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

• US 20030232326A1
• Filed: 02/21/2003
• Published: 12/18/2003
• Est. Priority Date: 02/21/2002
• Status: Abandoned Application

• Current Assignee: Erasmus University Medical Center Rotterdam
• Original Assignee: Medimmune LLC (Astrazeneca PLC)
• Inventors: Haller, Aurelia, Fouchier, Ronaldus Adrianus Maria, Van Den Hoogen, Bernadetta Gerarda, Tang, Roderick, Osterhaus, Albertus Dominicus Marcellinus Erasmus
• Granted Patent: US 7,449,324 B2
• Time in Patent Office: Days
• US Class Current: 435/5
• CPC Class Codes: A61K 2039/5254 avirulent or attenuated
• CPC Class Codes: A61K 2039/5256 expressing foreign proteins
• CPC Class Codes: A61K 2039/545 characterised by the dose, timing or administration schedule
• CPC Class Codes: A61K 2039/552 Veterinary vaccine
• CPC Class Codes: A61K 2039/58 raising an immune response against a target which is not the antigen used for immunisation
• CPC Class Codes: A61K 2039/70 Multivalent vaccine
• CPC Class Codes: A61K 2123/00 Preparations for testing in vivo
• 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 11/06 Antiasthmatics
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 19/00 Drugs for skeletal disorders
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 31/14 for RNA viruses
• CPC Class Codes: A61P 31/16 for influenza or rhinoviruses
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 specific for leukemia
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: A61P 37/04 : Immunostimulants
• CPC Class Codes: A61P 7/00 : Drugs for disorders of the blood or the extracellular fluid
• 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 2710/22034 : Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
• 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 2760/18522 : New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
• CPC Class Codes: C12N 2760/18534 : Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
• CPC Class Codes: C12N 2760/18621 : Viruses as such, e.g. new isolates, mutants or their genomic sequences
• 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 2760/18634 : Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
• CPC Class Codes: C12N 2760/18643 : viral genome or elements thereof as genetic vector
• 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 2333/08 : RNA viruses
• 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 33/5011 : for testing antineoplastic activity
• CPC Class Codes: G01N 33/502 : for testing non-proliferative effects
• CPC Class Codes: G01N 33/5088 : of vertebrates
• CPC Class Codes: G01N 33/56983 : Viruses
• CPC Class Codes: Y02A 90/10 : Information and communication technologies [ICT] supporting adaptation to climate change, e.g. for weather forecasting or climate simulation

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. An isolated 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, wherein the virus is phylogenetically more closely related to a virus isolate comprising the nucleotide sequence of SEQ ID NO:

18, SEQ ID NO;

19, SEQ ID NO;

20, or SEQ ID NO;

21 than it is related to turkey rhinotracheitis virus, the etiological agent of avian rhinotracheitis.

2. The isolated negative-sense single stranded RNA of claim 1, wherein the phylogenetic analysis uses 100 bootstraps and 3 jumbles.

3. An isolated negative-sense single stranded RNA metapneumovirus, wherein the genome of the virus comprises a nucleotide sequence of SEQ ID NO:

18.

4. An isolated negative-sense single stranded RNA metapneumovirus, wherein the genome of the virus comprises a nucleotide sequence of SEQ ID NO:

19.

5. An isolated negative-sense single stranded RNA metapneumovirus, wherein the genome of the virus comprises a nucleotide sequence of SEQ ID NO:

20.

6. An isolated negative-sense single stranded RNA metapneumovirus, wherein the genome of the virus comprises a nucleotide sequence of SEQ ID NO:

21.

7. An isolated nucleic acid, wherein the nucleic acid has a nucleotide sequence that is at least 70% identical to SEQ ID NO:

18, SEQ ID NO;

19, SEQ ID NO;

20 or SEQ ID NO;

21, wherein sequence identity is determined over the entire length of SEQ ID NO;

19, SEQ ID NO;

20, SEQ ID NO;

21 or SEQ ID NO;

22.

