A vaccine to protect a pig against actinobacillus pleuropneumoniae

Inactive Publication Date: 2021-03-18
INTERVET INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The patent describes a new vaccine that protects against a type of toxin called APP. The vaccine uses a modified virus called baculovirus, which is commonly used to infect insect cells. The vaccine was found to provide adequate protection, even at a level comparable to a commercial vaccine called Porcilis® APP. The patent also discusses the use of baculovirus to express recombinant proteins and the development of engineered baculovirus vectors to improve protein expression. Overall, the patent highlights the potential of baculovirus as a beneficial tool for vaccine development and protein expression.

Problems solved by technology

Although baculovirus expression as such is commonly known and has been used since many years to express immunogens of either viral or bacterial pathogens, it has not been used to provide an adequate vaccine against APP by expressing one or more RTX toxcins.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

nt Expression of ApxI

[0019]Construction of Transfervector pFastbac-ApxIA

[0020]The rApxIA gene was synthesized based on the ApxIA amino acid sequence of the Actinobacillus pleuropneumoniae strain 4074, Swiss prot accession number: P55128. The gene was codon optimized for baculovirus polyhedrin usage and a Kozak sequence (TATAAAT) and a 3′ hexahistidine tag were included. The rApxIA-His gene was cloned behind the polyhedrin promoter of plasmid pFastbac1 (Life Technologies, Carlsbad, USA) as a BamHI fragment, resulting in plasmid pFastbac-ApxIA TAT.

Generation of recombinant baculovirus BacdCCApxIA-TAT.

[0021]Recombinant baculovirus was generated using the plasmid as described here above in the Bac to Bac system (Life Technologies, Carlsbad, USA) according to manufacturer's protocol. E. coli cells used for transformation contained the parental baculovirus with a deletion of the chitinase and v-cathapsin genes (Kaba S A, Salcedo A M, Wafula P O, Vlak J M, van Oers M M. J Virol Methods. 20...

example 2

fficacy

Vaccine Formulation

[0024]Two different vaccines were made for the study. A first vaccine comprised purified baculovirus expressed ApxI as obtained with a method described under Example 1. A second vaccine for use as a positive control was comparable with the commercially available vaccine Porcilis® APP, comprising ApxI and ApxII purified from the culture supernatant of A. pleuropneumoniae (and thus complexed with LPS), also denoted as “native ApxI+ApxII”. The study vaccine different from the commercially available vaccine Porcilis® APP in that it did not contain the ApxIII toxin. However, for the challenge with a serotype 10 field isolate, this is not relevant (serotype 10 does not produce ApxIII). The antigens were mixed with a mineral oil-containing adjuvant (XSolve, available from MSD Animal Health, Boxmeer, The Netherlands) at a final concentration of 25 μg / ml for each antigen.

Vaccination Protocol

[0025]Three groups of eight piglets from an A. pleuropneumoniae free herd we...

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PUM

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Abstract

The present invention pertains to a vaccine to protect a pig against an infection with Actinobacillus pleuropneumoniae, the vaccine comprising an RTX toxin of Actinobacillus pleuropneumoniae recombinantly expressed by a baculovirus, and a pharmaceutically acceptable carrier.

Description

GENERAL FIELD OF THE INVENTION[0001]The invention in general pertains to a vaccine for protecting a pig against an infection with Actinobacillus pleuropneumoniae. BACKGROUND OF THE INVENTION[0002]The present invention pertains to a vaccine directed against porcine pleuropneumonia, a world-wide disease causing substantial economic loss to the swine industry. The etiological agent of porcine pleuropneumonia is Actinobacillus pleuropneumoniae (APP), a gram-negative bacterium belonging to the family Pasteurellaceae. The disease is transmitted by the aerosol route or direct contact with an infected pig, and is characterized by hemorrhagic, fibrinous and necrotic lung lesions. The clinical picture may range from peracute to chronic and asymptomatic carrier pigs can transmit the disease when introduced into uninfected herds. Based on capsular polysaccharides and lipopolysaccharide (LPS) O-chain components, 15 serovars have been described. Serotyping and other genetic typing methods for Act...

Claims

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Application Information

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IPC IPC(8): A61K39/102
CPCA61K39/102A61K2039/55544C12N15/866A61K2039/552A61P31/04A61K2039/54C07K14/285
Inventor WITVLIET, MAARTEN HENDRIKBIJLSMA, JOHANNA JACOBA ELISABETH
Owner INTERVET INC
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