Circovirus sequences associated with piglet weight loss disease (pwd)

Abstract

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 are further provided. Pharmaceutical, including vaccine, 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 - 16 . (canceled) 
     
     
         17 . A method for lessening the severity of or reducing the clinical symptoms caused by or associated with PCVB infection in a subject, comprising: administering to said subject at least one dose of an immunogenic composition comprising at least 2 μg of recombinant PCVB ORF′2 protein; or administering to said subject at least one dose of an immunogenic composition comprising at least 2 μg of recombinant PCVB ORF′2 protein and an immunogenic composition that comprises a PRRS antigen. 
     
     
         18 . The method of  claim 17 , wherein said PCVB ORF′2 protein is:
 i) a polypeptide comprising the sequence selected from the group consisting of SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:55, SEQ ID NO: 58, and combinations thereof;   ii) any polypeptide that is at least 80% homologous to the polypeptide of i);   iii) any polypeptide that is at least 90% homologous to the polypeptide of i);   iv) any polypeptide that is at least 95% homologous to the polypeptide of i);   v) any polypeptide that is at least 98% homologous to the polypeptide of i);   vi) any immunogenic portion of the polypeptides of i), ii), iii), iv) and/or v);   vii) the immunogenic portion of vi), comprising at least 10 contiguous amino acids included in a sequence selected from the group consisting of SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:55, SEQ ID NO:58, and combinations thereof;   viii) a polypeptide that is encoded by a DNA comprising the sequence of SEQ ID NO:15, SEQ ID NO:19; SEQ ID NO:23, SEQ ID NO:25; SEQ ID NO: 27, or combinations thereof;   ix) any polypeptide that is encoded by a polynucleotide that is at least 80% homologous to the polynucleotide of viii);   x) any polypeptide that is encoded by a polynucleotide that is at least 90% homologous to the polynucleotide of viii);   xi) any polypeptide that is encoded by a polynucleotide that is at least 95% homologous to the polynucleotide of viii);   xii) any polypeptide that is encoded by a polynucleotide that is at least 98% homologous to the polynucleotide of viii);   xiii) any immunogenic portion of the polypeptides encoded by the polynucleotide of viii), ix), x), xi) and/or xii); or   xiv) the immunogenic portion of xiii), wherein the polynucleotide coding for said immunogenic portion comprises at least 30 contiguous nucleotides included in the sequences of SEQ ID NO:15, SEQ ID NO:19; SEQ ID NO:23, SEQ ID NO:25; SEQ ID NO:27, or combinations thereof.   
     
     
         19 . The method of  claim 17 , wherein said composition further comprises an additional component selected from the group consisting of an inactivated viral vector, a cell culture supernate, BEI, sodium thiosulfate, a carrier, an adjuvant, medium, a viral inactivator, a diluent, an isotonic agent, an immunomodulatory agent, an antibiotic, a pharmaceutically acceptable salt, and combinations thereof. 
     
     
         20 . The method of  claim 19 , wherein said inactivated viral vector is a recombinant baculovirus coding for the PCVB ORF′2 protein. 
     
     
         21 . The method of  claim 19 , wherein said adjuvant is selected from the group consisting of acrylic acid, methacrylic acid, and any polymer thereof. 
     
     
         22 . The method of  claim 17 , wherein said composition further comprises a polymer of an acrylic acid or methacrylic acid which is cross-linked with polyalkenyl ethers of sugars or polyalcohols. 
     
     
         23 . The method of  claim 17 , wherein said composition further comprises a pharmaceutically acceptable salt, preferably saline. 
     
     
         24 . The method of  claim 17 , wherein said immunogenic composition comprises:
 (a) at least 4 μg of recombinant PCVB ORF′2 protein;   (b) at least 8 μg of recombinant PCVB ORF′2 protein; or   (c) at least 16 μg of recombinant PCVB ORF′2 protein.   
     
     
         25 . The method of  claim 17 , wherein said immunogenic composition is a vaccine. 
     
     
         26 . The method of  claim 17 , wherein said composition is effective after a one dose administration of said recombinant PCVB ORF′2 protein. 
     
     
         27 . The method of  claim 17 , wherein the clinical symptoms are selected from the group consisting of lung lesions, nasal shedding, cough, diarrhea, and combinations thereof. 
     
     
         28 . The method of  claim 17  or  claim 26 , wherein said one dose of said immunogenic composition has a volume of at least 1 ml. 
     
     
         29 . The method of  claim 17  or  claim 26 , wherein said one dose of said immunogenic composition has a volume of at least 2 ml. 
     