8. An isolated nucleic acid, wherein the nucleic acid encodes a protein comprising

(i) an amino acid sequence that is at least 66% identical to the G protein of a mammalian MPV variant B1 (SEQ ID NO:

324);

(ii) an amino acid sequence that is at least 98.5% identical to the N protein of a mammalian MPV variant Bi (SEQ ID NO;

368);

(iii) an amino acid sequence that is at least 96% identical the P protein of a mammalian MPV variant B1 (SEQ ID NO;

376);

(iv) an amino acid sequence that is identical the M protein of a mammalian MPV variant B1 (SEQ ID NO;

360);

(v) an amino acid sequence that is at least 99% identical the F protein of a mammalian MPV variant B1 (SEQ ID NO;

316);

(vi) an amino acid sequence that is at least 98% identical the M2-1 protein of a mammalian MPV variant B1 (SEQ ID NO;

340);

(vii) an amino acid sequence that is at least 99% identical the M2-2 protein of a mammalian MPV variant B1 (SEQ ID NO;

348);

(viii) an amino acid sequence that is at least 83% identical the SH protein of a mammalian MPV variant B1 (SEQ ID NO;

384);

or

(ix) an amino acid sequence that is at least 99% identical the L protein a mammalian MPV variant B1 (SEQ i) NO;

332).

16. A method for detecting a variant BI mammalian MPV in a sample, wherein the method comprises contacting the sample with the nucleic acid of claim 8.

9. An isolated nucleic acid, wherein the nucleic acid encodes a protein comprising

(i) an amino acid sequence that is at least 66% identical to the G protein of a mammalian MPV variant A1 (SEQ ID NO:

322);

(ii) an amino acid sequence that is at least 99.5% identical to the N protein of a mammalian MPV variant A1 (SEQ ID NO;

366);

(iii) an amino acid sequence that is at least 96% identical to the P protein of a mammalian MPV variant A1 (SEQ ID NO;

374);

(iv) an amino acid sequence that is at least 99% identical to the M protein of a mammalian MPV variant A1 (SEQ ID NO;

358);

(v) an amino acid sequence that is at least 98% identical to the F protein of a mammalian MPV variant A1 (SEQ ID NO;

314);

(vi) an amino acid sequence that is at least 99% identical to the M2-1 protein of a mammalian MPV variant A1 (SEQ ID NO;

338);

(vii) an amino acid sequence that is at least 96% identical to the M2-2 protein of a mammalian MPV variant A1 (SEQ ID NO;

346);

(viii) an amino acid sequence that is at least 84% identical to the SH protein of a mammalian MPV variant A1 (SEQ ID NO;

382);

or

(ix) an amino acid sequence that is at least 99% identical to the L protein of a virus of a mammalian MPV variant A1 (SEQ ID NO;

330).

17. A method for detecting a variant Al mammalian MPV in a sample, wherein the method comprises contacting the sample with the nucleic acid of claim 9.

10. An isolated nucleic acid, wherein the nucleic acid encodes a protein comprising

(i) an amino acid sequence that is at least 66% identical to the G protein of a mammalian MPV variant A2 (SEQ ID NO:

332);

(ii) an amino acid sequence that is at least 99.5% identical to the N protein of a mammalian MPV variant A2 (SEQ ID NO;

367);

(iii) an amino acid sequence that is at least 96% identical to the P protein of a mammalian MPV variant A2 (SEQ ID NO;

375);

(iv) an amino acid sequence that is at least 99% identical to the M protein of a mammalian MPV variant A2 (SEQ ID NO;

359);

(v) an amino acid sequence that is at least 98% identical to the F protein of a mammalian MPV variant A2 (SEQ ID NO;

315);

(vi) an amino acid sequence that is at least 99% identical to the M2-1 protein of a mammalian MPV variant A2 (SEQ ID NO;

     339);

(vii) an amino acid sequence that is at least 96% identical to the M2-2 protein of a mammalian MPV variant A2 (SEQ ID NO;

347);

(viii) an amino acid sequence that is at least 84% identical to the SH protein of a mammalian MPV variant A2 (SEQ ID NO;

383);

or

(ix) an amino acid sequence that is at least 99% identical to the L protein of a mammalian MPV variant A2 (SEQ ID NO;

331).

