Porcine circovirus type 3 (PCV3) vaccines and their production and use

Abstract

The present invention relates to the use of immunogenic compositions comprising porcine circovirus type 3 (PCV3) antigens for the treatment of several clinical conditions (diseases), preferably those associated with PCV3 infection.

Description

[Technical Field]

[0001] Related Applications and Incorporation by Reference This application claims priority to U.S. Provisional Application No. 62 / 829,400, filed April 4, 2019, the entire contents of which are incorporated herein by reference. Reference is also made to WO 2006 / 072065; and U.S. Patent Nos. 6,103,526; 9,610,345; 9,669,087; and 10,450,351, the disclosures of which are incorporated by reference in their entireties. The aforementioned applications, and all documents cited therein or during the prosecution of those applications ("application cited documents"), and all documents cited or referenced in the application cited documents, and all documents cited or referenced herein ("documents cited herein"), and all documents cited or referenced in the documents cited herein, together with any manufacturer's instructions, descriptions, product specifications, and product sheets for any products mentioned in this specification or any document incorporated by reference herein, are hereby incorporated by reference and may be used in the practice of this invention. More particularly, all referenced documents are incorporated by reference to the same extent as if each individual document was specifically and individually indicated to be incorporated by reference.

[0002] Sequence Listing Statement The sequence listing associated with this application is provided in text format in lieu of a printed copy and is incorporated herein by reference. The name of the text file containing the sequence listing is BI 19-AH009_ST25(sequence listing).txt. The text file is 188 KB; was created on April 6, 2020; and has been submitted electronically via EFS-Web contemporaneously with the filing of this application.

[0003] FIELD OF THE INVENTION Disclosed herein are recombinant baculovirus vectors containing a polynucleotide encoding porcine circovirus type 3 (PCV3) ORF2. Also disclosed herein are compositions and vaccines produced from the baculovirus-derived PCV3 ORF2 and BaculoG / PCV3 ORF2. Also disclosed are recombinant baculovirus vectors containing a mutant polynucleotide encoding porcine circovirus type 3 (PCV3) ORF2. Also disclosed are compositions and vaccines produced from the baculovirus-derived mutant PCV3 ORF2 and BaculoG / PCV3 ORF2. [Background technology]

[0004] Porcine circovirus type 3 (PCV3) is a non-enveloped, icosahedral, single-stranded DNA (ssDNA) virus belonging to the genus Circovirus within the family Circoviridae. Its genome encodes two major open reading frames (ORFs), with ORF1 encoding the replication-related protein (rep) and ORF2 encoding the viral capsid (cap) protein, which determines the virus's antigenic characteristics. PCV3 is significantly genetically distinct from porcine circovirus type 2 (PCV2); specifically, the amino acid identity between the two viruses is only 48% within the rep gene and only 26% within the cap gene. PCV3 was originally reported in the United States in 2016 by Palinski, Rachel, et al., "A Novel Porcine Circovirus Distantly Related to Known Circoviruses Is Associated with Porcine Dermatitis and Nephropathy Syndrome and Reproductive Failure," Journal of Virology, vol. 91, no. 1, 26 October 2016. The virus has since been identified worldwide, including Germany, Japan, North Korea, Russia, China, Thailand, Italy, Spain, Denmark, South Korea, Poland, Brazil, Colombia, India, Serbia, and Sweden. While testing is currently limited, findings about PCV3 from retrospective samples indicate that the virus likely circulated in swine populations worldwide before its first report in 2017. As testing increases, it is hypothesized that PCV3 will be identified in older samples from more countries.

[0005] Additionally, Chinese Patent Application No. 109207441, entitled "3 type Cap protein of recombinant baculovirus expression pig circular ring virus and its construction method and primer," claims priority to Chinese Patent Application No. 201810912587, filed on August 12, 2018. Chinese Patent Application No. 201810912587 describes the construction of baculovirus expression of PCV3 ORF2 to produce type 3 Cap protein of porcine circular ring virus.

[0006] Chinese Patent Application No. 109207441, entitled "3 type Cap protein of recombinant baculovirus expressing pig circular ring virus and its construction method and primer," claims priority to Chinese Patent Application No. 201810912587, filed on August 12, 2018. Chinese Patent Application No. 201810912587 describes the administration of baculovirus-expressed PCV3 ORF2 in mice and presents seroconversion data by ELISA.

[0007] Chinese Patent Application No. 109207522, entitled "It expresses 3 types of pig circular ring virus and truncates Cap protein of recombinant baculovirus and its construction method and primer," claims priority to Chinese Patent Application No. 201810912585, filed on August 12, 2018. Chinese Patent Application No. 201810912585 describes baculovirus-mediated truncated CAP / ORF2, administration in mice, and presents seroconversion data by ELISA.

[0008] Additionally, U.S. Patent No. 10,450,351 (i.e., U.S. Application No. 15 / 768,356), entitled "Porcine Circovirus Type 3 Immunogenic Compositions and Methods of Making and Using the Same," was first published on October 25, 2018, as U.S. Application Publication No. 2018 / 0305410. U.S. Patent No. 10,450,351 claims priority to U.S. Provisional Patent Application No. 62 / 242,866, filed October 16, 2015 (Inventor: Ben Hause, assigned to Kansas State University Research Foundation; see also Palinski, Rachel, et al. Journal of Virology, vol. 91, no. 1, 26 Oct. 2016, doi:10.1128 / jvi.01879-16, published online October 26, 2016). U.S. Provisional Patent Application No. 62 / 242,866 is directed to PCV3 derived from tissues "recovered from four sows from a farm experiencing chronic poor reproductive performance that had acute deaths with clinical signs consistent with PDNS." The patent application does not mention the location of the farms, but describes immunohistochemistry (IHC) and quantitative PCR (qPCR) negative results for PCV2, porcine reproductive and respiratory syndrome virus (PRRSV), and influenza A virus (IAV) in sows, as well as mummified, stillborn, and / or weak fetuses. The patent application describes virus isolation but not cell culture propagation.

[0009] The examples in the '351 patent describe detection of the PCV3 capsid gene by qPCR, virus isolation, cloning of the PCV3 capsid protein, development of anti-PCV3 capsid monoclonal antibodies, detection of PCV3, and development of an ELISA for the recombinant PCV3 capsid, but do not describe any vaccine studies or data.

[0010] Recently, a paper was published describing the evaluation of the pathogenesis of PCV3 by intranasal inoculation of 4- and 8-week-old specific pathogen-free piglets with an infectious PCV3 DNA clone, but no discussion was given of a vaccine to prevent PCV3 infection (Jiang, Haijun, et al. “Induction of Porcine Dermatitis and Nephropathy Syndrome in Piglets by Infection with Porcine Circovirus Type 3.” Journal of Virology, vol. 93, no. 4, 28 Nov. 2018, doi:10.1128 / jvi.02045-18). Citation or identification of any document in this application is not an admission that such document is available as prior art to the present invention. Summary of the Invention

[0011] PCV3 ORF2 antigenic proteins and variants thereof are disclosed that are useful for vaccinating or treating animals, particularly swine. Typically, the swine is a pig. In some aspects of the invention, the animal is a piglet. Typically, the piglet is a sow that is 15 weeks old or less, or 6 weeks old or less, or 3 weeks old or less, or 2 weeks old or less, or 1 week old or less. In some aspects of the invention, the swine is a sow or a gilt. In some embodiments of the invention, the pig is a sow or gilt (i.e., a female pig that has not farrowed) that is less than 1 year old, typically more than 4 months old and less than 1 year old, typically more than 5 months old and less than 1 year old, typically more than 6 months old and less than 1 year old, typically 4-8 months old, typically 5-8 months old, typically 5-7 months old, typically 5-6 months old. In some embodiments of the invention, the pigs are less than 1 year old, typically more than 4 months old, and less than 1 year old, typically more than 5 months old, and less than 1 year old, typically more than 6 months old, and , pregnant pigs that are less than one year old. In some aspects of the invention, the pigs are pre-breeding gilts that are less than 1 year old, typically greater than 4 months old and less than 1 year old, typically greater than 5 months old and less than 1 year old, typically greater than 6 months old and less than 1 year old, typically 4-8 months old, typically 5-8 months old, typically 5-7 months old, typically 5-6 months old.

[0012] Disclosed is the development of a baculovirus-derived PCV3 ORF2 expressed from "BaculoG / PCV3 ORF2," a composition, and three vaccines: BaculoG / PCV3 ORF2, P9; a live vaccine adjuvanted with 50% ISA 207VG; BaculoG / PCV3 ORF2, P9; a live vaccine adjuvanted with 20% carbopol; and a control BaculoG / no insert, P4; a live vaccine adjuvanted with 20% carbopol. Data demonstrating the efficacy of the vaccines in preventing PCV3 disease are presented. Also disclosed is the development of a baculovirus-derived PCV3 ORF2 derived from a killed virus. Also disclosed is the development of a baculovirus-derived PCV3 ORF2 derived from a mutant killed virus.

[0013] Therefore, in a first aspect, the present invention relates to a composition comprising a PCV3 ORF2 protein, preferably an antigenic PCV3 ORF2 protein (PCV3 ORF2 antigen). The composition is also referred to hereinafter as the "composition of the present invention." It is also understood that the term "composition of the present invention" as used herein is synonymous with the "composition of the present disclosure."

[0014] Preferably, the compositions of the present invention further comprise a veterinarily acceptable carrier selected from the group consisting of solvents, dispersion media, coatings, stabilizers, diluents, preservatives, antimicrobial agents, antifungal agents, isotonic agents, absorption delaying agents, adjuvants, cell culture supernatants, stabilizers, viral vectors, expression vectors, immunomodulators, and / or any combination thereof.

[0015] The present disclosure further relates to porcine circovirus type 3 (PCV3) ORF2 protein; and a veterinarily acceptable carrier comprising a solvent, dispersion medium, coating, stabilizer, diluent, preservative, antimicrobial agent, antifungal agent, isotonic agent, absorption delaying agent, adjuvant, cell culture supernatant, stabilizer, virus or expression vector, immunomodulator, and / or any combination thereof. In one embodiment, the veterinarily acceptable carrier comprises an adjuvant, an immunomodulatory agent, a cell culture supernatant, a virus or an expression vector, or any combination thereof, hi another embodiment, the veterinarily acceptable carrier comprises an adjuvant.

[0016] PCV3 ORF2 can be from group a1, b1, or b2 (using the subclassification designation of Fux et al., "Full genome characterization of porcine circovirus type 3 isolates reveals the existence of two distinct groups of virus strains," Virology Journal (2018) 15:25, DOI 10.1186 / s12985-018-0929-3 (incorporated herein by reference); see, e.g., Table 4). Thus, PCV3 referred to herein is any phylogenetic clade or combination of clades of PCV3, or is preferably selected from the group consisting of PCV3a and PCV3b, and most preferably selected from the group consisting of PCV3a1, PCV3b1, PCV3b2, and PCV3c. Therefore, the compositions of the present invention preferably comprise a PCV3 ORF2 protein selected from the group consisting of PCV3a ORF2 protein and PCV3b ORF2 protein, or most preferably, a PCV3 ORF2 protein of any phylogenetic clade or combination of clades of PCV3, or selected from the group consisting of PCV3a1 ORF2 protein, PCV3b1 ORF2 protein, and PCV3b2 ORF2 protein. In another embodiment, the PCV3 ORF2 protein comprises or consists of an amino acid sequence encoded by a polynucleotide sequence having at least 90%, or at least 91%, or at least 92%, or at least 93%, or at least 94%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity or homology to SEQ ID NO:1.Preferably, the PCV3 ORF2 protein comprises or consists of an amino acid sequence having at least 90%, or at least 91%, or at least 92%, or at least 93%, or at least 94%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity with the sequence of SEQ ID NO:4. According to certain preferred embodiments, the PCV3 ORF2 protein is a recombinant protein, or most preferably, a recombinant baculovirus-expressed protein. Thus, the composition preferably comprises a recombinant PCV3 ORF2 protein, or most preferably, a baculovirus-expressed PCV3 ORF2 protein.

[0017] In another embodiment, the PCV3 ORF2 protein is a recombinant PCV3 ORF2 protein derived from its expression by an expression vector comprising a polynucleotide sequence encoding the PCV3 ORF2 protein. Advantageously, the expression vector is a baculovirus. In yet another embodiment, the composition further comprises a PCV2 ORF protein that can be derived from expression by an expression vector comprising a polynucleotide sequence encoding the PCV2 ORF2 protein. Advantageously, the expression vector is a baculovirus. Additionally, the composition may further comprise at least one additional antigen of a further swine pathogen, wherein the additional one or more antigens of the swine pathogen comprise a PRRSV antigen, a Mycoplasma hyopneumoniae bacterin antigen, a Mycoplasma hyopneumoniae supernatant antigen, a Pseudorabies antigen or a Pseudorabies antigen, an IAV antigen, a swine fever antigen (classical swine fever antigen or African swine fever antigen, or a combination thereof), an Actinobacillus pleuropneumoniae antigen, an Escherichia coli antigen, a porcine parvovirus (PPV) antigen, a Pasteurella multocida antigen, an Erysipelothrix rhusiopathiae antigen, or a Mycoplasma hyorhinis antigen.

[0018] In another embodiment, the PCV3 ORF2 protein is at a concentration of 0.2 to about 400 μg / ml, or 2 to about 400 μg / ml, or 4 to about 400 μg / ml, or 8 to about 400 μg / ml, or about 0.3 to about 200 μg / ml, or 2 to about 200 μg / ml, or 4 to about 200 μg / ml, or 8 to about 200 μg / ml, or about 0.35 to about 100 μg / ml, or 2 to about 100 μg / ml, or 4 to about 100 μg / ml, or 8 to about 100 μg / ml, or is present in an amount of about 0.4 to about 50 μg / ml, or 2 to about 50 μg / ml, or 4 to about 50 μg / ml, or 8 to about 50 μg / ml, or about 0.45 to about 30 μg / ml, or about 0.6 to about 15 μg / ml, or about 0.75 to about 8 μg / ml, or about 1.0 to about 6 μg / ml, or about 1.3 to about 3.0 μg / ml, or about 1.4 to about 2.5 μg / ml, or about 1.5 to about 2.0 μg / ml, or about 1.6 μg / ml. In certain embodiments, the composition may have PCV3 ORF2 protein in an amount ranging from about 1.5 to about 2.0 μg per ml of composition. For example, in one embodiment, a 1 ml dose of the composition may contain about 1.6 μg of PCV3 ORF2 protein.

[0019] In another embodiment, the PCV3 ORF2 protein, or the total PCV2 and PCV3 ORF2 protein, is about 0.2 to about 400 μg per dose, or 2 to 400 μg per dose, or 4 to about 400 μg per dose, or 8 to about 400 μg per dose, or about 0.3 to about 200 μg per dose, or 2 to about 200 μg per dose, or 4 to about 200 μg per dose, or 8 to about 200 μg per dose, or about 0.35 to about 100 μg per dose, or 2 to about 100 μg per dose, or 4 to about 100 μg per dose, or 8 to about 100 μg per dose, or The PCV3 and PCV2 ORF2 proteins may be present in an amount ranging from about 0.4 to about 50 μg per dose, or from 2 to about 50 μg per dose, or from 4 to about 50 μg per dose, or from 8 to about 50 μg per dose, or from about 0.45 to about 30 μg per dose, or from about 0.6 to about 15 μg per dose, or from about 0.75 to about 8 μg per dose, or from about 1.0 to about 6 μg per dose, or from about 1.3 to about 3.0 μg per dose, or from about 1.4 to about 2.5 μg per dose, or from about 1.5 to about 2.0 μg per dose, or about 1.6 μg per dose. In certain embodiments, the composition may have PCV3 and PCV2 ORF2 total protein in an amount ranging from about 1.5 to about 2.0 μg per ml of composition. For example, in certain embodiments, a 1 ml dose of the composition may contain about 1.6 μg of a combination of PCV3 and PCV2 ORF2 protein.

[0020] In another embodiment, the adjuvant is aluminum hydroxide; aluminum phosphate; saponin; Quil A; QS-21; GPI-0100; a water-in-oil emulsion; an oil-in-water emulsion; a water-in-oil-in-water emulsion; an emulsion based on light liquid paraffin oil or a European Pharmacopoeia emulsion. Pharmacopea-type adjuvants; isoprenoid oils; squalane; squalene oils resulting from the oligomerization of alkenes or isobutene or decene; esters of acids or alcohols containing a linear alkyl group; vegetable oils; ethyl oleate; propylene glycol dicaprylate / caprate; glyceryl tricaprylate / caprate; propylene glycol dioleate; esters of branched fatty acids or alcohols; isostearate esters; nonionic surfactants; sorbitan or mannide or glycol or polyglycerol or propylene glycol, or oleic acid or isostearic acid or ricinoleic acid or hydroxystearic acid, esters of mannitol anhydride, optionally ethoxylated; polyoxypropylene-polyoxyethylene block copolymers, Pluronic products, Carbopol; Carbopol 974P; Carbopol 934P; Carbopol 971P; polymers of acrylic or methacrylic acid; copolymers of maleic anhydride and alkenyl derivatives; crosslinked polymers of acrylic or methacrylic acid; crosslinked polymers of acrylic or methacrylic acid with polyalkenyl ethers of sugars or polyalcohols; carbomers; acrylic polymers crosslinked with polyhydroxylated compounds having at least three and not more than eight hydroxyl groups, in which at least three of the hydroxyl hydrogen atoms may be replaced, or have been replaced, by unsaturated aliphatic radicals having at least two carbon atoms, such as vinyl, allyl, and other ethylenically unsaturated groups, containing 2 to 4 carbon atoms, which may themselves contain other substituents, such as methyl; RIBI adjuvant system; block copolymers; SAF-M; monophosphoryl lipid A; avridine lipid-amine adjuvant; Escherichia coli (E.These include heat-labile enterotoxins (recombinant or otherwise) derived from Escherichia coli; cholera toxin; IMS1314; or muramyl dipeptide.

[0021] In yet another embodiment, the adjuvant may be present in an amount of about 50 μg to about 2000 μg; or the adjuvant may be present in an amount of about 250 μg per ml of the dose of the composition; or the adjuvant may be present in an amount of about 100 μg to about 10 mg per dose; or the adjuvant may be present in an amount of about 500 μg to about 5 mg per dose; or the adjuvant may be present in an amount of about 750 μg to about 2.5 mg per dose; or the adjuvant may be present in an amount of about 1 mg per dose. In certain embodiments, the composition may include an adjuvant in the range of about 750 μg to about 2.5 mg per dose of the composition. For example, in certain embodiments, a dose of the composition may include about 1 mg of adjuvant. In one embodiment, the immunomodulatory agent comprises an interleukin, interferon, or other cytokine.

[0022] The dosage of antibiotic can be from about 1 μg / ml to about 60 μg / ml of antibiotic, or less than about 30 μg / ml of antibiotic. For example, embodiments of the composition can include less than about 30 μg / ml of antibiotic.

[0023] In one embodiment, the antibiotic comprises gentamicin.

[0024] The composition of the present disclosure may include (i) PCV3 ORF2 protein; (ii) at least a portion of a baculovirus expressing the PCV3 ORF2 protein; (iii) a portion of a cell culture of cells infected or transfected with a recombinant baculovirus expressing the PCV3 ORF2 protein; (iv) an inactivating agent or an inactivating agent including binary ethyleneimine (BEI); (v) sodium thiosulfate or an amount of sodium thiosulfate equivalent to the inactivating agent or BEI; (vi) an adjuvant or an adjuvant including Carbopol or Carbopol 971; and (vii) a physiologically acceptable concentration of phosphate. In one embodiment, about 90% of components (i) through (iii) may have a size smaller than 1 μm, and the pH of the composition is adjusted to about 6.5-7.5. In another embodiment, the BEI is derived from a cell culture treated with about 2-8 or about 5 mM BEI to inactivate the baculovirus. In another embodiment, the composition contains about 2-8 or about 5 mM BEI, and the composition may contain about 1 mg of Carbopol or Carbopol 971. For example, an embodiment of the composition may include a cell culture treated with a concentration of about 5 mM BEI to inactivate baculovirus. In some embodiments, a dose of the composition may include residual BEI and / or about 1 mg of Carbopol, Carbopol 971, or a combination thereof.

[0025] Any of the compositions of the present disclosure can be formulated and / or packaged for single dose or one-shot administration, as well as multiple dose regimens, where it is anticipated that a single dose may overcome the presence of maternal antibodies. In one embodiment, the composition may be PCV3 and PPV (advantageously packaged in VLPs) and / or PRRSV, advantageously for use in breeding sows / gilts. In such embodiments, doses for single or multiple administrations are envisaged. According to another embodiment, the composition of the invention is an immunogenic composition. The present invention further provides a composition of the invention for use as a pharmaceutical. Additionally, the compositions of the present invention are provided for use as vaccines. According to certain preferred embodiments, the compositions of the present invention are for use in a method for eliciting an immune or immunological response, or a protective immune or immunological response, against (i) PCV3, and / or (ii) PCV2 and PCV3, and / or (iii) PCV3 and other swine pathogens, and / or (iv) PCV3, PCV2, and other swine pathogens.

[0026] According to another preferred embodiment, the composition of the present invention is a composition for use in a method for reducing or preventing clinical signs or disease caused by PCV3 infection in an animal, preferably a porcine animal, or for use in a method for treating or preventing PCV3 infection in an animal. Additionally, compositions of the present invention are provided for use in methods for inducing an immune response against PCV3 in pigs, particularly preferably in pregnant pigs. According to yet another aspect, the compositions of the present invention are provided for use in a method for reducing or preventing clinical signs or disease caused by infection with PCV3 in piglets, wherein the piglets are nursed by a sow to which the composition has been administered.

[0027] Thus, the present invention further provides a composition of the present invention for use in a method for reducing or preventing clinical signs or disease caused by infection with PCV3 in piglets suckled by a sow to which the immunogenic composition has been administered, said sow being pregnant, particularly pregnant with piglets, or a gilt prior to breeding age.

[0028] Preferably, the composition of the invention for use in any one of the methods described above is administered to the sow intramuscularly or intradermally, in particular.

[0029] The present disclosure also encompasses methods for inducing an immune or immunological response or a protective immune or immunological response against (i) PCV3, and / or (ii) PCV2 and PCV3, and / or (iii) PCV3 and another swine pathogen, and / or (iv) PCV3, PCV2, and another swine pathogen, comprising administering any of the compositions disclosed herein to an animal. The animal may be a pig. Advantageously, the porcine may be a pig, a piglet, or a sow. The pig or piglet may be 15 weeks or less, or 6 weeks or less, or 3 weeks or less, or 2 weeks or less, or 1 week or less. Administration may occur within at least 1, 2, or 3 weeks of exposure to the virulent porcine circovirus. Administration may occur within at least 1, 2, or 3 weeks of exposure to the virulent porcine circovirus. For some embodiments, administration can include a single, one-shot or single-dose administration of the protein of the invention or composition of the invention; and may not include multiple shot or multiple dose regimens. For some embodiments, administration can include multiple shots or multiple dose regimens of the protein of the invention or composition of the invention. The present invention further provides a method of immunizing a subject, the method comprising the step of administering to the subject a composition of the present invention. The present invention further provides a method for immunizing pigs against clinical disease caused by at least one pathogen, comprising the step of administering to the animal a composition of the present invention, wherein the immunogenic composition is capable of inducing an immune response that does not cause clinical signs of infection but immunizes the animal against a pathogenic form of the at least one pathogen, and wherein the at least one pathogen is preferably PCV3. Furthermore, the present invention provides a method for inducing the production of PCV3-specific antibodies in sows, the method comprising the step of administering a composition of the present invention. The sow may be a pregnant pig. Alternatively, the sow may be a gilt (i.e., a female pig that has not given birth), preferably a gilt that is not yet in breeding season. The present invention further provides a method for reducing or preventing clinical signs or symptoms caused by PCV3 infection in piglets, comprising: - administering a composition of the present invention to a sow; - allowing the piglets to be suckled by said sow; Including, The sow is preferably a sow that is pregnant with piglets. A method is provided.

[0030] Preferably, the latter above-mentioned method comprises: - administering a composition of the present invention to a sow that is pregnant with piglets; - allowing the sow to give birth to the piglets; - allowing the piglet to be suckled by the sow; Includes: The present invention further provides a method for reducing clinical signs and / or disease in piglets caused by porcine epidemic diarrhea virus (PEDV) infection, wherein the piglets are nursed by a sow to which a composition of the present invention has been administered.

[0031] Preferably, where applicable, in any one of the above methods, the composition of the invention is administered to said sow intramuscularly or intradermally, in particular. According to another preferred embodiment, the immunogenic composition of the invention is administered twice to said sow, in particular intramuscularly or intradermally. In another preferred embodiment, the clinical signs referred to herein are selected from the group consisting of reduced average daily weight gain, and death. In a further preferred embodiment, the clinical signs referred to herein are selected from the group consisting of expulsion of a mummified fetus, a stillborn fetus, and / or a weak fetus. In yet another preferred embodiment, the clinical signs referred to herein are selected from the group consisting of gross lesions, histological lesions, replication of PCV3 in tissues, and PCV3 viremia. In an even more preferred embodiment, the clinical sign referred to herein is selected from the group consisting of the development or production of a mummified fetus, a stillborn fetus, and / or a weak fetus.

[0032] The present disclosure also encompasses the use of any of the compositions disclosed herein in any of the methods disclosed herein; or the use of PCV3 ORF2 protein, alone or in combination with any one of the compositions disclosed herein, for use in the preparation of a composition for inducing an immunological or immune response, or a protective immune or immunological response, against (i) PCV3, and / or (ii) PCV2 and PCV3, and / or (iii) PCV3 and another swine pathogen, and / or (iv) PCV3, PCV2, and another swine pathogen, or for use in a method for inducing an immunological or immune response, or a protective immune or immunological response, against (i) PCV3, and / or (ii) PCV2 and PCV3, and / or (iii) PCV3 and another swine pathogen, and / or (iv) PCV3, PCV2, and another swine pathogen.

[0033] In one embodiment, the composition may be PCV3 and PPV (advantageously packaged in VLPs) and / or PRRSV, advantageously for use in breeding sows / gilts. In such embodiments, doses for single or multiple administrations are envisioned. This particular embodiment encompasses the use of PCV3 ORF2 protein in combination with PPV protein, optionally in combination with PRRSV protein, for use in the preparation of a composition for inducing an immunological or immune response, or a protective immune or immunological response, against PCV3 and PPV, optionally against PSSRV, or for use in a method for inducing an immunological or immune response, or a protective immune or immunological response, against PCV3 and PPV, optionally against PSSRV.

[0034] In this embodiment, the composition may comprise (i) a porcine circovirus type 3 (PCV3) ORF2 protein, a parvovirus (PPV) protein, and optionally a PRRSV (porcine reproductive and respiratory syndrome virus) protein; and (ii) a veterinarily acceptable carrier selected from the group consisting of a solvent, dispersion medium, coating agent, stabilizer, diluent, preservative, antimicrobial agent, antifungal agent, isotonic agent, absorption delaying agent, adjuvant, cell culture supernatant, stabilizer, viral vector, expression vector, immunomodulator, and / or any combination thereof. The veterinarily acceptable carrier may comprise an adjuvant, immunomodulator, cell culture supernatant, virus, or expression vector, or any combination thereof. The veterinarily acceptable carrier may comprise an adjuvant. The composition may be utilized in a method for eliciting an immune or immunological response or a protective immune or immunological response against PCV3, PPV, and / or PRRSV. In one embodiment, the composition may be used in a method for inducing an immune response against PCV3 in pigs, particularly preferably in pregnant pigs. In another embodiment, the composition may be used in a method for reducing or preventing clinical signs or disease caused by PCV3 infection in piglets, wherein the piglets are nursed by a sow to which the composition has been administered. The composition may be administered intramuscularly or intradermally. Embodiments also relate to methods for inducing an immune or immunological response or a protective immune or immunological response against PCV3, PPV, and / or PRRSV, which may comprise administering any one of the above compositions to an animal. Embodiments also relate to methods for immunizing pigs against clinical disease caused in the animal by at least one pathogen, comprising administering any one of the above compositions to the animal, wherein the immunogenic composition is capable of inducing an immune response that does not cause clinical signs of infection but immunizes the animal against a pathogenic form of the at least one pathogen.

[0035] PPV is a self-replicating virus of the Parvovirinae subfamily of the Protoparvovirus genus within the Parvoviridae family, containing a single-stranded DNA molecule of approximately 5,100 nucleotides (Cotmore et al., 2014: Arch Virol.: 159(5): 1239-1247; Molitor et al., 1984: Virology: 137(2):241-54). Only the negative-strand DNA is packaged into virions. The viral genome encodes three capsid proteins (VP1, VP2, and VP3) and one nonstructural protein (NS1). Parvovirus capsids are approximately 22-25 nanometers in diameter and are composed of VP1 and VP2 subunits. These proteins are derived from alternative splice forms of the same RNA molecule, resulting in overlapping sequences. Furthermore, porcine parvovirus exhibits high levels of sequence similarity to feline panleukopenia virus, canine parvovirus, and rodent parvovirus (Ranz et al., 1989: J. gen. Virol: 70:2541-2553).

[0036] The PPV protein may be derived from inactivated or killed whole cells of PPV, or from inactivated or killed subunits of PPV. Advantageously, the PPV protein is a recombinant PPV protein.

[0037] EP0551449 discloses a method for producing a VP2 subunit vaccine against porcine parvovirus. Cadar D et al. (Infection, Genetics and Evolution 2012, 12: 1163-1171) describe the phylogenetic and evolutionary genetics of porcine parvovirus in wild boars. Streck AF et al. (Journal of General Virology 2011, 92: 2628-2636) describe rapid viral evolution in the capsid protein of porcine parvovirus. WO88 / 02026 relates to empty virus capsid vaccines. Martinez C et al. (Vaccine 1992, 10(10): 684-690) disclose the production of porcine parvovirus empty capsids with high immunogenic activity. Xu F et al. (Applied and Environmental Microbiology 2007, 73(21): 7041-7047) describe the induction of an immune response in mice after intragastric administration of Lactobacillus casei producing the porcine parvovirus VP2 protein. Furthermore, U.S. Patent No. 10,485,866 discloses immunogenic compositions comprising a type 2 PPV viral protein (VP2), advantageously a mutant PPV VP2 containing one or more mutations.

[0038] The term "porcine parvovirus" or "PPV" is well known to those skilled in the art. However, "porcine parvovirus" is a self-replicating virus of the genus Parvovirus within the family Parvoviridae that contains a single-stranded DNA molecule. The viral genome encodes three capsid proteins (VP1, VP2, and VP3) and one nonstructural protein (NS1). The disease caused by PPV in pigs is often referred to as SMEDI (an acronym for stillbirth, mummification, embryonic death, and infertility). The term "porcine parvovirus" encompasses all possible strains, genotypes, phenotypes, and serotypes of porcine parvovirus. The term "type 2 viral protein" or "VP2" refers to VP2, the capsid protein of porcine parvovirus. The term "type 2 viral protein" or "VP2" is well known to those skilled in the art.

[0039] Porcine reproductive and respiratory syndrome (PRRS) is currently considered by many to be the most important disease affecting the swine industry worldwide. PRRS virus (PRRSV) is an enveloped, single-stranded RNA virus classified in the Arteriviridae family. The antigenic characteristics of different PRRSV isolates vary widely, and effective measures to prevent infection are limited. Three major vaccine groups are available against PRRS: attenuated modified live virus (MLV) vaccines, killed virus vaccines, and recombinant vaccines. PRRSV viral envelope proteins are generally classified into major and minor proteins based on their abundance within the virion. The major viral envelope proteins, gp5 (ORF 5) and M (ORF 6), form dimers. The envelope minor proteins are gp2 (ORF2), gp3 (ORF3), gp4 (ORF4), and E (ORF2b), as well as a possibly newly identified viral protein, gp5a (ORF5a). The active antigenic component may include ORF4, ORF5, ORF6, or ORF7 from the PRRSV virus.

[0040] The recombinant PRRSV antigen can be expressed in a vectored PRRSV vaccine or vectored PRRSV composition, which comprises one or more engineered recombinant adenovirus vectors that carry and express a particular PRRSV antigen, and optionally a pharmaceutically or veterinarily acceptable carrier, adjuvant, excipient, or vehicle. Preferably, the vector is an adenovirus vector, although other vectors, such as baculovirus, are also contemplated.

