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EP-3868400-B1 - MULTIVALENT PCV2 IMMUNOGENIC COMPOSITIONS AND METHODS OF PRODUCING SUCH COMPOSITIONS

EP3868400B1EP 3868400 B1EP3868400 B1EP 3868400B1EP-3868400-B1

Inventors

  • Eichmeyer, Marc Allan
  • HAYES, PHILLIP WAYNE
  • NITZEL, GREG
  • ROOF, MICHAEL B.
  • Schaeffer, Merrill Lynn

Dates

Publication Date
20260513
Application Date
20061228

Claims (18)

  1. A multivalent combination vaccine comprising PCV2 ORF2 protein and at least one immunogenic active component against another disease-causing organism in swine for use in a method for lessening the severity of clinical symptoms associated with PCV2 infection in a pig, wherein the severity of the clinical symptoms associated with PCV2 infection is lessened by administering the vaccine once to the pig, and wherein said vaccine comprises 2 µg to 50 µg PCV2 ORF2 protein per dose.
  2. The multivalent combination vaccine according to claim 1 for the use of claim 1, wherein said PCV2 ORF2 protein is obtainable by (i) permitting infection of susceptible cells in culture with a recombinant baculovirus viral vector containing PCV2 ORF2 DNA coding sequences, wherein ORF2 protein is expressed by the recombinant baculovirus viral vector, and (ii) thereafter recovering said ORF2 in the supernate.
  3. The multivalent combination vaccine of any one of claims 1 to 2 for the use of any one of claims 1 to 2, wherein said vaccine comprises an inactivated baculovirus viral vector and cell culture supernate.
  4. The multivalent combination vaccine of claim 3 for the use of claim 3, wherein said viral vector is inactivated by addition of BEI.
  5. The multivalent combination vaccine of claim 4 for the use of claim 4, wherein said BEI is neutralized with sodium thiosulfate.
  6. The multivalent combination vaccine of any one of claims 1 to 5 for the use of any one of claims 1 to 5, wherein said vaccine comprises an adjuvant.
  7. The multivalent combination vaccine of claim 6 for the use of claim 6, wherein said adjuvant is selected from the group consisting of acrylic polymers and methacrylic acid polymers.
  8. The multivalent combination vaccine of claim 6 or 7 for the use of claim 6 or 7, wherein said adjuvant is a carbomer.
  9. The multivalent combination vaccine of any one of claims 1 to 8 for the use of any one of claims 1 to 8, wherein said vaccine comprises 500 µg to 5 mg adjuvant per dose.
  10. The multivalent combination vaccine of any one of claims 1 to 9 for the use of any one of claims 1 to 9, wherein said vaccine comprises 4 µg to 16 µg PCV2 ORF2 protein per dose.
  11. The multivalent combination vaccine of any one of claims 1 to 10 for the use of any one of claims 1 to 10, wherein said immunogenic active component is selected from the group consisting of an antigen of: Actinobacillus pleuropneumoniae; Adenovirus; Alphavirus such as Eastern equine encephalomyelitis viruses; Bordetella bronchiseptica; Brachyspira spp., preferably B. hyodysenteriae; B. piosicoli, Brucella suis, preferably biovars 1, 2, and 3; Classical swine fever virus; Clostridium spp., preferably Cl. difficile, Cl. perfringens types A, B, and C, Cl. novyi, Cl. septicum, Cl. tetani; Coronavirus, preferably Porcine Respiratory Corona virus; Eperythrozoonosis suis; Erysipelothrix rhusiopathiae; Escherichia coli; Haemophilus parasuis, preferably subtypes 1, 7 and 14; Hemagglutinating encephalomyelitis virus; Japanese Encephalitis Virus; Lawsonia intracellularis; Leptospira spp., preferably Leptospira australis; Leptospira canicola; Leptospira grippotyphosa; Leptospira icterohaemorrhagicae; and Leptospira interrogans; Leptospira pomona; Leptospira tarassovi; Mycobacterium spp., preferably M. avium, M. intracellulare and M.bovis; Mycoplasma hyopneumoniae (M hyo); Pasteurella multocida; Porcine cytomegalovirus; Porcine Parvovirus; Porcine Reproductive and Respiratory Syndrome (PRRS) Virus; Pseudorabies virus; Rotavirus; Salmonella spp., preferably S. thyphimurium and S. choleraesuis; Staph. hyicus; Staphylococcus spp., Streptococcus spp., preferably Strep. suis; Swine herpes virus; Swine Influenza Virus; Swine pox virus; Vesicular stomatitis virus; and Virus of vesicular exanthema of swine.
  12. The multivalent combination vaccine of any one of claims 1 to 11 for the use of any one of claims 1 to 11, wherein said further immunogenic active component is an immunological active component of Mycoplasma hyopneumoniae and an immunological active component of Porcine Reproductive and Respiratory Syndrome Virus.
  13. The multivalent combination vaccine of any one of claims 1 to 11 for the use of any one of claims 1 to 11, wherein said further immunogenic active component is an immunological active component of Porcine Reproductive and Respiratory Syndrome Virus.
  14. The multivalent combination vaccine of any one of claims 1 to 11 for the use of any one of claims 1 to 11, wherein said further immunogenic active component is an immunological active component of Mycoplasma hyopneumoniae.
  15. The multivalent combination vaccine of any one of claims 1 to 14 for the use of any one of claims 1 to 14, wherein said PCV2 ORF2 protein is recombinant PCV2 ORF2 protein.
  16. The multivalent combination vaccine of any one of claims 1 to 15 for the use of any one of claims 1 to 15, wherein the pig is a piglet.
  17. Use of PCV2 ORF2 protein in the manufacture of a combination vaccine for lessening the severity of clinical symptoms associated with PCV2 infection in a pig, wherein the vaccine comprises PCV2 ORF2 protein and at least one immunogenic active component of another disease-causing organism in swine, and wherein the severity of the clinical symptoms associated with PCV2 infection is lessened by administering the vaccine once to the pig, and wherein said vaccine comprises 2 µg to 50 µg PCV2 ORF2 protein per dose.
  18. The multivalent combination vaccine of any one of claims 1 to 16 for the use of any one of claims 1 to 16 or the use according to claim 17, wherein said lessening of the severity of clinical symptoms associated with PCV2 infection in a pig is a lowering of the viral titer in the infected pig.