12. An isolated nucleic acid, wherein the nucleic acid hybridizes specifically under high stringency conditions to the nucleic acid of claim 8, 9, 10 or 11.

13. An isolated nucleic acid, wherein the nucleic acid hybridizes under low stringency conditions to the nucleic acid of claim 8, 9, 10 or 11.

14. The isolated nucleic acid of claim 12, 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.

15. An isolated infectious virus, wherein the virus comprises the nucleic acid of claim 8, 9, 10 or 11.

18. A method for detecting a variant A2 mammalian MPV in a sample, wherein the method comprises contacting the sample with the nucleic acid of claim 10.

11. An isolated nucleic acid, wherein the nucleic acid encodes a protein comprising

(i) an amino acid sequence that is at least 66% identical to the G protein of a mammalian MPV variant B2 (SEQ ID NO:

325);

(ii) an amino acid sequence that is at least 97% identical to the N protein of a mammalian MPV variant B2 (SEQ ID NO;

369);

(iii) an amino acid sequence that is at least 96% identical to the P protein of a mammalian MPV variant B2 (SEQ ID NO;

377);

(iv) an amino acid sequence that is identical to the M protein of a mammalian MPV variant B2 (SEQ ID NO;

361);

(v) an amino acid sequence that is at least 99% identical to the F protein of a mammalian MPV variant B2 (SEQ ID NO;

317);

(vi) an amino acid sequence that is at least 98% identical to the M2-1 protein of a mammalian MPV variant B2 (SEQ ID NO;

341);

(vii) an amino acid sequence that is at least 99% identical to the M2-2 protein of a mammalian MPV variant B2 (SEQ ID NO;

349);

(viii) an amino acid sequence that is at least 84% identical to the SH protein of a mammalian MPV variant B2 (SEQ ID NO;

385);

or

(ix) an amino acid sequence that is at least 99% identical to the L protein of a mammalian MPV variant B2 (SEQ ID NO;

333).

19. A method for detecting a variant B2 mammalian MPV in a sample, wherein the method comprises contacting the sample with the nucleic acid of claim 11.

20. An isolated protein, wherein the protein comprises:

(i) an amino acid sequence that is at least 66% identical to the G protein of a mammalian MPV variant B1 (SEQ ID NO;

324);

(ii) an amino acid sequence that is at least 98.5% identical to the N protein of a mammalian MPV variant B1 (SEQ ID NO;

368);

(iii) an amino acid sequence that is at least 96% identical the P protein of a mammalian MPV variant B1 (SEQ ID NO;

376);

(iv) an amino acid sequence that is identical the M protein of a mammalian MPV variant B1 (SEQ ID NO;

360);

(v) an amino acid sequence that is at least 99% identical the F protein of a mammalian MPV variant B1 (SEQ ID NO;

316);

(vi) an amino acid sequence that is at least 98% identical the M2-1 protein of a mammalian MPV variant B1 (SEQ ID NO;

340);

(vii) an amino acid sequence that is at least 99% identical the M2-2 protein of a mammalian MPV variant B1 (SEQ ID NO;

348);

(viii) an amino acid sequence that is at least 83% identical the SH protein of a mammalian MPV variant B1 (SEQ ID NO;

384);

or

(ix) an amino acid sequence that is at least 99% identical the L protein a mammalian MPV variant B1 (SEQ ID NO;

332).

81. A method for serologically diagnosing a mammalian MPV infection of an animal, wherein the method comprises contacting a sample from the animal with the protein of claim 20, 21, 22 or 23.