[0041] The PRRSV can be any strain, as the novel compositions and methods disclosed herein are broadly applicable to all known and yet-to-be-discovered PRRSV strains. PRRSV viruses exist as two genotypes, designated "US" and "EU," which share approximately 50% sequence homology (Dea S et al. (2000). Arch Virol 145:659-88). These two genotypes can also be distinguished by their immune characteristics. Most sequencing information about various isolates is based on structural proteins, i.e., GP5, an envelope protein that accounts for only about 4% of the viral genome, while very little is known about nonstructural proteins (nsp). The isolation of PRRSV and the production of vaccines have been described in numerous publications (WO 92 / 21375, WO 93 / 06211, WO 93 / 03760, WO 93 / 07898, WO 96 / 36356, EP 0676467, EP 0732340, EP 0835930, U.S. Pat. No. 10,039,821). PRRSV antigens include PRRSV minor proteins (e.g., gp2, gp3, gp4, gp5a, gp5, or E) in any combination, and optionally additional PRRSV major proteins (e.g., gp5 or M). For example, PRRSV antigens could be displayed on the surface of virus-like particles (VLPs). In other embodiments, soluble forms of antigens containing oligomerized proteins with each other (including trimerization), or additionally with components of VSV-G, or other viral proteins, or any oligomerization (including trimerization motifs) (e.g., motifs derived from bacterial GCN4, etc.) can be administered to a host animal. Furthermore, it is envisioned that the TM / CT domains of type I viral surface glycoproteins serve the same purpose as, and are therefore interchangeable with, the corresponding domains derived from VSV-G.

[0042] In some embodiments, the one or more vectors comprise a nucleotide sequence encoding a PRRSV E antigen, a polypeptide thereof, an extracellular domain, or a variant thereof; or a nucleotide sequence encoding a modified PRRSV gp2 antigen, a modified PRRSV gp3 antigen, a modified PRRSV gp4 antigen, a modified PRRSV gp5a antigen, a modified PRRSV gp5 antigen, or a modified PRRSV M antigen, a polypeptide thereof, an extracellular domain, or a variant thereof, in which the existing intracellular localization sequence of gp2, gp3, gp4, gp5a, gp5, or M has been replaced with a cell surface expression determinant sequence derived from a heterologous gene. In some embodiments, the one or more vectors comprise a mixture of two vectors: a first vector expressing a retargeted PRRSV minor protein and a second vector expressing a retargeted PRRSV major protein.

[0043] In an advantageous embodiment, the immunogenic composition comprising PCV3, PPV, and / or PRRSV is administered to a subject in need thereof in two doses. However, the immunogenic composition comprising PCV3, PPV, and / or PRRSV may be administered in more than one dose, with the first dose being administered before the second (booster) dose. Preferably, the second dose is administered at least 15 days after the first dose. More preferably, the second dose is administered 15 to 40 days after the first dose. Even more preferably, the second dose is administered at least 17 days after the first dose. Even more preferably, the second dose is administered 17 to 30 days after the first dose. Even more preferably, the second dose is administered at least 19 days after the first dose. Even more preferably, the second dose is administered 19 to 25 days after the first dose. Most preferably, the second dose is administered at least 21 days after the first dose. Even more preferably, the second dose is administered about 21 days after the first dose or 21 days after the first dose. In a preferred embodiment of a two-dose regimen, the first and second doses of the immunogenic composition comprising PCV3, PPV, and / or PRRSV are administered in the same volume. Preferably, each dose is in the preferred volume specified above, with 1 ml or 2 ml doses being most preferred for the first and second doses. In addition to the first and second dose regimen, alternative embodiments include further subsequent doses. In these embodiments, for example, a third, fourth, or fifth dose may be administered. Preferably, the subsequent third, fourth, and fifth dose regimens are administered in the same volume as the first dose, with the time frame between administrations corresponding to the timing of the first and second doses described above.

[0044] The volume administered per subject depends on the route of vaccination and the age of the subject. Preferably, the total volume is about 0.2 ml to 5 ml, more preferably about 0.5 ml to 3.0 ml, even more preferably about 1.0 ml to 2.5 ml, and even more preferably about 1.0 ml to 2.0 ml. The most preferred volumes are 1 ml, 1.5 ml, 2 ml, or 2.5 ml per dose.

[0045] Immunogenic compositions containing PCV3, PPV, and / or PRRSV are preferably administered locally or systemically. Suitable administration routes commonly used include intranasal, intravenous, intradermal, transdermal, intramuscular, intraperitoneal, and subcutaneous administration, as well as oral or parenteral administration, such as inhalation. However, depending on the nature and mode of action of the compound, immunogenic compositions can also be administered by other routes. For example, such other routes include intradermal, intravenous, intravascular, intraarterial, intraperitoneal, intrathecal, intratracheal, intradermal, intracardiac, intralobar, intramedullary, intrapulmonary, intrarectal, and intravaginal routes. However, more preferably, immunogenic compositions containing PCV3, PPV, and / or PRRSV are administered subcutaneously or intramuscularly. Most preferably, the immunogenic composition comprising PCV3, PPV, and / or PRRSV is administered intramuscularly. In one embodiment, the immunogenic composition comprising PCV3, PPV, and / or PRRSV is administered intramuscularly. In one embodiment, the immunogenic composition comprising PCV3, PPV, and / or PRRSV is administered to gilts and / or sows.

[0046] Preferably, the immunogenic composition comprising PCV3, PPV, and / or PRRSV is administered to gilts and / or sows that are at least 3 months of age, more preferably at least 4 months of age, and most preferably at least 5 months of age. In one aspect, the immunogenic composition is administered to gilts and / or sows that are at least 3 months old. In one aspect, an immunogenic composition comprising PCV3, PPV, and / or PRRSV is administered to pre-pregnant gilts and / or sows.

[0047] In a two-shot regimen, the second dose of the immunogenic composition comprising PCV3, PPV, and / or PRRSV is advantageously administered to the gilts and / or sows 2, 3, 4, or 5 weeks prior to mating / insemination, most preferably about 3 weeks prior to mating / insemination. Preferably, the first dose of the immunogenic composition is administered to the gilts and / or sows 2, 3, 4, 5, or 6 weeks prior to administering the second dose, most preferably about 3 weeks prior to administering the second dose. However, after the two-shot regimen is applied, preferably the gilts and / or sows are re-vaccinated every 3, 4, 5, 6, 7, or 8 months, most preferably about every 6 months. In one aspect of the invention, the immunogenic composition is administered to pregnant and lactating gilts and / or sows and is safe for pregnant and lactating gilts and / or sows.

[0048] It is further claimed that the vaccine is capable of protecting breeding gilts and sows when challenged with PCV3 in all trimesters, or two trimesters, or at least one trimester throughout the 114 day gestation period. It is also claimed that the vaccine is capable of significantly reducing the incidence of mummification, stillbirth, and weak fetuses in vaccinated gilts and vaccinated sows when challenged with PCV3 in all trimesters, or two trimesters, or at least one trimester over the 114 day gestation period. In one aspect of the invention, the immunogenic composition is safe for gilts and / or sows from 30 days of gestation, preferably from 40 days of gestation.

[0049] Preferably, the immunogenic composition comprising PCV3, PPV, and / or PRRSV contains 0.1 μg to 150 μg, preferably 0.25 μg to 75 μg, more preferably 0.5 μg to 37.5 μg, even more preferably 0.5 μg to 15 μg, and most preferably 0.5 μg to 6 μg of PCV3, PPV, and / or PRRSV antigens. The immunogenic composition comprising PCV3, PPV, and / or PRRSV can be in an amount of about 0.25 μg, 0.5 μg, 0.75 μg, 1 μg, 1.25 μg, 1.5 μg, 1.75 μg, 2 μg, 2.25 μg, 2.5 μg, 2.75 μg, 3 μg, 3.5 μg, 4 μg, 4.5 μg, 5 μg, 5.5 μg, 6 μg, 6.5 μg, 7 μg, 7.5 μg, 8 μg, 8.5 μg, 9 μg, 9.5 μg, 10 μg, 10.5 μg, 11 μg, 11.5 μg, 12 μg, 12.5 μg, 13 μg, 13.5 μg, 14 μg, 14.5 μg, or 15 μg. In one aspect of the invention, the immunogenic composition comprises an immunogenic composition comprising 0.1 μg to 150 μg of PPV VP2 antigen, preferably 0.5 μg to 30 μg of PCV3, PPV, and / or PRRSV antigens. In one embodiment, the immunogenic composition protects against homologous and / or heterologous challenge.

[0050] The PCV3 ORF2 protein can be produced using a baculovirus expression system in cultured insect cells. The method can include a step of inactivating the baculovirus. Inactivation can be performed in a manner understood in the art. For example, in chemical inactivation, a suitable virus or serum sample containing the virus is treated with a sufficient amount or concentration of an inactivating agent for a sufficient length of time at a temperature (or a low temperature, depending on the inactivating agent) or high pH sufficient to inactivate the virus. Heat inactivation is performed at a temperature and for a sufficient length of time to inactivate the virus. Irradiation inactivation is performed using a wavelength of light or other energy source for a sufficient length of time to inactivate the virus. A virus is considered inactivated when it is unable to infect susceptible cells. The inactivation step can include heat treatment or the use of a virus inactivating agent. The inactivating agent can include an aziridine compound such as BEI.

[0051] The present disclosure also includes recombinant vectors comprising a polynucleotide sequence encoding a polypeptide sequence encoding a PCV3 ORF2 protein. PCV3 ORF2 can be from group a1, b1, or b2 (using the subclassification designation of Fux et al., "Full genome characterization of porcine circovirus type 3 isolates reveals the existence of two distinct groups of virus strains," Virology Journal (2018) 15:25, DOI 10.1186 / s12985-018-0929-3 (incorporated herein by reference)); see, e.g., Table 4). In another embodiment, the PCV3 ORF2 protein comprises or consists of an amino acid sequence encoded by a polynucleotide sequence having at least 90%, or at least 91%, or at least 92%, or at least 93%, or at least 94%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity or homology to SEQ ID NO:4. The recombinant vector may be a baculovirus. In another embodiment, the recombinant vector may comprise at least 90%, or at least 91%, or at least 92%, or at least 93%, or at least 94%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity or homology to SEQ ID NO:2.

[0052] It is noted that in this disclosure, particularly in the claims and / or sections, terms such as "comprises," "comprised," "comprising," and the like may have the meaning ascribed to them in U.S. patent law; e.g., may mean "includes," "included," "including," and the like; and terms such as "consisting essentially of" and "consisting essentially of" have the meaning ascribed to them in U.S. patent law, e.g., these terms permit elements not expressly recited, but exclude elements found in the prior art or that affect basic or novel characteristics of the disclosure.

[0053] The porcine, pig, or piglet to which the administration is given may have antibodies, eg, maternal antibodies, to a PCV, such as PCV2 and / or PCV3.

[0054] These and other embodiments are disclosed by or are obvious from and encompassed by the following detailed description. It is noted that in this disclosure, particularly in the claims and / or sections, terms such as "comprises," "comprised," "comprising," and the like may have the meaning ascribed to them in U.S. patent law; e.g., may mean "includes," "included," "including," and the like; and terms such as "consisting essentially of" and "consisting essentially of" have the meaning ascribed to them in U.S. patent law, e.g., these terms permit elements not expressly recited, but exclude elements found in the prior art or that affect the basic or novel characteristics of the invention.

[0055] These and other embodiments are disclosed by or are obvious from and encompassed by the following detailed description. The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee.

[0056] The following detailed description is given for illustrative purposes and is not intended to limit the invention to the specific embodiments merely described, which may be best understood in conjunction with the accompanying drawings. [Brief explanation of the drawings]

[0057] [Figure 1] This is the sequence of SEQ ID NO: 1, which is the PCV3 ORF2 nucleotide sequence within the recombinant baculovirus BaculoG / PCV3 ORF2. [Figure 2] This is the sequence of SEQ ID NO: 2, which is the sequence of recombinant baculovirus BaculoG / PCV3 ORF2. [Figure 3] FIG. 1 shows a map for a recombinant baculovirus (BaculoG / PCV3 ORF2 clone, 4B4-2E12 Pre-MSV p8) containing the PCV3 ORF2 gene under the control of the baculovirus polyhedrin promoter. [Figure 4] FIG. 1 shows the median log10 genome copy number of PCV3 DNA per mL in serum by group and study day in groups 1 to 5. [Figure 5] FIG. 1 shows the median log10 genome copy number of PCV3 DNA per mL in fecal samples by group and study day in groups 1 to 5. [Figure 6] FIG. 1 shows the log10 of the median PCV3 DNA genome copy number per mL in nasal samples by group and study day in groups 1 to 5. [Figure 7] FIG. 1 shows baseline-corrected least squares mean rectal temperature (°F) by group and study day. [Figure 8] FIG. 1 shows baseline-corrected least squares mean daily body weight (kg) for each group and day in Groups 1 to 5. [Figure 9] FIG. 1 shows mean body temperatures (°F) by group and day. [Figure 10] FIG. 1 shows sequence information (SEQ ID NOs: 3 to 5) for PCV3 PCR-positive tissue homogenates used for challenge materials. [Figure 11] FIG. 1 shows median PCR values ​​for groups 1 to 5 after 7 to 49 days. [Figure 12] FIG. 1 shows median PCR values ​​for groups 7-9 after 7-49 days. [Figure 13] FIG. 1 shows median PCR values ​​for fecal excretion in groups 1 to 5 after 7 to 49 days. [Figure 14] FIG. 1 shows median PCR values ​​for nasal discharge in groups 1-5 after 7-49 days. [Figure 15] FIG. 1 depicts arithmetic mean rectal temperature values ​​in Groups 1-6 over 14-49 days of evaluation. [Figure 16] FIG. 1 depicts least squares mean temperature values ​​by group and day for groups 1-5. [Figure 17] 1 is a line graph illustrating the mean rectal temperature (baseline corrected least squares) of animals by group and day for Groups 1-5. [Figure 18] FIG. 1 shows arithmetic mean body weights for Groups 1 to 6 after 14 to 49 days. [Figure 19] 1 is a line graph showing the (least squares) mean body weight for groups 1 to 5 by group and day. [Figure 20] 1 is a line graph depicting data for least squares means of body weight (baseline corrected) by group and day. [Figure 21] FIG. 1 shows a history plot for the production of pre-MSV+1. [Figure 22A] FIG. 22A shows cell counts. [Figure 22B] FIG. 22B shows the cell viability and size upon infection with BaculoG / PCV3 ORF2. [Figure 23] FIG. 1 shows the analysis of BaculoG / PCV3 ORF2 solution at the time of collection. [Figure 24] FIG. 1 shows images of inactivation after 72 hours. [Figure 25] FIG. 1 shows a comparison of the inactivation status of BaculoG / PCV3 ORF2 antigen after inactivation by Western blotting. [Figure 26] FIG. 1 shows a dot plot of fluorescence for PCV3 ORF2. [Figure 27] Figure 1 shows a plot of viremia observed within a sample population of pigs after challenge, based on log10 genome copies per mL. All control pigs were viremic as determined by PCR at each sampling time point during the challenge phase, and vaccination significantly reduced viral load at each sampling time point during the challenge phase (P<0.0050). [Figure 28] FIG. 1 shows a plot of mean rectal temperature (°F) measured pre-challenge (days 12, 13, 14) and post-challenge (days 14.5-20). [Figure 29] Figure 1 shows a plot of viremia observed in gilts challenged on day 83 (day 40 of gestation). Numbers indicate genome copies per mL. The Y-axis is shown on a linear scale to accurately represent values ​​at zero. Arrows indicate primary vaccine doses, booster doses, and challenge. [Figure 30]FIG. 1 shows a bar graph indicating the percent of affected piglets based on the observed number of autolyzed, mutilated, and mummified piglets born from farrowing sows in each treatment group. [Figure 31-1] FIG. 1 shows alignment of the amino acid sequence of the PCV3 capsid with the capsid of porcine PCV2 and the capsid of beak and feather disease virus (BFDV). [Figure 31-2] Figure 31-1 continued [Figure 32] FIG. 1 shows the structure of PCV3 ORF2 mutants in the FG loop with mutations in lysine and histidine. [Figure 33] FIG. 1 shows the structure of a PCV3 ORF2 mutant in which the natural stop codon of the PCV3 capsid protein has been mutated and the C-terminus has been extended to the next stop codon. [Figure 34] FIG. 1 depicts the nucleotide and amino acid sequences of PCV3 ORF2 mutants in the FG loop with mutations in lysine and histidine, and PCV3 ORF2 mutants in which the native stop codon for the PCV3 capsid protein has been mutated and the C-terminus has been extended to the next stop codon (SEQ ID NOs: 6-9). [Figure 35] FIG. 1 depicts the amino acid sequence of mutant PCV3 ORF2, "FG-PC" (SEQ ID NO: 10). DETAILED DESCRIPTION OF THE INVENTION

[0058] The present disclosure relates to PCV3 vaccines. Any sequence of PCV3 is contemplated. See, for example, Phan, Tung Gia, et al. "Detection of a Novel Circovirus PCV3 in Pigs with Cardiac and Multi-Systemic Inflammation," Virology Journal, vol. 13, no. 1, 2016, p. 184, doi:10.1186 / s12985-016-0642-z. Published November 11, 2016; and Fux et al., "Full genome characterization of porcine circovirus type 3 isolates reveals the existence of two distinct groups of virus strains," Virology Journal (2018) 15:25, DOI 10.1186 / s12985-018-0929-3, the disclosures of which are incorporated by reference.

[0059] PCV3 ORF2 and PCV3 genome sequences were derived from KT869077 (GenBank). In the examples, the entire PCV3 genome in a plasmid was used and described. ORF2 and the entire genome were synthesized using Genscript.

[0060] Two additional constructs, recircularized PCV3 genomes derived by two different methods, were used to rescue the virus in cell culture.

[0061] In the sequence listing, the following sequences are presented: [Table 1]

[0062] PCV3 ORF2 "FG" is an antigenic protein according to the present invention that contains an amino acid substitution in the FG loop of the native PCV3 ORF2 protein. PCV3 ORF2 "PC" is an antigenic protein according to the present invention that comprises an amino acid extension at the C-terminus of the native PCV3 ORF2 protein. In a preferred embodiment, the polypeptide of the present disclosure is a recombinant PCV3 ORF2 protein, such as a recombinant baculovirus-expressed PCV3 ORF2 protein. As used herein, the term "recombinant PCV3 ORF2 protein" particularly refers to a protein molecule expressed from a recombinant DNA molecule, such as a polypeptide produced by recombinant DNA techniques. Examples of such techniques include when DNA encoding the protein to be expressed is inserted into an appropriate expression vector, preferably a baculovirus expression vector, which is then used to transfect or, in the case of a baculovirus expression vector, infect a host cell to produce the protein or polypeptide encoded by the DNA. Thus, as used herein, the term "recombinant PCV3 ORF2 protein" particularly refers to a protein molecule expressed from a recombinant DNA molecule.

[0063] According to a specific example, recombinant PCV3 ORF2 protein is produced by a method involving the following steps: the gene for PCV3 ORF2 is cloned into a baculovirus transfer vector; the transfer vector is used to prepare a recombinant baculovirus containing the gene by homologous recombination in insect cells; and then, upon infection with the recombinant baculovirus, the PCV3 ORF2 protein is expressed in the insect cells.

[0064] It is further understood that the term "recombinant PCV3 protein consisting of a sequence" also relates to any one or more co-translational and / or post-translational modifications of the sequence, particularly those affected by the cell in which the polypeptide is expressed. Thus, the term "recombinant PCV3 ORF2 protein consisting of a sequence" as described herein also covers a sequence having one or more modifications made by the cell in which the polypeptide is expressed, particularly modifications of amino acid residues made during protein biosynthesis and / or protein processing, preferably selected from the group consisting of glycosylation, phosphorylation, and acetylation. Preferably, the recombinant PCV3 ORF2 protein according to the present disclosure is produced or obtained by a baculovirus expression system, especially in cultured insect cells. In an even more preferred embodiment, the polypeptide of the present disclosure is a PCV3 ORF2 protein comprising or consisting of an amino acid sequence having at least 90%, preferably at least 92%, more preferably at least 94%, even more preferably at least 96%, even more preferably at least 98%, or particularly 100% sequence identity to the amino acid sequence of SEQ ID NO:4.

[0065] " Sequence identity " as known in the art refers to the relationship between two or more polypeptide sequences or two or more polynucleotide sequences, that is, the relationship between a reference sequence and a given sequence that is compared with the reference sequence. Sequence identity is determined by comparing a given sequence with a reference sequence after the sequences are optimally aligned to produce the highest degree of sequence similarity determined by the match between the strings of such sequences. Once such alignment is performed, sequence identity is confirmed on a position-to-position basis; for example, if the nucleotide or amino acid residue at a particular position is the same, the sequence is "identical" at that position. Then, the total number of such position identities is divided by the total number of nucleotides or residues in the reference sequence to obtain the sequence identity percentage. Sequence identity was determined in Computational Molecular Biology, Lesk, AN, ed., Oxford University Press, New York (1988), Biocomputing: Informatics and Genome Projects, Smith, DW, ed., Academic Press, New York (1993); Computer Analysis of Sequence Data, Part I, Griffin, AM, and Griffin, HG, eds., Humana Press, New Jersey (1994); Sequence Analysis in Molecular Biology, von Heinge, G., Academic Press (1987); Sequence Analysis Primer, Gribskov, M. and Devereux, J., eds., M. Stockton Press, New York (1991); and Carillo, H., and Lipman, D., SIAM J. Applied Math., 48: 1073 (1988), the teachings of which are incorporated herein by reference.Preferred methods for determining sequence identity are designed to maximize the match between the sequences under test. Methods for determining sequence identity are codified in publicly available computer programs that determine sequence identity between given sequences. Examples of such programs include, but are not limited to, the GCG program package (Devereux, J., et al., Nucleic Acids Research, 12(1):387 (1984)), BLASTP, BLASTN, and FASTA (Altschul, SF et al., J. Molec. Biol., 215:403-410 (1990)). The BLASTX program is available from NCBI and other sources (see, for example, BLAST Manual, Altschul, S. et al., NCVI NLM NIH Bethesda, Md. 20894; Altschul, S. F. et al., J. Molec. Biol., 215:403-410 (1990)), the teachings of which are incorporated herein by reference. These programs optimally align sequences using default gap weights to achieve the highest level of sequence identity between a given sequence and a reference sequence. By way of example, a polynucleotide having a nucleotide sequence with at least, for example, 85%, preferably 90%, and even more preferably 95% "sequence identity" to a reference nucleotide sequence means that the nucleotide sequence of the given polynucleotide is identical to the reference sequence, except that the given polynucleotide sequence may contain up to 15, preferably up to 10, and even more preferably up to 5 point mutations per 100 nucleotides of the reference nucleotide sequence.In other words, in a polynucleotide having a nucleotide sequence that is at least 85%, preferably 90%, and even more preferably 95% identical to a reference nucleotide sequence, up to 15%, preferably 10%, and even more preferably 5% of the nucleotides in the reference sequence may be deleted or substituted with other nucleotides, and up to 15%, preferably 10%, and even more preferably 5% of the total nucleotides in the reference sequence may be inserted into the reference sequence. These mutations of the reference sequence may be made at the 5'-terminal or 3'-terminal position of the reference nucleotide sequence, or at any position between these terminal positions, either individually interspersed among nucleotides in the reference sequence or interspersed in one or more contiguous groups within the reference sequence. Similarly, by a polypeptide having a given amino acid sequence having at least, for example, 85%, preferably 90%, and even more preferably 95% sequence identity to a reference amino acid sequence, it is intended that the given amino acid sequence of the polypeptide is identical to the reference sequence, except that the given polypeptide sequence may contain up to 15, preferably up to 10, and even more preferably up to 5 amino acid changes per 100 amino acids in the reference amino acid sequence. In other words, to obtain a given polypeptide sequence having at least 85%, preferably 90%, and even more preferably 95% sequence identity with the reference amino acid sequence, up to 15%, preferably up to 10%, and even more preferably up to 5% of the amino acid residues in the reference sequence may be deleted or substituted with other amino acids, and up to 15%, preferably up to 10%, and even more preferably up to 5% of the total number of amino acid residues in the reference sequence may be inserted into the reference sequence. These alterations of the reference sequence may be made at the amino- or carboxy-terminal positions of the reference amino acid sequence, or anywhere between these terminal positions, either individually interspersed among residues in the reference sequence or interspersed in one or more contiguous groups within the reference sequence. Preferably, residue positions that are not identical differ by conservative amino acid substitutions.However, conservative substitutions are not counted as matches when determining sequence identity.

[0066] As used herein, "sequence homology" refers to a method for determining the relationship between two sequences. To determine sequence homology, two or more sequences are optimally aligned, and gaps are introduced, if necessary. However, in contrast to "sequence identity," conservative amino acid substitutions are counted as matches when determining sequence homology. In other words, to obtain a polypeptide or polynucleotide having 95% sequence homology with a reference sequence, 85%, preferably 90%, and even more preferably 95% of the amino acid residues or nucleotides in the reference sequence must match or contain conservative substitutions with other amino acids or nucleotides, or up to 15%, preferably up to 10%, and even more preferably up to 5% of the total number of amino acid residues or nucleotides in the reference sequence that do not contain conservative substitutions can be inserted into the reference sequence. Preferably, the homologous sequence comprises a stretch of at least 50 nucleotides, even more preferably 100 nucleotides, even more preferably 250 nucleotides, and even more preferably 500 nucleotides.

[0067] A "conservative substitution" refers to the substitution of an amino acid residue or nucleotide with another amino acid residue or nucleotide having similar characteristics or properties, including size, hydrophobicity, etc., such that the overall functionality does not change significantly.

[0068] The present invention also encompasses mutations in PCV3 proteins, including, but not limited to, mutations in the PCV3 capsid protein. Despite the diversity of capsid amino acid sequences between PCV2 and beak and feather disease virus (BFDV), the crystal structures are very similar. Mutations in PCV3 are advantageous if they can stabilize virus-like particles (VLPs). Although PCV3 capsid protein is believed to self-assemble into VLPs, the expression level of PCV3 protein is significantly lower than that of PCV2 capsid protein. In particular, only about 20% of the protein assembles into VLPs, while the remaining 80% of the protein aggregates into an insoluble fraction.

[0069] In some embodiments, the variant proteins of the invention are capable of higher VLP yields than the protein encoded by SEQ ID NO: 1. High yield is understood to particularly (for example) relate to higher molar yield. In other words, the variant proteins of the invention are capable of assembly of larger CAP (capsid (ORF2) protein) VLPs than the protein encoded by SEQ ID NO: 1. Examples of higher yield include at least a 5% higher yield, or at least a 10% higher yield, or at least a 15% higher yield, or at least a 20% higher yield, or at least a 25% higher yield, or at least a 30% higher yield, or at least a 35% higher yield, or at least a 40% higher yield, or at least a 50% higher yield. Thus, for example, without modification of the PCV3 ORF2 protein, baculovirus expression yields, for example, 20% PCV3 soluble protein (VLP) and 80% PCV3 insoluble protein, as determined by Western blot analysis. However, with modification, the expression yields 25%, 30%, 35%, 40%, 45%, 50%, 55%, or 60% or more PCV3 soluble protein (VLP) (an increase of 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, or 45%, etc., in PCV3 soluble protein (VLP)), which represents a high yield. Advantageously, modification of the PCV3 ORF2 protein results in a VLP yield (soluble PCV3 protein) that is at least 50% of the PCV3 protein expressed by the recombinant baculovirus system.

[0070] Suitable assays and techniques for use in the present invention include assays and techniques that have been used to track or quantify the assembly and disassembly of porcine circovirus capsid (ORF2) proteins into virus-like particles (VLPs), including enzyme-linked immunosorbent assays (ELISAs), SDS / PAGE, Western blots or immunoblots, which may involve silver or Coomassie dyes, size-exclusion chromatography (SEC), dynamic light scattering (DLS) or multi-angle light scattering (MALS), transmission electron microscopy (TEM), analytical ultracentrifugation, and fluorescence spectroscopy (FSA), which may be coupled to high-performance liquid chromatography (HPLC). Additional suitable techniques may also include agarose gel retardation tests for protein-nucleic acid complexes, immunodiffusion tests such as single radial immunodiffusion (SRID), nanoparticle tracking analysis (NTA) metabolic labeling-based assays, and chemiluminescent enzyme-based assays.Each of these assays is well known in the art, e.g., Fang, Mingli et al. "Detection of the Assembly and Disassembly of PCV2b Virus-Like Particles Using Fluorescence Spectroscopy Analysis" Intervirology vol. 58, 2015, pp. 318-323; Thompson, Christine et al. "Analytical technologies for influenza virus-like particle candidate vaccines: challenges and emerging approaches" Virology Journal vol. 10, 2013, p. 141; Steppert, Petra et al. "Quantification and characterization of virus-like particles by size-exclusion chromatography and nanoparticle tracking analysis" Journal of Chromatography A vol. 1487, 2017, pp. 89-99; Yadav, Shalini et al. "A facile quantitative assay for viral particle genesis reveals Cooperativity in virion assembly and saturation of an antiviral protein,” Virology, vol. 429, No. 2, 2012, pp. 155-162; and Zeltins, Andris “Construction and Characterization of Virus-Like Particles: A Review,” Molecular Biotechnology, vol. 53, 2013, pp. 92-107.

[0071] In one embodiment, the variant proteins of the invention are capable of higher VLP yields, as can be determined by Western blot analysis, than the protein encoded by SEQ ID NO: 1. In other words, the variant proteins of the invention are capable of assembly of larger CAP VLPs, as can be determined by Western blot analysis, than the protein encoded by SEQ ID NO: 1.

[0072] In various embodiments discussed herein, there are one or more mutations in the PCV3 ORF2 capsid protein, for example, that increase VLP yield. For example, in various embodiments, there may be one, two, three, or four mutations in the FG loop. In the present specification, an embodiment is exemplified and discussed that may involve the SKKK of the PCV3 ORF2 protein FG loop being replaced with a QPFS (e.g., PCV2 ORF2 protein motif). When making substitutions, those skilled in the art may practice the present invention by replacing only S with Q; or replacing only the first K with P; or replacing only the second K with F; or replacing only the third K with S; or any combination of these substitutions, such as replacing S with Q and replacing the first K with P; or replacing S with Q and replacing the second K with F; or replacing S with Q and replacing the third K with S; or replacing S with Q and replacing the first K with P and replacing the second K with F; or replacing S with Q and replacing the first K with P and replacing the second K with F; or replacing S with Q and replacing the first K with P and replacing the third K with S. Similarly, in these embodiments, in addition to or as an alternative to replacements or mutations within the FG loop, those skilled in the art may practice the present invention by adding amino acids to the C-terminus of the PCV3 ORF2 protein. Without a C-terminal extension or addition, the PCV3 ORF2 protein may be buried in the three-dimensional structure, as opposed to being exposed at the C-terminus of the ORF2 proteins or capsid proteins of other circoviruses. In embodiments in which the C-terminus of the PCV3 ORF2 protein is extended or added, it may be advantageous to extend or add to the C-terminus of the PCV3 ORF2 protein with a motif derived from another circovirus, such as PCV2.Thus, for example, one skilled in the art can extend or add to the C-terminus of the PCV3 ORF2 protein amino acids found at the C-terminus of the PCV2 ORF2 protein or PCV2 capsid protein, such as amino acids 215 to 234 or 215 to 233 of the PCV2 ORF2 protein or PCV2 capsid protein, or can extend or add to the PCV3 ORF2 protein or PCV3 capsid protein an epitope of the PCV2 ORF2 protein or PCV2 capsid protein. In this regard, reference is made to Trible et al., "Antibody Recognition of Porcine Circovirus Type 2 Capsid Protein Epitopes after Vaccination, Infection and Disease, Clinical and Vaccine Immunology 18(5): 749-757 (2011) doi:10.1128 / CVI.00418-10 (incorporated herein by reference). Within the PCV2 ORF2 (capsid) protein, immunoreactive regions have been reported to be residues 47-85, 165-200, and 200-233. Antibody-reactive regions of the PCV2 ORF2 (capsid) protein have been reported to be amino acids 23-43, 71-85, 117-131, and 171-202. The PCV2 ORF2 (capsid) protein at amino acids 117-131 has been reported to be immunoreactive. The ORF2 (capsid) protein region has been reported to be a major antibody recognition region, with amino acids 156-162, 175-192, 195-202, and 228-223 reported to be associated with antibody recognition. Another PCV2 ORF2 (capsid) protein epitope is 169-STIDYFQPNNKR, e.g., amino acids 169-180 (in which amino acid residues Y-173, F-174, Q-175, and K-179 may contribute to antibody recognition). Other PCV2 ORF2 (capsid) protein epitopes may be amino acids 43-233, 43-135, 43-160, 91-160, 43-180, 160-233, 135-233, and 91-233, as well as amino acids 169-188.Any of these PCV2 ORF2 epitopes, or any combination thereof, can be a C-terminal extension or addition to the PCV3 ORF2 (capsid) protein. In this regard, the C-terminal extension of PCV3 ORF2 can be at most about 200 amino acids, or at most about 190 amino acids, or at most about 185 amino acids, or at most about 180 amino acids, or at most about 175 amino acids, or at most about 170 amino acids, or at most about 165 amino acids, or at most about 160 amino acids, or at most about 155 amino acids, or at most about 150 amino acids, or at most about 145 amino acids, or at most about 140 amino acids, or at most about 135 amino acids, or at most about 130 amino acids, or at most about 125 ... It is mentioned that the length may be about 120 amino acids, or up to about 115 amino acids, or up to about 110 amino acids, or up to about 105 amino acids, or up to about 100 amino acids, or up to about 90 amino acids, or up to about 80 amino acids, or up to about 70 amino acids, or up to about 60 amino acids, or up to about 50 amino acids, or up to about 40 amino acids, or up to about 30 amino acids; for example, 1 to 50 amino acids, or 10 to 50 amino acids, or 10 to 40 amino acids, or 20 to 40 amino acids, or about 30 amino acids.

