PRRSV vaccines

• US 20030049274A1
• Filed: 09/07/2001
• Published: 03/13/2003
• Est. Priority Date: 03/08/1999
• Status: Abandoned Application

• Current Assignee: Boehringer Ingelheim Vetmedica GmbH (C.H. Boehringer Sohn AG & Co. KG)
• Original Assignee: Boehringer Ingelheim Vetmedica GmbH (C.H. Boehringer Sohn AG & Co. KG)
• Inventors: Verheije, Helene, Meulenberg, Janneke
• Application Number:
  US09/948,747
• Publication Number:
  US 20030049274A1
• Time in Patent Office: Days
• US Class Current: 424/204.1
• CPC Class Codes: A61K 2039/51 comprising whole cells, viruses or DNA/RNA
• CPC Class Codes: A61K 2039/5256 expressing foreign proteins
• CPC Class Codes: A61K 39/00 Medicinal preparations containing antigens or antibodies materials for immunoassay G01N33/53
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 37/04 Immunostimulants
• CPC Class Codes: C07K 14/005 from viruses
• CPC Class Codes: C12N 15/86 Viral vectors
• CPC Class Codes: C12N 2770/10022 New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
• CPC Class Codes: C12N 2770/10034 Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
• CPC Class Codes: C12N 2770/10043 viral genome or elements thereof as genetic vector
• CPC Class Codes: C12N 2770/10061 Methods of inactivation or attenuation
• 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

Abstract

The invention relates to the field of PRRS viruses and infectious clones obtained from PRRS viruses. Furthermore, the invention relates to vaccines and diagnostic assays obtainable by using and modifying such infectious clones of PRRS viruses. The invention provides a porcine reproductive and respiratory syndrom virus (PRRSV) replicon having at least some of its original PRRSV nucleic acid deleted, said replicon capable of in vivo RNA replication, said replicon further having been deprived of at least some of its original PRRSV nucleic acid and/or having been supplemented with nucleic acid derived from a heterologous microorganism.

Claims

1. A porcine reproductive and respiratory virus (PRRSV) replicon having at least some of its original PRRSV nucleic acid deleted, said replicon comprising essential elements from the PRRSV polymerase region and capable of in vivo RNA replication.

3. The PRRSV replicon of claim 1, further comprising nucleic acid derived from at least one heterologous microorganism.

4. The PRRSV replicon of any one of claims 1 to 3, wherein said replicon comprises RNA.

5. The PRRSV replicon of any one of claims 1 to 4, wherein said essential elements comprise essential nucleic acid sequence elements from the 5′

noncoding region, the ORF1a, the ORF 1b, the ORF7 and the 3′

noncoding region of PRRSV.

6. The PRRSV replicon of any one of claims 1 to 5, said essential elements comprise at least a nucleic acid corresponding to a 44 nucleotide region between nucleotides 14643 to 14687 in the ORF 7 region of PRRSV, as identified in the nucleic acid sequence of the Lelystad virus isolate.

7. The PRRSV replicon of any one of claims 1 to 6, further comprising at least one nucleic acid modification in the ORF7 gene encoding the N protein rendering it incapable of N-protein capsid formation during virus assembly.

8. The PRRSV replicon of claim 7, wherein said nucleic acid modification comprises a nucleic acid modification deleting a cysteine in the N-protein.

9. The PRRSV replicon of claim 7, wherein said nucleic acid modification comprises a nucleic acid modification leading to an amino acid change of the B-region comprising amino acids 25-30 of the N-protein (SEQ ID NO:

1).

10. The PRRSV replicon of claim 7, wherein said nucleic acid modification comprises a nucleic acid modification leading to an amino acid change of the D-region comprising amino acids 51-67 of the N-protein (SEQ ID NO:

2).

11. The PRRSV replicon of claim 7, wherein said nucleic acid modification comprises a nucleic acid modification leading to an amino acid change of the B-region comprising amino acids 80-90 of the N-protein (SEQ ID NO:

3).

12. The PRRSV replicon of any one of claims 1 to 11, further comprising a marker allowing serological discrimination.

13. The PRRSV replicon of claim 12, wherein said marker comprises a nucleic acid modification leading to an amino acid change of the original asparagine residue at position 62 of the N-protein.

14. The PRRSV replicon of claim 12, wherein said marker comprises a nucleic acid modification leading to an amino acid change in one or more ORFs selected from the group comprising ORF 2,3,4,5 and 6.

15. The PRRSV replicon of any one of claims 1 to 14, further comprising a nucleic acid modification in a virulence marker of PRRSV.

16. The PRRSV replicon of claim 15, wherein said nucleic acid modification comprises a modification in ORF 6 encoding the virion membrane spanning M-protein.

17. The PRRSV replicon of claim 16, wherein said nucleic acid modification modifies protein M in between its second and third virion membrane spanning fragment.

18. The PRRSV replicon of any one of claims 2 to 17, wherein said heterologous microorganism comprises a microorganism that is pathogenic to pigs.

19. The PRRSV replicon of claim 18, wherein said microorganism is a virus.

20. A vaccine comprising the PRRSV replicon of any one of claims 1 to 19.

21. A method of vaccinating pigs comprising:

administering to a pig, the vaccine of claim 20 in an amount sufficient to produce immunity.

2. A porcine reproductive and respiratory syndrome virus (PRRSV) replicon capable of in vivo RNA replication comprising PRRSV nucleic acid and nucleic acid derived from at least one heterologous microorganism.