83. A method for diagnosing an APV infection of a bird comprising contacting a sample from the animal with the protein of claim 20, 21, 22 or 23.

84. The method of claim 83, wherein said APV is APV-C.

21. An isolated protein, wherein the protein comprises:

(i) an amino acid sequence that is at least 66% identical to the G protein of a mammalian MPV variant A1 (SEQ ID NO;

322);

(ii) an amino acid sequence that is at least 99.5% identical to the N protein of a mammalian MPV variant A1 (SEQ ID NO;

366);

(iii) an amino acid sequence that is at least 96% identical to the P protein of a mammalian MPV variant A1 (SEQ ID NO;

374);

(iv) an amino acid sequence that is at least 99% identical to the M protein of a mammalian MPV variant A1 (SEQ ID NO;

358);

(v) an amino acid sequence that is at least 98% identical to the F protein of a mammalian MPV variant A1 (SEQ ID NO;

314);

(vi) an amino acid sequence that is at least 99% identical to the M2-1 protein of a mammalian MPV variant A1 (SEQ ID NO;

338);

(vii) an amino acid sequence that is at least 96% identical to the M2-2 protein of a mammalian MPV variant A1 (SEQ ID NO;

346);

(viii) an amino acid sequence that is at least 84% identical to the SH protein of a mammalian MPV variant A1 (SEQ ID NO;

382);

or

(ix) an amino acid sequence that is at least 99% identical to the L protein of a virus of a mammalian MPV variant A1 (SEQ ID NO;

330).

22. An isolated protein, wherein the protein comprises:

(i) an amino acid sequence that is at least 66% identical to the G protein of a mammalian MPV variant A2 (SEQ ID NO;

332);

(ii) an amino acid sequence that is at least 99.5% identical to the N protein of a mammalian MPV variant A2 (SEQ ID NO;

367);

(iii) an amino acid sequence that is at least 96% identical to the P protein of a mammalian MPV variant A2 (SEQ ID NO;

375);

(iv) an amino acid sequence that is at least 99% identical to the M protein of a mammalian MPV variant A2 (SEQ ID NO;

359);

(v) an amino acid sequence that is at least 98% identical to the F protein of a mammalian MPV variant A2 (SEQ ID NO;

315);

(vi) an amino acid sequence that is at least 99% identical to the M2-1 protein of a mammalian MPV variant A2 (SEQ ID NO;

     339);

(vii) an amino acid sequence that is at least 96% identical to the M2-2 protein of a mammalian MPV variant A2 (SEQ ID NO;

347);

(viii) an amino acid sequence that is at least 84% identical to the SH protein of a mammalian MPV variant A2 (SEQ ID NO;

383);

or

(ix) an amino acid sequence that is at least 99% identical to the L protein of a mammalian MPV variant A2 (SEQ ID NO;

331).

23. An isolated protein, wherein the protein comprises:

(i) an amino acid sequence that is at least 66% identical to the G protein of a mammalian MPV variant B2 (SEQ ID NO;

325);

(ii) an amino acid sequence that is at least 97% identical to the N protein of a mammalian MPV variant B2 (SEQ ID NO;

369);

(iii) an amino acid sequence that is at least 96% identical to the P protein of a mammalian MPV variant B2 (SEQ ifD NO;

377);

(iv) an amino acid sequence that is identical to the M protein of a mammalian MPV variant B2 (SEQ ID NO;

361);

(v) an amino acid sequence that is at least 99% identical to the F protein of a mammalian MPV variant B2 (SEQ ID NO;

317);

(vi) an amino acid sequence that is at least 98% identical to the M2-1 protein of a mammalian MPV variant B2 (SEQ ID NO;

341);

(vii) an amino acid sequence that is at least 99% identical to the M2-2 protein of a mammalian MPV variant B2 (SEQ ID NO;

349);

(viii) an amino acid sequence that is at least 84% identical to the SH protein of a mammalian MPV variant B2 (SEQ ID NO;

385);

or

(ix) an amino acid sequence that is at least 99% identical to the L protein of a mammalian MPV variant B2 (SEQ ID NO;

333).