[0073] In embodiments where the composition contains a PCV3 ORF2 (capsid) protein of the present invention, e.g., a mutated protein, e.g., where the mutation includes a C-terminal addition or extension, e.g., the C-terminal addition or extension includes an epitope of a PCV2 ORF2 (capsid) protein, and the composition also includes a PCV2 ORF2 (capsid) protein (e.g., both PCV2 and PCV3, e.g., alone or together with one or more antigens of a swine pathogen, such as an antigen disclosed throughout this disclosure or a swine pathogen, or for an indication or symptom or condition thereof, e.g., each derived from baculovirus expression, for one-shot administration), it may be advantageous for the PCV2 ORF2 (capsid) protein epitope to be the same or a different clade as the PCV2 ORF2 (capsid) protein incorporated into the composition. For example, if the PCV2 ORF2 (capsid) protein component is derived from a PCV2a strain (which may be based on Ingelvac CircoFlex), it may be advantageous to derive it from a different clade, such as the genotype PCV2b, PCV2c, or PCVd-mPCV2b, for addition or extension of the PCV3 ORF2 capsid protein (C-terminus). Regarding PCV2 genotypes, strains, or clades, see Franzo et al., "Revisiting the taxonomic classification of Porcine Circovirus type w (PCV2): still a real challenge," Virol J 12: 131 (2015) doi: 10.1186 / s12985-015-0361-x (incorporated herein by reference). The addition or extension at the C-terminus of the PCV3 ORF2 capsid protein may advantageously be an epitope of the PCV2 ORF2 capsid protein that provokes an immunological response against one or more of the clades, strains, or genotypes of PCV2, which may be of the same clade, strain, or genotype as the PCV2 ORF2 capsid protein component of the composition, or may be of a different clade, strain, or genotype.With regard to the foregoing, and more generally, with respect to the mutant PCV3 ORF2 capsid proteins of the invention discussed throughout this disclosure, the present invention provides nucleic acid molecules encoding such mutant PCV3 ORF2 capsid proteins, vectors, such as baculovirus vectors, containing such nucleic acid molecules (see EP2460821, incorporated herein by reference, along with the documents cited therein, for methods and materials for expressing PCV2 ORF2 capsid proteins via a baculovirus expression system, as well as PCV3 ORF2 capsid proteins, including mutant proteins disclosed herein, may also be employed in the practice of the present invention to express PCV2 ORF2 capsid proteins and, if desired, include such proteins in compositions of the invention), and vectors, such as baculovirus vectors, containing such nucleic acid molecules. Methods for producing or expressing the ORF2 capsid protein are encompassed, such as by infecting or transfecting relevant cells with a vector (e.g., when the vector is a baculovirus, the relevant cells can be insect cells, or Sf cells or Sf+ cells; see EP2460821, incorporated herein by reference, along with the documents cited therein). Following expression or production, it is advantageous to recover or isolate the protein, for example, by separating the solids and retaining the liquid or supernatant containing the soluble protein (e.g., VLPs). In addition to this paragraph, the compositions discussed throughout this disclosure can contain a mutant PCV3 ORF2 capsid protein (and optionally an additional PCV2 ORF2 capsid protein, and / or one or more additional antigens of a swine pathogen) in the amounts discussed throughout this disclosure, can be administered in the regimens discussed throughout this disclosure, such as one-shot or single-dose administration, and thus can be administered to pigs or piglets as discussed throughout this disclosure.

[0074] In the context of the present invention, the protein of the present invention, as an antigen in a composition such as an immunological composition, prevents or treats a PCV3 infection-associated disease or condition in a subject, for example, by inducing, stimulating, or enhancing an immune response against PCV3.

[0075] Previous studies have shown that expression of the full-length PCV3 cap gene and the NLS domain present within the N-terminal arginine-rich motif (ARM) causes protein misfolding and induces the formation of 10-12 nm circular viral complexes (Sarker et al. Nat Commun. 2016 Oct 4;7():13014). Wang et al. (AMB Expr 10, 3 (2020) https: / / doi.org / 10.1186 / s13568-019-0940-0) reported the ability of PCV3 VLPs to self-assemble, be successfully expressed in E. coli, and be applied to the development of an ELISA to test for specific antibodies in clinical swine sera. Specifically, to achieve high-level expression of recombinant PCV3 Cap in E. coli, the wild-type whole Cap (wt-eCap) gene was amplified from a clinical sample, and three optimized whole Cap nuclear localization signal (NLS) gene fragments (opti-eCap) and one optimized Cap-deleted nuclear localization signal (NLS) gene fragment (opti-dCap), encoding the same amino acid sequence as wt-eCap, were synthesized based on the codon bias of E. coli. Unlike the present invention, the region near the NLS of the PCV3 capsid is not targeted for VLP assembly and / or stability. Furthermore, removal of the NLS does not necessarily result in improved VLP assembly. However, embodiments of the present invention may include removal or modification of the PCV3 ORF2 capsid protein NLS, for example, in addition to one or more of the FG loop mutations and / or C-terminal extensions discussed herein.

[0076] In a preferred embodiment, the present invention encompasses mutations in regions encoding positively charged amino acids in PCV proteins, including, but not limited to, the PCV3 capsid protein. In particular, the PCV3 capsid contains a large amount of positive charge in the FG loop, located at the five-fold interface of the PCV3 capsid. The large amount of positive charge in this region can result in repulsion unless accompanied by the presence of nucleic acids predicted in the VLP. In one embodiment of the present invention, the positively charged amino acids are mutated to neutral and / or negatively charged amino acids. In a preferred embodiment, the lysines and histidines in this loop are mutated to amino acids derived from the PCV2 capsid (SEQ ID NO: 6). In one embodiment, the present invention provides an engineered PCV3 ORF2 protein that contains a reduced amount of positively charged amino acids compared to a non-engineered PCV3 ORF2 protein. The non-engineered protein may be a wild-type or naturally occurring PCV3 ORF2 protein, or it may be an ORF2 protein that has been previously modified for another purpose, where improved encapsidation activity, such as improved self-assembly, is desired in the presence or absence of a packageable polynucleotide.

[0077] In some embodiments, one or more positively charged amino acids are substituted, such as one or more lysines, arginines, or histidines, or a combination thereof. In some embodiments, two or more positively charged amino acids are substituted. In some embodiments, three or more positively charged amino acids are substituted. In certain embodiments, the charge associated with a region of the ORF2 protein, such as (but not limited to) the FG loop, is made more negative by substituting one or more negatively charged amino acids. In certain embodiments, a positively charged amino acid may be substituted with an amino acid that is less positively charged, and / or a non-positively charged amino acid may be substituted with an amino acid that is more negatively charged. That is, the charge of the ORF2 region may be altered by removing a positive charge, adding a negative charge, or both.

[0078] In an advantageous embodiment, the present invention encompasses the addition of additional amino acids to a PCV protein, such as, but not limited to, the PCV3 capsid protein. Without the presence of nucleic acid, a short hydrophobic chain at the C-terminus of the PCV3 capsid would result in the C-terminus being buried within the capsid, potentially resulting in VLP instability. In contrast, the C-termini of the PCV2 and BFDV capsid proteins protrude from the capsid. In one embodiment, the C-terminus of the PCV3 capsid is extended by about 1 to 50 amino acids, about 10 to 40 amino acids, or about 20 to 30 amino acids. In another embodiment, the C-terminus of the PCV3 capsid is extended by about 20, about 21, about 22, about 23, about 24, about 25, about 26, about 27, about 28, about 29, about 30, about 31, about 32, about 33, about 34, about 35, about 36, about 37, about 38, about 39, or about 40 amino acids. In an advantageous embodiment, the C-terminus of the PCV3 capsid protein is extended by mutating the stop codon. In a particularly advantageous embodiment, the native stop codon for the PCV3 capsid protein is mutated, and the C-terminus is extended to the next stop codon in the viral sequence (SEQ ID NO: 7). In another embodiment, the C-terminus of the PCV capsid may be extended and / or exchanged with the C-terminus of another porcine circovirus. The C-terminus of the PCV3 capsid protein can be extended by about 50 to about 200 amino acids, about 60 to about 190 amino acids, about 70 to about 180 amino acids, about 80 to about 170 amino acids, about 90 to about 160 amino acids, or about 100 to about 150 amino acids.

[0079] In certain embodiments, the C-terminal extension comprises the addition of amino acids at the C-terminus of the PCV3 capsid, for example, by mutation of a stop codon. The stop codon can be mutated by deletion, substitution, or insertion. In certain embodiments, the C-terminal extension comprises the insertion of amino acids near the C-terminus, including, but not limited to, the insertion of amino acids one residue from the C-terminus, two residues from the C-terminus, or three, four, five, six, seven, eight, or more residues upstream from the C-terminus. In one embodiment, the residues can be any set of negatively charged amino acids.

[0080] It is understood that the proteins of the present invention may differ from the exact sequences exemplified and described herein. Accordingly, the present invention contemplates deletions, additions, and substitutions to the sequences shown, so long as the sequences function according to the methods of the present invention. In this regard, particularly preferred substitutions will generally be conservative in nature, i.e., substitutions that occur within a family of amino acids. For example, amino acids are generally divided into four families: (1) acidic: aspartic acid and glutamic acid; (2) basic: lysine, arginine, histidine; (3) nonpolar: alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan; and (4) uncharged polar: glycine, asparagine, glutamine, cysteine, serine, threonine, and tyrosine. Phenylalanine, tryptophan, and tyrosine are sometimes classified as aromatic amino acids. It is reasonably expected that the isolated or unnatural substitution of leucine with isoleucine or valine, or vice versa; the isolated or unnatural substitution of aspartic acid with glutamic acid, or vice versa; the isolated or unnatural substitution of threonine with serine, or vice versa; or similar conservative substitution of amino acids with structurally related amino acids will not significantly affect biological activity.Therefore, proteins that have substantially the same amino acid sequence as the exemplified and described sequences but possess minor amino acid substitutions that do not substantially affect the immunogenicity of the protein are within the scope of the present invention.

[0081] The present invention further encompasses nucleotide sequences encoding functionally and / or antigenically equivalent variants and derivatives of the antigens of the present invention, as well as functionally equivalent fragments thereof. These functionally equivalent variants, derivatives, and fragments exhibit the ability to retain antigen activity. For example, DNA sequence changes that do not change the encoded amino acid sequence, as well as DNA sequence changes that result in conservative substitutions of amino acid residues, such as deletion or addition of one or a few amino acids, or substitution of amino acid residues with amino acid analogs, are DNA sequence changes that do not significantly affect the properties of the encoded polypeptide. Conservative amino acid substitutions include glycine / alanine; valine / isoleucine / leucine; asparagine / glutamine; aspartic acid / glutamic acid; serine / threonine / methionine; lysine / arginine; and phenylalanine / tyrosine / tryptophan. In one embodiment, a variant has at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% homology or identity to an antigen, epitope, immunogen, peptide, or polypeptide of interest.

[0082] In some embodiments, substitutions introduce conservative changes, replacing an amino acid with another amino acid having a similar chemical structure, similar chemical properties, or similar side chain volume. The introduced amino acids may have similar polarity, hydrophilicity, or hydrophobicity to the amino acids they replace. Conservative amino acid changes are well known in the art. If amino acids have similar polarity, this may also be determined by reference to the hydrophobicity scale for amino acid side chains.

[0083] Conservative amino acid changes may also be determined by reference to a scoring matrix for amino acid sequence conservation, such as the Point Accepted Mutation (PAM) family or the Blocks Substitution Matrix (BLOSUM) family. Thus, a conservative amino acid change may be a member of an equivalence group, which is a set of amino acids that have a mutually positive score in the similarity representation of a scoring matrix selected for use in aligning the reference and variant polypeptide chains.

[0084] It should be understood that nonpolar amino acids include amino acids with aliphatic side chains and amino acids with aromatic side chains. The amino acid proline is classified as nonpolar, but also has rigid properties that can cause changes in secondary structure. For example, proline is often found at the end of a helix. Depending on the specific context of the side chain of a given amino acid residue, for example, tyrosine, an amino acid generally classified as nonpolar due to its aromatic ring, can have a similar functional effect to polar amino acid residues such as threonine through its hydroxyl group. Therefore, tyrosine can be considered both a nonpolar and a polar amino acid for the purposes of the present invention. Furthermore, amino acids described as polar or hydrophilic can be uncharged, charged, basic, or acidic. It is well known that the amino acid histidine has a pKa value close to 7 at neutral pH, so that its side chain can be protonated or unprotonated, and therefore can carry or not carry a charge, depending on the protein environment. Thus, histidine can be considered to be both a polar charged and a polar uncharged amino acid residue for the purposes of the present invention.

[0085] The mutations discussed herein are generally introduced into protein by using methods known in the art, such as protein site-directed mutagenesis, PCR and gene shuffling, or by using multiple kinds of mutagenic oligonucleotides in the cycle of site-directed mutagenesis.Therefore, mutations can be introduced site-specifically or randomly.Therefore, mutagenesis method produces one or more polynucleotides that code for one or more different variants.

[0086] The development of a recombinant baculovirus containing the porcine circovirus type 3 ORF2 gene under the control of the baculovirus polyhedrin promoter (BaculoG / PCV3 ORF2 clone, 4B4-2E12 Pre-MSV p8; Lot No.: 3624-039) is described in Example 1. In some embodiments, the use of such recombinant baculoviruses in vaccines, as described in Example 1, may include killed and / or inactivated forms of the recombinant virus. Alternatively, in some vaccines, recombinant viruses similar to those described in Example 1 may be used as live modified viruses, for example.

[0087] Figure 1B provides the sequence of SEQ ID NO: 2, which is the sequence of the recombinant baculovirus BaculoG / PCV3 ORF2. Annotation for the backbone sequence is from Genbank accession number: NC_001623. One skilled in the art will appreciate that, given the complexity of DNA sequences, minor variations within the backbone between constructs are expected. A map of the construct is shown in Figure 2. The baculovirus expression vector, BaculoG / PCV3 ORF2, can be used to develop PCV3 vaccines and / or controls. Preferred adjuvants for a given vaccine and / or control may differ based on the type of expression vector used, e.g., live expression vector, live modified expression vector, inactivated expression vector, or killed expression vector. The effectiveness of an adjuvant may vary based on the state of the vector (e.g., virus) used. The amount of adjuvant used in a vaccine can be predetermined; for example, a predetermined percentage can be selected to be within a given range (e.g., mass percentage and / or volume percentage in the vaccine) for a given adjuvant and / or combination of adjuvants. In some cases, such as when using a live vaccine, multiple adjuvants can be used. For example, in some embodiments, a combination of adjuvants can be used, such as carbopol and Montanide ISA 207VG. Alternatively, a vaccine containing a live expression vector such as BaculoG / PCV3 ORF2 can be adjuvanted with ISA 207VG and / or carbopol. For example, the adjuvant can be present in the vaccine at a predetermined concentration. For example, a vaccine can contain ISA 207VG at a concentration of 50% by mass of the vaccine. Alternatively, another vaccine containing live BaculoG / PCV3 ORF2 can contain an adjuvant such as carbopol at 20% by volume of the vaccine.

[0088] Vaccines containing killed expression vectors, such as killed viruses, may contain carbopol as an adjuvant. For example, in some embodiments, vaccines containing killed BaculoG / PCV3 ORF2 may contain carbopol as an effective adjuvant. For example, such vaccines may contain a predetermined amount of adjuvant, e.g., a predetermined mass or volume percentage of the vaccine. In particular, a vaccine containing killed BaculoG / PCV3 ORF2 may contain carbopol at 20% by volume of the vaccine. Alternatively, a vaccine may contain killed BaculoG / PCV3 ORF2 and the adjuvant may be about 50% by mass of the vaccine solution. For example, a vaccine containing killed BaculoG / PCV3 ORF2 may contain ISA 207VG as an adjuvant at a predetermined mass percentage, such as 50 percent of the vaccine.

[0089] For example, a baculovirus expression vector, BaculoG / PCV3 ORF2, was used to develop two PCV3 vaccines and a control, outlined herein: Development of BaculoG / PCV3 ORF2, P9; live vaccine adjuvanted with 50% ISA 207VG (methods used to develop the vaccine are disclosed in Example 3); Development of BaculoG / PCV3 ORF2, P9; a live vaccine adjuvanted with 20% carbopol (the method used to develop the vaccine is disclosed in Example 4); Control: BaculoG / no insert, P4; development of a live vaccine adjuvanted with 20% carbopol (the method used to develop the vaccine is disclosed in Example 5); Development of BaculoG / PCV3 ORF2, P9; killed vaccine adjuvanted with 50% ISA 207VG (methods used to develop the vaccine are disclosed in Example 3); Development of BaculoG / PCV3 ORF2, P9; a killed vaccine adjuvanted with 20% carbopol (the method used to develop the vaccine is disclosed in Example 4); Control: BaculoG / no insert, P4; development of a killed vaccine adjuvanted with 20% carbopol (the method used to develop the vaccine is disclosed in Example 5).

[0090] Vaccine efficacy can be examined using whole PCV3 virus and PCR-positive tissues (low counts). Homogenates from tissues can be generated and sequenced. Homogenates and / or whole virus can be used to challenge vaccinated animals. For example, to test for vaccine efficacy, whole PCV3 virus and PCR-positive tissue (low counts) were provided. Homogenates from the tissues were generated and sequenced. The homogenates and whole virus were used to challenge vaccinated animals.

[0091] Subunit vaccines and / or immunogenic compositions based on the PCV3 recombinant ORF2 protein of the present disclosure can be produced using the method of Example 1 of WO 2006 / 072065, modified to express PCV3 ORF2 protein (rather than PCV2 ORF2 protein).

[0092] The PCV3 ORF2 coding sequence can be amplified from PCV3 genomic DNA and / or synthetic PCV3 ORF2 by polymerase chain reaction (PCR). Restriction sites can be used to insert the desired coding sequence into a transfer vector. For example, in some embodiments, the amplified PCV3 ORF2 coding sequence can include a Kozak consensus sequence (see, e.g., Kozak M (October 1987) Nucleic Acids Res. 15(20):8125-8148) immediately 5' to the start codon, along with flanking restriction enzyme sites.

[0093] In some embodiments, the amplified PCV3 ORF2 coding sequence can be subcloned into a baculovirus transfer vector using flanking restriction sites to create the desired transfer vector. For example, the amplified PCV3 ORF2 coding sequence can be subcloned into a baculovirus transfer vector using flanking restriction sites to create a transfer vector such as pVL1392-PCV3 ORF2 or pVL1393-PCV3 ORF2. Other transfer vectors commonly known in the art can also be used. Recombinant baculovirus can be produced by cotransfection of insect cells with the transfer vector and baculovirus DNA. The baculovirus DNA used can include linearized baculovirus DNA and / or circular baculovirus DNA. For example, in one embodiment, recombinant baculovirus can be generated by co-transfection of Sf9 (Spodoptera frugiperda) insect cells with a transfer vector (e.g., pVL1392-PCV3 ORF2 and / or pVL1393-PCV3) and linearized BaculoGold™ baculovirus DNA. Because linearized baculovirus DNA may be derived from Autographa californica nuclear polyhedrosis virus (AcNPV) and may contain a lethal deletion within the polyhedrin locus, rescue of viable baculovirus can be generated upon co-transfection with a transfer vector such as pVL1392-PCV3 ORF2 and / or pVL1393-PCV3 ORF2. The resulting recombinant baculovirus may contain the PCV3 ORF2 coding sequence under the control of the baculovirus polyhedrin promoter. The recombinant baculovirus may be amplified on Sf9 insect cells and then purified by limiting dilution cloning on Sf9 insect cells. In some embodiments, full-length circular baculovirus DNA may be used, such as Bac-to-Bac. For example, Bac-to-Bac may use transposon-mediated recombination to insert a gene of interest into the polyhedron locus. Other methods known in the art may also be used.In some embodiments, methods may be selected based on the potential stability of the method upon commercialization, for example, a baculovirus may be selected that confers increased vaccine stability.

[0094] In some embodiments, after the flasks are inoculated with the master cell culture, the flasks can be incubated at a predetermined temperature for a specific time period. For example, the cultures can be incubated at 27°C for 4 hours. Each flask can then be inoculated with a recombinant baculovirus containing the PCV3 ORF2 gene. For example, the pVL1392 plasmid containing the PCV3 ORF2 gene can be co-transfected with BaculoGold® (BD Biosciences Pharmingen) baculovirus DNA into Sf+ insect cells (Protein Sciences, Meriden, CT) to produce a recombinant baculovirus containing the PCV3 ORF2 gene. The recombinant baculovirus containing the PCV3 ORF2 gene can be plaque-purified and propagated on an SF+ cell line using a master seed virus (MSV), which can then be divided into aliquots and stored at -70°C. The MSV can be positively identified as a PCV3 ORF2 baculovirus by PCR-RFLP using baculovirus-specific primers. Insect cells infected with PCV3 ORF2 baculovirus to produce MSV or working seed virus can express the PCV3 ORF2 antigen, which can be detected by polyclonal serum or monoclonal antibody in an indirect fluorescent antibody assay. Additionally, the identity of the PCV3 ORF2 baculovirus can be confirmed by N-terminal amino acid sequencing. PCV3 ORF2 baculovirus MSV can also be inspected for purity in accordance with 9 C.F.R. sections 113.27(c), 113.28, and 113.55. Each recombinant baculovirus inoculated into spinner flasks can have a different multiplicity of infection (MOI).

[0095] After inoculation with baculovirus, the flasks can be incubated at 27±2°C for 7 days and can be agitated at 100 rpm during this period. The flasks can be fitted with vents to allow airflow. Samples from each flask can be taken every 24 hours for the next 7 days. Following extraction, each sample can be centrifuged, and both the pellet and supernatant are separated and then microfiltered through membranes with pore sizes of 0.45-1.0 μm.

[0096] The amount of ORF3 in the resulting sample can then be quantified via ELISA assay. The ELISA assay can be performed using anti-PCV3 antibody diluted 1:6000 in 0.05M carbonate buffer (pH 9.6). 100 μL of antibody can then be added to the wells of a microtiter plate, sealed, and incubated overnight at 37°C. The plate is then washed three times with a wash solution containing 0.5 mL of Tween 20 (Sigma, St. Louis, MO), 100 mL of 10x D-PBS (Gibco Invitrogen, Carlsbad, CA), and 899.5 mL of distilled water. Then, 250 μL of blocking solution (5 g of Carnation nonfat dry milk (Nestlé, Glendale, CA) in 10 mL of D-PBS QS, made up to 100 mL with distilled water) is added to each well. The next step is to wash the test plate and then add the pre-diluted antigen. Prediluted antigens are produced by adding 200 μL of diluent (0.5 mL of Tween 20 in 999.5 mL of D-PBS) to each well on the dilution plate. The samples are then diluted at 1:240 and 1:480 ratios, and 100 μL of each of these diluted samples is then added to one of the wells in the first row of the dilution plate (i.e., 100 μL of the 1:240 dilution is applied to the wells in the first row, and 100 μL of the 1:480 dilution is applied to the wells in the other row). Serial dilutions can then be performed across the remaining plates by removing 100 μL from each subsequent well and transferring it to the next well on the plate. Each well is mixed before the next transfer. Washing of the test plate involves washing the plate three times with wash buffer. The plate is then sealed and incubated at 37° C. for 1 hour, before being washed three more times with wash buffer. The detection antibody used is an antibody against PCV ORF2. The detection antibody is diluted 1:300 in diluent, and then 100 μL of the diluted detection antibody is added to the wells. The plate is then sealed and incubated at 37° C. for 1 hour, and then washed three times with wash buffer.The conjugate diluent is then prepared by adding normal rabbit serum (Jackson Immunoresearch, West Grove, PA) to the diluent to a concentration of 1%.

[0097] Conjugated antibody, goat anti-mouse (H+1)-HRP (Jackson Immunoresearch), is diluted 1:10,000 in conjugate diluent. 100 μL of the diluted conjugated antibody is then added to each well. The plate is then sealed and incubated at 37° C. for 45 minutes, and then washed three times with wash buffer. 100 μL of substrate (TMB Peroxidase Substrate, Kirkgaard and Perry Laboratories (KPL), Gaithersburg, MD) mixed with an equal volume of peroxidase substrate B (KPL) is added to each well. The plate is incubated at room temperature for 15 minutes. 100 μL of 1N HCl solution is then added to all wells to stop the reaction. The plate is then loaded into an ELISA reader.

[0098] Advantageously, insect cells can be cultured to produce PCV3 ORF2 protein under serum-free conditions, such as the serum-free insect cells of US Pat. No. 6,103,526 (expressSF+ cell line).

[0099] Adjuvants, cell culture supernatants, preservatives, stabilizers, viral vectors, immunomodulators, and dosages disclosed in U.S. Pat. Nos. 9,610,345 and 9,669,087, both of which are incorporated herein by reference, are contemplated.

[0100] The immunogenic composition used herein is effective for inducing an immune response against PCV3 and preventing, reducing, and / or alleviating the severity of clinical symptoms associated with PCV3 infection. The composition generally comprises at least one PCV3 antigen.

[0101] PCV3 in pigs can present with a wide variety of symptoms, and in many cases, individual animals exhibit only a small subset of potential symptoms. Symptoms associated with the presence of PCV3 include viremia, viral shedding, and the presence of viral nucleic acids in bodily excretions, such as colostrum, milk, feces, saliva, and eye swabs. For example, Jiang et al., "Induction of porcine dermatitis and nephropathy syndrome in piglets by infection with porcine circovirus type 3," J. Virol. doi:10.1128 / JVI.02045-18, the disclosure of which is incorporated by reference, relates to inoculating piglets with PCV3 and observing the resulting clinical signs. The present disclosure relates to treating and / or alleviating the symptoms of porcine dermatitis and nephropathy syndrome (PDNS)-like disease, lymphocyte metaplasia and necrosis, caused by PCV3, by administering a composition of the present disclosure. In particular, the mere presence of antibodies in young pigs or piglets, for example, pigs or piglets under 10 weeks of age, for example, under 6 weeks of age, for example, under 3, 2, or 1 week of age, or under 15 weeks of age, such as at birth, may not indicate exposure to PCV3 and / or disease. Pigs or piglets that have been exposed to PCV3 and / or have antibodies to PCV3 may still benefit from the compositions of the present disclosure, for example, by reducing, preventing, or alleviating the severity of symptoms.

[0102] Thus, the compositions of the present disclosure may be used in methods for inducing an immune response, which may be a protective immune response, as well as methods for reducing, preventing, or alleviating the severity of symptoms, and the dosages, formulations, etc. for reducing, preventing, or alleviating the severity of symptoms are similar to those for methods for inducing an immune response. Thus, methods described herein for inducing an immune response may be carried out to reduce, prevent, or alleviate the severity of symptoms; compositions described herein that are useful for inducing an immune response are similarly useful for reducing, preventing, or alleviating the severity of symptoms (not just being compositions for inducing an immune response).

[0103] Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this disclosure pertains. As used herein, the term "immunogenic composition" refers to any pharmaceutical composition containing a PCV3 antigen that can be used to prevent or treat a disease or condition associated with PCV3 infection in a subject. Preferred immunogenic compositions can induce, stimulate, or enhance an immune response to PCV3. Thus, the term "immunogenic composition" encompasses both subunit immunogenic compositions and compositions containing whole-killed PCV3, or attenuated and / or inactivated PCV3, as described below.

[0104] As used herein, the term "subunit immunogenic composition" refers to a composition containing at least one immunogenic polypeptide or antigen, but not all of the antigens are derived from or homologous to antigens derived from PCV3. Such compositions are substantially free of intact PCV3. Thus, a "subunit immunogenic composition" is prepared from immunogenic polypeptides that have been at least partially purified or fractionated (preferably substantially purified) from PCV3 or its recombinant analogs. A subunit immunogenic composition may contain one or more subunit antigens of interest that are substantially free of, or in fractionated form from, other antigens or polypeptides derived from PCV3. A preferred immunogenic subunit composition comprises the PCV3 ORF2 protein, described below.

[0105] An "immunological or immune response" to a composition or vaccine is the development in a host of a cell-mediated and / or antibody-mediated immune response to the composition or vaccine of interest. Typically, an "immune response" includes, but is not limited to, one or more of the following effects: the production or activation of antibodies, B cells, helper T cells, suppressor T cells, and / or cytotoxic T cells and / or γδ T cells specifically directed against one or more antigens contained in the composition or vaccine of interest. Preferably, the host will exhibit either a therapeutic or protective immunological response such that resistance to new infections is enhanced and / or the clinical severity of disease is reduced. Such protection will be demonstrated by either a reduction in the number, severity, or absence of one or more of the symptoms associated with PCV3 infection, as described above.

[0106] As used herein, the term "immunogenic" protein or polypeptide, or "antigen" refers to an amino acid sequence that elicits the immunological response described above. As used herein, "immunogenic" protein or polypeptide includes the full-length sequence of any PCV3 protein, analog, or immunogenic fragment thereof. The term "immunogenic fragment" refers to a fragment of a protein that contains one or more epitopes and thus elicits the immunological response described above. Such fragments can be identified using any number of epitope mapping methods known in the art. See, for example, "Epitope Mapping Protocols in Methods in Molecular Biology," Vol. 66 (Glenn E. Morris, Ed., 1996) Humana Press, Totowa, NJ. For example, linear epitopes can be determined by simultaneously synthesizing multiple peptides corresponding to portions of a protein molecule, e.g., on a solid support, and reacting the peptides with an antibody while the peptides are attached to the support. Such techniques are known in the art and are described, for example, in U.S. Patent No. 4,708,871; Geysen et al., (1984) Proc. Natl. Acad. Sci. USA 8:3998-4002; Geysen et al., (1986) Molec. Immunol. 23:709-715 (all of which are incorporated herein by reference).Similarly, conformational epitopes can be easily identified by determining the spatial conformation of amino acids, for example, by x-ray crystallography and two-dimensional nuclear magnetic resonance.See, for example, Epitope Mapping Protocols, supra.

[0107] Synthetic antigens, such as polyepitopes, flanking epitopes, and other recombinantly or synthetically derived antigens, are also included within the definition. See, e.g., Bergmann et al. (1993) Eur. J. Immunol. 23:2777-2781; Bergmann et al. (1996), J. Immunol. 157:3242-3249; Suhrbier, A. (1997), Immunol. and Cell Biol. 75:402-408; Gardner et al. (1998) 12th World AIDS Conference, Geneva, Switzerland, June 28-July 3, 1998.

