PORCINE PARVOVIRUS 5B, METHODS OF USE AND VACCINE
The present invention provides novel nucleotides sequences, protein sequences, immunogenic compositions, vaccines, and methods that relate to making and using new porcine parvovirus 5B (PPV5B) that infects, inter alia, domestic swine. The compositions and methods provide for the detection of infections by said new virus, monitoring genetic changes in the viral sequences in wild and domestic animals and herds, and making and using novel vaccines for protecting animals from infection by the virus.
1 . An isolated polynucleotide, comprising a polynucleotide
a) having the nucleic acid sequence of SEQ ID NO:1;
b) having a nucleic acid sequence that encodes a polypeptide of SEQ ID NO:3 or SEQ ID NO:4;
c) having a nucleic acid sequence at least 80% identical to SEQ ID NO:1, which encodes a polypeptide having a biological or immunologically-effective activity of a polypeptide of SEQ ID NO:2, SEQ ID NO:3 or SEQ ID NO:4;
d) that is a fragment of the nucleic acid sequence of SEQ ID NO:1, comprising at least 30 contiguous nucleotide sequences encoding SEQ ID NO:4, or
e) that is a fragment of the nucleic acid sequence of SEQ ID NO:1, comprising at least 30 contiguous nucleotide sequences of SEQ ID NO:1, and which encode an immunologically-effective activity of an amino acid sequence of SEQ ID NO:3 or SEQ ID NO:4.
2 . An isolated polypeptide, comprising a polypeptide
a) having the amino acid sequence of SEQ ID NO:3 or SEQ ID NO:4;
b) having an amino acid sequence at least 80% identical to SEQ ID NO:3 or SEQ ID NO:4 and having a biological or immunologically-effective activity of a polypeptide encoded by SEQ ID NO:3 or SEQ ID NO:4;
c) that is a fragment of the amino acid sequence of SEQ ID NO:3 or SEQ ID NO:4, comprising at least 13 contiguous amino acids of SEQ ID NO:3 or SEQ ID NO:4;
d) that is a fragment of the amino acid sequence of SEQ ID NO:3 or SEQ ID NO:4, comprising at least 13 contiguous amino acids of SEQ ID NO:3 or SEQ ID NO:4, and having an immunologically-effective activity; or
e) a protein fragment that is encoded by a polynucleotide that comprises at least 39 nucleotides included in the sequences of nucleotides 2161-2860 of SEQ ID NO:1, or nucleotides 2861-5014 of SEQ ID NO:1.
3 . An isolated porcine parvovirus 5B (PPV5B), comprising
a) a nucleic acid sequence of SEQ ID NO:1, or
b) a nucleic acid sequence at least 80% identical to SEQ ID NO:1, which encodes a polypeptide having a biological or immunologically-effective activity of a polypeptide of SEQ ID NO:3 or SEQ ID NO:4.
4 . The PPV5B of claim 3 , comprising an attenuated, non-virulent form of a PPV5B.
5 . A vaccine for treating or preventing an infection by a virulent PPV5B, comprising a killed or attenuated form of a PPV5B of claim 3 .
6 . A vaccine for treating or preventing an infection by a virulent PPV5B, comprising a subunit of a killed or attenuated form of a PPV5B of claim 3 .
7 . A vaccine of claim 6 , wherein the subunit is a capsid protein of SEQ ID NO:4.
8 . A vaccine of claim 6 , wherein the subunit is an immunologically-effective fragment of a polypeptide of SEQ ID NO:4.
9 . An immunogenic preparation comprising an immunologically-effective amount of a polypeptide of claim 2 , and a pharmaceutical- or veterinary-acceptable carrier, excipient or diluent.
10 . A method of generating an immune response in a mammal, comprising administering an immunologically-effective amount of a PPV5B of claim 3 .
11 . A method of generating an immune response in a mammal, comprising administering an immunologically-effective amount of a polypeptide of claim 2 .
12 . A method of generating an immune response in a mammal, comprising administering an immunologically-effective amount of a vaccine of claim 5 .
13 . A method of generating an immune response in a mammal, comprising administering an immunologically-effective amount of a vaccine of claim 6 .
14 . A method of generating an immune response in a mammal, comprising administering an immunologically-effective amount of a vaccine of claim 7 .
15 . A method of claim 10 , wherein the mammal is a swine, and the immune response provides protective immunity to disease caused by PPV5B infection.
16 . A method of claim 11 , wherein the mammal is a swine, and the immune response provides protective immunity to disease caused by PPV5B infection.
17 . A method of claim 12 , wherein the mammal is a swine, and the immune response provides protective immunity to disease caused by PPV5B infection.
18 . A method of claim 13 , wherein the mammal is a swine, and the immune response provides protective immunity to disease caused by PPV5B infection.
19 . A method of claim 14 , wherein the mammal is a swine, and the immune response provides protective immunity to disease caused by PPV5B infection.
20 . An isolated antibody that specifically binds to a PPV5B polypeptide encoded by a porcine parvovirus 5B polynucleotide having a sequence of SEQ ID NO:1, said polypeptide having an amino acid sequence of SEQ ID NO:2, SEQ ID NO:3 or SEQ ID NO:4, and wherein said antibody does not bind to a polypeptide encoded by a different parvovirus.
21 . A method of identifying the presence of PPV5B in a biological sample, comprising
a) contacting the biological sample with an antibody of claim 20 ,
b) detecting the formation of a complex between the antibody and a PPV5B polypeptide,
wherein the presence of said complex indicates the presence of PPV5B in the biological sample.
22 . A vector or plasmid that comprises a polynucleotide of claim 1 .
23 . A host cell comprising a vector of claim 22 .
24 . A hybridoma that expresses an antibody of claim 20 .
25 . A diagnostic kit for identifying the presence of PPV5B in a biological sample, comprising
a) an antibody of claim 20 , and
b) reagents for detecting the formation of a complex between the antibody and a PPV5B polypeptide.
26 . An immunogenic kit, comprising
a) at least one immunogenic PPV5B peptide which is effective to immunize an animal against at least one disease associated with PPV5B infection;
b) at least one carrier or adjuvant molecule;
c) a container for packaging the immunogenic composition;
d) a set of printed instructions; and
e) a dispenser capable of administering the immunogenic composition to an animal.
27 . An immunogenic kit of claim 26 , further comprising at least one immunogenic protein which is effective to immunize said animal against at least one other porcine pathogenic virus or bacteria which causes a disease associated with an infection with said virus or bacteria.