PRRSV Vaccines

• US 20060240041A1
• Filed: 06/08/2006
• Published: 10/26/2006
• Est. Priority Date: 10/30/1996
• Status: Active Grant

• Current Assignee: Johanna Jacoba Maria Meulenberg, Monique Helene Verheije
• Original Assignee: Johanna Jacoba Maria Meulenberg, Monique Helene Verheije
• Inventors: Verheije, Monique Helene, Meulenberg, Johanna Jacoba Maria
• Granted Patent: US 8,790,656 B2
• Time in Patent Office: Days
• US Class Current: 424/204.100
• 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 replicon capable of in vivo replication, the replicon derived from a Porcine Reproductive and Respiratory Virus (PRRSV) RNA that comprises an original 5′

noncoding region, original ORF1 a and ORF1b that encode RNA dependent RNA polymerase, original ORFs 2 to 6 that encode structural proteins, ORF7 that encodes nucleocapsid protein N and comprises a region essential for RNA replication, and an original 3′

noncoding region, the replicon comprising (i) the original 5′

noncoding region;

(ii) the original ORF1a and ORF1b;

(iii) a replicon-ORF7 comprising the region of the original ORF7 that is essential for RNA replication; and

(iv) the original 3′

noncoding region, wherein at least some of the original ORF2, ORF3, ORF4, ORF5, ORF6, or ORF7 is deleted.

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

3. The replicon of claim 1, wherein the region of the original ORF7 that is essential for RNA replication comprises nucleotides 14642 to 14686 as identified in the nucleic acid sequence of the Lelystad virus isolate of PRRSV.

4. The replicon of claim 1, wherein the replicon-ORF7 further comprises a mutation rendering the nucleocapsid protein N incapable of forming a proper capsid.

5. The replicon of claim 4, wherein the replicon-ORF7 further comprises a nucleic acid modification, whereby a cysteine in the nucleocapsid protein N is substituted or deleted.

6. The replicon of claim 1, further comprising a marker allowing serological discrimination.

7. The replicon of claim 6, wherein said marker comprises a nucleic acid modification leading to an amino acid change in one or more of ORFs 2, 3, 4, 5, and 6.

8. The replicon of claim 6, wherein the marker comprises a nucleic acid modification, whereby Asp-62 in nucleocapsid protein N is substituted or deleted.

9. The replicon of claim 1, further comprising a nucleic acid modification in a virulence marker of PRRSV.

10. The replicon of claim 9, wherein said nucleic acid modification comprises a modification in a replicon-ORF6 leading to a change in virion membrane spanning M-protein.

11. The replicon of claim 10, wherein the virion membrane spanning M-protein is changed between its second and third virion membrane spanning fragments.

12. The replicon of claim 1, wherein the replicon-ORF7 further comprises a nucleic acid modification leading to an amino acid change in an antigenic region of the nucleocapsid protein N, whereby the replicon is antigenically different from Lelystad isolate PRRSV or North American PRRSV.

13. The replicon of claim 12, wherein the antigenic region comprises amino acids selected from the group consisting of:

amino acids 25 to 30 of the nucleocapsid protein N (SEQ. ID. NO.

     1), amino acids 51 to 67 of the nucleocapsid protein N (SEQ. ID. NO.

     2), amino acids 80 to 90 of the nucleocapsid protein N (SEQ. ID. NO.

     3), and combinations thereof.

14. A vaccine comprising:

a replicon capable of in vivo replication, the replicon derived from a Porcine Reproductive and Respiratory Virus (PRRSV) RNA that comprises an original 5′

noncoding region, original ORF1 a and ORF1 b that encode RNA dependent RNA polymerase, original ORFs 2 to 6 that encode structural proteins, ORF7 that encodes nucleocapsid protein N and comprises a region essential for RNA replication, and an original 3′

noncoding region, the replicon comprising (i) the original 5′

noncoding region;

(ii) the original ORF1a and ORF1b;

(iii) a replicon-ORF7 comprising the region of the original ORF7 that is essential for RNA replication; and

(iv) the original 3′

noncoding region, wherein at least some of the original ORF2, ORF3, ORF4, ORF5, ORF6, or ORF7 is deleted.

15. The vaccine of claim 14, wherein the replicon-ORF7 further comprises a mutation leading to the nucleocapsid protein N being incapable of forming a proper capsid.

16. The vaccine of claim 14, further comprising a marker allowing serological discrimination.

17. The vaccine of claim 14, further comprising a nucleic acid modification in a virulence marker of PRRSV.

18. A method of vaccinating pigs comprising:

administering to a pig a vaccine comprising;

a replicon capable of in vivo replication, the replicon derived from a Porcine Reproductive and Respiratory Virus (PRRSV) RNA that comprises an original 5′

noncoding region, original ORF1a and ORF1b that encode RNA dependent RNA polymerase, original ORFs 2 to 6 that encode structural proteins, ORF7 that encodes nucleocapsid protein N and comprises a region essential for RNA replication, and an original 3′

noncoding region, the replicon comprising (i) the original 5′

noncoding region;

(ii) the original ORF1a and ORF1b;

(iii) a replicon-ORF7 comprising the region of the original ORF7 that is essential for RNA replication; and

(iv) the original 3′

noncoding region, wherein at least some of the original ORF2, ORF3, ORF4, ORF5, ORF6, or ORF7 is deleted.

19. The method of claim 18, wherein the replicon-ORF7 further comprises a mutation leading to the nucleocapsid protein N being incapable of forming a proper capsid.

20. The method of claim 18, further comprising a marker allowing serological discrimination.

21. The method of claim 18, further comprising a nucleic acid modification in a virulence marker of PRRSV.