     
         30 . The method of  claim 17 , wherein said composition is retained in a container. 
     
     
         31 . An immunogenic composition comprising at least 2 μg of recombinant PCVB ORF′2 protein, wherein said immunogenic composition is effective for lessening the severity of clinical symptoms associated with a PCVB infection. 
     
     
         32 . The immunogenic composition of  claim 31 , wherein said PCVB ORF′2 protein is:
 i) a polypeptide comprising the sequence selected from the group consisting of SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:55, SEQ ID NO: 58, and combinations thereof;   ii) any polypeptide that is at least 80% homologous to the polypeptide of i);   iii) any polypeptide that is at least 90% homologous to the polypeptide of i);   iv) any polypeptide that is at least 95% homologous to the polypeptide of i);   v) any polypeptide that is at least 98% homologous to the polypeptide of i);   vi) any immunogenic portion of the polypeptides of i), ii), iii), iv) and/or v);   vii) the immunogenic portion of vi), comprising at least 10 contiguous amino acids included in a sequence selected from the group consisting of SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:55, SEQ ID NO:58 and combinations thereof;   viii) a polypeptide that is encoded by a DNA comprising the sequence of SEQ ID NO:15, SEQ ID NO:19; SEQ ID NO:23, SEQ ID NO:25; SEQ ID NO: 27, or combinations thereof;   ix) any polypeptide that is encoded by a polynucleotide that is at least 80% homologous to the polynucleotide of viii);   x) any polypeptide that is encoded by a polynucleotide that is at least 90% homologous to the polynucleotide of viii);   xi) any polypeptide that is encoded by a polynucleotide that is at least 95% homologous to the polynucleotide of viii);   xii) any polypeptide that is encoded by a polynucleotide that is at least 98% homologous to the polynucleotide of viii);   xiii) any immunogenic portion of the polypeptides encoded by the polynucleotide of viii), ix), x), xi) and/or xii); or   xiv) the immunogenic portion of xiii), wherein the polynucleotide coding for said immunogenic portion comprises at least 30 contiguous nucleotides included in the sequences of SEQ ID NO:15, SEQ ID NO:19; SEQ ID NO:23, SEQ ID NO:25; SEQ ID NO:27, or combinations thereof.   
     
     
         33 . The immunogenic composition of  claim 31 , wherein said composition further comprises an additional component selected from the group consisting of an inactivated viral vector, a cell culture supernate, BEI, sodium thiosulfate, a carrier, an adjuvant, medium, a viral inactivator, a diluent, an isotonic agent, an immunomodulatory agent, an antibiotic, and combinations thereof. 
     
     
         34 . The immunogenic composition of  claim 33 , wherein said inactivated viral vector is a recombinant baculovirus coding for the PCVB ORF′2 protein. 
     
     
         35 . The immunogenic composition of  claim 33 , wherein said adjuvant is selected from the group consisting of acrylic acid, methacrylic acid, and any polymer thereof. 
     
     
         36 . The immunogenic composition of  claim 31 , wherein said composition further comprises a polymer of an acrylic acid or methacrylic acid which is cross-linked with polyalkenyl ethers of sugars or polyalcohols. 
     
     
         37 . The immunogenic composition of  claim 31 , wherein said composition further comprises a pharmaceutically acceptable salt, preferably saline. 
     
     
         38 . The immunogenic composition of  claim 31 , wherein said immunogenic composition comprises:
 (a) at least 4 μg of recombinant PCVB ORF′2 protein;   (b) at least 8 μg of recombinant PCVB ORF′2 protein; or   (c) at least 16 μg of recombinant PCVB ORF′2 protein.   
     
     
         39 . The immunogenic composition of  claim 31 , wherein said immunogenic composition is a vaccine. 
     
     
         40 . The immunogenic composition of  claim 31 , wherein said composition is effective after a one dose administration of said recombinant PCVB ORF′2 protein. 
     
     
         41 . The immunogenic composition of  claim 31 , wherein the clinical symptoms are selected from the group consisting of lung lesions, nasal shedding, cough, diarrhea, and combinations thereof. 
     
     
         42 . The immunogenic composition of  claim 40 , wherein said one dose of said immunogenic composition is formulated in at least 1 ml. 
     
     
         43 . The immunogenic composition of  claim 40 , wherein said one dose of said immunogenic composition is formulated in at least 2 ml. 
     
     
         44 . The immunogenic composition of  claim 40 , wherein said one dose of said composition is retained in a container. 
     
     
         45 . A kit comprising a container containing at least one dose of an immunogenic composition of at least 2 μg of recombinant PCVB ORF′2 protein. 
     
