Circovirus sequences associated with piglet weight loss disease (PWD)
View Patent ↗The genome sequences and the nucleotide sequences coding for the PWD circovirus polypeptides, such as the circovirus structural and non-structural polypeptides, vectors including the sequences, and cells and animals transformed by the vectors are provided. Methods for detecting the nucleic acids or polypeptides, and kits for diagnosing infection by a PWD circovirus, also are provided. Method for selecting compounds capable of modulating the viral infection is further provided. Pharmaceutical, including vaccines, compositions for preventing and/or treating viral infections caused by PWD circovirus and the use of vectors for preventing and/or treating diseases also are provided.
1. A vaccine comprising a recovered recombinant protein expressed by open reading frame 2 (ORF′2) of PCVB wherein recovering said protein comprises: A) cloning said recombinant ORF′2 from PCVB into a transfer vector; B) transferring the portion of said transfer vector containing said recombinant ORF′2 into a virus; C) infecting cells in media with said virus; D) causing said virus to express the protein from said ORF′2 and the cells to secrete said protein into the media; E) separating the cells from the media; and F) recovering said expressed ORF′2 protein from the media.
2. The vaccine according to claim 1 , wherein recovering said protein further includes the step of amplifying said ORF′2 from a strain of PCVB prior to cloning said ORF′2 into said transfer vector.
3. The vaccine according to claim 1 , wherein said PCVB ORF′2 comprises SEQ ID NO:25.
4. The vaccine according to claim 1 , wherein said cells comprise SF+ cells.
5. The vaccine according to claim 1 , wherein said virus comprises baculovirus.
6. The vaccine according to any one of claims 1 - 5 , further comprising an adjuvant.
7. A vaccine comprising a composition for inducing an immune response against PCVB, wherein preparing said composition comprises the steps of: a) transferring a recombinant DNA from ORF′2 of PCVB into a virus; b) infecting cells with said virus, said cells being in a growth media; c) causing said virus to express the protein from said ORF′2 and the cells to secrete said protein into the media; d) separating the cells from the media; e) recovering said recombinant protein from the media; and f) combining said recovered protein with a suitable adjuvant or other pharmaceutically acceptable carrier or excipient.
8. The vaccine according to claim 7 , wherein preparing said composition further includes the step of obtaining said DNA from a transfer vector.
9. The vaccine according to claim 7 , wherein preparing said composition further includes the step of amplifying said ORF′2 from a strain of PCVB prior to cloning said ORF′2 into said transfer vector.
10. The vaccine according to claim 7 , wherein said PCVB ORF′2 comprises SEQ ID NO:25.
11. The vaccine according to claim 7 , wherein said cells comprise SF+ cells.
12. The vaccine according to claim 7 , wherein said virus comprises baculovirus.
13. A vaccine comprising a recovered recombinant protein expressed by open reading frame 2 (ORF′2) of PCVB wherein recovering said protein comprises: A) cloning said recombinant ORF′2 from PCVB into a transfer vector; B) transferring the portion of said transfer vector containing said recombinant ORF′2 into a virus; C) infecting cells in media with said virus; D) causing said virus to express the protein from said ORF′2; and B) recovering said expressed ORF′2 protein.
14. The vaccine according to claim 13 , wherein recovering said protein further includes the step of amplifying said ORF′2 from a strain of PCVB prior to cloning said ORF′2 into said transfer vector.
15. The vaccine according to claim 13 , wherein said PCVB ORF′2 comprises SEQ ID NO:25.
16. The vaccine according to claim 13 , wherein said cells comprise SF+ cells.
17. The vaccine according to claim 13 , wherein said virus comprises baculovirus.
18. The vaccine according to any of one claims 13 - 17 further comprising an adjuvant.
19. A vaccine comprising a recovered protein expressed by ORF′2 from PCVB wherein the steps of recovering said protein comprises: a) infecting cells in growth media with a recombinant viral vector containing said ORF′2; b) causing said virus to express the protein from said ORF′2; and c) recovering said expressed protein.
20. The vaccine according to claim 19 , wherein said PCVB ORF′2 comprises SEQ ID NO:25.
21. The vaccine according to claim 19 , wherein said cells comprise SF+ cells.
22. The vaccine according to claim 19 , wherein said virus comprises baculovirus.
23. The vaccine according to any one of claims 19 - 22 , further comprising an adjuvant.
24. A vaccine comprising composition for inducing an immune response against PCVB, wherein preparing said composition comprises the steps of: a) transferring a recombinant DNA from ORF′2 of PCVB into a virus; b) infecting cells with said virus, said cells being in a growth media; c) causing said virus to express the protein from said ORF′2; d) recovering said recombinant protein; and e) combining said recovered protein with a suitable adjuvant or other pharmaceutically acceptable carrier or excipient.
25. The vaccine according to claim 24 , wherein preparing said composition further includes the step of obtaining said DNA from a transfer vector.
26. The vaccine according to claim 24 , wherein preparing said composition further includes the step of amplifying said ORF′2 from a strain of PCVB prior to cloning said ORF′2 into said transfer vector.
