US20080181910A1 - Use of a pcv2 immunogenic composition for lessening clinical symptoms in pigs - Google Patents
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- US20080181910A1 US20080181910A1 US11/617,435 US61743506A US2008181910A1 US 20080181910 A1 US20080181910 A1 US 20080181910A1 US 61743506 A US61743506 A US 61743506A US 2008181910 A1 US2008181910 A1 US 2008181910A1
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Definitions
- the present invention also relates to a method for the treatment of Pia like lesions, normally known to be associated with Lawsonia intracellularis infections, said method comprising the administration of an immunogenic composition comprising PCV2 antigen, preferably recombinant PCV2 antigen and more preferably PCV2 ORF2 protein as provided herein, to a pig.
- said treatment results in the lessening or reduction of the Pia like lesions, normally known to be associated with Lawsonia intracellularis infections.
- an “immunogenic composition” as used herein means a PCV2 ORF2 protein which elicits an “immunological response” in the host of a cellular and/or antibody-mediated immune response to PCV2 ORF2 protein.
- this immunogenic composition is capable of eliciting or enhancing an immune response against PCV2 thereby conferring protective immunity against PCV2 infection and a reduction in the incidence of, severity of, or prevention of one or more, and preferably all of the clinical signs associated therewith.
- Sequence Identity refers to a relationship between two or more polypeptide sequences or two or more polynucleotide sequences, namely a reference sequence and a given sequence to be compared with the reference sequence. Sequence identity is determined by comparing the given sequence to the reference sequence after the sequences have been optimally aligned to produce the highest degree of sequence similarity, as determined by the match between strings of such sequences. Upon such alignment, sequence identity is ascertained on a position-by-position basis, e.g., the sequences are “identical” at a particular position if at that position, the nucleotides or amino acid residues are identical.
- BLASTX program is publicly available from NCBI and other sources (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 in order to produce the highest level of sequence identity between the given and reference sequences.
- mutations of the reference sequence may occur at the 5′ or 3′ terminal positions of the reference nucleotide sequence or anywhere between those terminal positions, interspersed either individually among nucleotides in the reference sequence or in one or more contiguous groups within the reference sequence.
- a polypeptide having a given amino acid sequence having at least, for example, 85%, preferably 90%, 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 include up to 15, preferably up to 10, even more preferably up to 5 amino acid alterations per each 100 amino acids of the reference amino acid sequence.
- Sequence homology refers to a method of determining the relatedness of 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 a match when determining sequence homology.
- the immunogenic composition as used herein also refers to a composition that comprises i) any of the PCV2 ORF2 protein described above, preferably in concentrations described above, and ii) at least a portion of the viral vector expressing said PCV2 ORF2 protein, preferably of a recombinant baculovirus.
- the immunogenic composition can comprise i) any of the PCV2 ORF2 proteins described above, preferably in concentrations described above, ii) at least a portion of the viral vector expressing said PCV2 ORF2 protein, preferably of a recombinant baculovirus, and iii) a portion of the cell culture supernate.
- 2,909,462 which describes such acrylic polymers cross-linked with a polyhydroxylated compound having at least 3 hydroxyl groups, preferably not more than 8, the hydrogen atoms of at least three hydroxyls being replaced by unsaturated aliphatic radicals having at least 2 carbon atoms.
- the preferred radicals are those containing from 2 to 4 carbon atoms, e.g. vinyls, alkyls and other ethylenically unsaturated groups.
- the unsaturated radicals may themselves contain other substituents, such as methyl.
- the products sold under the name Carbopol; (BF Goodrich, Ohio, USA) are particularly appropriate. They are cross-linked with an allyl sucrose or with allyl pentaerythritol.
- the present invention also relates to an immunogenic composition that comprises i) any of the PCV2 ORF2 proteins described above, preferably in concentrations described above, ii) at least a portion of the viral vector expressing said PCV2 ORF2 protein, iii) a portion of the cell culture, iv) an inactivating agent to inactivate the recombinant viral vector preferably BEI, and v) an neutralization agent to stop the inactivation mediated by the inactivating agent, preferably sodium thiosulfate in equivalent amounts to BEI; and vi) a suitable adjuvant, preferably Carbopol 971 in amounts described above; wherein about 90% of the components i) to iii) have a size smaller than 1 ⁇ m.