[0108] In a preferred embodiment of the present disclosure, an immunogenic composition is provided that induces an immune response and, more preferably, confers protective immunity against the clinical signs of PCV3 infection. Most preferably, the composition comprises a polypeptide expressed by ORF2 of PCV3, or a fragment thereof, as the antigenic component of the composition. The PCV3 ORF2 DNA and protein used in the processes provided herein for preparing the compositions provided herein are highly conserved domains within PCV3 isolates, and therefore, any PCV3 ORF2 may be useful as a source of PCV3 ORF2 DNA and / or polypeptide for use herein. A preferred PCV3 ORF2 protein is translated from the nucleotide sequence of SEQ ID NO: 1. While a preferred PCV3 ORF2 polypeptide is provided herein, those skilled in the art will understand that this sequence may vary by up to 6-10% in sequence identity and still retain the antigenic characteristics that make it useful in immunogenic compositions. Furthermore, the antigenic characteristics of the modified antigen are still maintained if the modified antigen confers at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100% protective immunity compared to the PCV3 ORF2 protein encoded by the polynucleotide sequence of SEQ ID NO: 1. As used herein, an "immunogenic composition" refers to a PCV3 ORF2 protein that induces an "immunological response," which is a cell-mediated immune response and / or an antibody-mediated immune response, against the PCV3 ORF2 protein in a host. Preferably, the immunogenic composition induces or enhances an immune response against PCV3, thereby conferring protective immunity against PCV3 infection and reducing the incidence, severity, or prevention of one or more of the clinical symptoms associated therewith, preferably reducing the incidence, severity, or prevention of all of these.

[0109] In some embodiments, immunogenic portions of the PCV3 ORF2 protein are used as antigenic components in compositions. As used herein, the term "immunogenic portion" refers to truncated and / or substituted forms, or fragments of the PCV3 ORF2 protein and / or polynucleotide, respectively. Preferably, such truncated and / or substituted forms, or fragments will contain at least 6 consecutive amino acids from the full-length ORF2 polypeptide. More preferably, truncated or substituted forms, or fragments will contain at least 10, more preferably at least 15, and even more preferably at least 19 consecutive amino acids from the full-length ORF2 polypeptide. It is further understood that such sequences may be part of a larger fragment or truncated form.

[0110] A further preferred PCV3 ORF2 polypeptide provided herein is encoded by the nucleotide sequence of SEQ ID NO:1. However, it will be understood by those of skill in the art that this sequence may vary by up to 6-20% in sequence identity and still retain the antigenic characteristics that make it useful in immunogenic compositions. In some forms, truncated or substituted forms, or fragments of this PCV3 ORF2 polypeptide are used as antigenic components in compositions. Preferably, such truncated or substituted forms, or fragments will contain at least 18 contiguous nucleotides from the full-length ORF2 nucleotide sequence. More preferably, the truncated or substituted forms, or fragments will have at least 30, more preferably at least 45, and even more preferably at least 57 contiguous nucleotides of the full-length ORF2 nucleotide sequence, e.g., SEQ ID NO:1.

[0111] " Sequence identity " as known in the art refers to the relationship between two or more polypeptide sequences or two or more polynucleotide sequences, that is, the relationship between a reference sequence and a given sequence that is compared with the reference sequence. Sequence identity is determined by comparing a given sequence with a reference sequence after the sequences are optimally aligned to produce the highest degree of sequence similarity, as determined by the match between the strings of such sequences. Once such alignment is performed, sequence identity is confirmed on a position-to-position basis; for example, if the nucleotide or amino acid residue at a particular position is the same, the sequence is "identical" at that position.Then, to obtain the sequence identity percentage, the total number of such position identities is divided by the total number of nucleotides or residues in the reference sequence. Sequence identity was determined in Computational Molecular Biology, Lesk, AN, ed., Oxford University Press, New York (1988), Biocomputing: Informatics and Genome Projects, Smith, DW, ed., Academic Press, New York (1993); Computer Analysis of Sequence Data, Part I, Griffin, AM, and Griffin, HG, eds., Humana Press, New Jersey (1994); Sequence Analysis in Molecular Biology, von Heinge, G., Academic Press (1987); Sequence Analysis Primer, Gribskov, M. and Devereux, J., eds., M. Stockton Press, New York (1991); and Carillo, H., and Lipman, D., SIAM J. Applied Math., 48: 1073 (1988), the teachings of which are incorporated herein by reference.Preferred methods for determining sequence identity are designed to produce the largest match between the sequences under test. Methods for determining sequence identity are codified in publicly available computer programs that determine sequence identity between given sequences. Examples of such programs include, but are not limited to, the GCG program package (Devereux, J., et al., Nucleic Acids Research, 12(1):387 (1984)), BLASTP, BLASTN, and FASTA (Altschul, SF et al., J. Molec. Biol., 215:403-410 (1990)). BLASTX programs are publicly available from NCBI and other sources (the teachings of which are incorporated herein by reference; BLAST Manual, Altschul, S. et al., NCVI NLM NIH Bethesda, MD 20894; Altschul, S. F. et al., J. Molec. Biol., 215:403-410 (1990)). These programs optimally align sequences using default gap weights to produce the highest level of sequence identity between a given sequence and a reference sequence. By way of example, by a polynucleotide having a nucleotide sequence having at least, e.g., 85%, preferably 90%, and even more preferably 95% "sequence identity" to a reference nucleotide sequence, it is intended that the nucleotide sequence of the given polynucleotide is identical to the reference sequence, except that the given polynucleotide sequence may contain up to 15, preferably up to 10, and even more preferably up to 5 point mutations per every 100 nucleotides of the reference nucleotide sequence.In other words, in a polynucleotide having a nucleotide sequence that is at least 85%, preferably 90%, and even more preferably 95% identical to a reference nucleotide sequence, up to 15%, preferably 10%, and even more preferably 5% of the nucleotides in the reference sequence may be deleted or substituted with other nucleotides, or up to 15%, preferably 10%, and even more preferably 5% of the total nucleotides in the reference sequence may be inserted into the reference sequence. These mutations of the reference sequence may be made at the 5'-terminal or 3'-terminal position of the reference nucleotide sequence, or at any position between these terminal positions, either individually interspersed among nucleotides in the reference sequence or interspersed in one or more contiguous groups within the reference sequence. Similarly, by a polypeptide having a given amino acid sequence that has at least, e.g., 85%, preferably 90%, and even more preferably 95%, sequence identity to a reference amino acid sequence, it is intended that the given amino acid sequence of the polypeptide is identical to the reference sequence, except that the given polypeptide sequence may contain up to 15, preferably up to 10, and even more preferably up to 5 amino acid changes per 100 amino acids of the reference amino acid sequence. In other words, to obtain a given polypeptide sequence that has at least 85%, preferably 90%, and even more preferably 95% sequence identity with the reference amino acid sequence, up to 15%, preferably up to 10%, and even more preferably up to 5% of the amino acid residues in the reference sequence may be deleted or substituted with another amino acid, or up to 15%, preferably up to 10%, and even more preferably up to 5% of the total number of amino acid residues in the reference sequence may be inserted into the reference sequence. These alterations of the reference sequence may be made at the amino- or carboxy-terminal positions of the reference amino acid sequence, or at any position between these terminal positions, either individually interspersed among residues in the reference sequence or interspersed in one or more contiguous groups within the reference sequence. Preferably, residue positions that are not identical differ by conservative amino acid substitutions.However, conservative substitutions are not counted as matches when determining sequence identity.

[0112] As used herein, "sequence homology" refers to a method for determining the relationship between two sequences. To determine sequence homology, two or more sequences are optimally aligned, and gaps are introduced, if necessary. However, in contrast to "sequence identity," conservative amino acid substitutions are counted as matches when determining sequence homology. In other words, to obtain a polypeptide or polynucleotide having 95% sequence homology with a reference sequence, 85%, preferably 90%, and even more preferably 95% of the amino acid residues or nucleotides in the reference sequence must match or contain conservative substitutions with other amino acids or nucleotides, or up to 15%, preferably up to 10%, and even more preferably up to 5% of the total number of amino acid residues or nucleotides in the reference sequence that do not contain conservative substitutions can be inserted into the reference sequence. Preferably, the homologous sequence comprises a stretch of at least 50 nucleotides, even more preferably at least 100 nucleotides, even more preferably at least 250 nucleotides, and even more preferably at least 500 nucleotides.

[0113] A "conservative substitution" refers to the substitution of an amino acid residue or nucleotide with another amino acid residue or nucleotide having similar characteristics or properties, including size, hydrophobicity, etc., such that the overall functionality does not change significantly.

[0114] "Isolated" means altered "by artifice" from its natural state; that is, when something occurs in nature, it has been changed or removed from its original environment, or both. For example, as used herein, the term means that a polynucleotide or polypeptide that is naturally present in a living organism is not "isolated," but the same polynucleotide or polypeptide separated from the coexisting materials of its natural state is "isolated."

[0115] Therefore, the immunogenic composition as used herein also refers to a composition comprising a PCV3 ORF2 protein, wherein the PCV3 ORF2 protein is any one of the PCV3 ORF2 proteins described above. According to a further embodiment, the PCV3 ORF2 protein is provided in the immunological composition at an antigen loading level effective to induce a desired immune response, i.e., reduce the incidence of PCV3 infection, reduce the severity of PCV3 infection, or prevent one or more clinical symptoms resulting from PCV3 infection. Preferably, the incorporation level of PCV3 ORF2 protein is at least 0.2 μg of antigen per ml of the final immunogenic composition (μg / ml), more preferably about 0.2 to about 400 μg / ml, even more preferably about 0.3 to about 200 μg / ml, even more preferably about 0.35 to about 100 μg / ml, even more preferably about 0.4 to about 50 μg / ml, even more preferably about 0.45 to about 30 μg / ml, even more preferably about 0.6 to about 15 μg / ml, even more preferably about 0.75 to about 8 μg / ml, even more preferably about 1.0 to about 6 μg / ml, even more preferably about 1.3 to about 3.0 μg / ml, even more preferably about 1.4 to about 2.5 μg / ml, even more preferably about 1.5 to about 2.0 μg / ml, and most preferably about 1.6 μg / ml.

[0116] According to a further embodiment, the incorporation level of the ORF2 antigen is at least 0.2 μg of the above-described PCV3 ORF2 protein (μg per dose) per dose of the final antigenic composition, more preferably about 0.2 to about 400 μg per dose, even more preferably about 0.3 to about 200 μg per dose, even more preferably about 0.35 to about 100 μg per dose, even more preferably about 0.4 to about 50 μg per dose, even more preferably about 0.45 to about 30 μg per dose, even more preferably about 0.6 to about 15 μg per dose, even more preferably about 0.75 to about 8 μg per dose, even more preferably about 1.0 to about 6 μg per dose, even more preferably about 1.3 to about 3.0 μg per dose, even more preferably about 1.4 to about 2.5 μg per dose, even more preferably about 1.5 to about 2.0 μg per dose, and most preferably about 1.6 μg per dose. In one embodiment, the ORF2 antigen (e.g., PCV3 ORF2 protein) may be present in a range of about 1.3 to about 3 μg in a final composition administered. For example, the final antigenic composition may contain about 1.6 μg of PCV3 ORF2 protein per mL administered.

[0117] The PCV3 ORF2 polypeptide used in the immunogenic composition according to the present disclosure can be derived in any manner, including PCV3 ORF2 isolation and purification methods, standard protein synthesis methods, and recombinant methods. A preferred method for obtaining PCV3 ORF2 polypeptide is presented in U.S. Patent Application No. 11 / 034,797, the teachings and contents of which are incorporated herein by reference. Briefly, when susceptible cells are infected with a recombinant virus vector containing a PCV3 ORF2 DNA coding sequence, the PCV3 ORF2 polypeptide is expressed by the recombinant virus, and the expressed PCV3 ORF2 polypeptide is recovered from the supernatant by filtration and inactivated by any conventional method, preferably using binary ethyleneimine, which is then neutralized to stop the inactivation process.

[0118] The immunogenic composition as used herein also refers to a composition comprising i) any of the PCV3 ORF2 proteins described above, preferably at the concentrations described above, and ii) at least a portion of a viral vector, preferably a recombinant baculovirus, that expresses the PCV3 ORF2 protein. Furthermore, the immunogenic composition may comprise i) any of the PCV3 ORF2 proteins described above, preferably at the concentrations described above, ii) at least a portion of a viral vector, preferably a recombinant baculovirus, that expresses the PCV3 ORF2 protein, and iii) a portion of a cell culture supernatant. As used herein, the immunogenic composition also refers to a composition comprising: i) any of the PCV3 ORF2 proteins described above, preferably at the concentrations described above; ii) at least a portion of a viral vector, preferably a recombinant baculovirus, that expresses the PCV3 ORF2 protein; and iii) a portion of a cell culture; wherein approximately 90% of the components may have a size smaller than 1 μm.

[0119] As used herein, the term "immunogenic composition" also refers to a composition comprising: i) any of the PCV3 ORF2 proteins described above, preferably at the concentrations described above; ii) at least a portion of a viral vector expressing the PCV3 ORF2 protein; iii) a portion of a cell culture; and iv) an inactivating agent, preferably BEI, that inactivates the recombinant viral vector, wherein approximately 90% of the components in i) to iii) may have a size smaller than 1 μm. Preferably, the BEI is present at a concentration effective to inactivate the baculovirus. The effective concentration is described above. The immunogenic composition as used herein also refers to a composition comprising: i) any of the PCV3 ORF2 proteins described above, preferably at the concentrations described above; ii) at least a portion of a viral vector expressing the PCV3 ORF2 protein; iii) a portion of a cell culture; iv) an inactivating agent, preferably BEI, that inactivates the recombinant viral vector; and v) a neutralizing agent that stops the inactivation mediated by the inactivating agent, wherein approximately 90% of the components in i) to iii) may have a size smaller than 1 μm. Preferably, when the inactivating agent is BEI, the composition comprises sodium thiosulfate in an amount equal to that of BEI.

[0120] The polypeptide is incorporated into a composition that can be administered to an animal susceptible to PCV3 infection. In a preferred form, the composition may also contain additional components known to those skilled in the art (see also Remington's Pharmaceutical Sciences. (1990). 18th ed. Mack Publ., Easton). In addition, the composition may contain one or more veterinarily acceptable carriers. As used herein, "veterinarily acceptable carrier" includes any and all solvents, dispersion media, coating agents, adjuvants, stabilizers, diluents, preservatives, antibacterial and antifungal agents, isotonic agents, absorption delaying agents, and the like. In a preferred embodiment, the immunogenic composition comprises the PCV3 ORF2 protein provided herein, preferably at the concentrations described above, mixed with an adjuvant, preferably Carbopol, and saline.

[0121] Those skilled in the art will understand that the compositions used herein may incorporate known, injectable, physiologically acceptable sterile solutions. For example, aqueous isotonic solutions, such as saline or corresponding plasma protein solutions, are readily available for preparing ready-to-use solutions for parenteral injection or infusion. Additionally, the immunogenic and vaccine compositions of the present disclosure may contain diluents, isotonic agents, stabilizers, or adjuvants. Diluents may include water, saline, dextrose, ethanol, glycerol, etc. Isotonic agents may include, among others, sodium chloride, dextrose, mannitol, sorbitol, and lactose. Stabilizers include, among others, albumin and alkali salts of ethylenediaminetetraacetic acid.

[0122] As used herein, "adjuvants" may include aluminum hydroxide and aluminum phosphate, saponins such as Quil A, QS-21 (Cambridge Biotech Inc., Cambridge Mass.), and GPI-0100 (Galenica Pharmaceuticals, Inc., Birmingham, Ala.), water-in-oil emulsions, oil-in-water emulsions, and water-in-oil-in-water emulsions. Emulsions may be based, inter alia, on light liquid paraffin oil (European Pharmacopoeia type); isoprenoid oils such as squalane or squalene obtained from the oligomerization of alkenes, especially isobutene or decene; esters of acids or alcohols containing linear alkyl groups, more particularly esters of vegetable oils, ethyl oleate, propylene glycol di(caprylic / capric) acid, glyceryl tri(caprylic / capric) acid, or propylene glycol dioleate; or esters of branched fatty acids or alcohols, especially isostearate. Oil is used in combination with emulsifier to form emulsion.Emulsifier is preferably nonionic surfactant, especially sorbitan, mannide (for example, anhydrous mannitol oleate), glycol, polyglycerol, propylene glycol, and oleic acid, isostearic acid, ricinoleic acid or hydroxystearic acid ester, optionally ethoxylated ester, and polyoxypropylene-polyoxyethylene block copolymer, especially Pluronic product, especially Pluronic L121.See Hunter et al., The Theory and Practical Application of Adjuvants (Ed.Stewart-Tull, DES).John Wiley and Sons, NY, pp51-94 (1995); and Todd et al., Vaccine 15:564-570 (1997).

[0123] For example, it is possible to use SPT emulsion described on page 147 of "Vaccine Design, The Subunit and Adjuvant Approach" edited by M. Powell and M. Newman, Plenum Press, 1995, and emulsion MF59 described on page 183 of the same book.

[0124] Further examples of adjuvants are compounds selected from polymers of acrylic acid or methacrylic acid and copolymers of maleic anhydride and alkenyl derivatives. Advantageous adjuvant compounds are, inter alia, polymers of acrylic acid or methacrylic acid crosslinked with polyalkenyl ethers of sugars or polyalcohols. These compounds are known by the term carbomer (Phameuropa Vol. 8, No. 2, June 1996). Those skilled in the art can also refer to U.S. Pat. No. 2,909,462, which describes such acrylic polymers crosslinked with polyhydroxylated compounds, having at least three, preferably no more than eight, hydroxyl groups, in which at least three hydroxyl hydrogen atoms are replaced by unsaturated aliphatic radicals having at least two carbon atoms. Preferred radicals are those containing two to four carbon atoms, such as vinyl, allyl, and other ethylenically unsaturated groups. The unsaturated radicals themselves may contain other substituents, such as methyl. Particularly suitable are products sold under the Carbopol trademark (BF Goodrich, Ohio, USA). Carbopol is crosslinked with allyl sucrose or allyl pentaerythritol. Among these, Carbopol 974P, Carbopol 934P, and Carbopol 971P may be mentioned. Most preferred is the use of Carbopol, especially Carbopol 971P, preferably in an amount of about 500 μg to about 5 mg per dose, even more preferably in an amount of about 750 μg to about 2.5 mg per dose, and most preferably in an amount of about 1 mg per dose. In particular, the dose of the final composition may contain Carbopol or Carbopol 971 in the range of about 750 μg to about 2.5 mg. For example, in some embodiments, the dose of the final composition may contain about 1 mg of Carbopol 971.

[0125] Additional suitable adjuvants include, but are not limited to, the RIBI adjuvant system (Ribi Inc.), block copolymers (CytRx, Atlanta, Ga.), SAF-M (Chiron, Emeryville, Calif.), monophosphoryl lipid A, avridine lipid-amine adjuvant, heat-labile enterotoxin from Escherichia coli (recombinant or other forms), cholera toxin, IMS 1314, or muramyl dipeptide, among others.

[0126] Preferably, the adjuvant is added in an amount of about 100 μg to about 10 mg per dose. Even more preferably, the adjuvant is added in an amount of about 100 μg to about 10 mg per dose. Even more preferably, the adjuvant is added in an amount of about 500 μg to about 5 mg per dose. Even more preferably, the adjuvant is added in an amount of about 750 μg to about 2.5 mg per dose. Most preferably, the adjuvant is added in an amount of about 1 mg per dose.

[0127] In addition, the composition may contain one or more pharmaceutically acceptable carriers. As used herein, "pharmaceutically acceptable carriers" includes any and all solvents, dispersion media, coating agents, stabilizers, diluents, preservatives, antibacterial and antifungal agents, isotonic agents, absorption delaying agents, etc. Most preferably, the composition provided herein contains PCV3 ORF2 protein recovered from the supernatant of in vitro cultured cells, the cells being infected with a recombinant viral vector containing PCV3 ORF2 DNA and expressing the PCV3 ORF2 protein, and the cell culture treated with about 2 to about 8 mM BEI, preferably about 5 mM BEI, to inactivate the viral vector, and an equal concentration of a neutralizing agent, preferably a sodium thiosulfate solution to a final concentration of about 2 to about 8 mM, preferably about 5 mM.

[0128] The present disclosure also relates to an immunogenic composition comprising: i) any of the PCV3 ORF2 proteins described above, preferably at the concentrations described above; ii) at least a portion of a viral vector expressing the PCV3 ORF2 protein; iii) a portion of a cell culture; iv) an inactivating agent, preferably BEI, that inactivates the recombinant viral vector; v) a neutralizing agent that stops inactivation mediated by the inactivating agent, preferably sodium thiosulfate, in an amount equivalent to the BEI; and vi) a suitable adjuvant, preferably Carbopol 971, in the amount described above; wherein approximately 90% of the components in i) to iii) have a size smaller than 1 μm. According to a further embodiment, the immunogenic composition further comprises a pharmaceutically acceptable salt, preferably a phosphate salt, at a physiologically acceptable concentration. Preferably, the pH of the immunogenic composition is adjusted to physiological pH, meaning approximately 6.5 to 7.5.

[0129] In some embodiments, an immunogenic composition may refer to a composition comprising, in a 1 ml dose, i) at least a portion of a PCV3 ORF2 protein, ii) a baculovirus expressing the PCV3 ORF2 protein, iii) a cell culture, iv) an inactivating agent (e.g., BEI) having a concentration ranging from about 2 to about 8 mM, v) a neutralizing agent (e.g., sodium thiosulfate) in an amount equivalent to the inactivating agent, and vi) a predetermined amount of an adjuvant (e.g., Carbopol 971), and vii) a physiologically acceptable concentration of phosphate. In some embodiments, the components may be selected such that 90% of the combined components, including i) the PCV3 ORF2 protein, ii) the baculovirus expressing the protein, and iii) the cell culture, have a size smaller than 1 μm. Furthermore, in some embodiments, one or more components of the immunogenic composition may be selected such that the immunogenic composition has a pH ranging from about 6.5 to about 7.5. Decisions regarding the selection of components and / or amounts and / or concentrations may relate to a variety of factors affecting the stability of the immunogenic composition, ease of manufacture, availability of materials, the age, size, and / or condition of the animal to be treated, and / or the desired result.

[0130] For example, the immunogenic composition as used herein also refers to a composition containing, per ml: i) at least 1.6 μg of the PCV3 ORF2 protein described above; ii) at least a portion of a baculovirus expressing the PCV3 ORF2 protein; iii) a portion of a cell culture; iv) about 2 to 8 mM BEI; v) sodium thiosulfate in an amount equivalent to the BEI; and vi) about 1 mg of Carbopol 971; and vii) a physiologically acceptable concentration of phosphate; wherein about 90% of components i) to iii) have a size smaller than 1 μm, and the pH of the immunogenic composition is adjusted to about 6.5 to 7.5.

[0131] The immunogenic composition may further comprise one or more other immunomodulatory agents, such as, for example, interleukins, interferons, or other cytokines (such as, but not limited to, IL-1, IL-2, IL-7, IFN-alpha, IFN-beta, and IFN-gamma). The immunogenic composition may also comprise gentamicin and merthiolate. While the amounts and concentrations of adjuvants and additives useful in the context of the present disclosure can be readily determined by one of skill in the art, the present disclosure contemplates compositions comprising about 50 μg to about 2000 μg of adjuvant. In some embodiments, it may be preferable to use an adjuvant in an amount of about 250 μg of adjuvant per milliliter dose of vaccine composition. In some embodiments, the immunogenic composition may comprise an antibiotic at a concentration ranging from about 1 μg / mL to about 60 μg / mL. For example, the immunogenic composition may comprise less than about 30 μg / mL of antibiotic.

[0132] The immunogenic composition as used herein also refers to a composition comprising: i) any of the PCV3 ORF2 proteins described above, preferably at the concentrations described above; ii) at least a portion of a viral vector expressing the PCV3 ORF2 protein; iii) a portion of a cell culture; iv) an inactivating agent, preferably BEI, that inactivates the recombinant viral vector; and v) a neutralizing agent that stops the inactivation mediated by the inactivating agent, preferably sodium thiosulfate, in an amount equivalent to BEI; vi) a suitable adjuvant, preferably Carbopol 971, in the amount described above; vii) a physiological saline buffer, preferably phosphate, in a pharmaceutically acceptable concentration; and viii) an antimicrobial active agent; wherein approximately 90% of the components in i) to iii) have a size smaller than 1 μm.

[0133] The compositions according to the present disclosure may be applied intradermally, intratracheally, or intravaginally. Preferably, the compositions may be applied intramuscularly or intranasally, most preferably intramuscularly. It may prove advantageous to apply the pharmaceutical compositions described above intravenously or by direct injection into the target tissue within the animal body. For systemic application, intravenous, intravascular, intramuscular, intranasal, intraarterial, intraperitoneal, oral, esophagogastric, or intrathecal routes are preferred. More localized application may also be performed subcutaneously, intradermally, intracutaneously, intracardially, intralobarly, intramedullary, intrapulmonary, or directly within or adjacent to the tissue to be treated (connective tissue, bone tissue, muscle tissue, nerve tissue, epithelial tissue). Depending on the desired duration and effectiveness of treatment, the compositions according to the present disclosure may be administered in a single dose, multiple doses, or intermittently, for example, daily, over several days, weeks, or months, in different dosages. Single or multiple doses are also contemplated. Vaccines in combination with other antigens of swine pathogens are also contemplated. Preferred combination compositions contain PCV3 ORF2 protein and PPV, PRRSV antigen, M. hyopneumoniae antigen (supernatant or bacterin), or PRRSV antigen and M. hyopneumoniae antigen (supernatant or bacterin), or any combination with the aforementioned PCV2 ORF2 protein.

[0134] In some embodiments, a dosing regimen can be developed to deliver an amount of PCV3 ORF2 effective to induce a desired effect, such as an immune response in the animal and / or its progeny. Decisions regarding the dosing regimen can relate to the desired outcome, the components selected for use in the immunogenic composition, the route of administration, such as parenteral and / or subcutaneous administration, the number or doses delivered, e.g., single or multiple doses, and / or the specific characteristics of the animal or animal population being treated, such as the age, size, and / or condition of the animal. The condition of the animal can refer, for example, to health status, pregnancy status, size, etc. Thus, sows and piglets may require different effective doses. As stated above, treatment methods may vary based on the desired outcome, for example, sows may be treated to inhibit and / or prevent conditions associated with porcine circovirus, and sows may be treated to inhibit and / or prevent the negative effects of infection by porcine circovirus in their piglets.

[0135] A dosing regimen may include one or more administrations of an immunogenic composition containing a predetermined amount of PCV3 ORF2 protein. For example, a dosing regimen may include administration of about 2 micrograms to about 400 micrograms of PCV3 ORF2 protein. In certain embodiments, a dosing regimen for a particular immunogenic composition may include more than about 2 micrograms of PCV3 ORF2 protein. In some cases, each administration of a particular immunogenic composition may include more than about 4 micrograms of PCV3 ORF2 protein. Some dosing regimen embodiments for an immunogenic composition may include the immunogenic composition at a dose of at least about 8 micrograms of PCV3 ORF2 protein. For example, some dosing regimens for the immunogenic compositions disclosed herein may be constructed such that at least one administration includes more than about 16 micrograms of the desired PCV3 ORF2 protein.

[0136] In some embodiments, a dosing regimen can be selected based on the desired expression of a specific PCV3 ORF2 protein in an animal. For example, given an immunogenic composition comprising a vector and / or expression system suitable for swine, it may be desirable for the vector delivered by the immunogenic composition to be capable of delivering an amount of PCV3 ORF2 protein in vivo ranging from about 2 micrograms to about 400 micrograms. In some embodiments, a dosing regimen for a particular immunogenic composition is configured to deliver greater than about 2 micrograms of PCV3 ORF2 protein to an animal. In some instances, a dosing regimen for a particular immunogenic composition is configured to deliver greater than about 4 micrograms of PCV3 ORF2 protein to an animal. Some dosing regimen embodiments for immunogenic compositions are configured to deliver greater than about 8 micrograms of PCV3 ORF2 protein to an animal. For example, some dosing regimens for the immunogenic compositions disclosed herein can be constructed such that greater than about 16 micrograms of the desired PCV3 ORF2 protein can be delivered to an animal.

[0137] The dosing regimen may also include guidelines for the route of administration and / or the number of doses. For example, depending on the immunogenic composition and the desired outcome, it may be desirable to deliver a dose of the immunogenic composition to piglets at a specific age, particularly about 1 week, 2 weeks, or 3 weeks of age. In some cases, piglets may be administered the immunogenic composition at an age ranging from about 7 days to about 28 days of age. In dosing regimen embodiments, pigs may be administered the immunogenic composition at an age ranging from about 14 days to about 26 days of age. For example, the dosing window for piglets may be selected from an age ranging from about 16 days to about 26 days of age. Some dosing regimen embodiments may include administering the immunogenic composition to piglets at an age ranging from about 18 days to about 24 days of age.

[0138] The immunogenic composition may comprise a recombinant PCV3 ORF2 protein. In particular, the immunogenic composition may comprise a recombinant PCV3 ORF2 protein expressed from a baculovirus. Furthermore, in some cases, immunogenic compositions containing recombinant PCV3 ORF2 proteins can be administered in combination with one or more doses of additional antigens, such as antigens derived from PCV2 ORF2, PPV, PRRSV, and / or M. hyopneumoniae ("M. Hyo"). The PRRSV antigen can be a live attenuated vaccine. The M. Hyo antigen can be a bacterin, a supernatant, or a combination of a bacterin and a supernatant. Multiple doses of the immunogenic composition can be administered in a dosing regimen. For example, the dosing regimen can include administering an immunogenic composition containing a recombinant PCV3 ORF2 protein and an immunogenic composition containing a recombinant PCV2 ORF2 protein. In some cases, the administration can include approximately equal amounts of recombinant PCV3 ORF2 protein and recombinant PCV2 ORF2 protein. An embodiment of the dosing regimen can include administering an immunogenic composition containing a recombinant PCV3 ORF2 protein and a recombinant PCV2 ORF2 protein, both of which can be expressed using a baculovirus-based expression system.

[0139] Embodiments of recombinant PCV3 ORF2 immunogenic compositions can include additional antigens, such as recombinant proteins derived from PCV3 ORF2, as well as antigens such as live attenuated PRRSV and / or M.Hyo bacterin, supernatant, or a combination of bacterin and supernatant. Some embodiments of immunogenic compositions can include baculovirus-expressed recombinant proteins from PCV3 ORF2 and PCV2 ORF2, as well as PRRSV antigens (e.g., live attenuated vaccine) and / or M.Hyo (e.g., bacterin and / or supernatant). Furthermore, in some cases, immunogenic compositions can include PCV3 ORF2 protein in combination with PCV2 ORF2 protein, live attenuated PRRSV, and / or M.Hyo bacterin and / or supernatant.

[0140] The immunogenic composition may comprise a recombinant PCV3 ORF2 protein and a recombinant PCV2 ORF2 protein. In some cases, a dose may comprise approximately equal amounts of the recombinant PCV3 ORF2 protein and the recombinant PCV2 ORF2 protein. An embodiment of a dosing regimen may include administration of an immunogenic composition comprising a recombinant PCV3 ORF2 protein and a recombinant PCV2 ORF2 protein, either of which may be expressed using a baculovirus expression system.

[0141] Some embodiments of the immunogenic composition may include PRRSV and / or M.Hyo antigens in addition to baculovirus-expressed recombinant proteins from PCV3 ORF2. Additionally, baculovirus-expressed recombinant proteins from PCV3 ORF2 and PCV2 ORF2 may be combined with PRRSV antigens and / or M.Hyo to form an immunogenic composition. As disclosed above, the additional antigens may include live-attenuated PRRSV and / or M.Hyo bacterin and / or supernatant. For example, the immunogenic composition may include a recombinant PCV3 ORF2 protein and a recombinant PCV2 ORF2 protein. In some cases, the immunogenic composition includes approximately equal amounts of the recombinant PCV3 ORF2 protein and the recombinant PCV2 ORF2 protein. Some embodiments of the immunogenic composition may include a combination of baculovirus-expressed recombinant proteins from PCV3 ORF2 and PCV2 ORF2, as well as PRRSV and / or M.Hyo.

