PRRS virus variant, European PRRS virus cDNA clone, and uses thereof

• US 10,010,601 B2
• Filed: 12/19/2014
• Issued: 07/03/2018
• Est. Priority Date: 12/20/2013
• Status: Active Grant

• 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: Gallei, Andreas, Keller, Christoph, Schacht, Erik, Herrel, Marieke
• Primary Examiner(s): Foley, Shanon A.
• Application Number:
  US15/105,760
• Publication Number:
  US 20160317642A1
• Time in Patent Office: 1,292 Days
• Field of Search: None
• CPC Class Codes: A61K 2039/5254 avirulent or attenuated
• CPC Class Codes: A61K 2039/552 Veterinary vaccine
• CPC Class Codes: A61K 39/12 Viral antigens
• 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: C12N 15/86 Viral vectors
• CPC Class Codes: C12N 2770/10021 Viruses as such, e.g. new isolates, mutants or their genomic sequences
• 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 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 1/02 involving viable microorganisms
• CPC Class Codes: G01N 2333/08 RNA viruses

Abstract

The present invention belongs to the field of animal health and provides means to study Porcine Reproductive and Respiratory Syndrome (PRRS), a viral disease affecting swine, and for the development of vaccines, therapeutics and diagnostics for the prophylaxis, treatment and diagnosis of PRRS. In a first consideration, the invention relates to a new PRRS virus variant, and, in a second consideration, to a nucleic acid sequence which comprises the genome of an infectious genotype I (EU) PRRS virus clone. Based on this, new PRRS vaccine candidates with improved properties are provided.

Claims

1. A DNA molecule which encodes a genotype I PRRS virus and which is capable of producing live virus when transfected into cells, wherein said molecule comprises a nucleic acid sequence having at least 99.1% sequence identity with the nucleic acid sequence of SEQ ID NO:

48, wherein maximally 0.8% of the differences in sequence identity are located within the first 110 nucleotides of the 5′

end of SEQ ID NO;

48 and within the the last 97 nucleotides of the 3′

end of SEQ ID NO;

48 and wherein maximally 0.1% of the differences in sequence identity are located in the remaining nucleotides of SEQ ID NO;

48.

2. The DNA molecule of claim 1, wherein said virus is attenuated and/or wherein said virus is able to induce a protective immune response against respiratory and/or reproductive signs of disease after infection with Porcine Reproductive and Respiratory Syndrome (PRRS) virus in swine;

and/or wherein said virus is able to reach titers of at least 5×

10⁵ to 1×

10⁶ tissue culture infectious dose 50 (TCID₅₀) per milliliter (ml) within 24 hours post infection of MA104 cells, preferably at an MOI (multiplicity of infection) of 0.001 to 0.1, and/or wherein said virus is able to reach titers of at least 5×

10⁶ to 1×

10⁷ tissue culture infectious dose 50 (TCID₅₀) per milliliter (ml) within 48 hours post infection of MA104 cells, at an MOI (multiplicity of infection) of 0.001 to 0.1.

3. An RNA transcript of the DNA construct of claim 1.

4. A cell transfected with the DNA construct of claim 1 or with the RNA transcript of claim 3.

5. A genotype I PRRS virus produced by the cell of claim 4.

6. A genotype I PRRS virus whose genome comprises a nucleic acid molecule according to claim 1.

7. A method for producing a genotype I PRRS virus comprising transfecting a cell with the DNA construct of claim 1.

8. A composition comprising a nucleic acid molecule of claim 1 suspended in a suitable amount of a pharmaceutically acceptable diluent or excipient.

9. A method of prophylaxis or treatment of Porcine Reproductive and Respiratory Syndrome comprising administering the virus according to claim 5 to an animal.

10. A kit comprising the PRRS virus or a fragment thereof according to claim 5 as a detection marker for the differentiation between infected and vaccinated animals (DIVA).

11. A nucleic acid molecule of claim 1, wherein said nucleic acid sequence of SEQ ID NO:

48 comprises an nsp2 region from 1380 to 3808 and an ORF4 region encoding GP4 from 12805 to 13356.