3, and wherein the viral genome comprises, from upstream to downstream, a BPIV3 genomic promoter followed by BPIV3 N, P, and M genes, HPIV3 F and HN genes, and a BPIV3 L gene, and wherein the heterologous gene encoding the recombinant RSV F ectodomain is located between the genes encoding the N and P proteins, and wherein the RSV F ectodomain linked to the TM and CT of the parainfluenza virus F protein is encoded by the nucleotide sequence set forth as SEQ ID NO: 22 or 23 ;

the recombinant B/HPIV3, and wherein the viral genome comprises, from upstream to downstream, a BPIV3 genomic promoter followed by BPIV3 N, P, and M genes, HPIV3 F and HN genes, and a BPIV3 L gene, and wherein the heterologous gene encoding the recombinant RSV F ectodomain is located between the genomic promoter and the gene encoding the N protein, and wherein the RSV F ectodomain linked to the TM and CT of the parainfluenza virus F protein is encoded by the nucleotide sequence set forth as SEQ ID NO: 22 or 23 ;

the recombinant B/HPIV3, and wherein the viral genome comprises, from upstream to downstream, a BPIV3 genomic promoter followed by BPIV3 N, P, and M genes, HPIV3 F and HN genes, and a BPIV3 L gene, and wherein the heterologous gene encoding the recombinant RSV F ectodomain is located between the genes encoding the N and P proteins, and wherein the RSV F ectodomain linked to the TM and CT of the parainfluenza virus F protein is encoded by the nucleotide sequence set forth as SEQ ID NO: 22 or 23 .

9. The recombinant paramyxovirus of claim 1 , wherein:

the HPIV3 is a HPIV3 JS strain;

the HPIV3 comprises I263T and T370P substitutions in the HN protein; or

the B/HPIV3 comprises I263T and T370P substitutions in the HN protein.

10. The recombinant paramyxovirus of claim 1 , wherein at least 90% of viral particles produced by a host cell infected with the recombinant paramyxovirus or viral vector comprise a viral envelope comprising the ectodomain encoded by the heterologous gene.

11. The recombinant paramyxovirus of claim 1 , wherein the recombinant paramyxovirus is an infectious, attenuated, and self-replicating virus.

12. The recombinant paramyxovirus of claim 1 , wherein the RSV F ectodomain is present on the viral envelope of the paramyxovirus.

13. An immunogenic composition comprising the recombinant paramyxovirus of claim 1 and a pharmaceutically acceptable carrier.

14. A method of eliciting an immune response to respiratory syncytial virus and parainfluenza virus in a subject comprising administering a therapeutically effective amount of the immunogenic composition of claim 13 to the subject.

15. A nucleic acid molecule comprising the genome of the recombinant paramyxovirus of claim 1 .

16. The recombinant paramyxovirus of claim 1 , comprising the recombinant B/HPIV3, wherein:

the viral genome comprises, from upstream to downstream, a BPIV3 genomic promoter, a gene encoding a BPIV3 N protein, the heterologous gene encoding the recombinant RSV F ectodomain, a gene encoding a BPIV3 P protein, a gene encoding a BPIV3 M protein, a gene encoding a HPIV3 F protein, a gene encoding a HPIV3 HN protein, and a gene encoding and a BPIV3 L protein; and

the RSV F ectodomain is linked to the TM and CT of a BPIV3 F protein.

17. The recombinant paramyxovirus of claim 16 , wherein

the BPIV3 N protein, the recombinant RSV F ectodomain linked to the TM and CT from a BPIV3 F protein, the BPIV3 P protein, the BPIV3 M protein, the HPIV3 F protein, the HPIV3 HN protein, and the BPIV3 L protein comprise amino acid sequences at least 90% identical to SEQ ID NOs: 47, 21, 48, 49, 43, 101, and 52, respectively; and

the HPIV3 HN protein further comprises I263T and T370P substitutions.

18. The recombinant paramyxovirus of claim 17 , wherein

the BPIV3 N protein, the recombinant RSV F ectodomain linked to the TM and CT from a BPIV3 F protein, the BPIV3 P protein, the BPIV3 M protein, the HPIV3 F protein, the HPIV3 HN protein, and the BPIV3 L protein comprise amino acid sequences set forth as SEQ ID NOs: 47, 21, 48, 49, 43, 101, and 52, respectively; and

the HPIV3 HN protein further comprises I263T and T370P substitutions.

