Use this URL to cite or link to this record in EThOS: http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.659926
Title: Pulmonary colonisation of patients with cystic fibrosis by Pseudomonas aeruginosa
Author: Nelson, J. W.
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 1991
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Abstract:
Chronic respiratory colonisation by the adaptable opportunistic pathogen Pseudomonas aeruginosa is a major debilitating feature of the inherited disease cystic fibrosis (CF). This thesis considers various aspects of the pathogenesis of P.aeruginosa in CF, including the serological response to bacterial colonisation, and possible factors involved in early colonisation. Anti-P.aeruginosa lipopolysaccharide (LPS) antibodies in sera, saliva and sputa from patients with CF were measured by enzyme-linked immunosorbent assay (ELISA) incorporating either a polyvalent pseudomonas smooth LPS extract vaccine, or P.aeruginosa core, rough LPS. Elevated levels of anti-LPS IgG antibodies in serum, and IgA antibodies in saliva and sputum were demonstrated in patients chronically colonised by P.aeruginosa. Low levels of serum anti-LPS IgG antibodies were detected in some patients intermittently colonised by P.aeruginosa, but not in non-P.aeruginosa colonised patients. Anti-LPS IgA antibodies were detected in some of both intermittently and non-colonised patients. Immunoblot analysis of serum IgG and sputum IgA antibodies to P.aeruginosa LPS revealed a response directed towards O-antigenic LPS in the initial stages of pulmonary colonisation with non-mucoid P.aeruginosa and a response towards common core LPS during subsequent chronic infection with mucoid P.aeruginosa. Flagellar preparations from P.aeruginosa strains were characterised and used in ELISA and immunoblot studies to detect anti-P.aeruginosa flagellar antibodies in sera, saliva and sputum. Serum anti-flagellar IgG antibodies were detected, particularly in those CF patients intermittently or chronically colonised by P.aeruginosa. Antibodies to both type -a and -b flagella were detected; in some patients a pronounced antibody response to only one of the flagellar types was evident. Anti-P.aeruginosa LPS monoclonal antibodies (MAbs) were produced for use in a sandwich ELISA for the detection of P.aeruginosa in respiratory secretions of patients with CF. LPS defective mutants expressing only common core LPS were used to immunise mice for preparation of MAbs. Antibodies were screened in ELISA and the antigenic component(s) of LPS recognised by the most cross-reactive MAbs was checked by immunoblotting. Five IgG MAbs were characterised and found to recognise the core component of P.aeruginosa LPS. Two of the MAbs were particularly reactive against core LPS from all O-antigenic serotypes of P.aeruginosa and were included in the sandwich ELISA for detection of P.aeruginosa LPS. A biotin-streptavidin amplification system was used to increase assay sensitivity. The sensitivity of the assay was 0.1 ng/ml P.aeruginosa LPS; the assay was able to detect P.aeruginosa LPS in the respiratory secretions from patients with CF.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID: uk.bl.ethos.659926  DOI: Not available
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