[0142] The administration regimen can be used to improve the economics of pig farming. For example, an immunogenic composition such as a vaccine can be administered to sows and / or piglets in an effort to protect the sows, piglets, or both. In particular, vaccination of sows prior to conception can reduce the number of mummified, stillborn, and / or weak piglets at farrowing when the sows are challenged by exposure to PCV3. PCV3 is generally considered a reproductive disease. Furthermore, the use of an inactivated baculovirus-expressed PCV3 ORF2 vaccine can reduce and / or inhibit viral replication in sows. This reduction in replication can reduce the number of mummified pigs at farrowing to a rate of approximately 4% for vaccinated sows. Such a reduction can have a significant economic impact on pig farmers.

[0143] It is further claimed that the vaccine is capable of protecting breeding gilts and sows when challenged with PCV3 in all trimesters, or two trimesters, or at least one trimester throughout the 114 day gestation period. It is also claimed that the vaccine is capable of significantly reducing the incidence of mummification, stillbirth, and weak fetuses in vaccinated gilts and vaccinated sows when challenged with PCV3 in all trimesters, or two trimesters, or at least one trimester over the 114 day gestation period.

[0144] The administration regimen may involve vaccinating young sows (i.e., 5 months of age or younger) with at least one dose of the immunogenic composition described herein prior to breeding. The dose of the immunogenic composition described herein may be administered intramuscularly as a 1 mL dose prior to breeding. In some embodiments, one or more administrations of the vaccine are administered to the sow. For example, a first vaccine may be administered, followed by a booster vaccine 21 days later and prior to breeding. In some embodiments, the sow may be bred between 14 and 21 days after the booster vaccination. This time frame may allow the sow to mount an immune response. Utilizing such an administration regimen may reduce and / or prevent the number of mummifications at farrowing. Furthermore, use of an administration regimen comprising administration of a 1 mL dose of an immunogenic composition comprising a PCV3 antigen (i.e., recombinant PCV3 ORF2) can reduce, decrease, and / or inhibit lymphadenopathy, lymphodepletion, and / or multinucleated / megahistiocytosis in pigs infected with PCV3.

[0145] In some embodiments, a dosing regimen for vaccinating piglets at approximately 3 weeks of age using a baculovirus-expressed PCV3 ORF2 vaccine can reduce the amount of virus when the piglets are subsequently challenged with PCV3. For example, the amount of virus replication in the tissues of vaccinated piglets can be reduced compared to unvaccinated piglets. Furthermore, vaccination of piglets with a PCV3 ORF2 vaccine can reduce death, clinical signs, gross lesions, and / or histological lesions in vaccinated and subsequently exposed piglets to PCV3 compared to unvaccinated piglets.

[0146] The term "immune stimulant" or "immunostimulant" as used herein preferably refers to any agent or composition capable of eliciting an immune response without initiating or enhancing a specific immune response, e.g., an immune response against a specific pathogen. It is further indicated that the immune stimulant is administered in an appropriate dose. Advantageously, the immune stimulant is keyhole limpet hemocyanin (KLH) and / or incomplete Freund's adjuvant (IFA). The role of the immune stimulant as used herein is not that of an adjuvant, but rather as a challenge enhancer. Advantageously, KLH can be emulsified in IFA containing 1 mg of KLH per mL and administered intramuscularly two days before and two days after challenge. In accordance with further discussion, there is provided a porcine circovirus type 3 (PCV3) antigenic protein which is a functional antigenic variant of the PCV3 ORF2 protein, and which is also referred to hereinafter as the "protein according to further discussion." Preferably, the protein under further consideration is a functional antigenic variant of the PCV3 ORF2 protein encoded by SEQ ID NO:1. In one preferred embodiment, the protein under further investigation comprises a substitution and / or extension of PCV3 ORF2.

[0147] In another preferred embodiment, the protein under further investigation is a functional antigenic variant of the protein encoded by SEQ ID NO:1 and / or the functional antigenic variant is capable of higher yields of virus-like particles (VLPs) than the protein encoded by SEQ ID NO:1. Preferably, the functional antigenic variant is capable of producing a higher yield of VLPs than the protein encoded by SEQ ID NO: 1, as can be determined by Western blot analysis. According to one preferred embodiment, said functional antigenic variant has fewer positively charged amino acid residues than the protein encoded by SEQ ID NO:1.

[0148] According to another preferred embodiment, the functional antigenic variant has one or more substitutions in the FG loop of the protein encoded by SEQ ID NO: 1, in which case preferably these substitutions include one or more of the S and / or K and / or H residues of the motif SKKKH of the FG loop of the protein encoded by SEQ ID NO: 1. According to yet another preferred embodiment, the functional antigenic variant has one or more substitutions in the FG loop of the protein encoded by SEQ ID NO:1, including one or more substitutions of the S and / or K residues of the motif SKKKH of the FG loop of the protein encoded by SEQ ID NO:1. According to an even more preferred embodiment, the functional antigenic variant has one or more substitutions in the FG loop of the protein encoded by SEQ ID NO:1, including substitutions of all of the S or H and K residues in the motif SKKKH of the FG loop of the protein encoded by SEQ ID NO:1.

[0149] In yet another preferred embodiment, the functional antigenic variant has one or more substitutions in the FG loop of the protein encoded by SEQ ID NO: 1, including substitution of at least S and / or H, and any K in the motif SKKKH of the FG loop of the protein encoded by SEQ ID NO: 1, with Q or P or F or S. In an even more preferred embodiment, the functional antigenic variant has one or more substitutions in the FG loop of the protein encoded by SEQ ID NO: 1, including a substitution of the motif SKKK within the motif SKKKH in the FG loop of the protein encoded by SEQ ID NO: 1 with QPFS, or a substitution of the motif KKKH within the motif SKKKH in the FG loop of the protein encoded by SEQ ID NO: 1 with QPFS.

[0150] In yet another even more preferred embodiment, the functional antigenic variant is encodable by all or part of SEQ ID NO: 1, 2, 5, 6, or 7. In an even more preferred embodiment, the functional antigenic variant is encoded by all or part of SEQ ID NO: 1, 2, 5, 6, or 7. According to a particularly preferred embodiment, the functional antigenic variant has a C-terminus extending beyond the terminal SVL sequence of the protein encoded by SEQ ID NO:1, preferably the extension is or comprises all of a sequence derived from a virus of the Circoviridae family, and preferably at least part of the extension replaces the terminal SVL sequence of the protein encoded by SEQ ID NO:1. According to another preferred embodiment, said functional antigenic variant has a C-terminus extending beyond the terminal SVL sequence of the protein encoded by SEQ ID NO: 1; said extension is between 1 and 100 amino acids in length. According to a further preferred embodiment, said functional antigenic variant has a C-terminus extending beyond the terminal SVL sequence of the protein encoded by SEQ ID NO: 1; said extension is between 1 and 50 amino acids in length.

[0151] According to yet another preferred embodiment, the functional antigenic variant has a C-terminus extending beyond the terminal SVL sequence of the protein encoded by SEQ ID NO: 1; the extension is between 1 and 30 amino acids in length. In one particularly preferred embodiment, said functional antigenic variant has a C-terminus extending beyond the terminal SVL sequence of the protein encoded by SEQ ID NO:1. Preferably, the extension is between 1 and 30 amino acids in length and / or the extension comprises the entire sequence VKININLTPPVATSRVPSRALPLRFGCGHR. In a further preferred embodiment, the functional antigenic variant is encodable by all or part of SEQ ID NO: 1, 2, 5, 6, or 7.

[0152] In a preferred embodiment, the variant protein comprises or consists of an amino acid sequence having at least about 80%, or at least about 85%, or at least about 86%, or at least about 87%, or at least about 88%, or at least about 89%, in the range of about 83% to about 89%, such as 84%, or 85%, or 86%, or 87%, or 88%, or 89% sequence identity and / or sequence homology to the sequence of SEQ ID NO: 3, 4, 8, 9, or 10, and / or wherein the protein is a recombinant protein, and wherein the protein has one or more substitutions in the FG loop.

[0153] In a preferred embodiment, the variant protein comprises or consists of an amino acid sequence having at least about 80%, or at least about 85%, or at least about 86%, or at least about 87%, or at least about 88%, or at least about 89%, in the range of about 83% to about 89%, such as 84%, or 85%, or 86%, or 87%, or 88%, or 89% sequence identity and / or sequence homology to the sequence of SEQ ID NO: 3, 4, 8, 9, or 10, and / or wherein the protein is a recombinant protein and the protein has a C-terminus extending beyond the terminal SVL sequence of the protein encoded by SEQ ID NO: 1.

[0154] In a preferred embodiment, the variant protein comprises an FG loop with one or more substitutions within the FG loop and further comprises a C-terminus extending beyond the terminal SVL sequence of the protein encoded by SEQ ID NO: 1, wherein the sequence of the variant protein comprises or consists of an amino acid sequence having at least about 80%, or at least about 85%, or at least about 86%, or at least about 87%, or at least about 88%, or at least about 89%, in the range of about 83% to about 89%, such as 84%, or 85%, or 86%, or 87%, or 88%, or 89% sequence identity and / or sequence homology to the sequence of SEQ ID NO: 3, 4, 8, 9, or 10, and / or the protein is a recombinant protein. In another preferred embodiment, the protein under further consideration is a recombinant protein, prepared by recombinant DNA methods. In yet another preferred embodiment, the protein under further investigation is a baculovirus-expressed protein. Preferably, said PCV3 is any phylogenetic clade or combination of clades of PCV3. Preferably, said PCV3 is selected from the group consisting of PCV3a and PCV3b. In particular, said PCV3 is preferably selected from the group consisting of PCV3a1, PCV3b1, and PCV3b2.

[0155] PCV3 may also be selected from PCV3c (BMC Vet Res. 2019 Jul 15;15(1):244. doi: 10.1186 / s12917-019-1977-7). More specifically, the PCV3 ORF2 is preferably from group a1, b1, or b2 (using the subclassification notation of Fux et al., "Full genome characterization of porcine circovirus type 3 isolates reveals the existence of two distinct groups of virus strains," Virology Journal (2018) 15:25, DOI 10.1186 / s12985-018-0929-3 (incorporated herein by reference); see, for example, Table 4).

[0156] In a preferred embodiment, the PCV3 ORF2 protein comprises or consists of an amino acid sequence encoded by a polynucleotide sequence having at least 90%, or at least 91%, or at least 92%, or at least 93%, or at least 94%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity to or sequence homology with SEQ ID NO:1. In another preferred embodiment, the variant protein comprises or consists of an amino acid sequence encoded by a polynucleotide sequence having at least 90%, or at least 91%, or at least 92%, or at least 93%, or at least 94%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 1 or sequence homology to SEQ ID NO: 6.

[0157] In yet another preferred embodiment, the variant protein comprises or consists of an amino acid sequence encoded by a polynucleotide sequence having at least 90%, or at least 91%, or at least 92%, or at least 93%, or at least 94%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 1 or sequence homology to SEQ ID NO: 7. In an even more preferred embodiment, the PCV3 ORF2 protein comprises or consists of an amino acid sequence having at least 90%, or at least 91%, or at least 92%, or at least 93%, or at least 94%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity to the sequence of SEQ ID NO: 3, 4, 8, 9, or 10, and / or the protein is a recombinant protein.

[0158] In yet another preferred embodiment, the variant protein comprises or consists of an amino acid sequence having at least 90%, or at least 91%, or at least 92%, or at least 93%, or at least 94%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity to the sequence of SEQ ID NO: 3, 4, 8, 9, or 10, and / or the protein is a recombinant protein. In yet another preferred embodiment, the variant protein comprises or consists of an amino acid sequence having at least 90%, or at least 91%, or at least 92%, or at least 93%, or at least 94%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 3, 4, 8, 9, or 10, and / or wherein the protein is a recombinant protein, and wherein the protein has one or more substitutions in the FG loop.

[0159] In a preferred embodiment, the variant protein comprises or consists of an amino acid sequence having at least 90%, or at least 91%, or at least 92%, or at least 93%, or at least 94%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity to the sequence of SEQ ID NO: 3, 4, 8, 9, or 10, and / or the protein is a recombinant protein. In another preferred embodiment, the variant protein comprises or consists of an amino acid sequence having at least 90%, or at least 91%, or at least 92%, or at least 93%, or at least 94%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity to the sequence of SEQ ID NO: 3, 4, 8, 9, or 10, and / or the protein is a recombinant protein, and the protein has a C-terminus extending beyond the terminal SVL sequence of the protein encoded by SEQ ID NO: 1.

[0160] According to a preferred embodiment, said protein is a recombinant protein resulting from its expression by an expression vector comprising a polynucleotide sequence encoding the protein. According to a preferred embodiment, said protein is a recombinant protein resulting from its expression by a baculovirus expression vector comprising a polynucleotide sequence encoding the protein.

[0161] In another preferred embodiment, a nucleotide sequence is provided which encodes a protein according to further discussion, hereinafter also referred to as a "nucleotide sequence according to further discussion." In a further preferred embodiment, a vector is provided which comprises the further discussed nucleotide sequence, hereinafter also referred to as a "further discussed vector." Also provided are recombinant vectors comprising the nucleotide sequences according to the further discussion. Additionally, there is provided an expression host that is transformed or transfected with the further discussed nucleotide sequence, hereinafter also referred to as an "expression host according to the further discussion." Also provided are baculovirus expression hosts transformed or transfected with the further discussed nucleotide sequences, also referred to hereinafter as "further discussed baculovirus expression hosts."

[0162] Additionally provided are methods of preparing the further discussed proteins comprising expressing the further discussed nucleotide sequences. Also provided is a method of preparing a protein according to the further studies, comprising expressing a vector according to the further studies. Additionally provided is a method of preparing a protein according to the further discussion, comprising expressing a recombinant vector according to the further discussion. Also provided is a method of preparing a further investigated protein, comprising culturing a further investigated expression host to cause expression of the protein.

[0163] Additionally provided is a method of preparing a protein according to the further discussion, comprising transfecting an expression host with a nucleotide sequence according to the further discussion, or a vector according to the further discussion, and culturing the expression host to cause expression of the protein. Also provided is a method of preparing a further investigated protein, comprising culturing a further investigated baculovirus expression host to cause expression of the protein. Also provided is a method of preparing a further investigated protein, comprising transfecting a baculovirus expression host with a further investigated nucleotide sequence or a further investigated vector, and culturing the baculovirus expression host to cause expression of the protein.

[0164] Preferably, in any of the above methods of preparing a protein according to further consideration, an inactivating agent is used when a sufficient level of protein expression is achieved, in which case the inactivating agent is preferably binary ethyleneimine. Preferably, any of the above methods for preparing a protein according to further consideration comprises transfecting a baculovirus expression host with the nucleotide sequence of the vector and culturing the baculovirus expression host in a medium to cause expression of the protein; wherein the medium after expression of the protein comprises: (i) the protein; (ii) at least a portion of the baculovirus expressing the protein; and (iii) a portion of a cell culture of cells infected or transfected with a recombinant baculovirus expressing the protein. Preferably, any of the above methods for preparing a protein according to further consideration comprises transfecting a baculovirus expression host with the nucleotide sequence of the vector and culturing the baculovirus expression host in a medium to cause expression of the protein; wherein the medium after expression of the protein comprises: (i) the protein; (ii) at least a portion of the baculovirus expressing the protein; and (iii) a portion of a cell culture of cells infected with or transfected with a recombinant baculovirus expressing the protein; and about 90% of components (i)-(iii) have a size less than 1 μm.

[0165] Preferably, any of the above methods for preparing a protein according to further consideration comprises transfecting a baculovirus expression host with the nucleotide sequence of the vector and culturing the baculovirus expression host in a medium to cause expression of the protein; wherein the medium after expression of the protein comprises: (i) the protein; (ii) at least a portion of the baculovirus expressing the protein; and (iii) a portion of a cell culture of cells infected with or transfected with a recombinant baculovirus expressing the protein; and about 90% of components (i)-(iii) have a size less than 1 μm, and the pH of the composition is adjusted to about 6.5-7.5.

[0166] Preferably, any of the above methods of preparing a protein according to further discussion comprises producing the protein by a baculovirus expression system in cultured insect cells. Preferably, any of the above methods of preparing a protein according to further discussion comprises producing the protein by a baculovirus expression system in cultured insect cells; the method comprises inactivating the baculovirus. Preferably, any of the above methods of preparing a protein according to further discussion comprises producing the protein by a baculovirus expression system in cultured insect cells; the method comprises inactivating the baculovirus; and the inactivating comprises heat treatment or the use of a virus-inactivating agent.

[0167] Preferably, any of the above methods of preparing a protein according to further consideration comprises producing the protein by a baculovirus expression system in cultured insect cells; the method comprises inactivating the baculovirus; the inactivating comprises heat treatment or the use of a virus-inactivating agent; and the virus-inactivating agent comprises an aziridine compound. Preferably, any of the above methods of preparing a protein according to further considerations comprises producing the protein by a baculovirus expression system in cultured insect cells; the method comprises inactivating the baculovirus; the inactivating comprises heat treatment or the use of a virus-inactivating agent; the virus-inactivating agent comprises an aziridine compound; and the aziridine compound comprises BEI. Further provided are proteins obtainable by any of the above methods for preparing a protein according to further discussion. Also provided are compositions comprising a protein obtainable by any of the above methods for preparing a protein according to further consideration, preferably comprising a carrier, diluent, or excipient. Additionally provided are compositions obtainable by any of the above methods for preparing a protein according to further consideration, wherein the composition preferably comprises a carrier, diluent, or excipient.

[0168] In particular, hereinafter any of the compositions will also be referred to as a "composition according to further considerations." Further studies have shown that the protein in the composition is preferably present at a concentration of 0.2 to about 400 μg / ml, or 2 to about 400 μg / ml, or 4 to about 400 μg / ml, or 8 to about 400 μg / ml, or about 0.3 to about 200 μg / ml, or 2 to about 200 μg / ml, or 4 to about 200 μg / ml, or 8 to about 200 μg / ml, or about 0.35 to about 100 μg / ml, or 2 to about 100 μg / ml, or is present in an amount of 4 to about 100 μg / ml, or 8 to about 100 μg / ml, or about 0.4 to about 50 μg / ml, or about 0.45 to about 30 μg / ml, or about 0.6 to about 15 μg / ml, or about 0.75 to about 8 μg / ml, or about 1.0 to about 6 μg / ml, or about 1.3 to about 3.0 μg / ml, or about 1.4 to about 2.5 μg / ml, or about 1.5 to about 2.0 μg / ml, or about 1.6 μg / ml.

[0169] Preferably, compositions under further consideration include any one or more of a solvent, dispersion medium, coating agent, stabilizer, diluent, preservative, antimicrobial agent, antifungal agent, isotonic agent, absorption delaying agent, adjuvant, cell culture supernatant, stabilizer, viral vector, expression vector, and / or immunomodulator.

[0170] Preferably, compositions according to further consideration are provided wherein the carrier, diluent, or excipient is any one or more of an adjuvant, an immunomodulator, a cell culture supernatant, a virus or an expression vector, or any combination thereof. Preferably, compositions according to further consideration are provided wherein the carrier, diluent, or excipient comprises an adjuvant.

[0171] Preferably, the carrier, diluent, or excipient comprises an adjuvant; the adjuvant is selected from the group consisting of polymers of acrylic or methacrylic acid, copolymers of maleic anhydride and alkenyl derivatives, crosslinked polymers of acrylic or methacrylic acid, crosslinked polymers of acrylic or methacrylic acid with polyalkenyl ethers of sugars or polyalcohols, carbomers, acrylic polymers crosslinked with polyhydroxylated compounds having at least three and up to eight hydroxyl groups, in which at least three hydroxyl hydrogen atoms may be replaced, or be replaced, by unsaturated aliphatic radicals having at least two carbon atoms, such as vinyl, allyl, and other ethylenically unsaturated groups, containing 2 to 4 carbon atoms, which may themselves contain other substituents, such as methyl, carbopol, Carbopol 974P, Carbopol 934P, Carbopol 971P, aluminum hydroxide, aluminum phosphate, saponin, Quill A;QS-21;GPI-0100;Water-in-oil emulsions;Oil-in-water emulsions;Water-in-oil-in-water emulsions;Emulsions based on light liquid paraffin oil or European Pharmacopoeia-type adjuvants;Isoprenoid oils;Squalane;Squalene oil obtained as a result of the oligomerization of alkenes or isobutene or decene;Esters of acids or alcohols containing linear alkyl groups;Vegetable oils;Ethyl oleate;Propylene glycol dicaprylate / caprate;Caprylic / capric triglyceride;Propylene glycol dioleate;Esters of branched fatty acids or alcohols;Isostearin acid esters; nonionic surfactants; sorbitan or mannide or glycol or polyglycerol or propylene glycol, or oleic acid or isostearic acid or ricinoleic acid or hydroxystearic acid, esters of mannitol oleate anhydride which may be ethoxylated; polyoxypropylene-polyoxyethylene block copolymers, Pluronic products, RIBI adjuvant system; block copolymers; SAF-M; monophosphoryl lipid A; avridine lipid-amine adjuvants; heat-labile enterotoxin derived from Escherichia coli (recombinant or other forms);Further contemplated compositions are provided that include one or more of cholera toxin; IMS1314; or muramyl dipeptide. Preferably, compositions according to further consideration are provided wherein the carrier, diluent, or excipient comprises an adjuvant; and wherein the adjuvant comprises Carbopol or Carbopol 971.

[0172] Further consideration provides compositions in which the carrier, diluent, or excipient preferably comprises an adjuvant; the adjuvant is present in an amount of about 50 μg to about 2000 μg per ml of the composition dose, or the adjuvant is present in an amount of about 250 μg per ml of the composition dose, or the adjuvant is present in an amount of about 100 μg to about 10 mg per ml of the composition dose; or the adjuvant is present in an amount of about 500 μg to about 5 mg per ml of the composition dose; the adjuvant is present in an amount of about 750 μg to about 2.5 mg per ml of the composition dose, or the adjuvant is present in an amount of about 1 mg per ml of the composition dose.

[0173] Preferably, compositions according to further discussion are provided which include an immunomodulatory agent. Preferably, compositions according to further consideration are provided which include an immunomodulatory agent; the immunomodulatory agent being any one or more of an interleukin, an interferon, or other cytokine. Preferably, compositions according to further discussion are provided that include an antibiotic. Preferably, compositions according to further discussion are provided which comprise an antibiotic; the antibiotic comprises gentamicin. Preferably, compositions according to further consideration are provided that include an antibiotic; the antibiotic being about 1 μg / ml to about 60 μg / ml. Preferably, compositions according to further considerations are provided that include an antibiotic; that include from about 1 μg / ml to less than about 30 μg / ml of antibiotic. Preferably, compositions according to further discussion are provided which include additional antigens. Preferably, compositions according to further consideration are provided which comprise an additional antigen; wherein said additional antigen is not a PCV3 ORF2 antigen. Preferably, compositions according to further consideration are provided which comprise an additional antigen; wherein said additional antigen is not a PCV3 antigen.

[0174] Preferably, compositions according to further considerations are provided which include additional antigens of further swine pathogens. Preferably, compositions according to further consideration are provided which further comprise an antigen of a further swine pathogen; said pathogen being any one or more of PCV2, PRRSV (Porcine Reproductive and Respiratory Syndrome Virus) antigen, Mycoplasma hyopneumoniae bacterin antigen, Mycoplasma hyopneumoniae supernatant antigen, Aujeszky's disease antigen or Pseudorabies antigen, swine influenza antigen, swine fever antigen (classical swine fever antigen or African swine fever antigen, or a combination thereof), Actinobacillus pleuropneumoniae antigen, Escherichia coli antigen, porcine parvovirus (PPV) antigen, or Pasteurella multocida antigen.

[0175] Compositions according to further considerations are provided which preferably further comprise an antigen of a further swine pathogen, further comprising one or more of a PCV2 antigen, a PRRSV antigen, and a PPV antigen. Preferably, compositions according to further considerations are provided which further comprise a PCV2 antigen. Preferably, compositions according to further consideration are provided which further comprise a PCV2 antigen, wherein the PCV2 antigen is PCV2 ORF2 protein. Preferably, compositions according to further consideration are provided which further comprise a PCV2 antigen, wherein the PCV2 antigen is a recombinant PCV2 ORF2 protein.

[0176] Preferably, a composition according to further consideration is provided, further comprising a PCV2 antigen, wherein the PCV2 antigen is a recombinant baculovirus-expressed PCV2 ORF2 protein. Preferably, compositions according to further discussion are provided that are in dosage form. Compositions according to further discussion are provided that are preferably formulated and / or packaged for single dose or one-shot administration.

[0177] Compositions according to further discussion are provided that are preferably formulated and / or packaged for multiple dose regimens. Compositions according to further discussion are provided which are preferably formulated and / or packaged for a two-dose regimen. Further considerations provide compositions that are preferably in dosage form; that are delivered from a container containing a large quantity of the composition; and that are capable of delivering a dosage form of the composition from the container.

[0178] Further consideration provides a composition that is preferably in a dosage form; that is delivered from a container containing a large quantity of the composition, and that is capable of delivering a dosage form of the composition from the container; and that contains at least 10 doses of the composition. Further consideration provides a composition that is preferably in a dosage form; that is delivered from a container containing a large quantity of the composition; that is capable of delivering a dosage form of the composition from the container; and that contains at least 50 doses of the composition. Further consideration provides compositions that are preferably in dosage form; that are delivered from a container containing a large quantity of the composition; that are capable of delivering a dosage form of the composition from the container; and that contain at least 100 doses of the composition. Further consideration provides a composition that is preferably in a dosage form; that is delivered from a container containing a large quantity of the composition; that is capable of delivering a dosage form of the composition from the container; and that contains at least 200 doses of the composition.

[0179] Further consideration provides a composition that is preferably in a dosage form; that is delivered from a container containing a large quantity of the composition; that is capable of delivering a dosage form of the composition from the container; and that contains at least 250 doses of the composition. Preferably, the composition contains a PCV2 antigen; the PCV2 antigen is a recombinant baculovirus-expressed PCV2 ORF2 protein; and the total amount of one of the PCV3 ORF protein and the PCV2 ORF protein, or a combination thereof, is about 0.2 to about 400 μg per dose, or 2 to about 400 μg per dose, or 4 to about 400 μg per dose, or 8 to about 400 μg per dose, or about 0.3 to about 200 μg per dose, or 2 to about 200 μg per dose, or 4 to about 200 μg per dose, or 8 to about 200 μg per dose, or about 0.35 to about 100 μg per dose, or 2 to about 100 μg per dose, or Further investigated compositions are provided wherein the soluble ... Preferably, compositions according to further consideration are provided that include a salt. Preferably, compositions according to further discussion are provided which comprise an inactivated viral vector and / or a cell culture supernatant. Preferably, a composition according to further discussion is provided which comprises an inactivated viral vector and a cell culture supernatant.

[0180] Further discussed compositions are provided, preferably comprising: (i) a protein; (ii) at least a portion of a baculovirus expressing said protein; (iii) a portion of a cell culture of cells infected with or transfected with a recombinant baculovirus expressing said protein; (iv) an inactivating agent or an inactivating agent comprising binary ethyleneimine (BEI); (v) sodium thiosulfate or an amount of sodium thiosulfate equivalent to the inactivating agent or BEI; (vi) an adjuvant or an adjuvant comprising Carbopol or Carbopol 971; and (vii) a physiologically acceptable concentration of phosphate.

[0181] (ii) at least a portion of a baculovirus expressing said protein; (iii) a portion of a cell culture of cells infected with or transfected with a recombinant baculovirus expressing said protein; (iv) an inactivating agent or an inactivating agent comprising binary ethyleneimine (BEI); (v) sodium thiosulfate or an amount of sodium thiosulfate equivalent to the inactivating agent or BEI; (vi) an adjuvant or an adjuvant comprising Carbopol or Carbopol 971; and (vii) a physiologically acceptable concentration of phosphate; wherein the BEI is derived from a cell culture treated with about 2-8 or about 5 mM BEI to inactivate the baculovirus, and / or the composition contains about 2-8 or about 5 mM BEI, and / or the composition contains about 1 mg of Carbopol or Carbopol 971.

[0182] There is provided a composition according to further considerations that is preferably an immunogenic composition comprising a protein according to further considerations and a carrier, diluent, or excipient. There is provided a composition according to further considerations that is preferably an immunogenic composition comprising a protein according to further considerations and a carrier, diluent, or excipient; and an additional antigen as described above. Also provided is a process for making a composition according to the further considerations, wherein a protein according to the further considerations is mixed with a carrier, diluent, or excipient. Additionally provided is a process for making a composition according to the further considerations, wherein a protein according to the further considerations is mixed with a carrier, diluent, or excipient; and an additional antigen. Additionally, further investigations of proteins for use as pharmaceuticals are provided.

[0183] Also provided is a protein according to the further investigations, or a composition according to the further investigations, for use as a vaccine. Also provided is a protein according to the further studies, or a composition according to the further studies, for use in a method for inducing an immune or immunological response, or a protective immune or immunological response, against PCV3 in an animal. Also provided is a further investigated protein or a further investigated composition for use in a method for inducing an immune or immunological response, or a protective immune or immunological response, against PCV3 in pigs.

[0184] Also provided is a further investigated protein or a further investigated composition for use in a method for inducing an immune or immunological response, or a protective immune or immunological response, against PCV3 in pigs. Also provided is a protein according to the further studies, or a composition according to the further studies, for use in a method for inducing an immune or immunological response, or a protective immune or immunological response, against PCV3 in piglets. Also provided is a protein according to further considerations or a composition according to further considerations for use in a method for inducing an immune or immunological response, or a protective immune or immunological response, against PCV3 in piglets suckled by a sow to which the protein according to further considerations or composition according to further considerations has been administered. Also provided is a further investigated protein or a further investigated composition for use in a method for inducing an immune or immunological response, or a protective immune or immunological response, against PCV3 in a sow. Also provided is a further investigated protein or a further investigated composition for use in a method for inducing an immune or immunological response, or a protective immune or immunological response, against PCV3 in a pregnant pig, a gilt, or a pre-breeding gilt.

[0185] Also provided is a further investigated protein or a further investigated composition for use in inducing an immune response against PCV3 in an animal. Also provided is a further investigated protein or a further investigated composition for use in inducing an immune response against PCV3 in pigs. Also provided is a further investigated protein or a further investigated composition for use in inducing an immune response against PCV3 in pigs. Also provided is a further investigated protein or a further investigated composition for use in inducing an immune response against PCV3 in piglets. Also provided is a further investigated protein or a further investigated composition for use in inducing an immune response against PCV3 in piglets nursed by a sow to which the further investigated protein or the further investigated composition has been administered. Also provided is a further investigated protein or a further investigated composition for use in inducing an immune response against PCV3 in sows.

[0186] Also provided is a further investigated protein or a further investigated composition for use in inducing an immune response against PCV3 in a pregnant sow, a gilt, or a pre-breeding gilt. Also provided are further studied proteins or further studied compositions for use in methods for reducing or preventing clinical signs or symptoms or disease caused by PCV3 infection in an animal, or for use in methods for treating or preventing PCV3 infection in an animal.

[0187] Also provided are further studied proteins or further studied compositions for use in methods for reducing or preventing clinical signs or symptoms or disease caused by PCV3 infection in porcine animals, or for use in methods for treating or preventing PCV3 infection in animals. Also provided are further studied proteins or further studied compositions for use in methods for reducing or preventing clinical signs or symptoms or disease caused by PCV3 infection in porcine animals, or for use in methods for treating or preventing PCV3 infection in animals. Also provided is a further studied protein or a further studied composition for use in a method for reducing or preventing clinical signs or symptoms or disease caused by infection with PCV3 in an animal that is a piglet, or for use in a method for treating or preventing infection with PCV3 in an animal.

[0188] Also provided is a protein according to further considerations, or a composition according to further considerations, for use in a method for reducing or preventing clinical signs or symptoms or disease caused by infection with PCV3 in an animal that is a piglet nursed by a sow to which the protein according to further considerations, or the composition according to further considerations, has been administered, or for use in a method for treating or preventing infection with PCV3 in an animal. Also provided is a further studied protein or a further studied composition for use in a method for reducing or preventing clinical signs or disease caused by infection with PCV3 in an animal that is a sow, or for use in a method for treating or preventing infection with PCV3 in an animal that is a sow. Also provided are further studied proteins or further studied compositions for use in methods for reducing or preventing clinical signs or symptoms or disease caused by infection with PCV3 in animals that are pregnant pigs, gilts, or pre-breeding gilts, or for use in methods for treating or preventing infection with PCV3 in animals.