27. The vaccine according to claim 24 , wherein said PCVB ORF′2 comprises SEQ ID NO:25.
28. The vaccine according to claim 24 , wherein said cells comprise SF+ cells.
29. The vaccine according to claim 24 , wherein said virus comprises baculovirus.
30. A vaccine comprising a baculovirus-expressed immunogenic polypeptide encoded by a baculovirus containing an exogenous nucleotide sequence wherein the nucleotide sequence encodes said immunogenic polypeptide and the nucleotide sequence has at least 90% identity to the nucleotide sequence encoding ORF′2 (SEQ ID NO:25).
31. A vaccine comprising a virus vector containing and expressing an exogenous nucleotide sequence, wherein the nucleotide sequence encodes an immunogenic polypeptide whose nucleotide sequence has at least 90% identity to the nucleotide sequence encoding ORF′2 (SEQ ID NO:25).
32. The vaccine according to claim 30 or claim 31 , wherein the nucleotide sequence that encodes the immunogenic polypeptide has at least 95% identity to the nucleotide sequence encoding ORF′2 (SEQ ID NO:25).
33. The vaccine according to claim 30 or claim 31 , wherein the nucleotide sequence that encodes the immunogenic polypeptide has at least 98% identity to the nucleotide sequence encoding ORF′2 (SEQ ID NO:25).
34. The vaccine according to any one of claims 30 - 33 , further comprising an adjuvant.
35. The vaccine comprising a host cell transformed with the baculovirus of any one of claims 19 , 21 , or 22 .
36. The vaccine according to claim 35 , further comprising an adjuvant.
37. A vaccine comprising an immunogenic polypeptide having at least 90% identity to the nucleotide sequence encoded ORF′2 (SEQ ID NO:25), wherein the immunogenic polypeptide has been expressed from a host cell transformed with the baculovirus of claim 30 .
38. The vaccine according to claim 37 , wherein the nucleotide sequence that encodes the immunogenic polypeptide has at least 95% identity to the nucleotide sequence encoding ORF′2 (SEQ ID NO:25).
39. The vaccine according to claim 37 , wherein the nucleotide sequence that encodes the immunogenic polypeptide has at least 98% identity to the nucleotide sequence encoding ORF′2 (SEQ ID NO:25).
40. The vaccine according to claim 37 , wherein the immunogenic polypeptide is further isolated from a cell lysate of the transformed host cell.
41. The vaccine according to claim 40 , further comprising purifying the immunogenic polypeptide.
42. The vaccine according to claim 37 , further comprising purifying the immunogenic polypeptide from media in which the transformed host cell was cultured.
43. A vaccine comprising a baculovirus expressed immunogenic polypeptide encoded by a baculovirus containing an exogenous nucleotide sequence, wherein the nucleotide sequence encodes an immunogenic polypeptide comprising at least 5 contiguous amino acids of ORF′2 (SEQ ID NO:26).
44. The vaccine according to claim 43 , wherein the immunogenic polypeptide comprises at least 10 contiguous amino acids of ORF′2 (SEQ ID NO:26).
45. The vaccine according to claim 43 , wherein the immunogenic polypeptide comprises at least 15 contiguous amino acids of ORF′2 (SEQ ID NO:26).
46. The vaccine according to claim 43 , wherein the immunogenic polypeptide comprises at least 25 contiguous amino acids of ORF′2 (SEQ ID NO:26).
47. A vaccine comprising a host cell transformed with the baculovirus of claim 43 .
48. The vaccine according to any one of claims 43 - 47 , further comprising an adjuvant.
49. A vaccine comprising an immunogenic polypeptide comprising at least 5 contiguous amino acids of ORF′2 (SEQ ID NO:26) expressed from a host cell transformed with the baculovirus of claim 43 , said transformed host cell cultured under conditions whereby the immunogenic polypeptide is expressed.
50. The vaccine according to claim 49 , wherein the immunogenic polypeptide comprises at least 10 contiguous amino acids of ORF′2 (SEQ ID NO:26).
51. The vaccine according to claim 49 , wherein the immunogenic polypeptide comprises at least 15 contiguous amino acids of ORF′2 (SEQ ID NO:26).
52. The vaccine according to claim 49 , wherein the immunogenic polypeptide comprises at least 25 contiguous amino acids of ORF′2 (SEQ ID NO:26).
53. The vaccine according to claim 49 , wherein the immunogenic polypeptide is further isolated from a cell lysate of the transformed host cell.
54. The vaccine according to claim 53 , wherein the immunogenic polypeptide is further purified.
55. The vaccine according to claim 53 , wherein the immunogenic polypeptide is further purified from media in which the transformed host cell was cultured.
56. The vaccine according to any one of claims 49 - 55 , further comprising an adjuvant.
57. A vaccine comprising a baculovirus-expressed immunogenic polypeptide expressed from a baculovirus containing an exogenous nucleotide sequence, wherein the nucleotide sequence encodes an immunogenic polypeptide having at least 90% identity to ORF′2 (SEQ ID NO:26).
58. The vaccine according to claim 57 , wherein the immunogenic polypeptide has at least 95% identity to ORF′2 (SEQ ID NO:26).
59. The vaccine according to claim 57 , wherein the immunogenic polypeptide has at least 98% identity to ORF′2 (SEQ ID NO:26).
60. The vaccine according to any one of claims 57 - 59 , further comprising an adjuvant.