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Conserved inner core lipopolysaccharide epitopes as multi-species vaccine candidates

A lipopolysaccharide and conservative technology, applied in the direction of multivalent vaccines, vaccines, sugar derivatives, etc., can solve the problems that a single component cannot provide protection and has no bactericidal effect

Inactive Publication Date: 2007-08-08
NAT RES COUNCIL OF CANADA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Additionally, immunization of mice with the LPS-protein complex provided 100% protection when mice were challenged with a homologous strain, however when used separately the single components of the complex did not confer protection
LPS-induced MAbs from Pm confer only partial protection in mouse models, although they are opsonophagocytic but not bactericidal in the presence of complement

Method used

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  • Conserved inner core lipopolysaccharide epitopes as multi-species vaccine candidates
  • Conserved inner core lipopolysaccharide epitopes as multi-species vaccine candidates
  • Conserved inner core lipopolysaccharide epitopes as multi-species vaccine candidates

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0174] This example forms the basis for the publication in Can. J. Chem. 80, 1715 (2002). Study Mandellium haemolyticus serotype A1. Analysis of the LPS of M. haemolytica serotype A1 showed that Hex 3 Hep 5 Kdo is the main core oligosaccharide component. This was determined by mild acid hydrolysis (1% HOAc, 100° C., 3 h) of LPS samples and 1D 1H NMR and FAB-MS analysis of the obtained oligosaccharide fractions. GLC-MS analysis of sugar alcohol acetate and 2-butyl glycoside derivatives of core oligosaccharides showed that the main components of core oligosaccharides contained D-glucose, D-galactose, L-D-heptose and DD- Heptose, its molar ratio is 2:1:3:2. The D-Gal residue was found to be the terminal non-reducing part from the methylation analysis, and in the presence of Hex 2 Hep 5 Kdo is missing in a minority of components of the core oligosaccharide component. The presence of Kdo in LPS was confirmed by colorimetric analysis.

[0175] The LPS of M. hemolyticus is fi...

Embodiment 2

[0229] This example forms Carbohydr.Res. 339 : Basis for publications in 1973 (2004). Actinobacillus pleuropneumoniae serotypes 5a, b, 2 and 1 were studied.

[0230] Sugar analysis of column-fractionated LPS showed glucose (Glc), galactose (Gal), N-acetyl-glucosamine (GlcNAc), D-glycerol-D-mannose-heptose (DD-Hep) and L- The respective ratios of glycerol-D-mannose-heptose (LD-Hep) are approximately 2:1.5:1:2:3. Core OS was purified by gel filtration chromatography of the acid hydrolyzate as described in Materials and methods to obtain a fraction enriched in core OS and relatively free of O-antigen, and subjected to sugar analysis, which revealed; Glc , Gal, DD-Hep, and LD-Hep in a ratio of 2:1:2:3. Since the capsular polysaccharides of serotype 5b contain N-acetyl-D-glucosamine and ketose residues, we suspected some capsular (CPS) adulteration of LPS, the absence of CPS in the fractionated core OS samples was due to Ketosidic bonds in CPS are sensitive to the acid hydrolys...

Embodiment 3

[0282] Example 3 This example formed the basis for a publication in Glycobiology 15:323 (2005). Studying the Pasteurella multocida strain Pm70

[0283] Carbohydrate analysis of purified LPS indicated that the respective ratios of glucose (Glc), galactose (Gal) and L-glycerol-D-mannose-heptose (LD-Hep) were approximately 4:2:3. Small amounts of N-acetyl-glucosamine (GlcNAc) and N-acetyl-galactosamine (GalNAc) were also identified, and, in contrast to other recently studied pathogens of mammals, no D-glycerol-D- Manna-heptose (DD-Hep) (Brisson, et al, 2002; St. Michael et al, 2004). GLC analysis of core oligosaccharide (OS) derived butyl-glycosides indicated that Glc, Gal and GalNAc residues were present in their D-isomer form.

[0284] O-deacylated LPS (LPS-OH) was prepared and fractionated by gel filtration chromatography and analyzed by CE-MS (Example 3, Table 1). In CE-MS analysis, a simple mass spectrum was observed with the main peak being a triple charged ion at m / z 11...

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Abstract

A conserved inner-core oligosaccharide epitope expressed on the lipopolysaccharide (LPS) of a range of disease causing pathogenic bacterial isolates, including Actinobacillus pleuropneumoniae (Ap), Mannheimia haemolytica (Mh) and Pasteurella multocida (Pm), is disclosed. Construction of a mutant bacterial strain exclusively expressing the conserved inner core OS epitope as a terminally exposed structure has allowed the identification, production and isolation of an inner core LPS which is common to all three organisms. Further provided are associated vaccines, antibodies raised against the conserved LPS inner core and glycoconjugates comprising the LPS inner core linked to an immunogenic carrier.

Description

technical field [0001] The present invention relates to lipopolysaccharide (LPS) of a veterinary bacterial pathogen comprising one or more epitopes of the endo-oligosaccharide portion of said LPS. The present invention identifies conserved epitopes expressed on the LPS of some pathogenic isolates of veterinary bacterial pathogens, including but not limited to Actinobacillus pleuropneumoniae (Ap), hemolytic Mannheimia haemolytica (Mh) and Pasteurella multocida (Pm). Background technique [0002] Bacterial pathogens are a large cause of economic loss in commercial farm operations, as well as of health problems in a very wide range of animal populations, including humans. The three most common bacterial pathogens associated with veterinary disease are Mannella hemolytica (Pasteurella haemolytica, Mh), Actinobacillus pleuropneumoniae (Ap), and Pasteurella multocida (Pm). Mh is mainly a pathogen of cattle and sheep, Ap is a pathogen of pigs, and Pm is a pathogen of multiple spe...

Claims

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Application Information

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IPC IPC(8): C12N1/20A61K31/739C07K16/12G01N33/569A61K39/102A61K39/02A61P31/04C12P19/04A61K47/48A61K39/116C08B37/00C07H13/04A61K39/00A61K39/05
CPCA61K39/05A61K39/116A61K39/0001A61K2039/522A61K2039/70C07H13/04A61K39/102A61P31/04
Inventor 安德鲁·考克斯詹姆斯·C.·理查兹
Owner NAT RES COUNCIL OF CANADA
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