[0189] Also provided is a further investigated protein or a further investigated composition for use in immunizing an animal against PCV3. Also provided is a further investigated protein or a further investigated composition for use in immunizing a porcine animal against PCV3. Also provided is a further investigated protein or a further investigated composition for use in immunizing a porcine animal against PCV3. Also provided is a further investigated protein or a further investigated composition for use in immunizing an animal, which is a piglet, against PCV3. Also provided is a further investigated protein or a further investigated composition for use in immunizing against PCV3 an animal, which is a piglet nursed by a sow to which the further investigated protein or the further investigated composition has been administered.

[0190] Also provided is a further investigated protein or a further investigated composition for use in immunizing an animal, that is a sow, against PCV3. Also provided is a further investigated protein or a further investigated composition for use in immunizing an animal, which is a pregnant sow, a gilt, or a pre-breeding gilt, against PCV3. Also provided is a further investigated protein or a further investigated composition for use in a method of reducing, eliminating, or suppressing PCV3 viral expression in an animal. Also provided is a further investigated protein or a further investigated composition for use in a method of reducing, eliminating or suppressing PCV3 viral expression in an animal that is a porcine. Also provided is a further investigated protein or a further investigated composition for use in a method of reducing, eliminating or suppressing PCV3 viral expression in an animal that is a porcine.

[0191] Also provided is a further investigated protein or a further investigated composition for use in a method of reducing, eliminating or suppressing PCV3 viral expression in an animal, the animal being a piglet. Also provided is a further investigated protein or a further investigated composition for use in a method of reducing, eliminating or suppressing expression of PCV3 virus in an animal, the animal being a piglet nursing by a sow to which the further investigated protein or the further investigated composition has been administered. Also provided is a further investigated protein or a further investigated composition for use in a method of reducing, eliminating or suppressing PCV3 viral expression in an animal that is a sow. Also provided is a further investigated protein or a further investigated composition for use in a method for reducing, eliminating, or suppressing expression of PCV3 virus in an animal that is a pregnant pig, a gilt, or a pre-breeding gilt. Also provided is a further investigated protein or a further investigated composition for use in inducing the production of antibodies specific to PCV3 in an animal. Also provided is a further investigated protein or a further investigated composition for use in inducing the production of antibodies specific for PCV3 in an animal that is a porcine.

[0192] Also provided is a further investigated protein or a further investigated composition for use in inducing the production of antibodies specific for PCV3 in an animal that is a porcine.

[0193] Also provided is a further investigated protein or a further investigated composition for use in inducing the production of antibodies specific to PCV3 in an animal, the animal being a piglet. Also provided is a further investigated protein or a further investigated composition for use in inducing the production of antibodies specific to PCV3 in an animal, which is a piglet nursed by a sow to which the further investigated protein or the further investigated composition has been administered. Also provided is a further investigated protein or a further investigated composition for use in inducing the production of antibodies specific for PCV3 in an animal that is a sow. Also provided is a further investigated protein or a further investigated composition for use in inducing the production of antibodies specific for PCV3 in an animal that is a pregnant sow, a gilt, or a pre-breeding gilt. Preferably, the protein according to the further considerations or the composition according to the further considerations is administered to the animal intramuscularly or intradermally. Preferably, the protein according to said further considerations, or the composition according to said further considerations, is administered to said animal together with another antigen, preferably in this case the other pathogen is an antigen derived from a swine pathogen.

[0194] Preferably, the protein according to the further considerations or the composition according to the further considerations is administered to the animal together with another antigen; in this case, the other antigen is not a PCV3 ORF2 antigen, and preferably the other pathogen is an antigen derived from a swine pathogen. Preferably, the protein according to the further considerations, or the composition according to the further considerations, is administered to the animal together with another antigen; in this case, the other antigen is not a PCV3 antigen, and preferably the other pathogen is an antigen derived from a swine pathogen. Also provided is a protein according to further considerations, or a composition according to further considerations, for any of the above uses, wherein the animal is a sow that is pregnant with piglets.

[0195] Also provided is a protein according to further considerations or a composition according to further considerations for any of the above uses, wherein the animal is a sow that is pregnant with piglets; and the piglets are nursed by the sow to which the protein according to further considerations or composition according to further considerations has been administered. Also provided is a protein according to further considerations, or a composition according to further considerations, for any of the above uses, wherein the animal is a sow; and the protein or composition is administered twice to the sow. Preferably, said animal is a sow; and said protein according to said further considerations, or said composition according to said further considerations, is administered to said sow only twice.

[0196] Also provided is a protein according to further considerations, or a composition according to further considerations, for any of the above uses, wherein the animal is a piglet; and wherein the protein or composition is administered once to the piglet. Also provided is a protein according to further considerations, or a composition according to further considerations, for any of the above uses, wherein the animal is a piglet; and wherein the protein or composition is administered only once to the piglet. Also provided is a protein according to further studies or a composition according to further studies for any of the above uses, wherein the animal is a sow; the sow is administered twice; and the use does not include administration of any other PCV3 antigen to the animal before or at the time of administration of the protein according to further studies or the composition according to further studies.

[0197] Also provided is a protein according to further studies or a composition according to further studies for any of the above uses, wherein the animal is a sow; the sow is administered twice; and the use does not include administration of any other PCV3 antigen to the animal before or at the time of administration of the protein according to further studies or the composition according to further studies. Also provided is a protein according to further investigations or a composition according to further investigations for any of the above uses, wherein the animal is a pig; the administration is to the sow only twice; and the use does not include administration of any other PCV3 antigen to the animal before or at the time of administration of the protein according to further investigations or the composition according to further investigations. Also provided is a protein according to further studies or a composition according to further studies for any of the above uses, wherein the animal is a piglet; is administered once to the piglet; and the use does not include administration of any other PCV3 antigen to the animal before or at the time of administration of the protein according to further studies or the composition according to further studies.

[0198] Also provided is a protein according to further studies or a composition according to further studies for any of the above uses, wherein the animal is a piglet; is administered once to the piglet; and the use does not include administration of any other PCV3 antigen to the animal before or at the time of administration of the protein according to further studies or the composition according to further studies. Also provided is a protein according to further investigations or a composition according to further investigations for any of the above uses, wherein the animal is a piglet; is administered only once to the piglet; and the use does not include administration of any other PCV3 antigen to the animal before or at the time of administration of the protein according to further investigations or the composition according to further investigations.

[0199] Also provided is a protein according to further considerations, or a composition according to further considerations, for any of the above uses, wherein the administration to the animal in the use consists of a single, one-shot administration or a single, one-dose administration of said protein according to further considerations, or composition according to further considerations. Also provided is a protein according to further considerations, or a composition according to further considerations, for any of the above uses, wherein the administration to the animal in the use consists of a multiple shot or multiple dose regimen of said protein according to further considerations, or composition according to further considerations.

[0200] Also provided is a protein according to further considerations, or a composition according to further considerations, for any of the above uses, wherein the administration to the animal in the use consists of two shots or two doses of said protein according to further considerations, or composition according to further considerations. Also provided is a protein according to the further investigations, or a composition according to the further investigations, for any of the above uses, wherein administration to the animal is within at least 1, 2 or 3 weeks of exposure to the virulent porcine circovirus. Also provided is a further considered protein or a further considered composition wherein the animal is a piglet 15 weeks of age or less, or 6 weeks of age or less, or 3 weeks of age or less, or 2 weeks of age or less, or 1 week of age or less. Also provided is a protein according to further considerations or a composition according to further considerations for any of the above uses, wherein said protein according to further considerations is a protein for any of the above uses.

[0201] Also provided is a protein according to further considerations, or a composition according to further considerations, for any of the above uses, wherein said protein according to further considerations is a protein for two or more of the uses described above. Also provided is a protein according to further considerations or a composition according to further considerations for any of the above uses, wherein said composition according to further considerations is a composition for any of the above uses. Also provided is a protein according to further considerations, or a composition according to further considerations, for any of the above uses, wherein a second antigen is administered to the animal prior to administration of the protein according to further considerations, or a composition according to further considerations.

[0202] Also provided is a protein according to further considerations, or a composition according to further considerations, for any of the above uses, wherein a second antigen is administered to the animal simultaneously with administration of the protein according to further considerations, or a composition according to further considerations. Also provided is a protein according to further considerations, or a composition according to further considerations, for any of the above uses, wherein a second antigen is administered to the animal simultaneously with, and in the same composition as, the administration of the protein according to further considerations, or the composition according to further considerations. Also provided is a protein according to further considerations, or a composition according to further considerations, for any of the above uses, wherein a second antigen is administered to the animal simultaneously with administration of the protein according to further considerations, or a composition according to further considerations, and in a different composition.

[0203] Also provided is a protein according to further considerations, or a composition according to further considerations, for any of the above uses, wherein a second antigen is administered to the animal after administration of the protein according to further considerations, or a composition according to further considerations. Also provided is a further investigated protein or a further investigated composition for use in vaccinating pigs to reduce the severity of clinical signs or symptoms resulting from PCV3 infection in pigs, wherein the protein is present in an immunogenic composition that is administered to the pig in a single dose. Also provided is a further investigated protein or a further investigated composition for use in vaccinating pigs to reduce the severity of clinical signs or symptoms resulting from PCV3 infection in pigs, wherein the protein is present in an immunogenic composition that is administered to the pig in only one dose.

[0204] Also provided is a further investigated protein or a further investigated composition for use in vaccinating pigs to reduce the severity of clinical signs or symptoms resulting from PCV3 infection in pigs, wherein the protein is present in an immunogenic composition that is administered to the pig in two doses. Also provided is a further investigated protein or a further investigated composition for use in vaccinating pigs to reduce the severity of clinical signs or symptoms resulting from PCV3 infection in pigs, wherein the protein is present in an immunogenic composition that is administered to the pig in only two doses. Also provided is a further investigated protein, or a further investigated composition, for use as a single PCV3 antigen for vaccination of pigs to reduce the severity of clinical signs or symptoms resulting from PCV3 infection in pigs, wherein the protein is present in an immunogenic composition that is administered to pigs in a single dose. Also provided is a further investigated protein, or a further investigated composition, for use as a single PCV3 antigen for vaccination of pigs to reduce the severity of clinical signs or symptoms resulting from PCV3 infection in pigs, wherein the protein is present in an immunogenic composition that is administered to pigs in only one dose.

[0205] Also provided is a further investigated protein, or a further investigated composition, for use as a single PCV3 antigen for vaccination of pigs to reduce the severity of clinical signs or symptoms resulting from PCV3 infection in pigs, wherein the protein is present in an immunogenic composition that is administered to pigs in two doses. Also provided is a further investigated protein, or a further investigated composition, for use as a single PCV3 antigen for vaccination of pigs to reduce the severity of clinical signs or symptoms resulting from PCV3 infection in pigs, wherein the protein is present in an immunogenic composition that is administered to pigs in only two doses.

[0206] Also, A dose of the immunogenic composition is administered to a pig in a vaccination regimen that reduces the severity of clinical signs or symptoms resulting from PCV3 infection in the pig; administration of one dose of the immunogenic composition to a pig in a vaccination method reduces the severity of clinical signs or symptoms resulting from PCV3 infection in the pig; Further contemplated proteins are antigenic components in one dose of an immunogenic composition in a vaccination method that reduces the severity of clinical signs or symptoms resulting from PCV3 infection in pigs; Preferably, the protein is present in an amount of at least 2 μg per dose of the immunogenic composition; The protein is an antigenic component in a vaccination method that reduces the severity of clinical signs or symptoms resulting from PCV3 infection in pigs. Also provided is a further investigated protein or a further investigated composition for reducing the severity of clinical signs or symptoms resulting from PCV3 infection in pigs.

[0207] Also herein, only one dose of the immunogenic composition is administered to the pig in a vaccination regimen that reduces the severity of clinical signs or clinical symptoms resulting from PCV3 infection in the pig; administration of one dose of the immunogenic composition to a pig in a vaccination method reduces the severity of clinical signs or symptoms resulting from PCV3 infection in the pig; Further contemplated proteins are antigenic components in one dose of an immunogenic composition in a vaccination method that reduces the severity of clinical signs or symptoms resulting from PCV3 infection in pigs; Preferably, the protein is present in an amount of at least 2 μg per dose of the immunogenic composition; The protein is an antigenic component in a vaccination method that reduces the severity of clinical signs or symptoms resulting from PCV3 infection in pigs. Further investigational immunogenic compositions for reducing the severity of clinical signs or symptoms resulting from PCV3 infection in pigs are also presented.

[0208] Also, two doses of the immunogenic composition are administered to pigs in a vaccination regimen that reduces the severity of clinical signs or clinical symptoms resulting from PCV3 infection in pigs; administration of two doses of the immunogenic composition to pigs in a vaccination method reduces the severity of clinical signs or symptoms resulting from PCV3 infection in pigs; Further discussed is a protein that is an antigenic component in two doses of an immunogenic composition in a vaccination method that reduces the severity of clinical signs or symptoms resulting from PCV3 infection in pigs; Preferably, the protein is present in an amount of at least 2 μg per dose of the immunogenic composition; The protein is an antigenic component in a vaccination method that reduces the severity of clinical signs or symptoms resulting from PCV3 infection in pigs. Also provided is a further investigated protein or a further investigated composition for reducing the severity of clinical signs or symptoms resulting from PCV3 infection in pigs.

[0209] Also, only two doses of the immunogenic composition are administered to pigs in a vaccination regimen that reduces the severity of clinical signs or clinical symptoms resulting from PCV3 infection in pigs; administration of two doses of the immunogenic composition to pigs in a vaccination method reduces the severity of clinical signs or symptoms resulting from PCV3 infection in pigs; Further contemplated proteins are antigenic components in two doses of an immunogenic composition in a vaccination method that reduces the severity of clinical signs or clinical symptoms resulting from PCV3 infection in pigs; Preferably, the protein is present in an amount of at least 2 μg per dose of the immunogenic composition; The protein is an antigenic component in a vaccination method that reduces the severity of clinical signs or symptoms resulting from PCV3 infection in pigs. Further investigated immunogenic compositions for reducing the severity of clinical signs or symptoms resulting from PCV3 infection in pigs are also provided. Preferably, in any of the above uses, said clinical signs or symptoms are selected from the group consisting of reduced average daily weight gain, and death. Preferably, in any of the above-mentioned uses, the clinical signs or symptoms are selected from the group consisting of macroscopic lesions, histological lesions, replication of PCV3 in tissues, and PCV3 viremia. Preferably, in any of the above uses, said clinical signs or symptoms are selected from the group consisting of the occurrence or production of a mummified fetus, a stillborn fetus, and / or a weak fetus. Preferably, in any of the above uses, said clinical signs or symptoms are or include expulsion of a mummified fetus, a stillborn fetus, and / or a weak fetus.

[0210] The present invention is now described by the following clause sets. For ease of reference, these clause sets are designated as Clause Set A, Clause Set B, etc. The disclosure in each clause set is equally applicable to the present invention. Similarly, the disclosure in each clause set is equally applicable to every other clause set.

[0211] Clause Set A: Clause Set A: The present invention is now described by the following numbered set of clauses (Clause Set A). The disclosure in this set of clauses is equally applicable to the present invention. Similarly, the disclosure in this set of clauses is equally applicable to each of the other sets of clauses.

[0212] with porcine circovirus type 1.3 (PCV3) ORF2 protein; a veterinarily acceptable carrier, including a solvent, dispersion medium, coating, stabilizer, diluent, preservative, antimicrobial agent, antifungal agent, isotonic agent, absorption delaying agent, adjuvant, cell culture supernatant, stabilizer, virus or expression vector, and immunomodulator, or any combination thereof; A composition comprising: 2. The composition of clause 1, wherein the veterinarily acceptable carrier comprises an adjuvant, an immunomodulator, a cell culture supernatant, a virus or an expression vector, or any combination thereof. 3. The composition of clause 1, wherein the veterinarily acceptable carrier comprises an adjuvant. 4. The composition of any one of clauses 1 to 3, wherein PCV3 ORF2 is from group a1, b1, or b2 (using the subclassification designation of Fux et al., "Full genome characterization of porcine circovirus type 3 isolates reveals the existence of two distinct groups of virus strains," Virology Journal (2018) 15:25, DOI 10.1186 / s12985-018-0929-3 (incorporated herein by reference); see, e.g., Table 4). 5. The composition of any one of clauses 1 to 3, wherein the PCV3 ORF2 protein comprises or consists of an amino acid sequence encoded by a polynucleotide sequence having at least 90%, or at least 91%, or at least 92%, or at least 93%, or at least 94%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity or sequence homology to SEQ ID NO:1. 6. The composition of any one of clauses 1 to 5, wherein the PCV3 ORF2 protein is a recombinant PCV3 ORF2 protein derived from its expression by an expression vector comprising a polynucleotide sequence encoding the PCV3 ORF2 protein. 7. The composition of clause 6, wherein the expression vector is a baculovirus. 8. The composition of any one of clauses 1 to 7, further comprising PCV2 ORF2 protein. 9. The composition of clause 8, wherein the PCV2 ORF2 protein is derived from expression by an expression vector comprising a polynucleotide sequence encoding the PCV2 ORF2 protein. 10. The composition of clause 9, wherein the expression vector is a baculovirus. 11. The composition of any one of clauses 1-10, further comprising an additional antigen of a further swine pathogen. 12. The composition of clause 11, wherein the additional antigen of a further swine pathogen comprises a PRRSV (Porcine Reproductive and Respiratory Syndrome Virus) antigen, a Mycoplasma hyopneumoniae bacterin antigen, a Mycoplasma hyopneumoniae supernatant antigen, a Pseudorabies antigen or a swine influenza antigen, a swine fever antigen (classical swine fever antigen or African swine fever antigen, or a combination thereof), an Actinobacillus pleuropneumoniae antigen, an Escherichia coli antigen, or a Pasteurella multocida antigen. 13. The composition of any one of clauses 1 to 12, wherein the PCV3 ORF2 protein is present in an amount of 0.2 to about 400 μg / ml, or about 0.3 to about 200 μg / ml, or about 0.35 to about 100 μg / ml, or about 0.4 to about 50 μg / ml, or about 0.45 to about 30 μg / ml, or about 0.6 to about 15 μg / ml, or about 0.75 to about 8 μg / ml, or about 1.0 to about 6 μg / ml, or about 1.3 to about 3.0 μg / ml, or about 1.4 to about 2.5 μg / ml, or about 1.5 to about 2.0 μg / ml, or about 1.6 μg / ml. 14. The composition of any one of clauses 1 to 12, wherein the PCV3 ORF2 protein, or the combined PCV2 and PCV3 ORF2 protein, is present in an amount of about 0.2 to about 400 μg per dose, or about 0.3 to about 200 μg per dose, or about 0.35 to about 100 μg per dose, or about 0.4 to about 50 μg per dose, or about 0.45 to about 30 μg per dose, or about 0.6 to about 15 μg per dose, or about 0.75 to about 8 μg per dose, or about 1.0 to about 6 μg per dose, or about 1.3 to about 3.0 μg per dose, or about 1.4 to about 2.5 μg per dose, or about 1.5 to about 2.0 μg per dose, or about 1.6 μg per dose. 15. Adjuvants include aluminum hydroxide, aluminum phosphate, saponin, and Quinoa. A; QS-21; GPI-0100; Water-in-oil emulsions; Oil-in-water emulsions; Water-in-oil-in-water emulsions; Emulsions based on light liquid paraffin oil or European Pharmacopoeia-type adjuvants; Isoprenoid oils; Squalane; Squalene oil obtained as a result of the oligomerization of alkenes or isobutene or decene; Esters of acids or alcohols containing a linear alkyl group; Vegetable oils; Ethyl oleate; Propylene glycol dicaprylate / caprate; Glyceryl tricaprylate / caprate; Propylene glycol dioleate; Esters of branched fatty acids or alcohols; Isostearate esters; Nonionic surfactants; Sorbitan or mannide or glycol or polyglycerol or propylene glycol, or oleic acid or isostearic acid or ricinoleic acid or hydroxystearic acid, esters of mannitol anhydride oleate, optionally ethoxylated; Polyoxypropylene-polyoxyethylene block copolymers, Pluronic products, Carbopol; Carbopol 974P; Carbopol 934P; Carbopol 971P; polymers of acrylic or methacrylic acid; copolymers of maleic anhydride and alkenyl derivatives; crosslinked polymers of acrylic or methacrylic acid; crosslinked polymers of acrylic or methacrylic acid with polyalkenyl ethers of sugars or polyalcohols; carbomers; acrylic polymers crosslinked with polyhydroxylated compounds having at least three and not more than eight hydroxyl groups, in which at least three hydroxyl hydrogen atoms may be replaced, or have been replaced, by unsaturated aliphatic radicals having at least two carbon atoms, such as vinyl, allyl, and other ethylenically unsaturated groups, containing 2 to 4 carbon atoms, which may themselves contain other substituents, such as methyl; RIBI adjuvant system; block copolymers; SAF-M; monophosphoryl lipid A; avridine lipid-amine adjuvants; heat-labile enterotoxin from Escherichia coli (recombinant or otherwise); cholera toxin;15. The composition according to any one of clauses 1 to 14, comprising IMS1314 or muramyl dipeptide; 16. The composition of any one of clauses 1 to 15, comprising about 50 μg to about 2000 μg of adjuvant; or the adjuvant is present in an amount of about 250 μg per ml of dose of the composition, or the adjuvant is present in an amount of about 100 μg to about 10 mg per dose, or the adjuvant is present in an amount of about 500 μg to about 5 mg per dose, or the adjuvant is present in an amount of about 750 μg to about 2.5 mg per dose, or the adjuvant is present in an amount of about 1 mg per dose. 17. The composition of any one of clauses 1-16, wherein the immunomodulator comprises an interleukin, interferon, or other cytokine, or keyhole limpet hemocyanin (KLH), or KLH emulsified with incomplete Freund's adjuvant (KLH / ICFA). 18. The composition of any one of clauses 1-17, comprising from about 1 μg / ml to about 60 μg / ml of antibiotic, or less than about 30 μg / ml of antibiotic. 19. The composition of any one of clauses 1 to 18, wherein the antibiotic comprises gentamicin. 20. The composition of any one of clauses 1 to 19, comprising: (i) a PCV3 ORF2 protein; (ii) at least a portion of a baculovirus expressing the PCV3 ORF2 protein; (iii) a portion of a cell culture of cells infected with or transfected with a recombinant baculovirus expressing the PCV3 ORF2 protein; (iv) an inactivating agent or an inactivating agent comprising binary ethyleneimine (BEI); (v) sodium thiosulfate or an amount of sodium thiosulfate equivalent to the inactivating agent or BEI; (vi) an adjuvant or an adjuvant comprising Carbopol or Carbopol 971; and (vii) a physiologically acceptable concentration of phosphate. 21. The composition of clause 20, wherein about 90% of components (i) to (iii) have a size smaller than 1 μm, and the pH of the composition is adjusted to about 6.5 to 7.5. 22. The composition of clause 20 or 21, wherein the BEI is derived from a cell culture treated with about 2-8 or about 5 mM BEI to inactivate baculovirus, and / or the composition contains about 2-8 or about 5 mM BEI, and / or the composition contains about 1 mg of Carbopol or Carbopol 971. 23. The composition of any one of clauses 1 to 22, which is formulated and / or packaged for single dose or one shot administration rather than a multiple dose regimen. 24. A method for inducing an immune or immunological response, or a protective immune or immunological response, against (i) PCV3, and / or (ii) PCV2 and PCV3, and / or (iii) PCV3 and another swine pathogen, and / or (iv) PCV3, PCV2, and another swine pathogen, said method comprising the step of administering to an animal a composition described in any one of clauses 1 to 23. 25. The method of clause 25, wherein the animal is a porcine. 26. The method according to clause 25, wherein the porcine is a pig or piglet. 27. The method of clause 26, wherein the pig or piglet is 15 weeks old or less, or 6 weeks old or less, or 3 weeks old or less, or 2 weeks old or less, or 1 week old or less. 28. The method of clause 26, wherein the administration occurs within at least 1, 2, or 3 weeks of exposure to the highly virulent porcine circovirus. 29. The method of any one of clauses 24-28, wherein administering comprises a single, one-shot administration or a single, one-dose administration; does not include multiple shot or multiple dose regimens. 30. Use of the composition of any one of clauses 1 to 23 in the method of any one of clauses 24 to 29; or use of PCV3 ORF2 protein, alone or in combination with any one of the compositions of any one of clauses 1 to 23, for use in the preparation of a composition for inducing an immunological or immune response, or a protective immune or immunological response, against (i) PCV3, and / or (ii) PCV2 and PCV3, and / or (iii) PCV3 and another swine pathogen, and / or (iv) PCV3, PCV2, and another swine pathogen, or for use in a method for inducing an immunological or immune response, or a protective immune or immunological response, against (i) PCV3, and / or (ii) PCV2 and PCV3, and / or (iii) PCV3 and another swine pathogen, and / or (iv) PCV3, PCV2, and another swine pathogen. 31. A method for preparing the composition according to any one of clauses 1 to 23, comprising producing PCV3 ORF2 protein in cultured insect cells using a baculovirus expression system. 32. The method of clause 31, comprising the step of inactivating the baculovirus. 33. The method of clause 32, wherein the inactivating step comprises heat treatment or the use of a viral inactivating agent. 34. The method of clause 25, wherein the virus inactivating agent comprises an aziridine compound. 35. The method of clause 26, wherein the aziridine compound comprises BEI. 36. A recombinant vector comprising a polynucleotide sequence encoding a polypeptide sequence encoding the PCV3 ORF2 protein. 37. The recombinant vector of clause 36, wherein PCV3 ORF2 is derived from group a1, b1, or b2 (using the subclassification designation of Fux et al., "Full genome characterization of porcine circovirus type 3 isolates reveals the existence of two distinct groups of virus strains," Virology Journal (2018) 15:25, DOI 10.1186 / s12985-018-0929-3 (incorporated herein by reference); see, e.g., Table 4). 38. The recombinant vector according to clause 36, wherein the PCV3 ORF2 protein comprises or consists of an amino acid sequence encoded by a polynucleotide sequence having at least 90%, or at least 91%, or at least 92%, or at least 93%, or at least 94%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity or sequence homology to SEQ ID NO:1. 39. The recombinant vector of any one of clauses 36 to 38, which is a baculovirus. 40. The recombinant vector of clause 39, comprising at least 90%, or at least 91%, or at least 92%, or at least 93%, or at least 94%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity or homology with SEQ ID NO:2.

[0213] Clause Set B: Clause Set B: The present invention is now described by the following set of numbered clauses (Clause Set B). The disclosure in this set of clauses is equally applicable to the present invention. Similarly, the disclosure in this set of clauses is equally applicable to each of the other sets of clauses.