24. An antibody, wherein the antibody binds specifically to a protein consisting of:

(i) an amino acid sequence that is at least 66% identical to the G protein of a mammalian MPV variant B1 (SEQ ID NO;

324);

(ii) an amino acid sequence that is at least 98.5% identical to the N protein of a mammalian MPV variant Bi (SEQ ID NO;

368);

(iii) an amino acid sequence that is at least 96% identical the P protein of a mammalian MPV variant B1 (SEQ ID NO;

376);

(iv) an amino acid sequence that is identical the M protein of a mammalian MPV variant B1 (SEQ ID NO;

360);

(v) an amino acid sequence that is at least 99% identical the F protein of a mammalian MPV variant B1 (SEQ ID NO;

316);

(vi) an amino acid sequence that is at least 98% identical the M2-1 protein of a mammalian MPV variant B1 (SEQ ID NO;

340);

(vii) an amino acid sequence that is at least 99% identical the M2-2 protein of a mammalian MPV variant B1 (SEQ ID NO;

348);

(viii) an amino acid sequence that is at least 83% identical the SH protein of a mammalian MPV variant B1 (SEQ ID NO;

384);

or

(ix) an amino acid sequence that is at least 99% identical the L protein a mammalian MPV variant B1 (SEQ ID NO;

332).

28. A method for detecting a variant B1 mammalian MPV in a sample, wherein said method comprises contacting the sample with the antibody of claim 24.

32. A method for identifying a viral isolate as a mammalian MPV, wherein said method comprises contacting said isolate or a component thereof with the antibody of claim 24, 25, 26 or 27.

33. A method for virologically diagnosing a MPV infection of a mammal comprising determining in a sample of said mammal the presence of a viral isolate or component thereof by contacting the sample with the antibody of claim 24, 25, 26 or 27.

34. A method for virologically diagnosing a mammalian MPV infection of a subject, wherein said method comprises:

(a) obtaining a sample from the subject;

(b) contacting the sample with the antibody of claim 24, 25, 26 or 27,

wherein if the antibody binds to the sample the subject is infected with mammalian MPV.

25. An antibody, wherein the antibody binds specifically to a protein consisting of:

(i) an amino acid sequence that is at least 66% identical to the G protein of a mammalian MPV variant A1 (SEQ ID NO;

322);

(ii) an amino acid sequence that is at least 99.5% identical to the N protein of a mammalian MPV variant A1 (SEQ ID NO;

366);

(iii) an amino acid sequence that is at least 96% identical to the P protein of a mammalian MPV variant A1 (SEQ ID NO;

374);

(iv) an amino acid sequence that is at least 99% identical to the M protein of a mammalian MPV variant A1 (SEQ ID NO;

358);

(v) an amino acid sequence that is at least 98% identical to the F protein of a mammalian MPV variant A1 (SEQ ID NO;

314);

(vi) an amino acid sequence that is at least 99% identical to the M2-1 protein of a mammalian MPV variant A1 (SEQ ID NO;

338);

(vii) an amino acid sequence that is at least 96% identical to the M2-2 protein of a mammalian MPV variant A1 (SEQ ID NO;

346);

(viii) an amino acid sequence that is at least 84% identical to the SH protein of a mammalian MPV variant A1 (SEQ ID NO;

382);

or

(ix) an amino acid sequence that is at least 99% identical to the L protein of a virus of a mammalian MPV variant A1 (SEQ ID NO;

330).

29. A method for detecting a variant Al mammalian MPV in a sample, wherein said method comprises contacting the sample with the antibody of claim 25.

