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Title: Novel immunisation strategies against Salmonella
Author: Carpenter, Zoe Karen
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 2003
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This thesis presents two novel immunisation strategies against Salmonella enterica serovar Typhimurium (S. typhimurium) in the mouse model. Firstly, mice immunised with multiple antigens in the form of a DNA vaccine have been shown to develop specific humoral and cellular responses to proteins encoded within the vaccine. Secondly, it has been shown that immunisation with multiple cytosolic antigens (CA) of S. typhimurium SL1344, formulated with the adjuvant dimethyl dioctadecyl ammoniumbromide (DDA), induces strong humoral and cellular responses. These responses have been characterised, and their ability to confer protection on mice challenged with a lethal dose of S. typhimurium SL1344 has been investigated. The aim of DNA vaccination is to induce immune responses to protein antigens expressed in vivo by injection plasmid DNA encoding the antigen sequence. Expression library immunisation (ELI) is a new technique that draws on DNA immunisation and can be developed to identify the key antigens of a pathogen that confer protection. ELI has previously been used in a number of disease models in mice and has been used in this thesis as a means of inducing immune responses against an unspecified subset of bacterial antigens. An expression library (EL) of S. typhimurium SL1344 was constructed using a mammalian expression vector encoding EGFP. The expression of S. typhimurium SL1344 antigens forming fusion proteins with EGFP was visualised as green fluorescence in mouse fibroblast cells in vitro. The EL consisted of 140,000 clones of which 14,000 were used for immunisation. The library was administered to both BALB/c and CBA mice to examine the effect of mouse strain and was administered both intradermally (ID) and intramuscularly (IM) to examine the effect of immunisation route. Analysis by Western blot and ELISA showed that humoral responses were induced in both BALB/c and CBA mice. ID and IM inoculation produced similar results in BALB/c mice. Responses included a significant Thl component as judged by the presence of IgG2a in the serum and the secretion of IFN-γ when T cells from peripheral lymph nodes were stimulated by CA in culture.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available