[0214] A composition comprising a porcine circovirus type 1.3 (PCV3) ORF2 protein, preferably an antigenic PCV3 ORF2 protein (PCV3 ORF2 antigen). 2. The composition of clause 1, further comprising a veterinarily acceptable carrier selected from the group consisting of solvents, dispersion media, coatings, stabilizers, diluents, preservatives, antimicrobial agents, antifungal agents, isotonic agents, absorption delaying agents, adjuvants, cell culture supernatants, stabilizers, viral vectors, expression vectors, immunomodulators, and / or any combination thereof. A composition comprising porcine circovirus type 3.3 (PCV3) ORF2 protein; and a veterinarily acceptable carrier, including a solvent, dispersion medium, coating agent, stabilizer, diluent, preservative, antimicrobial agent, antifungal agent, isotonic agent, absorption delaying agent, adjuvant, cell culture supernatant, stabilizer, virus or expression vector, immunomodulator, and / or any combination thereof, particularly a composition described in clause 1 or 2. 4. The composition of any one of clauses 1-3, wherein the veterinarily acceptable carrier comprises an adjuvant, an immunomodulator, a cell culture supernatant, a virus or an expression vector, or any combination thereof. 5. The composition of any one of clauses 1 to 4, wherein the veterinarily acceptable carrier comprises an adjuvant. 6. The composition according to any one of clauses 1 to 5, wherein the PCV3 is selected from the group consisting of PCV3a and PCV3b. 7. The composition of any one of clauses 1-6, wherein the PCV3 is any phylogenetic clade of PCV3 or is selected from the group consisting of PCV3a1, PCV3b1, PCV3b2, and PCV3c. 8. The composition of any one of clauses 1 to 7, wherein PCV3 ORF2 is from group a1, b1, or b2. 9. The composition of any one of clauses 1 to 8, wherein the PCV3 ORF2 protein comprises or consists of an amino acid sequence encoded by a polynucleotide sequence having at least 90%, or at least 91%, or at least 92%, or at least 93%, or at least 94%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity to or sequence homology with SEQ ID NO:1. 10. The composition according to any one of clauses 1 to 9, wherein the PCV3 ORF2 protein comprises or consists of an amino acid sequence having at least 90% sequence identity with the sequence of SEQ ID NO:4. 11. The composition of any one of clauses 1 to 10, wherein the PCV3 ORF2 protein is a recombinant PCV3 ORF2 protein. 12. The composition according to any one of clauses 1 to 11, wherein the PCV3 ORF2 protein is a recombinant PCV3 ORF2 protein derived from its expression by an expression vector comprising a polynucleotide sequence encoding the PCV3 ORF2 protein. 13. The composition of clause 12, wherein the expression vector is a baculovirus. 14. The composition of any one of clauses 1 to 13, wherein the PCV3 ORF2 protein is recombinant baculovirus-expressed PCV3 ORF2. 15. The composition according to any one of clauses 1 to 14, further comprising a PCV2 ORF2 protein, preferably an antigenic PCV2 ORF2 protein (PCV2 ORF2 antigen). 16. The composition of clause 15, wherein the PCV2 ORF2 protein is derived from expression by an expression vector comprising a polynucleotide sequence encoding the PCV2 ORF2 protein. 17. The composition of clause 16, wherein the expression vector is a baculovirus. 18. The composition of any one of clauses 1-17, further comprising an additional antigen of a further swine pathogen. 19. The composition of clause 18, wherein the additional antigen of a further swine pathogen comprises a PRRSV (Porcine Reproductive and Respiratory Syndrome Virus) antigen, a Mycoplasma hyopneumoniae bacterin antigen, a Mycoplasma hyopneumoniae supernatant antigen, a Pseudorabies antigen or a swine influenza antigen, a swine fever antigen (classical swine fever antigen or African swine fever antigen, or a combination thereof), an Actinobacillus pleuropneumoniae antigen, an Escherichia coli antigen, a porcine parvovirus (PPV) antigen, or a Pasteurella multocida antigen, or a combination thereof. 20. The composition of any one of clauses 1 to 19, wherein the PCV3 ORF2 protein is present in an amount of 0.2 to about 400 μg / ml, or about 0.3 to about 200 μg / ml, or about 0.35 to about 100 μg / ml, or about 0.4 to about 50 μg / ml, or about 0.45 to about 30 μg / ml, or about 0.6 to about 15 μg / ml, or about 0.75 to about 8 μg / ml, or about 1.0 to about 6 μg / ml, or about 1.3 to about 3.0 μg / ml, or about 1.4 to about 2.5 μg / ml, or about 1.5 to about 2.0 μg / ml, or about 1.6 μg / ml. 21. The composition of any one of clauses 1 to 20, wherein the PCV3 ORF2 protein, or the combined PCV2 and PCV3 ORF2 protein, is present in an amount of about 0.2 to about 400 μg per dose, or about 0.3 to about 200 μg per dose, or about 0.35 to about 100 μg per dose, or about 0.4 to about 50 μg per dose, or about 0.45 to about 30 μg per dose, or about 0.6 to about 15 μg per dose, or about 0.75 to about 8 μg per dose, or about 1.0 to about 6 μg per dose, or about 1.3 to about 3.0 μg per dose, or about 1.4 to about 2.5 μg per dose, or about 1.5 to about 2.0 μg per dose, or about 1.6 μg per dose. 22. The adjuvant is selected from the group consisting of polymers of acrylic or methacrylic acid, copolymers of maleic anhydride and alkenyl derivatives, crosslinked polymers of acrylic or methacrylic acid, crosslinked polymers of acrylic or methacrylic acid with polyalkenyl ethers of sugars or polyalcohols, carbomers, acrylic polymers crosslinked with polyhydroxylated compounds having at least three and not more than eight hydroxyl groups, in which at least three hydroxyl hydrogen atoms may be replaced, or have been replaced, by unsaturated aliphatic radicals having at least two carbon atoms, such as vinyl, allyl, and other ethylenically unsaturated groups, containing 2 to 4 carbon atoms, which may themselves contain other substituents, such as methyl, carbopol, Carbopol 974P, Carbopol 934P, Carbopol 971P, aluminum hydroxide, aluminum phosphate, saponin, Quill A;QS-21;GPI-0100;Water-in-oil emulsions;Oil-in-water emulsions;Water-in-oil-in-water emulsions;Emulsions based on light liquid paraffin oil or European Pharmacopoeia-type adjuvants;Isoprenoid oils;Squalane;Squalene oil obtained as a result of the oligomerization of alkenes or isobutene or decene;Esters of acids or alcohols containing linear alkyl groups;Vegetable oils;Ethyl oleate;Propylene glycol di(caprylic / capric)ate;Caprylic / capric tri(glyceride);Propylene glycol dioleate;Esters of branched fatty acids or alcohols;Isostearate esters; non-ionic surfactants; sorbitan or mannide or glycol or polyglycerol or propylene glycol, or oleic acid or isostearic acid or ricinoleic acid or hydroxystearic acid, esters of mannitol oleate anhydride, which may be ethoxylated; polyoxypropylene-polyoxyethylene block copolymers, Pluronic products, RIBI adjuvant system; block copolymers; SAF-M; monophosphoryl lipid A; avridine lipid-amine adjuvants; heat-labile enterotoxin from Escherichia coli (recombinant or other forms); cholera toxin;22. The composition according to any one of clauses 2 to 21, comprising IMS1314 or muramyl dipeptide; 23. The composition of any one of clauses 2 to 22, comprising about 50 μg to about 2000 μg of adjuvant; or the adjuvant is present in an amount of about 250 μg per ml of dose of the composition, or the adjuvant is present in an amount of about 100 μg to about 10 mg per dose, or the adjuvant is present in an amount of about 500 μg to about 5 mg per dose; the adjuvant is present in an amount of about 750 μg to about 2.5 mg per dose, or the adjuvant is present in an amount of about 1 mg per dose. 24. The composition of any one of clauses 2-23, wherein the immunomodulator comprises an interleukin, interferon, or other cytokine. 25. The composition of any one of clauses 1-24, comprising from about 1 μg / ml to about 60 μg / ml of antibiotic, or less than about 30 μg / ml of antibiotic. 26. The composition of any one of clauses 1 to 25, wherein the antibiotic comprises gentamicin. 27. The composition of any one of clauses 1 to 26, comprising: (i) a PCV3 ORF2 protein; (ii) at least a portion of a baculovirus expressing the PCV3 ORF2 protein; (iii) a portion of a cell culture of cells infected with or transfected with a recombinant baculovirus expressing the PCV3 ORF2 protein; (iv) an inactivating agent or an inactivating agent comprising binary ethyleneimine (BEI); (v) sodium thiosulfate or an amount of sodium thiosulfate equivalent to the inactivating agent or BEI; (vi) an adjuvant or an adjuvant comprising Carbopol or Carbopol 971; and (vii) a physiologically acceptable concentration of phosphate. 28. The composition of clause 27, wherein about 90% of components (i) to (iii) have a size smaller than 1 μm, and the pH of the composition is adjusted to about 6.5 to 7.5. 29. The composition of clause 27 or 28, wherein the BEI is derived from a cell culture treated with about 2-8 or about 5 mM BEI to inactivate baculovirus, and / or the composition contains about 2-8 or about 5 mM BEI, and / or the composition contains about 1 mg of Carbopol or Carbopol 971. 30. The composition of any one of clauses 1 to 29, which is formulated and / or packaged for single dose or one-shot administration of the composition rather than a multiple dose regimen; or which is formulated and / or packaged for a multiple dose regimen of the composition. 31. The composition of any one of clauses 1 to 30, which is an immunogenic composition. 32. A composition according to any one of clauses 1 to 31 for use as a medicament. 33. The composition according to any one of clauses 1 to 31 for use as a vaccine. 34. The composition of any one of clauses 1 to 31 for use in a method for inducing an immune or immunological response, or a protective immune or immunological response, against (i) PCV3, and / or (ii) PCV2 and PCV3, and / or (iii) PCV3 and another swine pathogen, and / or (iv) PCV3, PCV2, and another swine pathogen. 35. A composition according to any one of clauses 1 to 31 for use in a method for reducing or preventing clinical signs or disease caused by infection with PCV3 in an animal, preferably a pig, or for use in a method for treating or preventing infection with PCV3 in an animal. 36. A composition according to any one of clauses 1 to 31 for use in a method for inducing an immune response against PCV3 in pigs, in particular preferably in pregnant pigs. 37. A composition according to any one of clauses 1 to 31 for use in a method for reducing or preventing clinical signs or disease caused by infection with PCV3 in piglets nursed by a sow to which the composition has been administered. 38. A composition for use according to clause 37, wherein the sow to which the composition has been administered is a sow to which an immunogenic composition has been administered, and wherein the sow is pregnant, in particular pregnant with the piglet, or is a gilt prior to breeding season. 39. The composition for use according to any one of clauses 32 to 38, which is administered intramuscularly or intradermally. 40. The composition for use according to any one of clauses 36 to 39, which is administered intramuscularly or intradermally to sows. 41. A method for inducing an immune or immunological response, or a protective immune or immunological response, against (i) PCV3, and / or (ii) PCV2 and PCV3, and / or (iii) PCV3 and another swine pathogen, and / or (iv) PCV3, PCV2, and another swine pathogen, said method comprising the step of administering to an animal a composition described in any one of clauses 1 to 31. 42. The method according to clause 41, wherein the animal is a porcine. 43. The method according to clause 42, wherein the porcine is a pig or piglet. 44. The method according to clause 42 or 43, wherein the pig is a sow. 45. A method of immunizing a subject, the method comprising the step of administering to the subject a composition according to any one of clauses 1 to 31. 46. ​​A method of immunizing swine against clinical disease caused in said animal by at least one pathogen, said method comprising the step of administering to said animal a composition according to any one of clauses 1 to 31, said immunogenic composition being capable of inducing an immune response which does not cause clinical signs of infection but which immunizes the animal against a pathogenic form of said at least one pathogen. 47. The method according to clause 46, wherein at least one pathogen is PCV3. 48. A method for inducing the production of antibodies specific to PCV3 in a sow, the method comprising administering to the sow a composition described in any one of clauses 1 to 31. 49. A method for reducing or preventing clinical signs or symptoms caused by PCV3 infection in piglets, the method comprising the steps of administering a composition described in any one of clauses 1 to 31 to a sow, and allowing the piglets to be suckled by the sow. 50. The method according to clause 49, wherein the sow is in particular a sow that is pregnant with said piglets or a gilt before the breeding season. 51. A method according to clause 49 or 50, comprising administering a composition according to any one of clauses 1 to 31 to a sow that is pregnant with the piglet, allowing the sow to give birth to the piglet, and allowing the piglet to be suckled by the sow. 52. A method for reducing clinical signs and / or symptoms caused by infection with PEDV in piglets, wherein the piglets are nursed by a sow to which the composition of any one of clauses 1 to 31 has been administered. 53. The method according to any one of clauses 45 to 52, wherein the immunogenic composition or said vaccine or pharmaceutical composition is administered intramuscularly or intradermally to said sow. 54. The method according to any one of clauses 45 to 53, wherein the immunogenic composition or said vaccine or pharmaceutical composition is administered twice to said sow. 55. The method according to any one of clauses 45 to 54, wherein the immunogenic composition or said vaccine or pharmaceutical composition is administered twice transmucosally, preferably twice intranasally, to said sow. 56. The composition for use according to any one of clauses 32 to 40 or the method according to any one of clauses 41 to 55, wherein the clinical signs are selected from the group consisting of reduced average daily weight gain, and death. 57. The composition for use according to any one of clauses 32 to 40 or the method according to any one of clauses 41 to 55, wherein the clinical signs are selected from the group consisting of expulsion of a mummified fetus, a stillborn fetus, and / or a weak fetus. 58. The composition for use according to any one of clauses 32 to 40, or the method according to any one of clauses 41 to 55, wherein the clinical symptoms are selected from the group consisting of macroscopic lesions, histological lesions, replication of PCV3 in tissues, and PCV3 viremia. 59. The composition for use according to any one of clauses 32 to 40 or the method according to any one of clauses 41 to 55, wherein the clinical symptoms are selected from the group consisting of the occurrence or production of a mummified fetus, a stillborn fetus, and / or a weak fetus. 60. The composition for use according to any one of clauses 32 to 40, or the method according to any one of clauses 41 to 55, wherein the pig or piglet is 15 weeks old or less, or 6 weeks old or less, or 3 weeks old or less, or 2 weeks old or less, or 1 week old or less. 61. The method of clause 60, wherein the administration occurs within at least 1, 2 or 3 weeks of exposure to the highly virulent porcine circovirus. 62. A composition for use according to any one of clauses 32 to 41 or a method according to any one of clauses 42 to 55, wherein the administration comprises a single, one-shot administration of the composition; or a single, one-dose administration but does not comprise a multiple-shot or multiple-dose regimen; or the administration comprises a single, one-shot administration; or a single, one-dose administration but does not comprise a multiple-shot or multiple-dose regimen; or the administration comprises a multiple-shot or multiple-dose regimen of the composition; or the administration comprises a two-shot or two-dose regimen of the composition, or the administration consists of a two-shot or two-dose regimen of the composition. 63. Use of a composition according to any one of clauses 1 to 31 in a method according to any one of clauses 42 to 55; or use of PCV3 ORF2 protein, alone or in combination with any one of the compositions according to clauses 1 to 31, for use in the preparation of a composition for inducing an immunological or immune response, or a protective immune or immunological response, against (i) PCV3, and / or (ii) PCV2 and PCV3, and / or (iii) PCV3 and another swine pathogen, and / or (iv) PCV3, PCV2, and another swine pathogen, or for use in a method for inducing an immunological or immune response, or a protective immune or immunological response, against (i) PCV3, and / or (ii) PCV2 and PCV3, and / or (iii) PCV3 and another swine pathogen, and / or (iv) PCV3, PCV2, and another swine pathogen. 64. A method for preparing the composition according to any one of clauses 1 to 31, comprising producing PCV3 ORF2 protein in cultured insect cells using a baculovirus expression system. 65. The method of clause 64, comprising the step of inactivating the baculovirus. 66. The method of clause 65, wherein the inactivating step comprises heat treatment or the use of a virus inactivating agent. 67. The method of clause 66, wherein the virus inactivating agent comprises an aziridine compound. 68. The method of clause 67, wherein the aziridine compound comprises BEI. 69. A recombinant vector comprising a polynucleotide sequence encoding a polypeptide sequence encoding the PCV3 ORF2 protein. 70. The recombinant vector according to clause 69, wherein PCV3 ORF2 is from group a1, b1, or b2. 71. A composition comprising (i) a porcine circovirus type 3 (PCV3) ORF2 protein, a parvovirus (PPV) protein, and optionally a PRRSV (porcine reproductive and respiratory syndrome virus) protein, and (ii) a veterinarily acceptable carrier selected from the group consisting of a solvent, dispersion medium, coating agent, stabilizer, diluent, preservative, antimicrobial agent, antifungal agent, isotonic agent, absorption delaying agent, adjuvant, cell culture supernatant, stabilizer, viral vector, expression vector, immunomodulator, and / or any combination thereof. 72. The composition of clause 71, wherein the veterinarily acceptable carrier comprises an adjuvant, an immunomodulator, a cell culture supernatant, a virus or an expression vector, or any combination thereof. 73. The composition according to clause 71 or 72, wherein the PPV protein is the PPV VP2 capsid protein. 74. The composition of any one of clauses 71-73, wherein the PRRSV protein is PRRSV ORF4, ORF5, ORF6, or ORF7. 75. The composition of clause 73 or 74, wherein the PPV protein and / or the PRRSV protein is expressed in a vector. 76. The composition of any one of clauses 71 to 75, which is an immunogenic composition administered to pigs in two doses. 77. The composition according to clause 76, wherein the pig is a gilt or sow. 78. The composition according to clause 76 or 77, wherein administration is before mating / insemination, before conception, during pregnancy, or during lactation. 79. The composition of any one of clauses 76 to 78, wherein the immunogenic composition comprises 0.1 μg to 150 μg, preferably 0.25 μg to 75 μg, more preferably 0.5 μg to 37.5 μg, even more preferably 0.5 μg to 15 μg, and most preferably 0.5 μg to 6 μg of PCV3, PPV, and / or PRRSV antigen. 80. The composition of any one of clauses 76 to 79, wherein the immunogenic composition is administered intramuscularly. 81. A method for inducing an immune or immunological response, or a protective immune or immunological response, against porcine circovirus 3 (PCV3), comprising parenterally or subcutaneously administering to a pig a single shot, single administration, or single dose of (i) at least 2 μg to about 400 μg of PCV3 ORF2 recombinant protein expressed by a baculovirus system, and (ii) a veterinarily acceptable carrier comprising a solvent, dispersion medium, coating agent, stabilizer, diluent, preservative, antimicrobial agent, antifungal agent, isotonic agent, absorption delaying agent, adjuvant, cell culture supernatant, stabilizer, virus or expression vector, immunomodulator, and / or any combination thereof. 82. The method according to clause 81, wherein the porcine is a piglet, a pig or a sow, or a gilt before breeding age. 83. The method of clause 81 or clause 82, wherein the pig is about 1 week or 2 weeks or 3 weeks old or 7 to 28 or 7 to 22 or 14 to 22 or 16 to 22 or 21±5 days old. 84. The method of any one of clauses 81 to 83, wherein the veterinarily acceptable carrier comprises an adjuvant, an immunomodulator, a cell culture supernatant, a virus or an expression vector, or any combination thereof. 85. The method of any one of clauses 81 to 84, wherein PCV3 ORF2 is any phylogenetic clade of PCV3 or is from the group PCV3a, PCV3a1, PCV3b, PCV3b1, or PCV3b. 86. The method of any one of clauses 81 to 85, wherein the PCV3 ORF2 protein comprises or consists of an amino acid sequence encoded by a polynucleotide sequence having at least 90%, or at least 91%, or at least 92%, or at least 93%, or at least 94%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO:1, or sequence homology to SEQ ID NO:1, SEQ ID NO:6, or SEQ ID NO:7. 87. The method of any one of clauses 81 to 86, wherein the single shot, single administration, or single dose further comprises PCV2 ORF2 protein, or an additional antigen of a further swine pathogen. 88. The method of clause 87, wherein the additional antigen of a further swine pathogen comprises a PRRSV (porcine reproductive and respiratory syndrome virus) antigen, a Mycoplasma hyopneumoniae bacterin antigen, a Mycoplasma hyopneumoniae supernatant antigen, a Pseudorabies antigen or a swine influenza antigen, a swine fever antigen (classical swine fever antigen or African swine fever antigen, or a combination thereof), an Actinobacillus pleuropneumoniae antigen, an Escherichia coli antigen, a porcine parvovirus (PPV) antigen, or a Pasteurella multocida antigen, or a combination thereof. 89. The adjuvant is selected from the group consisting of polymers of acrylic or methacrylic acid; copolymers of maleic anhydride and alkenyl derivatives; crosslinked polymers of acrylic or methacrylic acid; crosslinked polymers of acrylic or methacrylic acid with polyalkenyl ethers of sugars or polyalcohols; carbomers; acrylic polymers crosslinked with polyhydroxylated compounds having at least three and not more than eight hydroxyl groups, in which at least three hydroxyl hydrogen atoms may be replaced, or have been replaced, by unsaturated aliphatic radicals having at least two carbon atoms, such as vinyl, allyl, and other ethylenically unsaturated groups, containing 2 to 4 carbon atoms, which may themselves contain other substituents, such as methyl; Carbopol; Carbopol 974P; Carbopol 934P; Carbopol 971P; aluminum hydroxide; aluminum phosphate; saponin; Quill A;QS-21;GPI-0100;Water-in-oil emulsions;Oil-in-water emulsions;Water-in-oil-in-water emulsions;Emulsions based on light liquid paraffin oil or European Pharmacopoeia-type adjuvants;Isoprenoid oils;Squalane;Squalene oil obtained as a result of the oligomerization of alkenes or isobutene or decene;Esters of acids or alcohols containing linear alkyl groups;Vegetable oils;Ethyl oleate;Propylene glycol di(caprylic / capric)ate;Caprylic / capric tri(glyceride);Propylene glycol dioleate;Esters of branched fatty acids or alcohols;Isostearate esters; non-ionic surfactants; sorbitan or mannide or glycol or polyglycerol or propylene glycol, or oleic acid or isostearic acid or ricinoleic acid or hydroxystearic acid, esters of mannitol oleate anhydride, which may be ethoxylated; polyoxypropylene-polyoxyethylene block copolymers, Pluronic products, RIBI adjuvant system; block copolymers; SAF-M; monophosphoryl lipid A; avridine lipid-amine adjuvants; heat-labile enterotoxin from Escherichia coli (recombinant or other forms); cholera toxin;89. The method of any one of clauses 81 to 88, comprising IMS1314 or muramyl dipeptide; 90. The method of any one of clauses 81 to 89, wherein the PCV3 ORF2 protein is present in an amount of 0.2 to about 400 μg / ml, or about 0.3 to about 200 μg / ml, or about 0.35 to about 100 μg / ml, or about 0.4 to about 50 μg / ml, or about 0.45 to about 30 μg / ml, or about 0.6 to about 15 μg / ml, or about 0.75 to about 8 μg / ml, or about 1.0 to about 6 μg / ml, or about 1.3 to about 3.0 μg / ml, or about 1.4 to about 2.5 μg / ml, or about 1.5 to about 2.0 μg / ml, or about 1.6 μg / ml. 91. The method of clause 87, wherein the PCV3 ORF2 protein, or the PCV2 and PCV3 ORF2 combined protein, is present in an amount of about 0.2 to about 400 μg per dose, or about 0.3 to about 200 μg per dose, or about 0.35 to about 100 μg per dose, or about 0.4 to about 50 μg per dose, or about 0.45 to about 30 μg per dose, or about 0.6 to about 15 μg per dose, or about 0.75 to about 8 μg per dose, or about 1.0 to about 6 μg per dose, or about 1.3 to about 3.0 μg per dose, or about 1.4 to about 2.5 μg per dose, or about 1.5 to about 2.0 μg per dose, or about 1.6 μg per dose. 92. The method of any one of clauses 81 to 91, comprising about 50 μg to about 2000 μg of adjuvant; or the adjuvant is present in an amount of about 250 μg per ml of dose of the composition, or the adjuvant is present in an amount of about 100 μg to about 10 mg per dose; or the adjuvant is present in an amount of about 500 μg to about 5 mg per dose; or the adjuvant is present in an amount of about 750 μg to about 2.5 mg per dose, or the adjuvant is present in an amount of about 1 mg per dose. 93. The method of any one of clauses 82-92, wherein the immunomodulator comprises an interleukin, interferon, or other cytokine. 94. The method of any one of clauses 81-93, wherein the single shot, single administration, or single dose further comprises between about 1 μg / ml and about 60 μg / ml of an antibiotic, or less than about 30 μg / ml of an antibiotic. 95. The method according to clause 84, wherein the antibiotic comprises gentamicin. 96. The method of any one of clauses 81 to 95, wherein the single shot, single administration, or single dose comprises: (i) PCV3 ORF2 protein; (ii) at least a portion of a baculovirus expressing said PCV3 ORF2 protein; (iii) a portion of a cell culture of cells infected with or transfected with a recombinant baculovirus expressing said PCV3 ORF2 protein; (iv) an inactivating agent or an inactivating agent comprising binary ethyleneimine (BEI); (v) sodium thiosulfate or an amount of sodium thiosulfate equivalent to the inactivating agent or BEI; (vi) an adjuvant or an adjuvant comprising Carbopol or Carbopol 971; and (vii) a physiologically acceptable concentration of phosphate. 97. The method of claim 96, wherein about 90% of components (i) to (iii) have a size smaller than 1 μm, and the pH of the composition is adjusted to about 6.5 to 7.5. 98. The method of clause 96 or 97, wherein the BEI is derived from a cell culture treated with about 2-8 or about 5 mM BEI to inactivate baculovirus, and / or the composition contains about 2-8 or about 5 mM BEI, and / or the composition contains about 1 mg of Carbopol or Carbopol 971. 99. The method of any one of clauses 81 to 98, further comprising the step of reducing or preventing clinical signs or disease caused by PCV3 or porcine epidemic diarrhea virus (PEDV) infection in pregnant sows or piglets. 100. The method of clause 99, wherein the step of reducing or preventing clinical signs or disease in piglets comprises nursing piglets to a sow that has been administered a single shot, single administration, or single dose. 101. The method of clause 99, wherein the step of reducing or preventing clinical signs or disease in piglets comprises administering a single shot, single administration, or single dose to the pregnant pig. 102. The method of clause 101, further comprising allowing the sow to nurse the piglets after the sow has given birth to the piglets. 103. The method according to any one of clauses 99 to 102, wherein the clinical signs are a reduction in average daily weight gain, death, the occurrence, production or expulsion of mummified, stillborn and / or weak fetuses, gross lesions, histological lesions, replication of PCV3 in tissues, or PCV3 viremia. 104. The method according to any one of clauses 81 to 103, wherein parenteral or subcutaneous administration is intramuscular or intradermal administration. 105. A non-native PCV3 ORF2 protein comprising an engineered FG loop, wherein the FG loop comprises no more than three positively charged amino acids. 106. The PCV3 ORF2 protein according to clause 105, wherein the FG loop comprises two positively charged amino acids. 107. The PCV3 ORF2 protein according to clause 105, wherein the FG loop comprises one positively charged amino acid. 108. The PCV3 ORF2 protein according to clause 105, wherein the FG loop lacks positively charged amino acids. 109. The PCV3 ORF2 protein according to clause 105, wherein the FG loop lacks arginine and lysine residues. 110. The PCV3 ORF2 protein according to clause 105, wherein the FG loop lacks arginine, lysine, and histidine residues. 111. The PCV3 ORF2 protein of clause 105, wherein the FG loop comprises QPFSYH, LSRGF, or MASGF. 112. Non-native PCV3 ORF2 protein containing an engineered C-terminal extension. 113. The PCV3 ORF2 protein of clause 112, wherein the C-terminal extension comprises about 1 to about 10, about 5 to about 20, or about 10 to about 30 amino acids. 114. The PCV3 ORF2 protein of clause 112, wherein the C-terminal extension comprises about 1 to about 10, or about 5 to about 20, or about 10 to 30 amino acids, about 50 to about 200 amino acids, about 60 to about 190 amino acids, about 70 to about 180 amino acids, about 80 to about 170 amino acids, about 90 to about 160 amino acids, or about 100 to about 150 amino acids. 115. The PCV3 ORF2 protein according to clause 112, wherein the C-terminal extension comprises C-terminal amino acids derived from a different capsid protein. 116. The PCV3 ORF2 protein of clause 115, wherein the C-terminal extension comprises C-terminal amino acids derived from a PCV2 capsid, a BFDV capsid, or a CaCV capsid. 117. The PCV3 ORF2 protein of clause 112, wherein the C-terminal extension comprises EFNLKDPPLN, PK, or QFAPNNPSTEFDYETGRQL. 118. A method for producing a self-assembling PCV3 ORF2 capsid protein, the method comprising substituting one or more arginine, lysine, or histidine amino acids in the FG loop with a non-positively charged amino acid. 119. A method for enhancing the self-assembly of PCV3 ORF2 capsid protein, the method comprising the step of adding or inserting an amino acid residue at the C-terminus of the protein. 120. The method of clause 118, comprising adding or inserting 1 to 10, or about 5 to about 20, or about 10 to about 30 amino acids, about 50 to about 200 amino acids, about 60 to about 190 amino acids, about 70 to about 180 amino acids, about 80 to about 170 amino acids, about 90 to about 160 amino acids, or about 100 to about 150 amino acids. 121. The method of clause 119, comprising adding or inserting amino acids from a different capsid protein. 122. The method of clause 121, wherein the added or inserted amino acids are derived from a PCV2 capsid, a BFDV capsid, or a CaCV capsid. 123. The method of clause 121, wherein the added or inserted amino acids comprise EFNLKDPPLN, PK, or QFAPNNPSTEFDYETGRQL. 124. A composition comprising a PCV protein according to any one of clauses 105 to 117, or a protein produced by the method of any one of clauses 118 to 123, in an amount that induces an immune response or a protective immune response against PCV3 and / or its clinical symptoms from a single administration, and a veterinarily acceptable carrier comprising a solvent, dispersion medium, coating agent, stabilizer, diluent, preservative, antimicrobial agent, antifungal agent, isotonic agent, absorption delaying agent, adjuvant, cell culture supernatant, stabilizer, virus or expression vector, immunomodulator, and / or any combination thereof. 125. The composition according to clause 124, wherein the PCV3 ORF2 protein is encoded by SEQ ID NO:6 or SEQ ID NO:7. 126. The composition of clause 124 or 125, wherein the veterinarily acceptable carrier comprises an adjuvant, an immunomodulator, a cell culture supernatant, a virus or an expression vector, or any combination thereof. 127. The composition according to any one of clauses 124 to 126, further comprising a PCV2 ORF2 protein, preferably an antigenic PCV2 ORF2 protein (PCV2 ORF2 antigen), or an additional antigen of a further swine pathogen. 128. The composition of clause 127, wherein the additional antigen of a further swine pathogen comprises a PRRSV (Porcine Reproductive and Respiratory Syndrome Virus) antigen, a Mycoplasma hyopneumoniae bacterin antigen, a Mycoplasma hyopneumoniae supernatant antigen, a Pseudorabies antigen or a swine influenza antigen, a swine fever antigen (classical swine fever antigen or African swine fever antigen, or a combination thereof), an Actinobacillus pleuropneumoniae antigen, an Escherichia coli antigen, a porcine parvovirus (PPV) antigen, or a Pasteurella multocida antigen, or a combination thereof. 129. The adjuvant is selected from the group consisting of polymers of acrylic or methacrylic acid, copolymers of maleic anhydride and alkenyl derivatives, crosslinked polymers of acrylic or methacrylic acid, crosslinked polymers of acrylic or methacrylic acid with polyalkenyl ethers of sugars or polyalcohols, carbomers, acrylic polymers crosslinked with polyhydroxylated compounds having at least three and not more than eight hydroxyl groups, in which at least three hydroxyl hydrogen atoms may be replaced, or replaced, by unsaturated aliphatic radicals having at least two carbon atoms, such as vinyl, allyl, and other ethylenically unsaturated groups, containing 2 to 4 carbon atoms, which may themselves contain other substituents, such as methyl, carbopol, Carbopol 974P, Carbopol 934P, Carbopol 971P, aluminum hydroxide, aluminum phosphate, saponin, Quill A;QS-21;GPI-0100;Water-in-oil emulsions;Oil-in-water emulsions;Water-in-oil-in-water emulsions;Emulsions based on light liquid paraffin oil or European Pharmacopoeia-type adjuvants;Isoprenoid oils;Squalane;Squalene oil obtained as a result of the oligomerization of alkenes or isobutene or decene;Esters of acids or alcohols containing linear alkyl groups;Vegetable oils;Ethyl oleate;Propylene glycol di(caprylic / capric)ate;Caprylic / capric tri(glyceride);Propylene glycol dioleate;Esters of branched fatty acids or alcohols;Isostearate esters; non-ionic surfactants; sorbitan or mannide or glycol or polyglycerol or propylene glycol, or oleic acid or isostearic acid or ricinoleic acid or hydroxystearic acid, esters of mannitol oleate anhydride, which may be ethoxylated; polyoxypropylene-polyoxyethylene block copolymers, Pluronic products, RIBI adjuvant system; block copolymers; SAF-M; monophosphoryl lipid A; avridine lipid-amine adjuvants; heat-labile enterotoxin from Escherichia coli (recombinant or other forms); cholera toxin;129. The composition of any one of clauses 124 to 128, comprising IMS 1314 or muramyl dipeptide; 130. The composition of any one of clauses 124 to 129, wherein the PCV3 ORF2 protein is present in an amount of 0.2 to about 400 μg / ml, or about 0.3 to about 200 μg / ml, or about 0.35 to about 100 μg / ml, or about 0.4 to about 50 μg / ml, or about 0.45 to about 30 μg / ml, or about 0.6 to about 15 μg / ml, or about 0.75 to about 8 μg / ml, or about 1.0 to about 6 μg / ml, or about 1.3 to about 3.0 μg / ml, or about 1.4 to about 2.5 μg / ml, or about 1.5 to about 2.0 μg / ml, or about 1.6 μg / ml. 131. The composition according to any one of clauses 124 to 130, wherein the PCV3 ORF2 protein, or the PCV2 and PCV3 ORF2 combined protein, is present in an amount of about 0.2 to about 400 μg per dose, or about 0.3 to about 200 μg per dose, or about 0.35 to about 100 μg per dose, or about 0.4 to about 50 μg per dose, or about 0.45 to about 30 μg per dose, or about 0.6 to about 15 μg per dose, or about 0.75 to about 8 μg per dose, or about 1.0 to about 6 μg per dose, or about 1.3 to about 3.0 μg per dose, or about 1.4 to about 2.5 μg per dose, or about 1.5 to about 2.0 μg per dose, or about 1.6 μg per dose. 132. The composition of any one of clauses 124 to 131, comprising about 50 μg to about 2000 μg of adjuvant; or the adjuvant is present in an amount of about 250 μg per ml of dose of the composition, or the adjuvant is present in an amount of about 100 μg to about 10 mg per dose, or the adjuvant is present in an amount of about 500 μg to about 5 mg per dose; the adjuvant is present in an amount of about 750 μg to about 2.5 mg per dose; or the adjuvant is present in an amount of about 1 mg per dose. 133. The composition of any one of clauses 125-132, wherein the immunomodulator comprises an interleukin, interferon, or other cytokine. 134. A vector containing and expressing a PCV protein according to any one of clauses 105 to 117, or a protein produced by the method according to any one of clauses 118 to 123. 135. The vector according to clause 134, wherein the PCV protein is expressed according to SEQ ID NO: 6 or SEQ ID NO: 7. 136. The vector according to clause 134 or 135, which is a baculovirus. 137. A method for preparing the composition according to any one of clauses 125 to 133, said method comprising the step of producing PCV3 ORF2 protein in cultured insect cells using a baculovirus expression system. 138. The method of clause 137, further comprising the step of inactivating the baculovirus. 139. The method of clause 138, wherein the inactivating step comprises heat treatment or the use of a virus inactivating agent. 140. The method of clause 139, wherein the virus inactivating agent comprises an aziridine compound. 141. The method of clause 140, wherein the aziridine compound comprises BEI.

[0215] Clause Set C: Clause Set C: The present invention is now described by the following numbered set of clauses (Clause Set C). The disclosure in this set of clauses is equally applicable to the present invention. Similarly, the disclosure in this set of clauses is equally applicable to each of the other sets of clauses.