26. An antibody, wherein the antibody binds specifically to a protein consisting of:

(i) an amino acid sequence that is at least 66% identical to the G protein of a mammalian MPV variant A2 (SEQ ID NO;

332);

(ii) an amino acid sequence that is at least 96% identical to the N protein of a mammalian MPV variant A2 (SEQ ID NO;

367);

(iii) an amino acid sequence that is at least 96% identical to the P protein of a mammalian MPV variant A2 (SEQ ID NO;

375);

(iv) an amino acid sequence that is at least 99% identical to the M protein of a mammalian MPV variant A2 (SEQ ID NO;

359);

(v) an amino acid sequence that is at least 98% identical to the F protein of a mammalian MPV variant A2 (SEQ ID NO;

315);

(vi) an amino acid sequence that is at least 99% identical to the M2-1 protein of a mammalian MPV variant A2 (SEQ ID NO;

     339);

(vii) an amino acid sequence that is at least 96% identical to the M2-2 protein of a mammalian MPV variant A2 (SEQ ID NO;

347);

(viii) an amino acid sequence that is at least 84% identical to the SH protein of a mammalian MPV variant A2 (SEQ ID NO;

383);

or

(ix) an amino acid sequence that is at least 99% identical to the L protein of a mammalian MPV variant A2 (SEQ ID NO;

331).

30. A method for detecting a variant A2 mammalian MPV in a sample, wherein said method comprises contacting the sample with the antibody of claim 26.

27. An antibody, wherein the antibody binds specifically to a protein consisting of:

(i) an amino acid sequence that is at least 66% identical to the G protein of a mammalian MPV variant B2 (SEQ ID NO;

325);

(ii) an amino acid sequence that is at least 97% identical to the N protein of a mammalian MPV variant B2 (SEQ ID NO;

369);

(iii) an amino acid sequence that is at least 96% identical to the P protein of a mammalian MPV variant B2 (SEQ ID NO;

377);

(iv) an amino acid sequence that is identical to the M protein of a mammalian MPV variant B2 (SEQ ID NO;

361);

(v) an amino acid sequence that is at least 99% identical to the F protein of a mammalian MPV variant B2 (SEQ ID NO;

317);

(vi) an amino acid sequence that is at least 98% identical to the M2-1 protein of a mammalian MPV variant B2 (SEQ ID NO;

341);

(vii) an amino acid sequence that is at least 99% identical to the M2-2 protein of a mammalian MPV variant B2 (SEQ ID NO;

349);

(viii) an amino acid sequence that is at least 84% identical to the SH protein of a mammalian MPV variant B2 (SEQ ID NO;

385);

or

(ix) an amino acid sequence that is at least 99% identical to the L protein of a mammalian MPV variant B2 (SEQ ID NO;

333).

31. A method for detecting a variant B2 mammalian MPV in a sample, wherein said method comprises contacting the sample with the antibody of claim 27.

35. An infectious recombinant virus, wherein the recombinant virus comprises the genome of a mammalian MPV and further comprises a non-native MPV sequence.

48. The infectious chimeric virus of claim 35, 42, 43, 46 or 47, wherein the avian MPV is APV type A, APV type B, APV type C or APV type D.

49. The infectious chimeric virus of claim 35, 42, 43, 46 or 47, wherein the mammalian MPV is variant A1, variant A2, variant BI or variant B2.

50. The infectious virus of claim 35, 42, 43, 46 or 47, wherein the virus further comprises a heterologous nucleotide sequence.

51. The infectious virus of claim 50, wherein the heterologous nucleotide sequence is inserted at position 1, 2, 3, 4, 5, or 6 of the metapneumovirus genome.

52. The infectious virus of claim 50, wherein a nucleotide sequence of the genome of the virus is substituted with the heterologous nucleotide sequence.

53. The virus of claim 50, wherein the heterologous sequence is derived from a negative stand RNA virus.

54. The virus of claim 50, wherein the heterologous sequence is derived from a RSV, PIV, APV, or from a mammalian MPV.

55. The virus of claim 54, wherein the RSV is a respiratory syncytial virus type A, a respiratory syncytial virus type B, a bovine respiratory syncytial virus or an ovine respiratory syncytial virus.

