Pcv 2-based methods and compositions for the treatment of pigs

A porcine adenovirus and expression vector technology, applied in the field of recombinant expression vectors and porcine virus vectors, can solve problems such as failure to produce vaccines

Inactive Publication Date: 2012-01-25
VECTOGEN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, although several attempts have been made to elicit an appropriate protective immune response against PCVAD using the PCV2 ORF2 gene inserted into and expressed by a viral vector, none of these attempts have resulted in an industrially viable vaccine
It has been found that although ORF2 of PCV2 is a serological marker of associated disease, when PCV2 ORF2 is expressed by the viral host for vaccination purposes, such vaccines fail to elicit a sufficiently adequate immune response to protect pigs from disease

Method used

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  • Pcv 2-based methods and compositions for the treatment of pigs
  • Pcv 2-based methods and compositions for the treatment of pigs
  • Pcv 2-based methods and compositions for the treatment of pigs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0075] figure 1 Exemplary protocols for the preparation of recombinant viral vectors used herein are shown. The truncated PCV2 ORF2 gene was PCR amplified from the full-length PCV2 ORF2 gene cloned in a plasmid as template using 5' and 3' gene-specific primers. The 5' PCR primers were specifically designed to bind 127 bp downstream of the start of the PCV2 ORF2 gene (thus allowing deletion of the NLS) and introduce a signal sequence that was integrated in-frame at the 5' end of the final PCR product. To aid product cloning, the 5' and 3' primers also introduced BglII and HindIII restriction sites, respectively, into the final PCR product.

[0076] The PCR amplification product containing the truncated PCV2 ORF2 gene with signal sequence was then cloned into the BglII and HindIII sites of the expression cassette in the PAV3RHE plasmid. The recombinant PAV3RHE and PAV3LHE plasmids were then linearized with restriction enzymes that cut specifically within the plasmid backbone s...

Embodiment 2

[0081] In order to test the efficacy of the vaccine of the present invention, groups of piglets were given two doses of the following vaccines: a vaccine based on the modified PCV2 ORF2 described herein, or a vaccine comprising unmodified PCV2 ORF2, and the piglets' response to PCV2 challenge was determined. susceptibility. In addition, the modified vaccine was tested for its ability to induce neutralizing antibodies and confer protection when administered by the oral route.

[0082] This example describes a study to evaluate the protective effect of two doses of three different candidate recombinant porcine adenovirus serotype 3 vaccines in post-weaning piglets containing synthetically derived Consensus of open reading frame 2 from porcine circovirus 2 (PCV2). The parental recombinant was named rPAV-3PCV2mORF2. Inoculated piglets were challenged with the American Type Culture Collection (ATCC) PCV2 isolate TBA and measured for viremia (measured by virus isolation), body wei...

Embodiment 3

[0096] Embodiment 3: test data

[0097] A trial was performed to evaluate the protection conferred by vaccines based on modified PCV2 ORF2 in weaned piglets. In this trial, two doses of three different candidate recombinant porcine adenovirus serotype 3 vaccine candidates containing PCV2 ORF2 derived from a synthetic consensus sequence were used. The parental recombinant was named rPAV-3PCV2mORF2. The vaccinated piglets were challenged with PCV2 and the effect on viremia (as measured by virus isolation) and clinical signs was measured, followed by an assessment of protection.

[0098] The three vaccine candidates are:

[0099] (1) The full length of PAdV3-PCV2ORF2, wherein PCV2ORF 3 is not modified;

[0100] (2) PAdV3-PCV2ORF2 truncated form in which the PCV2ORF2 nuclear localization signal has been removed, and

[0101] (3) PAdV3-OCV2ORF2 secreted form, in which the NLS of PCV2ORF2 has been removed and replaced with a hydrophobic signal sequence and a cleavage site.

[0...

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Abstract

The present invention relates to methods and compositions for vaccinating pigs against porcine circovirus type 2 (PCV2) associated diseases. In particular, the invention relates to recombinant expression vectors allowing for secretion or cell membrane expression of a truncated form of the PCV2 open reading frame 2 (ORF2) protein.

Description

Background of the invention [0001] Porcine circoviruses (PCVs) are animal pathogens of the circoviridae family (circoviridae), and are some of the smallest viruses that replicate autonomously in mammalian cells. Virus particles are icosahedral, non-enveloped, and 17nm in diameter. There are two recently recognized types of PCV, porcine circovirus type I (PCV1) and porcine circovirus type II (PCV2). PCV1 is non-pathogenic, whereas PCV2 has been associated with a variety of diseases and syndromes, including but not limited to postweaning multisystemic wasting syndrome (PMWS), porcine dermatitis and nephrotic syndrome (PDNS), and congenital tremor, which These can be collectively referred to as porcine circovirus-associated disease (PCVAD). It is now recognized that the disease caused by PCV2 has a severe economic impact in many swine producing regions of the world. [0002] PMWS is of particular industrial importance, causing considerable levels of mortality in many herds and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/86A61K39/00
CPCA61K2039/53A61K2039/552A61K39/12C12N2750/10032C12N2750/10034A61P31/12A61P31/20C12N15/86C12N15/11
Inventor M.G.谢波德S.T.莱
Owner VECTOGEN
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