- this immunogenic composition further comprises a pharmaceutical acceptable salt, preferably a phosphate salt in physiologically acceptable concentrations.
- the pH of said immunogenic composition is adjusted to a physiological pH, meaning between about 6.5 and 7.5.
- the immunogenic compositions can further include one or more other immunomodulatory agents such as, e.g., interleukins, interferons, or other cytokines.
- the immunogenic compositions can also include Gentamicin and Merthiolate. While the amounts and concentrations of adjuvants and additives useful in the context of the present invention can readily be determined by the skilled artisan, the present invention contemplates compositions comprising from about 50 ⁇ g to about 2000 ⁇ g of adjuvant and preferably about 250 ⁇ g/ml dose of the vaccine composition.
- the immunogenic composition as used herein also refers to a composition that comprises from about 1 ug/ml to about 60 ⁇ g/ml of antibiotics, and more preferably less than about 30 ⁇ g/ml of antibiotics.
- FIGS. 2 a and 2 b are each schematic flow diagrams of how to produce one of the compositions used in accordance with the present invention.
- a blocking solution (5 g Carnation Non-fat dry milk (Nestle, Glendale, Calif.) in 10 mL of D-PBS QS to 100 mL with distilled water) was added to each of the wells.
- the next step was to wash the test plate and then add pre-diluted antigen.
- the pre-diluted antigen was produced by adding 200 ⁇ L of diluent solution (0.5 mL Tween 20 in 999.5 mL D-PBS) to each of the wells on a dilution plate.
- the flask was then incubated at 27° C. for a total of 6 days. After incubation, the flask was then centrifuged and three samples of the resulting supernatant were harvested and inactivated. The supernatant was inactivated by bringing its temperature to 37 ⁇ 2° C.
- a 0.4M solution of 2-bromoethyleneamine hydrobromide which had been cyclized to 0.2M binary ethylenimine (BEI) in 0.3N NaOH was added to the supernatant to give a final concentration of BEI of 5 mM.
- BEI binary ethylenimine
- no BEI was added to the supernatant.
- nasal shedding of PCV2 was assessed as an efficacy parameter and the previous PCV2 efficacy parameters from previous studies were reconfirmed. Results from this study indicate that nasal shedding of PCV2 occurs following intra nasal challenge and that PCV2 vaccines reduce nasal shedding of PCV2 post-challenge. Furthermore, results from this study and reports in the literature indicate that IHC should continue to be evaluated in future PCV2 vaccine trials as well.
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Priority Applications (12)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US11/617,435 US20080181910A1 (en) | 2005-12-29 | 2006-12-28 | Use of a pcv2 immunogenic composition for lessening clinical symptoms in pigs |
US12/137,067 US7838214B2 (en) | 2005-12-29 | 2008-06-11 | Use of a PCV2 immunogenic composition for lessening clinical symptoms in pigs |
US12/137,086 US20080267995A1 (en) | 2005-12-29 | 2008-06-11 | Use of a pcv2 immunogenic composition for lessening clinical symptoms in pigs |
US12/136,942 US7838213B2 (en) | 2005-12-29 | 2008-06-11 | Use of a PCV2 immunogenic composition for lessening clinical symptoms in pigs |
US12/137,105 US20080279889A1 (en) | 2005-12-29 | 2008-06-11 | Use of a pcv2 immunogenic composition for lessening clinical symptoms in pigs |
US12/137,141 US7829273B2 (en) | 2005-12-29 | 2008-06-11 | Use of a PCV2 immunogenic composition for lessening clinical symptoms in pigs |
US12/136,923 US7968285B2 (en) | 2005-12-29 | 2008-06-11 | Use of a PCV2 immunogenic composition for lessening clinical symptoms in pigs |