56. The virus of claim 54, wherein said parainfluenza virus is a parainfluenza virus type 1, a parainfluenza virus type 2, a parainfluenza virus type 3, a parainfluenza virus type 4 or a bovine parainfluenza virus.

57. The virus of anyone of claims 50-56, wherein the heterologous sequence encodes a F protein or a G protein.

58. The virus of claim 50, wherein said heterologous sequence is derived from a virus that causes Acquired Immune Deficiency Syndrome.

59. The virus of claim 35, 40, 42, 43, 46, or 47, wherein the virus is an attenuated virus.

60. The virus of claim 35, 40, 42, 43, 46, or 47, wherein the genome of said virus contains mutations or modifications, in addition to said heterologous nucleotide sequences, that result in a chimeric virus having a phenotype more suitable for use in vaccine formulations such an attenuated phenotype or a phenotype with enhanced antigenicity or enhanced immunogenicity.

61. The virus of claim 35, 40, 42, 43, 46, or 47, wherein the genome of the mammalian MPV is a mammalian MPV comprising the nucleotide sequence of SEQ ID NO:

18, SEQ ID NO;

19, SEQ ID NO;

20 or SEQ ID NO;

21.

62. An immunogenic composition, wherein the immunogenic composition comprises the infectious recombinant virus of claim 35, 40, 42, 43, 46, or 47.

63. A pharmaceutical composition, wherein the pharmaceutical composition comprises the infectious recombinant virus of claim 35, 40, 42, 43, 46, or 47.

65. A method for treating or preventing a respiratory tract infection in a mammal, said method comprising administering a vaccine comprising the recombinant mammalian metapneumovirus of claim 35, 40, 42, 43, 46, or 47.

68. A method for treating or preventing a respiratory tract infection in a subject, said method comprising administering to the subject the composition of claim 62 or 63.

36. A recombinant nucleic acid, wherein the recombinant nucleic acid comprises (i) a nucleic acid encoding a G polypeptide of an MPV Al variant;

and (ii) a nucleic acid encoding a non-native MPV polypeptide.

40. An isolated infectious recombinant virus, wherein the recombinant virus is encoded by the nucleic acid of claim 36, 37, 38 or 39.

41. The isolated infectious recombinant virus of claim 40, wherein the nucleic acid of claim 36, 37, 38 or 39 further comprises a heterologous sequence.

37. A recombinant nucleic acid, wherein the recombinant nucleic acid comprises (i) a nucleic acid encoding a G polypeptide of an MPV A2 variant;

and (ii) a nucleic acid encoding a non-native MPV polypeptide.

38. A recombinant nucleic acid, wherein the recombinant nucleic acid comprises (i) a nucleic acid encoding a G polypeptide of an MPV B I variant;

and (ii) a nucleic acid encoding a non-native MPV polypeptide.

39. A recombinant nucleic acid, wherein the recombinant nucleic acid comprises (i) a nucleic acid encoding a G polypeptide of an MPV B2 variant;

and (ii) a nucleic acid encoding a non-native MPV polypeptide.

42. An infectious chimeric virus, wherein the chimeric virus comprises the genome of a mammalian MPV of a first variant, wherein one or more of the open reading frames in the genome of the mammalian MPV of the first variant have been replaced by the analogous open reading frame from a mammalian MPV of a second variant.

44. The infectious chimeric virus of claim 42 or 43, wherein

(i) if the first variant is A1, the second variant is A2, B1 or B2;

(ii) if the first variant is A2, the second variant is A1, B1 or B2;

(iii) if the first variant is B1, the second variant is A1, A2 or B2;

(iv) if the first variant is B2, the second variant is A1, A2, or B I.

45. The infectious chimeric virus of claim 42 or 43, wherein the analogous open reading frame encodes a F protein or a G protein.