Description

SEQUENCE LISTING This application contains a sequence listing in paper format and in computer readable format BACKGROUND OF THE INVENTION Field of the Invention a combination vaccine for use as claimed comprising One aspect of the present invention is concerned with a protein expressed by open reading frame 2 (ORF2) of porcine circovirus type 2 (PCV2). More particularly, the protein is a recombinant protein expressed by a transfected virus containing recombinant coding sequences for porcine circovirus type 2, open reading frame 2. Still more particularly, the transfected virus is permitted to infect cells in growth media and the protein expressed by open reading frame 2 is recovered in the supernate, rather than from inside the cells. Even more particularly, the method involves the steps of amplifying the open reading frame 2 gene from porcine circovirus type 2, cloning this amplified portion into a first vector, excising the open reading frame 2 portion from this first vector and cloning it into a transfer vector, cotransfecting the transfer vector with a viral vector into cells in growth media, causing the cells to become infected by the viral vector and thereby express open reading frame 2, and recovering the expressed recombinant protein coded for by open reading frame 2 in the supernate. a multivalent combination vaccine for use as claimed In another aspect, the present invention is concerned with effective for inducing an immune response against PCV2 The multivalent combination vaccine is effective for providing an immune response that protects an animal receiving the composition and reduces, or lessens the severity, of the clinical symptoms associated with PCV2 infection. Still more particularly, the present multivalent combination vaccine invention is concerned with a protein-based that confers effective protection against infection by PCV2. Even more particularly, the present invention is multivalent combination vaccine concerned with a comprising ORF2 of PCV2, wherein administration of PCV2-ORF2 results in protection against infection by PCV2. Most multivalent combination vaccine particularly, the present invention is concerned with effective multivalent combination vaccine for conferring effective immunity to a swine receiving the , and multivalent combination vaccine wherein the comprises the protein expressed by ORF2 of PCV2. In the present invention, combination vaccines or multivalent vaccines are provided. More particularly, the present invention provides immunogenic compositions effective at inducing an immune response against infection by PCV2 and at least one other disease-causing organism for swine. Description of the Prior Art Porcine circovirus type 2 (PCV2) is a small (17 -22 nm in diameter), icosahedral, non-enveloped DNA virus, which contains a single-stranded circular genome. PCV2 shares approximately 80% sequence identity with porcine circovirus type 1 (PCV1). However, in contrast with PCV1, which is generally non-virulent, swine infected with PCV2 exhibit a syndrome commonly referred to as Post-weaning Multisystemic Wasting Syndrome (PMWS). PMWS is clinically characterized by wasting, paleness of the skin, unthriftiness, respiratory distress, diarrhea, icterus, and jaundice. In some affected swine, a combination of all symptoms will be apparent while other swine will only have one or two of these symptoms. During necropsy, microscopic and macroscopic lesions also appear on multiple tissues and organs, with lymphoid organs being the most common site for lesions. A strong correlation has been observed between the amount of PCV2 nucleic acid or antigen and the severity of microscopic lymphoid lesions. Mortality rates for swine infected with PCV2 can approach 80%. In addition to PMWS, PCV2 has been associated with several other infections including pseudorabies, porcine reproductive and respiratory syndrome (PRRS), Glasser's disease, streptococcal meningitis, salmonellosis, postweaning colibacillosis, dietetic hepatosis, and suppurative bronchopneumonia. Open reading frame 2 (ORF2) protein of PCV2, having an approximate molecular weight of 30 kDa when run on SDS-PAGE gel, has been utilized in the past as an antigenic component in vaccines for PCV2. Typical methods of obtaining ORF2 for use in such vaccines generally consist of amplifying the PCV2 DNA coding for ORF2, transfecting a viral vector with the ORF2 DNA, infecting cells with the viral vector containing the ORF2 DNA, permitting the virus to express ORF2 protein within the cell, and extracting the ORF2 protein from the cell via cell lysis. These procedures generally take up to about four days after infection of the cells by the viral vector. However, these procedures have a disadvantage in that the extraction procedures are both costly and time-consuming. Additionally, the amount of ORF2 recovered from the cells is not very high; consequently, a large number of cells need to be infected by a large number of viral vectors in or