19. The recombinant paramyxovirus of claim 1 , wherein the recombinant RSV F ectodomain linked to the TM and CT of the parainfluenza virus protein is encoded by the nucleic acid sequence set forth as SEQ ID NO: 23.

20. A recombinant paramyxovirus, comprising:

a viral genome comprising genes encoding parainfluenza virus N, P, M, F, HN, and L proteins, and further comprising a heterologous gene encoding a type I membrane protein comprising a recombinant respiratory syncytial virus (RSV) F ectodomain linked to a transmembrane domain (TM) and cytoplasmic tail (CT) of a parainfluenza virus F protein;

wherein the recombinant paramyxovirus is a recombinant human/bovine parainfluenza virus 3 (B/HPIV3); and

the recombinant RSV F ectodomain linked to the TM and CT from a BPIV3 F protein is encoded by the nucleic acid sequence set forth as SEQ ID NO: 23.

21. The recombinant paramyxovirus of claim 20 , wherein the viral genome comprises, from upstream to downstream, a BPIV3 genomic promoter, a gene encoding a BPIV3 N protein, the heterologous gene encoding the recombinant RSV F ectodomain, a gene encoding a BPIV3 P protein, a gene encoding a BPIV3 M protein, a gene encoding a HPIV3 F protein, a gene encoding a HPIV3 HN protein, and a gene encoding a BPIV3 L protein.

22. The recombinant paramyxovirus of claim 21 , wherein

the BPIV3 N protein, the BPIV3 P protein, the BPIV3 M protein, the HPIV3 F protein, the HPIV3 HN protein, and the BPIV3 L protein comprise amino acid sequences at least 90% identical to SEQ ID NOs: 47, 21, 48, 49, 43, 101, and 52, respectively; and

the HPIV3 HN protein further comprises I263T and T370P substitutions.

23. The recombinant paramyxovirus of claim 22 , wherein

the BPIV3 N protein, the BPIV3 P protein, the BPIV3 M protein, the HPIV3 F protein, the HPIV3 HN protein, and the BPIV3 L protein comprise amino acid sequences set forth as SEQ ID NOs: 47, 21, 48, 49, 43, 101, and 52, respectively; and

the HPIV3 HN protein further comprises the I263T and T370P substitutions.

24. The recombinant paramyxovirus of claim 20 , wherein the heterologous gene encoding the recombinant RSV F ectodomain is the first or second gene downstream of a genomic promoter of the viral genome.

25. The recombinant paramyxovirus of claim 20 , wherein at least 90% of viral particles produced by a host cell infected with the recombinant paramyxovirus or viral vector comprise a viral envelope comprising the ectodomain encoded by the heterologous gene.

26. The recombinant paramyxovirus of claim 20 , wherein the recombinant paramyxovirus is an infectious, attenuated, and self-replicating virus.

27. The recombinant paramyxovirus of claim 20 , wherein the RSV F ectodomain is present on the viral envelope of the paramyxovirus.

28. An immunogenic composition comprising the recombinant paramyxovirus of claim 20 and a pharmaceutically acceptable carrier.

29. A method of eliciting an immune response to respiratory syncytial virus and parainfluenza virus in a subject comprising administering a therapeutically effective amount of the immunogenic composition of claim 28 to the subject.

30. A nucleic acid molecule comprising the genome of the recombinant paramyxovirus of claim 20 .

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Jan 16, 2020
From: COLLINS, PETER; LIANG, BO; MUNIR, SHIRIN; NUTT, ANNE SCHAAP; BUCHHOLZ, URSULA; MACKOW, NATALIE; KWONG, PETER; GRAHAM, BARNEY; MCLELLAN, JASON
To: THE UNITED STATES OF AMERICA, AS REPRESENTED BY THE SECRETARY, DEPARTMENT OF HEALTH AND HUMAN SERVICES
Reel/Frame 051539/0951 →
Continuity (2)
Provisional Application 62105667 · Jan 20, 2015
Related Publication 20180312544A1 · Nov 1, 2018
Cited By (1)
US 12,643,925