Metapneumovirus strains and their use in vaccine formulations and as vectors for expression of antigenic sequences

• US 20100143407A1
• Filed: 12/31/2008
• Published: 06/10/2010
• Est. Priority Date: 01/19/2001
• Status: Active Grant

• Current Assignee: Erasmus University Medical Center Rotterdam
• Original Assignee: Medimmune Vaccines Incorporated (Astrazeneca PLC)
• Inventors: Haller, Aurelia, Osterhaus, Albertus Dominicus Marcellinus Erasmus, Tang, Roderick, Van Den Hoogen, Benadetta Gerarda, Fouchier, Ronaldus Adrianus Maria
• Granted Patent: US 8,722,341 B2
• Time in Patent Office: Days
• US Class Current: 424/204.100
• CPC Class Codes: A61K 2039/5254 avirulent or attenuated
• CPC Class Codes: A61K 2039/5256 expressing foreign proteins
• CPC Class Codes: A61K 39/12 Viral antigens
• CPC Class Codes: A61K 39/155 Paramyxoviridae, e.g. parainfluenza virus
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 31/14 for RNA viruses
• CPC Class Codes: A61P 37/04 Immunostimulants
• CPC Class Codes: C07K 14/005 from viruses
• CPC Class Codes: C07K 16/1027 Paramyxoviridae, e.g. respiratory syncytial virus
• CPC Class Codes: C12N 15/86 Viral vectors
• CPC Class Codes: C12N 2760/18321 Viruses as such, e.g. new isolates, mutants or their genomic sequences
• CPC Class Codes: C12N 2760/18322 New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
• CPC Class Codes: C12N 2760/18334 Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
• CPC Class Codes: C12N 2760/18343 viral genome or elements thereof as genetic vector
• CPC Class Codes: C12N 2760/18622 New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
• CPC Class Codes: C12N 2840/203 having an IRES
• CPC Class Codes: C12N 7/00 Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00
• CPC Class Codes: C12Q 1/701 Specific hybridization probes
• CPC Class Codes: C12Q 2600/158 Expression markers
• CPC Class Codes: G01N 33/5008 for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
• CPC Class Codes: G01N 33/502 : for testing non-proliferative effects
• CPC Class Codes: G01N 33/5091 : for testing the pathological state of an organism
• CPC Class Codes: G01N 33/56983 : Viruses

Abstract

The present invention provides an isolated mammalian negative strand RNA virus, metapneumovirus (MPV), within the sub-family Pneumoviridae, of the family Paramyxoviridae. The invention also provides isolated mammalian negative strand RNA viruses identifiable as phylogenetically corresponding or relating to the genus Metapneumovirus and components thereof. In particular the invention provides a mammalian MPV, subgroups and variants thereof. The invention relates to genomic nucleotide sequences of different isolates of mammalian metapneumoviruses, in particular human metapneumoviruses. The invention relates to the use of the sequence information of different isolates of mammalian metapneumoviruses for diagnostic and therapeutic methods. The present invention relates to nucleotide sequences encoding the genome of a metapneumovirus or a portion thereof, including both mammalian and avian metapneumovirus. The invention further encompasses chimeric or recombinant viruses encoded by said nucleotide sequences. The invention also relates to chimeric and recombinant mammalian MPV that comprise one or more non-native or heterologous sequences. The invention further relates to vaccine formulations comprising mammalian or avian metapneumovirus, including recombinant and chimeric forms of said viruses. The vaccine preparations of the invention encompass multivalent vaccines, including bivalent and trivalent vaccine preparations.

Claims

1-14. -14. (canceled)

15. An isolated negative-sense single stranded RNA metapneumovirus, wherein the negative-sense single stranded RNA metapneumovirus encodes:

(i) a N protein with at least 90% amino acid sequence identity to SEQ ID NO;

366;

(ii) a P protein with at least 70% amino acid sequence identity to SEQ ID NO;

374;

(iii) a M protein with at least 90% amino acid sequence identity to SEQ ID NO;

358;

(iv) a F protein with at least 82% amino acid sequence identity to SEQ ID NO;

314;

(v) a M2-1 protein with at least 85% amino acid sequence identity to SEQ ID NO;

338;

or

(vi) a M2-2 protein with at least 60% amino acid sequence identity to SEQ ID NO;

346,

wherein sequence identity is determined over the entire length of the respective protein.

16. The virus of claim 15, wherein the genome of the virus comprises a nucleotide sequence of SEQ ID NO:

19.

17. The virus of claim 15, wherein the virus is an attenuated virus.

18. An immunogenic composition, wherein the immunogenic composition comprises the virus of claim 15 or 16.

19. (canceled)

20. An isolated nucleic acid, wherein the nucleic acid encodes a protein selected from the group consisting of a protein comprising an amino acid sequence that is at least 90% identical to SEQ ID NO:

366;

a protein comprising an amino acid sequence that is at least 70% identical to SEQ ID NO;

374;

a protein comprising an amino acid sequence that is at least 90% identical to SEQ ID NO;

358;

a protein comprising an amino acid sequence that is at least 82% identical to SEQ ID NO;

314;

a protein comprising an amino acid sequence that is at least 85% identical to SEQ ID NO;

338; and

a protein comprising an amino acid sequence that is at least 60% identical to SEQ ID NO;

346.

26. An isolated nucleic acid, wherein the isolated nucleic acid hybridizes specifically under high stringency conditions with the nucleic acid of claim 20.

27. The isolated nucleic acid of claim 26, wherein said high stringency conditions comprise hybridization in a buffer consisting of 6×

SSC, 50 mM Tris-HCl (pH=7.5), 1 mM EDTA, 0.02% PVP, 0.02% Ficoll, 0.02% BSA and 100 μ

g/ml denatured salmon sperm DNA, for 48 hours at 65°

C., washing in a buffer consisting of 2×

SSC, 0.01% PVP, 0.01% Ficoll and 0.01% BSA, for 45 minutes at 37°

C., and washing in a buffer consisting of 0.1×

SSC, for 45 minutes at 50°

C.

28. A method of detecting a mammalian metapneumovirus in a sample, wherein the method comprises contacting the sample with the nucleic acid of claim 20 or 26.

29. The method of claim 28, wherein the mammalian MPV is a human MPV.

30. The nucleic acid of claim 20, wherein the nucleic acid further comprises a vector.

31. A host cell comprising the nucleic acid of claim 20.

43. The isolated nucleic acid of claim 20, wherein

(i) the nucleic acid encodes the genome of a mammalian negative-sense single stranded RNA virus MPV belonging to the sub-family Pneumovirinae of the family Paramyxoviridae and identifiable as phylogenetically corresponding to the genus Metapneumovirus;

and

(ii) the nucleic acid further encodes a heterologous sequence.

21-25. -25. (canceled)

32. An isolated protein, wherein the protein comprises an amino acid sequence selected from the group consisting of that is at least 90% identical to SEQ ID NO:

366;

an amino acid sequence that is at least 70% identical to SEQ ID NO;

374;

an amino acid sequence that is at least 90% identical to SEQ ID NO;

358;

an amino acid sequence that is at least 82% identical to SEQ ID NO;

314;

an amino acid sequence that is at least 85% identical to SEQ ID NO;

338; and

an amino acid sequence that is at least 60% identical to SEQ ID NO;

346.

38. An antibody, wherein the antibody binds specifically to the protein of claim 32.

39. A method of detecting a mammalian metapneumovirus in a sample, wherein the method comprises contacting the sample with the antibody of claims 38.

33-37. -37. (canceled)

40-42. -42. (canceled)

44-48. -48. (canceled)