[0216] 1. A porcine circovirus type 3 (PCV3) antigenic protein, which is PCV3 ORF2 protein or a functional antigenic variant thereof. 2. The protein of clause 1, wherein the PCV3 ORF2 protein is the protein encoded by SEQ ID NO:1. 3. The protein of clause 1 or clause 2, which is a functional antigenic variant of PCV3 ORF2. 4. The protein of any one of clauses 1 to 3, which is a functional antigenic variant of the protein encoded by SEQ ID NO:1. 5. The protein of any one of clauses 1 to 4, wherein the functional antigenic variant is capable of higher yields of virus-like particles (VLPs) than the protein encoded by SEQ ID NO:1. 6. The protein of any one of clauses 1 to 5, wherein the functional antigenic variant allows for a higher yield of VLPs than the protein encoded by SEQ ID NO: 1, as can be determined by Western blot analysis. 7. The protein of any one of clauses 1 to 6, wherein the functional antigenic variant has fewer positively charged amino acid residues than the protein encoded by SEQ ID NO:1. 8. The protein of any one of clauses 1-7, wherein the functional antigenic variant has one or more substitutions in the FG loop of the protein encoded by SEQ ID NO:1. 9. The protein of clause 8, wherein the functional antigenic variant has one or more substitutions in the FG loop of the protein encoded by SEQ ID NO:1, wherein these substitutions include substitution of one or more of the S and / or K and / or H residues of the motif SKKKH in the FG loop of the protein encoded by SEQ ID NO:1. 10. The protein of clause 8, wherein the functional antigenic variant has one or more substitutions in the FG loop of the protein encoded by SEQ ID NO:1, wherein these substitutions include substitution of one or more of the S and / or K residues of the motif SKKKH in the FG loop of the protein encoded by SEQ ID NO:1. 11. The protein of clause 8, wherein the functional antigenic variant has one or more substitutions in the FG loop of the protein encoded by SEQ ID NO:1, and these substitutions include substitutions of S or H residues and all of the K residues in the motif SKKKH in the FG loop of the protein encoded by SEQ ID NO:1. 12. The protein of clause 8, wherein the functional antigenic variant has one or more substitutions in the FG loop of the protein encoded by SEQ ID NO:1, wherein these substitutions include replacement of at least S and / or H, and any K, with Q or P or F or S in the motif SKKKH of the FG loop of the protein encoded by SEQ ID NO:1. 13. The protein of clause 8, wherein the functional antigenic variant has one or more substitutions in the FG loop of the protein encoded by SEQ ID NO:1, comprising a substitution of the motif SKKK within the motif SKKKH in the FG loop of the protein encoded by SEQ ID NO:1 with QPFS, or a substitution of the motif KKKH within the motif SKKKH in the FG loop of the protein encoded by SEQ ID NO:1 with QPFS. 14. The protein of any one of clauses 1-13, wherein the functional antigenic variant is encodable by all or part of SEQ ID NO: 1, 2, 5, 6, or 7. 15. The protein of any one of clauses 1-13, wherein the functional antigenic variant is encoded by all or part of SEQ ID NO: 1, 2, 5, 6, or 7. 16. The protein of any one of clauses 1 to 13, wherein the functional antigenic variant has a C-terminus extending beyond the terminal SVL sequence of the protein encoded by SEQ ID NO:1, preferably said extension is or comprises all of a sequence derived from a virus of the Circoviridae family, and preferably at least part of said extension replaces the terminal SVL sequence of the protein encoded by SEQ ID NO:1. 17. The protein of clause 16, wherein the functional antigenic variant has a C-terminus extending beyond the terminal SVL sequence of the protein encoded by SEQ ID NO: 1; and said extension is 1 to 100 amino acids in length. 18. The protein of clause 16, wherein the functional antigenic variant has a C-terminus extending beyond the terminal SVL sequence of the protein encoded by SEQ ID NO: 1; and said extension is 1 to 50 amino acids in length. 19. The protein of clause 16, wherein the functional antigenic variant has a C-terminus extending beyond the terminal SVL sequence of the protein encoded by SEQ ID NO: 1; and said extension is 1 to 30 amino acids in length. 20. The protein of clause 19, wherein the functional antigenic variant has a C-terminus extending beyond the terminal SVL sequence of the protein encoded by SEQ ID NO: 1; said extension is 1 to 30 amino acids in length, and said extension comprises all or part of the sequence VKININLTPPVATSRVPSRALPLRFGCGHR. 21. The protein of clause 19, wherein the functional antigenic variant has a C-terminus extending beyond the terminal SVL sequence of the protein encoded by SEQ ID NO: 1; said extension is 1 to 30 amino acids in length, and said extension comprises the entire sequence VKININLTPPVATSRVPSRALPLRFGCGHR. 22. The protein of any one of clauses 1-21, wherein the functional antigenic variant is encodable by all or part of SEQ ID NO: 1, 2, 5, 6, or 7. 23. The protein of any one of clauses 1-21, wherein the functional antigenic variant is encoded by all or part of SEQ ID NO: 1, 2, 5, 6, or 7. 24. The protein of any one of clauses 1 to 23, which is a recombinant protein prepared by recombinant DNA techniques. 25. The protein of any one of clauses 1 to 24, which is a baculovirus-expressed protein. 26. The protein according to any one of clauses 1 to 25, wherein PCV3 is selected from the group consisting of PCV3a and PCV3b. 27. The protein of any one of clauses 1-25, wherein PCV3 is any phylogenetic clade of PCV3 or is selected from the group consisting of PCV3a1, PCV3b1, PCV3b2, and PCV3c. 28. The protein of any one of clauses 1 to 27, wherein PCV3 ORF2 is from group a1, b1, or b2 (using the subclassification designation of Fux et al., "Full genome characterization of porcine circovirus type 3 isolates reveals the existence of two distinct groups of virus strains," Virology Journal (2018) 15:25, DOI 10.1186 / s12985-018-0929-3 (incorporated herein by reference); see, e.g., Table 4). 29. The protein of any one of clauses 1 to 28, wherein the PCV3 ORF2 protein comprises or consists of an amino acid sequence encoded by a polynucleotide sequence having at least 90%, or at least 91%, or at least 92%, or at least 93%, or at least 94%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity to or sequence homology with SEQ ID NO:1. 30. The protein of any one of clauses 1 to 28, wherein the variant protein comprises or consists of an amino acid sequence encoded by a polynucleotide sequence having at least 90%, or at least 91%, or at least 92%, or at least 93%, or at least 94%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 1, or sequence homology to SEQ ID NO: 6. 31. The protein of any one of clauses 1 to 28, wherein the variant protein comprises or consists of an amino acid sequence encoded by a polynucleotide sequence having at least 90%, or at least 91%, or at least 92%, or at least 93%, or at least 94%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 1, or sequence homology to SEQ ID NO: 7. 32. The protein of any one of clauses 1-31, wherein the PCV3 ORF2 protein comprises or consists of an amino acid sequence having at least 90%, or at least 91%, or at least 92%, or at least 93%, or at least 94%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity to the sequence of SEQ ID NO: 3, 4, 8, 9, or 10, and / or wherein the protein is a recombinant protein. 33. The protein of any one of clauses 1 to 32, wherein the variant protein comprises or consists of an amino acid sequence having at least 90%, or at least 91%, or at least 92%, or at least 93%, or at least 94%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity to the sequence of SEQ ID NO: 3, 4, 8, 9, or 10, and / or wherein the protein is a recombinant protein; or wherein said variant protein comprises or consists of an amino acid sequence having at least 90%, or at least 91%, or at least 92%, or at least 93%, or at least 94%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity to the sequence of SEQ ID NO: 3, 4, 8, 9, or 10, and / or wherein the protein is a recombinant protein; 33. The protein of any one of clauses 1 to 32, comprising or consisting of an amino acid sequence having at least 90%, or at least 91%, or at least 92%, or at least 93%, or at least 94%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity with sequence numbered 3, 4, 8, 9, or 10, and / or wherein the protein is a recombinant protein, and wherein the protein has one or more substitutions in the FG loop. 34. The protein of any one of clauses 1 to 32, wherein the variant protein comprises or consists of an amino acid sequence having at least 90%, or at least 91%, or at least 92%, or at least 93%, or at least 94%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity to the sequence of SEQ ID NO: 3, 4, 8, 9, or 10, and / or wherein the protein is a recombinant protein; or wherein said variant protein is a recombinant protein 33. The protein of any one of clauses 1 to 32, comprising or consisting of an amino acid sequence having at least 90%, or at least 91%, or at least 92%, or at least 93%, or at least 94%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity with any one of the sequences of SEQ ID NO: 1, and / or wherein the protein is a recombinant protein, and wherein the protein has a C-terminus extending beyond the SVL sequence at the end of the protein encoded by SEQ ID NO: 1. 35. The protein of any one of clauses 1 to 34, which is a recombinant protein derived from its expression by an expression vector comprising a polynucleotide sequence encoding the protein. 36. The protein of any one of clauses 1 to 35, which is a recombinant protein derived from its expression by a baculovirus expression vector comprising a polynucleotide sequence encoding the protein. 37. A nucleotide sequence encoding a protein according to any one of clauses 1 to 36. 38. A vector comprising a nucleotide sequence according to clause 37. 39. A recombinant vector comprising a nucleotide sequence according to clause 37. 40. An expression host transformed or transfected with a nucleotide sequence according to clause 37. 41. A baculovirus expression host transformed or transfected with a nucleotide sequence according to clause 37. 42. A method for preparing a protein according to any one of clauses 1 to 36, comprising expressing a nucleotide sequence according to clause 37. 43. A method for preparing a protein according to any one of clauses 1 to 36, comprising expressing a vector according to any one of clauses 35 to 39. 44. A method for preparing a protein according to any one of clauses 1 to 36, comprising expressing a recombinant vector according to clause 39. 45. A method for preparing a protein according to any one of clauses 1 to 36, comprising culturing an expression host according to any one of clauses 40 to 41 to cause expression of the protein. 46. ​​A method for preparing a protein according to any one of clauses 1 to 36, comprising transfecting an expression host with the nucleotide sequence of a vector according to any one of clauses 35 to 39, and culturing the expression host to cause expression of the protein. 47. A method for preparing a protein according to any one of clauses 1 to 36, comprising culturing a baculovirus expression host according to clause 41 to cause expression of the protein. 48. A method for preparing a protein according to any one of clauses 1 to 36, comprising transfecting a baculovirus expression host with the nucleotide sequence of a vector according to any one of clauses 35 to 39, and culturing the baculovirus expression host to cause expression of the protein. 49. The method of any one of clauses 42 to 48, wherein an inactivating agent is used when a sufficient level of protein expression has been achieved. 50. The method of any one of clauses 42 to 49, wherein an inactivation agent comprising binary ethyleneimine (BEI) is used when a sufficient level of protein expression is achieved. 51. A method for preparing a protein according to any one of clauses 1 to 36, comprising the steps of transfecting a baculovirus expression host with the nucleotide sequence of a vector according to any one of clauses 35 to 39, and culturing the baculovirus expression host in a culture medium to cause expression of the protein; wherein the culture medium after expression of the protein comprises: (i) the protein; (ii) at least a portion of a baculovirus expressing the protein; and (iii) a portion of a cell culture of cells infected with or transfected with a recombinant baculovirus expressing the protein. 52. A method for preparing a protein according to any one of clauses 1 to 36, comprising the steps of transfecting a baculovirus expression host with the nucleotide sequence of a vector according to any one of clauses 35 to 39, and culturing the baculovirus expression host in a culture medium to cause expression of the protein; wherein the culture medium after expression of the protein contains: (i) the protein; (ii) at least a portion of a baculovirus expressing the protein; and (iii) a portion of a cell culture of cells infected with or transfected with a recombinant baculovirus expressing the protein; and wherein about 90% of components (i) to (iii) have a size less than 1 μm. 53. A method for preparing a protein according to any one of clauses 1 to 36, comprising the steps of transfecting a baculovirus expression host with the nucleotide sequence of the vector according to any one of clauses 35 to 39, and culturing the baculovirus expression host in a medium to cause expression of the protein; wherein the medium after expression of the protein contains (i) the protein; (ii) at least a portion of a baculovirus expressing the protein; and (iii) a portion of a cell culture of cells infected with or transfected with a recombinant baculovirus expressing the protein; wherein about 90% of components (i) to (iii) have a size less than 1 μm, and wherein the pH of the composition is adjusted to about 6.5 to 7.5. 54. A method for preparing a protein according to any one of clauses 1 to 36, comprising producing the protein by a baculovirus expression system in cultured insect cells. 55. A method for preparing a protein according to any one of clauses 1 to 36, comprising producing the protein by a baculovirus expression system in cultured insect cells; and inactivating the baculovirus. 56. A method for preparing a protein according to any one of clauses 1 to 36, comprising producing the protein by a baculovirus expression system in cultured insect cells; and inactivating the baculovirus; wherein the inactivating step comprises heat treatment or the use of a virus-inactivating agent. 57. A method for preparing a protein according to any one of clauses 1 to 36, comprising producing the protein using a baculovirus expression system in cultured insect cells; inactivating the baculovirus; the inactivating step comprising heat treatment or the use of a virus inactivating agent; and the virus inactivating agent comprising an aziridine compound. 58. A method for preparing a protein according to any one of clauses 1 to 36, comprising producing the protein using a baculovirus expression system in cultured insect cells; comprising inactivating the baculovirus; the inactivating step comprises heat treatment or the use of a virus inactivating agent; the virus inactivating agent comprises an aziridine compound; and the aziridine compound comprises BEI. 59. A protein obtainable by the method according to any one of clauses 42 to 58. 60. A composition comprising a protein obtained by the method according to any one of clauses 42 to 58. 61. A composition obtainable by the method according to any one of clauses 42 to 58. 62. A composition comprising a protein according to any one of clauses 1 to 36 and a carrier, diluent or excipient. 63. A composition comprising a protein according to any one of clauses 1 to 36 and a veterinarily acceptable carrier, diluent or excipient. 64. The protein is 0.2 to about 400 μg / ml, or 2 to about 400 μg / ml, or 4 to about 400 μg / ml, or 8 to about 400 μg / ml, or about 0.3 to about 200 μg / ml, or 2 to about 200 μg / ml, or 4 to about 200 μg / ml, or 8 to about 200 μg / ml, or about 0.35 to about 100 μg / ml, or 2 to about 100 μg / ml, or 4 to about 100 μg / ml, or 8 to about 1 64. The composition of any one of clauses 60 to 63, wherein the soluble ... 65. A composition comprising a protein according to any one of clauses 1-36, comprising any one or more of a solvent, dispersion medium, coating, stabilizer, diluent, preservative, antimicrobial agent, antifungal agent, isotonic agent, absorption delaying agent, adjuvant, cell culture supernatant, stabilizer, viral vector, expression vector, and / or immunomodulator. 66. The composition of any one of clauses 60-65, wherein the carrier, diluent, or excipient is any one or more of an adjuvant, an immunomodulator, a cell culture supernatant, a virus or an expression vector, or any combination thereof. 67. The composition of any one of clauses 60 to 65, wherein the carrier, diluent, or excipient comprises an adjuvant. 68. The carrier, diluent, or excipient comprises an adjuvant; the adjuvant is selected from the group consisting of polymers of acrylic or methacrylic acid, copolymers of maleic anhydride and alkenyl derivatives, crosslinked polymers of acrylic or methacrylic acid, crosslinked polymers of acrylic or methacrylic acid with polyalkenyl ethers of sugars or polyalcohols, carbomers, acrylic polymers crosslinked with polyhydroxylated compounds having at least three and not more than eight hydroxyl groups, in which at least three hydroxyl hydrogen atoms may be replaced, or replaced, by unsaturated aliphatic radicals having at least two carbon atoms, such as vinyl, allyl, and other ethylenically unsaturated groups, containing 2 to 4 carbon atoms, which may themselves contain other substituents, such as methyl, carbopol, Carbopol 974P, Carbopol 934P, Carbopol 971P, aluminum hydroxide, aluminum phosphate, saponin, Quill A;QS-21;GPI-0100;Water-in-oil emulsions;Oil-in-water emulsions;Water-in-oil-in-water emulsions;Emulsions based on light liquid paraffin oil or European Pharmacopoeia-type adjuvants;Isoprenoid oils;Squalane;Squalene oil obtained as a result of the oligomerization of alkenes or isobutene or decene;Esters of acids or alcohols containing linear alkyl groups;Vegetable oils;Ethyl oleate;Propylene glycol di(caprylic / capric)ate;Caprylic / capric tri(glyceride);Propylene glycol dioleate;Esters of branched fatty acids or alcohols;Isostearate esters; non-ionic surfactants; sorbitan or mannide or glycol or polyglycerol or propylene glycol, or oleic acid or isostearic acid or ricinoleic acid or hydroxystearic acid, esters of mannitol oleate anhydride, which may be ethoxylated; polyoxypropylene-polyoxyethylene block copolymers, Pluronic products, RIBI adjuvant system; block copolymers; SAF-M; monophosphoryl lipid A; avridine lipid-amine adjuvants; heat-labile enterotoxin from Escherichia coli (recombinant or other forms); cholera toxin;68. The composition of claim 67, comprising one or more of IMS1314, or muramyl dipeptide; 69. The composition of clause 67, wherein the carrier, diluent, or excipient comprises an adjuvant; and the adjuvant comprises Carbopol or Carbopol 971. 70. The composition of clause 67, wherein the carrier, diluent, or excipient comprises an adjuvant; the adjuvant is present in an amount of about 50 μg to about 2000 μg per ml of dose of the composition; or the adjuvant is present in an amount of about 250 μg per ml of dose of the composition, or the adjuvant is present in an amount of about 100 μg to about 10 mg per ml of dose of the composition; or the adjuvant is present in an amount of about 500 μg to about 5 mg per ml of dose of the composition; or the adjuvant is present in an amount of about 750 μg to about 2.5 mg per ml of dose of the composition, or the adjuvant is present in an amount of about 1 mg per ml of dose of the composition. 71. A composition according to any one of clauses 60-70, comprising an immunomodulatory agent. 72. The composition of clause 71, comprising an immunomodulator; wherein the immunomodulator is any one or more of an interleukin, an interferon, or other cytokine. 73. A composition according to any one of clauses 60 to 72, comprising an antibiotic. 74. The composition according to clause 73, comprising an antibiotic; said antibiotic comprising gentamicin. 75. The composition of clause 73, comprising from about 1 μg / ml to about 60 μg / ml of the antibiotic. 76. The composition of clause 73, comprising from about 1 μg / ml to less than about 30 μg / ml of the antibiotic. 77. The composition of any one of clauses 60 to 76, comprising an additional antigen. 78. The composition of clause 77, comprising an additional antigen; and said additional antigen is not a PCV3 ORF2 antigen. 79. The composition of clause 77, comprising an additional antigen; and said additional antigen is not a PCV3 antigen. 80. The composition of clause 77, comprising an additional antigen of a further swine pathogen. 81. The composition of clause 80, further comprising an antigen of a further swine pathogen, wherein the pathogen is any one or more of PCV2, PRRSV (Porcine Reproductive and Respiratory Syndrome Virus) antigen, Mycoplasma hyopneumoniae bacterin antigen, Mycoplasma hyopneumoniae supernatant antigen, Aujeszky's disease antigen or Pseudorabies antigen, swine influenza antigen, swine fever antigen (classical swine fever antigen or African swine fever antigen, or a combination thereof), Actinobacillus pleuropneumoniae antigen, Escherichia coli antigen, porcine parvovirus (PPV) antigen, or Pasteurella multocida antigen. 82. The composition of clause 80, further comprising an antigen of a further swine pathogen, further comprising one or more of a PCV2 antigen, a PRRSV antigen, and a PPV antigen. 83. The composition according to clause 82, further comprising a PCV2 antigen. 84. The composition according to clause 83, further comprising a PCV2 antigen, wherein the PCV2 antigen is PCV2 ORF2 protein. 85. The composition according to clause 83, further comprising a PCV2 antigen, wherein the PCV2 antigen is a recombinant PCV2 ORF2 protein. 86. The composition of clause 83, further comprising a PCV2 antigen, wherein the PCV2 antigen is a recombinant baculovirus-expressed PCV2 ORF2 protein. 87. A composition according to any one of clauses 60 to 86, which is in a dosage form. 88. The composition according to clause 87, formulated and / or packaged for single dose or one-shot administration. 89. The composition according to clause 87, formulated and / or packaged for a multiple dose regimen. 90. The composition of clause 87, formulated and / or packaged for a two-dose regimen. 91. The composition according to clause 87, which is a dosage form; said dosage form is delivered from a container containing a large quantity of said composition, and said dosage form of said composition is capable of being delivered from said container. 92. The composition of claim 87, which is a dosage form; said dosage form is delivered from a container containing a large quantity of said composition, and wherein a dosage form of said composition is capable of being delivered from said container; and said container contains at least 10 doses of said composition. 93. The composition of clause 87, which is a dosage form; said dosage form is delivered from a container containing a large quantity of said composition, and wherein a dosage form of said composition is capable of being delivered from said container; and said container contains at least 50 doses of said composition. 94. The composition of clause 87, which is a dosage form; said dosage form is delivered from a container containing a large quantity of said composition, and wherein a dosage form of said composition is capable of being delivered from said container; and said container contains at least 100 doses of said composition. 95. The composition of clause 87, which is a dosage form; said dosage form is delivered from a container containing a large quantity of said composition, and wherein a dosage form of said composition is capable of being delivered from said container; and said container contains at least 200 doses of said composition. 96. The composition of clause 87, which is a dosage form; said dosage form is delivered from a container containing a large quantity of said composition, and wherein a dosage form of said composition is capable of being delivered from said container; and said container contains at least 250 doses of said composition. 97. The vaccine comprises a PCV2 antigen; the PCV2 antigen is a recombinant baculovirus-expressed PCV2 ORF2 protein; and one of the PCV3 ORF2 protein and the PCV2 ORF protein, or the total amount of the combined PCV3 ORF2 protein and the PCV2 ORF protein, is about 0.2 to about 400 μg per dose, or 2 to about 400 μg per dose, or 4 to about 400 μg per dose, or 8 to about 400 μg per dose, or about 0.3 to about 200 μg per dose, or 2 to about 200 μg per dose, or 4 to about 200 μg per dose, or 8 to about 200 μg per dose, or about 0.35 to about 100 μg per dose, or 2 to about 100 μg per dose, or 97. The composition of any one of clauses 60-96, wherein the IL-14 is present in an amount of from 4 to about 100 μg per dose, or from 8 to about 100 μg per dose, or from about 0.4 to about 50 μg per dose, or from about 0.45 to about 30 μg per dose, or from about 0.6 to about 15 μg per dose, or from about 0.75 to about 8 μg per dose, or from about 1.0 to about 6 μg per dose, or from about 1.3 to about 3.0 μg per dose, or from about 1.4 to about 2.5 μg per dose, or from about 1.5 to about 2.0 μg per dose, or about 1.6 μg per dose. 98. The composition of any one of clauses 60 to 97, comprising a salt. 99. The composition of any one of clauses 60 to 98, comprising an inactivated viral vector and / or a cell culture supernatant. 100. A composition according to any one of clauses 60 to 98, comprising an inactivated viral vector and a cell culture supernatant. 101. The composition of any one of clauses 60 to 100, comprising: (i) a protein; (ii) at least a portion of a baculovirus expressing said protein; (iii) a portion of a cell culture of cells infected with or transfected with a recombinant baculovirus expressing said protein; (iv) an inactivating agent or an inactivating agent comprising binary ethyleneimine (BEI); (v) sodium thiosulfate or an amount of sodium thiosulfate equivalent to the inactivating agent or BEI; (vi) an adjuvant or an adjuvant comprising Carbopol or Carbopol 971; and (vii) a physiologically acceptable concentration of phosphate. 102. The composition of clause 101, comprising: (i) a protein; (ii) at least a portion of a baculovirus expressing the protein; (iii) a portion of a cell culture of cells infected with or transfected with a recombinant baculovirus expressing the protein; (iv) an inactivating agent or an inactivating agent comprising binary ethyleneimine (BEI); (v) sodium thiosulfate or an amount of sodium thiosulfate equivalent to the inactivating agent or BEI; (vi) an adjuvant or an adjuvant comprising Carbopol or Carbopol 971; and (vii) a physiologically acceptable concentration of phosphate; wherein the BEI is derived from a cell culture treated with about 2 to 8 or about 5 mM BEI to inactivate the baculovirus, and / or the composition contains about 2 to 8 or about 5 mM BEI, and / or the composition contains about 1 mg of Carbopol or Carbopol 971. 103. A composition according to any one of clauses 60 to 102 which is an immunogenic composition comprising a protein according to any one of clauses 1 to 36 and a carrier, diluent or excipient. 104. A composition according to clause 103 which is an immunogenic composition comprising a protein according to any one of clauses 1 to 36 and a carrier, diluent or excipient; and an additional antigen according to any one of clauses 77 to 80. 105. A process for making a composition according to any one of clauses 1 to 104, wherein the protein according to any one of clauses 1 to 36 is mixed with a carrier, diluent or excipient. 106. A process for making a composition according to any one of clauses 1 to 104, wherein the protein according to any one of clauses 1 to 36 is mixed with a carrier, diluent, or excipient; and an additional antigen. 107. A protein according to any one of clauses 1 to 36, or a nucleotide sequence according to clause 37, or an expression vector according to any one of clauses 38 to 39, or an expression host according to any one of clauses 40 to 41, or a composition according to any one of clauses 60 to 104, for use as a medicament. 108. A protein according to any one of clauses 1 to 36, or a nucleotide sequence according to clause 37, or an expression vector according to any one of clauses 38 to 39, or an expression host according to any one of clauses 40 to 41, or a composition according to any one of clauses 60 to 104, for use as a vaccine. 109. A protein according to any one of clauses 1 to 36, or a nucleotide sequence according to clause 37, or an expression vector according to any one of clauses 38 to 39, or an expression host according to any one of clauses 40 to 41, or a composition according to any one of clauses 60 to 104, for use in a method for inducing an immune or immunological response, or a protective immune or immunological response, against PCV3 in an animal. 110. A protein according to any one of clauses 1 to 36, or a nucleotide sequence according to clause 37, or an expression vector according to any one of clauses 38 to 39, or an expression host according to any one of clauses 40 to 41, or a composition according to any one of clauses 60 to 104, for use in a method for inducing an immune or immunological response, or a protective immune or immunological response, against PCV3 in a pig. 111. A protein according to any one of clauses 1 to 36, or a nucleotide sequence according to clause 37, or an expression vector according to any one of clauses 38 to 39, or an expression host according to any one of clauses 40 to 41, or a composition according to any one of clauses 60 to 104, for use in a method for inducing an immune or immunological response, or a protective immune or immunological response, against PCV3 in a pig. 112. A protein according to any one of clauses 1 to 36, or a nucleotide sequence according to clause 37, or an expression vector according to any one of clauses 38 to 39, or an expression host according to any one of clauses 40 to 41, or a composition according to any one of clauses 60 to 104, for use in a method for inducing an immune or immunological response, or a protective immune or immunological response, against PCV3 in a piglet. 113. A protein according to any one of clauses 1 to 36, or a nucleotide sequence according to clause 37, or an expression vector according to any one of clauses 38 to 39, or an expression host according to any one of clauses 40 to 41, or a composition according to any one of clauses 60 to 104, for use in a method for inducing an immune or immunological response against PCV3, or a protective immune or immunological response, in piglets suckled by a sow to which has been administered the protein according to any one of clauses 1 to 36, or the nucleotide sequence according to clause 37, or the expression vector according to any one of clauses 38 to 39, or the expression host according to any one of clauses 40 to 41, or the composition according to any one of clauses 60 to 104. 114. A protein according to any one of clauses 1 to 36, or a nucleotide sequence according to clause 37, or an expression vector according to any one of clauses 38 to 39, or an expression host according to any one of clauses 40 to 41, or a composition according to any one of clauses 60 to 104, for use in a method for inducing an immune or immunological response, or a protective immune or immunological response, against PCV3 in a sow. 115. A protein according to any one of clauses 1 to 36, or a nucleotide sequence according to clause 37, or an expression vector according to any one of clauses 38 to 39, or an expression host according to any one of clauses 40 to 41, or a composition according to any one of clauses 60 to 104, for use in a method for inducing an immune or immunological response, or a protective immune or immunological response, against PCV3 in a pregnant pig, a gilt, or a pre-breeding gilt. 116. A protein according to any one of clauses 1 to 36, or a nucleotide sequence according to clause 37, or an expression vector according to any one of clauses 38 to 39, or an expression host according to any one of clauses 40 to 41, or a composition according to any one of clauses 60 to 104, for use in inducing an immune response against PCV3 in an animal. 117. A protein according to any one of clauses 1 to 36, or a nucleotide sequence according to clause 37, or an expression vector according to any one of clauses 38 to 39, or an expression host according to any one of clauses 40 to 41, or a composition according to any one of clauses 60 to 104, for use in inducing an immune response against PCV3 in a pig. 118. A protein according to any one of clauses 1 to 36, or a nucleotide sequence according to clause 37, or an expression vector according to any one of clauses 38 to 39, or an expression host according to any one of clauses 40 to 41, or a composition according to any one of clauses 60 to 104, for use in inducing an immune response against PCV3 in a pig. 119. A protein according to any one of clauses 1 to 36, or a nucleotide sequence according to clause 37, or an expression vector according to any one of clauses 38 to 39, or an expression host according to any one of clauses 40 to 41, or a composition according to any one of clauses 60 to 104, for use in inducing an immune response against PCV3 in piglets. 120. A protein according to any one of clauses 1 to 36, or a nucleotide sequence according to clause 37, or an expression vector according to any one of clauses 38 to 39, or an expression host according to any one of clauses 40 to 41, or a composition according to any one of clauses 60 to 104, for use in inducing an immune response against PCV3 in piglets suckled by a sow to which the protein according to any one of clauses 1 to 36, or the nucleotide sequence according to clause 37, or the expression vector according to any one of clauses 38 to 39, or the expression host according to any one of clauses 40 to 41, or the composition according to any one of clauses 60 to 104 has been administered. 121. A protein according to any one of clauses 1 to 36, or a nucleotide sequence according to clause 37, or an expression vector according to any one of clauses 38 to 39, or an expression host according to any one of clauses 40 to 41, or a composition according to any one of clauses 60 to 104, for use in inducing an immune response against PCV3 in a sow. 122. A protein according to any one of clauses 1 to 36, or a nucleotide sequence according to clause 37, or an expression vector according to any one of clauses 38 to 39, or an expression host according to any one of clauses 40 to 41, or a composition according to any one of clauses 60 to 104, for use in inducing an immune response against PCV3 in a pregnant pig, a gilt, or a pre-breeding gilt. 123. A protein according to any one of clauses 1 to 36, or a nucleotide sequence according to clause 37, or an expression vector according to any one of clauses 38 to 39, or an expression host according to any one of clauses 40 to 41, or a composition according to any one of clauses 60 to 104, for use in a method for alleviating or preventing clinical signs or symptoms or disease caused by infection with PCV3 in an animal, or for use in a method for treating or preventing infection with PCV3 in an animal. 124. A protein according to any one of clauses 1 to 36, or a nucleotide sequence according to clause 37, or an expression vector according to any one of clauses 38 to 39, or an expression host according to any one of clauses 40 to 41, or a composition according to any one of clauses 60 to 104, for use in a method for reducing or preventing clinical signs or symptoms or disease caused by infection with PCV3 in an animal that is a swine, or for use in a method for treating or preventing infection with PCV3 in an animal. 125. A protein according to any one of clauses 1 to 36, or a nucleotide sequence according to clause 37, or an expression vector according to any one of clauses 38 to 39, or an expression host according to any one of clauses 40 to 41, or a composition according to any one of clauses 60 to 104, for use in a method for reducing or preventing clinical signs or symptoms or disease caused by infection with PCV3 in an animal that is a swine, or for use in a method for treating or preventing infection with PCV3 in an animal. 126. A protein according to any one of clauses 1 to 36, or a nucleotide sequence according to clause 37, or an expression vector according to any one of clauses 38 to 39, or an expression host according to any one of clauses 40 to 41, or a composition according to any one of clauses 60 to 104, for use in a method for reducing or preventing clinical signs or symptoms or disease caused by infection with PCV3 in an animal, which is a piglet, or for use in a method for treating or preventing infection...

Claims

1. A method for preventing PCV3 viremia in piglets, comprising administering a composition to a gilt or sow before it becomes pregnant with piglets, allowing the gilt or sow to become pregnant with the piglets, and allowing the pregnant gilt or sow to give birth to the piglets, wherein the composition comprises a PCV3 ORF2 protein having an amino acid sequence having at least 97% sequence identity to SEQ ID NO:4, the PCV3 ORF2 protein being PCV3 ORF2 protein expressed by a recombinant baculovirus, and the administration to the pig is before mating / insemination of the pig.

2. 10. The method of claim 1, wherein the administration comprises a multiple shot or multiple dose regimen of the composition, or wherein the administration comprises a two shot or two dose regimen of the composition.

3. 3. The method of claim 1 or 2, comprising allowing the sow to suckle the piglets.

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