43. An infectious chimeric virus, wherein the chimeric virus comprises the genome of a mammalian MPV of a first variant, wherein one or more of open reading frames of a mammalian MPV of a second variant are inserted into the genome of the mammalian MPV of the first variant.

46. An infectious chimeric virus, wherein the chimeric virus comprises the genome of a mammalian MPV, wherein one or more of the open reading frames in the genome of the mammalian MPV have been replaced by an ORF which encodes one or more of

(i) an avian MPV F protein;

(ii) an avian MPV G protein;

(iii) an avian MPV SH protein;

(iv) an avian MPV N protein;

(v) an avian MPV P protein;

(vi) an avian MPV M2 protein;

(vii) an avian MPV M2-1 protein;

(viii) an avian MPV M2-2 protein;

or

(ix) an avian MPV L protein.

47. An infectious chimeric virus, wherein the chimeric virus comprises the genome of an avian MPV, wherein one or more of the open reading frames in the genome of the avian MPV have been replaced by an ORF which encodes one or more of

(i) a mammalian MPV F protein;

(ii) a mammalian MPV G protein;

(iii) a mammalian MPV SH protein;

(iv) a mammalian MPV N protein;

(v) a mammalian MPV P protein;

(vi) a mammalian MPV M2 protein;

(vii) a mammalian MPV M2-1 protein;

(viii) a mammalian MPV M2-2 protein;

or

(ix) a mammalian MPV L protein.

64. A method for treating or preventing a respiratory tract infection in a mammal, said method comprising administering a vaccine comprising a mammalian metapneumovirus.

69. The method of claim 64, 65, 66, 67 or 68, wherein the respiratory tract infection is a MPV infection.

70. The method of claim 64, 65, 66, 67 or 68, wherein the respiratory tract infection is an infection with MPV and RSV.

71. The method of claim 64, 65, 66, 67 or 68, wherein the subject is a human.

72. The method of claim 71, wherein the human subject is less than 5 years of age.

73. The method of claim 71, wherein the human subject is less than 2 years of age.

74. The method of claim 71, wherein the human subject suffers from a disease or a condition in addition to the respiratory tract infection.

75. The method of claim 71, wherein the disease or condition is selected from a group consisting of cystic fibrosis, leukaemia, non-Hodgkin lymphoma, Asthma, and bone marrow transplantation and kidney transplantation.

76. The method of claim 71, wherein the human subject is an immunocompromised individual.

77. The method of claim 71, wherein the human subject is an elderly.

66. A method for treating or preventing a respiratory tract infection in a mammal, said method comprising administering a vaccine comprising avian metapneumovirus.

67. A method for treating or preventing a respiratory tract infection in a human, said method comprising administering a vaccine comprising avian metapneumovirus.

78. A method for identifying a compound useful for the treatment of infections with mammalian MPV, wherein the method comprises:

(a) infecting an animal with a mammalian MPV;

(b) administering to the animal a test compound; and

(c) determining the effect of the test compound on the infection of the animal,

wherein a test compound that reduces the extent of the infection or that ameliorates the symptoms associated with the infection is identified as a compound useful for the treatment of infections with mammalian MPV.

79. A method for identifying a compound useful for the treatment of infections with mammalian MPV, wherein the method comprises:

(a) infecting a cell culture with a mammalian MPV;

(b) incubating the cell culture with a test compound; and

(c) determining the effect of the test compound on the infection of the cell culture,

wherein a test compound that reduces the extent of the infection is identified as a compound useful for the treatment of infections with mammalian MPV.

80. A method for diagnosing a mammalian MPV infection of an animal, wherein the method comprises determining in a sample of said animal the presence of a viral isolate or component thereof by reacting said sample with a nucleic acid or an antibody reactive with a component of an aviant pneumovirus, said nucleic acid or antibody being cross-reactive with a component of MPV.

82. A method for serologically diagnosing a mammalian MPV infection of an animal, wherein the method comprises contacting a sample from the animal with a protein of an APV.