     
         46 . The kit of  claim 45 , further comprising a container containing an immunogenic composition comprising a PRRS antigen. 
     
     
         47 . The kit of  claim 45 , further comprising an instruction manual wherein said instruction manual includes information for the intramuscular application of at least one dose of the immunogenic composition into a piglet to lessen the severity of clinical symptoms associated with PCVB infection. 
     
     
         48 . The kit of  claim 45 , wherein said PCVB ORF′2 protein is:
 i) a polypeptide comprising the sequence selected from the group consisting of SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:55, SEQ ID NO: 58, and combinations thereof;   ii) any polypeptide that is at least 80% homologous to the polypeptide of i);   iii) any polypeptide that is at least 90% homologous to the polypeptide of i);   iv) any polypeptide that is at least 95% homologous to the polypeptide of i);   v) any polypeptide that is at least 98% homologous to the polypeptide of i);   vi) any immunogenic portion of the polypeptides of i), ii), iii), iv) and/or v);   vii) the immunogenic portion of vi), comprising at least 10 contiguous amino acids included in a sequence selected from the group consisting of SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:55, SEQ ID NO:58, and combinations thereof;   viii) a polypeptide that is encoded by a DNA comprising the sequence of SEQ ID NO:15, SEQ ID NO:19; SEQ ID NO:23, SEQ ID NO:25; SEQ ID NO: 27, or combinations thereof;   ix) any polypeptide that is encoded by a polynucleotide that is at least 80% homologous to the polynucleotide of viii);   x) any polypeptide that is encoded by a polynucleotide that is at least 90% homologous to the polynucleotide of viii);   xi) any polypeptide that is encoded by a polynucleotide that is at least 95% homologous to the polynucleotide of viii);   xii) any polypeptide that is encoded by a polynucleotide that is at least 98% homologous to the polynucleotide of viii);   xiii) any immunogenic portion of the polypeptides encoded by the polynucleotide of viii), ix), x), xi) and/or xii); or   xiv) the immunogenic portion of xiii), wherein the polynucleotide coding for said immunogenic portion comprises at least 30 contiguous nucleotides included in the sequences of SEQ ID NO:15, SEQ ID NO:19; SEQ ID NO:23, SEQ ID NO:25; SEQ ID NO:27, or combinations thereof.   
     
     
         49 . The kit of  claim 45 , wherein said composition further comprises an additional component selected from the group consisting of an inactivated viral vector, a cell culture supernate, BEI, sodium thiosulfate, a carrier, an adjuvant, medium, a viral inactivator, a diluent, an isotonic agent, an immunomodulatory agent, an antibiotic, or combinations thereof 
     
     
         50 . The kit of  claim 49 , wherein said inactivated viral vector is a recombinant baculovirus coding for the PCVB ORF′2 protein. 
     
     
         51 . The kit of  claim 49 , wherein said adjuvant is selected from the group consisting of acrylic acid, methacrylic acid, and any polymer thereof. 
     
     
         52 . The kit of  claim 45 , wherein said composition further comprises a polymer of an acrylic acid or methacrylic acid which is cross-linked with polyalkenyl ethers of sugars or polyalcohols. 
     
     
         53 . The kit of  claim 45 , wherein said composition further comprises a pharmaceutically acceptable salt, preferably saline. 
     
     
         54 . The kit of  claim 45 , wherein said immunogenic composition comprises:
 (a) at least 4 μg of recombinant PCVB ORF′2 protein;   (b) at least 8 μg of recombinant PCVB ORF′2 protein; or   (c) at least 16 μg of recombinant PCVB ORF′2 protein.   
     
     
         55 . The kit of  claim 45 , wherein said immunogenic composition is a vaccine. 
     
     
         56 . The kit of  claim 45 , wherein said composition is effective after a one dose administration of said recombinant PCVB ORF′2 protein. 
     
     
         57 . The kit of  claim 45 , wherein the clinical symptoms are selected from the group consisting of lung lesions, nasal shedding, cough, diarrhea, and combinations thereof. 
     
     
         58 . The kit of  claim 45 , wherein said one dose has a volume of at least 1 ml. 
     
     
         59 . The kit of  claim 45 , wherein said one dose has a volume of at least 2 ml. 
     
     
         60 . The kit of  claim 46 , further comprising an instruction manual including instructions indicating that both compositions should be administered to a pig. 
     
     
         61 . The kit of  claim 46 , further comprising an instruction manual including instructions indicating that the PCVB ORF′2 protein-containing composition should be administered before the PRRS antigen-containing composition.