Novel prrs virus inducing type i interferon in susceptible cells

Abstract

The present invention relates to the field of attenuated live viruses useful as vaccine or medicament for preventing or treating Porcine Reproductive and Respiratory Syndrome (PRRS) in swine, and is based on the surprising finding of a PRRS virus which is able to induce the interferon type I response of a cell infected by said virus. In one embodiment, the PRRS virus according to the invention is a PRRS virus mutant comprising, in comparison with the genome of a wild type strain, a mutation in the gene encoding the non structural protein 1 (nsp1) of said virus.

1 . A replication-competent Porcine Reproductive and Respiratory Syndrome (PRRS) virus comprising a deletion of at least three consecutive nucleotide triplets in the gene sequence encoding the N-terminal domain (NTD) of the nsp1β subunit of the nsp1 protein of said virus, wherein the NTD starts with the N-terminal amino acid sequence SXXY (SEQ ID NO: 60) of nsp1β and ends with the serine (S) residue within the amino acid sequence SFP (SEQ ID NO: 62), wherein said deletion of at least three consecutive nucleotide triplets comprises a deletion of the nucleotide triplet encoding the first arginine residue (R) located at least 21 amino acid residues in the C-terminal direction from the N-terminal amino acid residue of nsp1β and a deletion of the two nucleotide triplets coding for the two amino acid residues flanking said R residue in the N-terminal direction, and wherein the modified PRRS virus induces production and secretion of interferon type I by a cell infected with said virus. 
     
     
         2 . The PRRS virus according to  claim 1 , wherein said mutation comprises a deletion of three consecutive nucleotide triplets coding for the amino acid sequence RXR (SEQ ID NO: 66) of the nsp1β subunit, or wherein said mutation comprises a deletion of four consecutive nucleotide triplets coding for the amino acid residues GRXR (SEQ ID NO: 68) of the nsp1β subunit, wherein X is a genetically encoded amino acid residue. 
     
     
         3 . An immunogenic composition comprising the PRRS virus according to  claim 1 . 
     
     
         4 . The immunogenic composition according to  claim 3 , comprising an amount of 10 1  to 10 7  viral particles per dose 
     
     
         5 . The immunogenic composition according to  claim 3 , comprising an amount of 10 3  to 10 6  particles per dose. 
     
     
         6 . The immunogenic composition according to  claim 3 , comprising an amount of 10 4  to 10 6  particles per dose. 
     
     
         7 . The immunogenic composition according to  claim 3 , comprising an amount of said PRRS virus which is equivalent to a virus titer of at least about 10 3  TCID50/mL per dose. 
     
     
         8 . The immunogenic composition according to  claim 3 , comprising an amount of said PRRS virus which is equivalent to a virus titer of between 10 3  to 10 5  TCID50/mL per dose. 
     
     
         9 . The immunogenic composition according to  claim 3 , further comprising one or more pharmaceutically acceptable carriers or excipients. 
     
     
         10 . The immunogenic composition according to  claim 9 , wherein said one or more pharmaceutically acceptable carriers or excipients are selected from the group consisting of solvents, dispersion media, adjuvants, stabilizing agents, diluents, preservatives, antibacterial and antifungal agents, isotonic agents, and adsorption delaying agents. 
     
     
         11 . The PRRS virus according to  claim 1 , wherein said PPRS virus has increased sensitivity to interferon type I. 
     
     
         12 . The PRRS virus according to  claim 1 , wherein said PRRS virus is an attenuated PRRS virus. 
     
     
         13 . The PRRS virus according to  claim 1 , wherein said PPRS virus is a genotype I virus. 
     
     
         14 . The PRRS virus according to  claim 1 , wherein said interferon type I is Interferon-α, and/or Interferon-β or combinations thereof. 
     
     
         15 . The PRRS virus according to  claim 1 , wherein said cell is a mammalian cell, and wherein said mammalian cell is a cell of a primary or secondary cell line or a host cell. 
     
     
         16 . The PRRS virus according to  claim 15 , wherein said mammalian cell is a porcine cell or a simian cell. 
     
     
         17 . The PRRS virus according to  claim 16 , wherein said porcine cell is a porcine macrophage. 
     
     
         18 . The PRRS virus according to  claim 16 , wherein said simian cell is a MA-104 or a MARC-145 cell. 
     
     
         19 . A polynucleotide comprising the genome of the PRRS virus according to  claim 1 . 
     
     
         20 . A DNA Vector comprising a copy of the polynucleotide of  claim 19 . 
     
     
         21 . A virus particle comprising the polynucleotide of  claim 19 . 
     
     
         22 . A cell comprising the polynucleotide of  claim 19 . 
     
     
         23 . A cell comprising the DNA-vector of  claim 20 . 
     
     
         24 . A method of preparing a PRRS virus composition comprising the cultivation of the PRRS virus according to  claim 1  in cell culture. 
     
     
         25 . A method for the treatment of Porcine Reproductive and Respiratory Syndrome comprising administering an effective amount of the PRRS virus according to  claim 1  to a swine, pig, piglet or a sow in need of said treatment. 
     
     
         26 . A method for inducing an immune response against PRRSV comprising administering an effective amount of the immunogenic composition according to  claim 1  to a swine, pig, piglet or a sow in need thereof. 
     
     
         27 . A method for the detection of interferon type I and/or the mRNA coding for interferon type I, by the cell infected with the modified PRRS virus according to  claim 1 , wherein the interferon type I and/or the mRNA coding for the interferon type I is detected by a bioassay, wherein said bioassay is preferably selected from the group consisting of ELISA, PCR, GLISA, IFA, biosensoric measurement, Surface Plasmon Resonance (SPR) measurement, selective media, lateral flow, biochip measurement, immunomagnetic separation, electrochemiluminescence, chromogenic media, immunodiffusion, DNA hybridization, staining, colorimetric detection, luminescence, and combinations thereof. 
     
     
         28 . The method according to  claim 24 , wherein the immunogenic composition is administered in a single dose or in two doses.