US13/106,606 US9011868B2 (en) | 2005-12-29 | 2011-05-12 | Use of a PCV2 immunogenic composition for lessening clinical symptoms in pigs |
US14/661,969 US9669087B2 (en) | 2005-12-29 | 2015-03-18 | Use of a PCV2 immunogenic composition for lessening clinical symptoms in pigs |
US15/076,381 US9987349B2 (en) | 2005-12-29 | 2016-03-21 | Use of a PCV2 immunogenic composition for lessening clinical symptoms in pigs |
US15/249,158 US9610345B2 (en) | 2005-12-29 | 2016-08-26 | Use of a PCV2 immunogenic composition for lessening clinical symptoms in pigs |
US15/957,663 US10568955B2 (en) | 2005-12-29 | 2018-04-19 | Use of a PCV2 immunogenic composition for lessening clinical symptoms in pigs |
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US75501605P | 2005-12-29 | 2005-12-29 | |
US82980906P | 2006-10-17 | 2006-10-17 | |
US11/617,435 US20080181910A1 (en) | 2005-12-29 | 2006-12-28 | Use of a pcv2 immunogenic composition for lessening clinical symptoms in pigs |
Related Child Applications (6)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US12/136,923 Division US7968285B2 (en) | 2005-12-29 | 2008-06-11 | Use of a PCV2 immunogenic composition for lessening clinical symptoms in pigs |
US12/137,067 Division US7838214B2 (en) | 2005-12-29 | 2008-06-11 | Use of a PCV2 immunogenic composition for lessening clinical symptoms in pigs |
US12/136,942 Division US7838213B2 (en) | 2005-12-29 | 2008-06-11 | Use of a PCV2 immunogenic composition for lessening clinical symptoms in pigs |
US12/137,086 Division US20080267995A1 (en) | 2005-12-29 | 2008-06-11 | Use of a pcv2 immunogenic composition for lessening clinical symptoms in pigs |
US12/137,105 Division US20080279889A1 (en) | 2005-12-29 | 2008-06-11 | Use of a pcv2 immunogenic composition for lessening clinical symptoms in pigs |
US12/137,141 Division US7829273B2 (en) | 2005-12-29 | 2008-06-11 | Use of a PCV2 immunogenic composition for lessening clinical symptoms in pigs |
Publications (1)
Publication Number | Publication Date |
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US20080181910A1 true US20080181910A1 (en) | 2008-07-31 |
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Family Applications (12)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US11/617,435 Abandoned US20080181910A1 (en) | 2005-12-29 | 2006-12-28 | Use of a pcv2 immunogenic composition for lessening clinical symptoms in pigs |
US12/137,086 Abandoned US20080267995A1 (en) | 2005-12-29 | 2008-06-11 | Use of a pcv2 immunogenic composition for lessening clinical symptoms in pigs |
US12/136,923 Active US7968285B2 (en) | 2005-12-29 | 2008-06-11 | Use of a PCV2 immunogenic composition for lessening clinical symptoms in pigs |
US12/137,105 Abandoned US20080279889A1 (en) | 2005-12-29 | 2008-06-11 | Use of a pcv2 immunogenic composition for lessening clinical symptoms in pigs |
US12/136,942 Active US7838213B2 (en) | 2005-12-29 | 2008-06-11 | Use of a PCV2 immunogenic composition for lessening clinical symptoms in pigs |
US12/137,141 Active US7829273B2 (en) | 2005-12-29 | 2008-06-11 | Use of a PCV2 immunogenic composition for lessening clinical symptoms in pigs |
US12/137,067 Active US7838214B2 (en) | 2005-12-29 | 2008-06-11 | Use of a PCV2 immunogenic composition for lessening clinical symptoms in pigs |
US13/106,606 Active US9011868B2 (en) | 2005-12-29 | 2011-05-12 | Use of a PCV2 immunogenic composition for lessening clinical symptoms in pigs |
US14/661,969 Active US9669087B2 (en) | 2005-12-29 | 2015-03-18 | Use of a PCV2 immunogenic composition for lessening clinical symptoms in pigs |
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