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Title: Interaction of Bordetella bronchiseptica and different Bordetella factors with sheep bone marrow mast cells and other cell types
Author: Al-Turkestany, Ismail M. A.
ISNI:       0000 0001 3409 2810
Awarding Body: University of Glasgow
Current Institution: University of Glasgow
Date of Award: 2006
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A number of studies have reported that mast cells (MCs) play a critical role in host immune defence against bacterial infections. These studies have shown that mast cells have the ability to recognize and engulf bacteria. This project aimed to investigate the effect of MCs on selected microbial pathogens and the effect of these pathogen and their products on MCs and other cell types (RBL-2H3 and J774A.1 cell lines). The phagocytic activity of sheep BMMC was studied by using bioluminescent strains of B. bronchiseptica and E. coli, and degranulation was followed by estimating beta-hexosaminidase release. The bioluminescent method is based on the assumption that the level of light output from bacteria is directly proportional to the number of viable bacterial cells. Comparison between light output of bioluminescent E. coli and B. bronchiseptica (Lux 95) in a short-run experiment (< 20 h) suggested that the correlation between bioluminescence output and optical density of the bioluminescent E. coli was not as good as with B. bronchiseptica (Lux 95). The difference is probably due to the fact that E. coli is a much more rapidly growing organism than B. bronchiseptica and will therefore reach stationary phase in a much shorter time. A problem encountered in this study with B. bronchiseptica (Lux 95) was that it had been derived from a laboratory strain that may have partially lost its virulence attributes. Comparison of B. bronchiseptica (Lux 95) strain with B. bronchiseptica (Bvg+) and B. bronchiseptica (Bvg-) strains revealed that it had more properties in common with the Bvg- than the Bvg+ mode and may not, therefore, have been an ideal strain for this study. Construction of further bioluminescent derivatives of a wild-type (Byg+) and avirulent (Bvg-) strains should be considered for future work. The bioluminescent method however, was shown to offer an easy and simple method of measuring the viability of the bacteria in real-time. Based on in vitro invasion assays, the kinetics of interaction between sheep BMMC and B. bronchiseptica (Lux 95) was followed. The data indicated for the first time, the presence of viable bioluminescent B. bronchiseptica within sheep BMMC, and this observation was confirmed by both viable count and TEM. For the first time, viable intracellular B. bronchiseptica (Lux 95) were shown within sheep BMMC. These results establish baseline data for individual and age-related variation in sheep, BMMC phagocytic function, and form a basis for further evaluation of other functions such as chemotaxis, oxidative burst and clearance of bacterial infection by mast cells, either in sheep or other animal models.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral