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Preparation and Immunogenicity Analysis of a Lipopolysaccharide-protein Conjugate Vaccine for Haemophilus Parasuis

Author: RenHuiJun
Tutor: ZhouRui
School: Huazhong Agricultural University
Course: Preventive Veterinary Medicine
Keywords: Parasuis Lipopolysaccharide PilA protein Lipopolysaccharide - protein conjugate vaccine Immunogenicity
CLC: S858.28
Type: Master's thesis
Year: 2011
Downloads: 60
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Abstract


Parasuis (Haemophilus parasuis, HPS) is the presence of pathogens in the upper respiratory tract in normal pigs leather Glaser's disease (Glasser's disease), caused to the cellulose polyserositis inflammation, arthritis and meningitis is characterized by serious systemic disease. HPS at least 15 kinds of antigens and virulence of different serotypes, and its the vaccines lack of significant effect of cross-protective immunity. Lipopolysaccharide as a pathogen of a serotype-specific immunogen can stimulate the body to produce a protective immune response, but its immunogenicity is weak. If the polysaccharide and carrier protein is chemically bonded to form the conjugate vaccine can be greatly improved immunogenicity. Unlike other literature, this paper is not commonly used proteins, such as bovine serum albumin (BSA), tetanus toxoid, etc., instead of using the H. parasuis lipopolysaccharide this experiment real study found having good immunogenicity parasuis II secretion system IV pilus proteins PilA covalent coupling, to obtain a better immune effects and greater protection force. In this study, 68 ° C hot phenol water law from parasuis place four type 0322 strains and five type 0165 strain were extracted lipopolysaccharide (lipopolysaccharide, LPS). The main components of the reduction experiment, and UV spectroscopy to verify the extracted product i.e. reduction of the polysaccharide. Removed by acid hydrolysis with the toxicity of the lipid A portion, SephadexG-75 column The glucan gel chromatography preloaded condensate column purified lipopolysaccharide main structural units - O-polysaccharide side chains (O- specific polysaccharide, O-SP). Adipoyl dihydrazide (ADH) as a coupling agent, with the PilA protein covalently coupled lipopolysaccharide after activation of carbodiimide (EDCA), and then by the glucan gel chromatography preloaded condensate SephadexG-75 The column column purification, were prepared for parasuis strains OSP0322-PilA 0322 combined product, combined parasuis strains OSP0165-PilA 0165 products for both mixed product OSP0322-PilA OSP0165-PilA . The combination of these three products were immune females BalB / C mice, and do parasuis inactivated vaccine and adjuvant control group. Respectively in the first and the 14th day after the second immunization mice were tail lancing, OD630 value of serum antibody was detected by ELISA. Results show that immunization of the adjuvant control group, serum anti-LPS IgG antibodies while three groups OSP-PilA after immunization, the mouse serum were generated anti-LPS IgG antibodies, and three groups of OSP-PilA immune groups in the first 14 days after the second immunization IgG antibody than after the first immunization, the level of 14 days were significantly improved, and shows that the The OSP-PilA combination product has good immunogenicity. Were used in the first 14 days after the second immunization of 2 × LD50 and 5 × LD50 the HPS0322 and HPS0165 challenge experiments in mice, the results show OSP0322-PilA and OSP0165-PilA two groups combined product in the attack toxic dose of 5 × LD50 high performance protection force. Product group of mice mortality OSP0322-PilA OSP0165-PilA binding large do not have good protection.

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CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Animal Medicine ( Veterinary Medicine) > Livestock, poultry, wildlife diseases > Livestock > Pig
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