Aspergillus fumigatus is increasingly recognised as a cystic fibrosis (CF) pathogen with reported infection rates up to 50% and associated with increased hospitalisations and lung deterioration. Research investigating immune responses of CF phagocytes against A. fumigatus has been scarce and the role of the cystic fibrosis transmembrane conductance regulator (CFTR) protein in anti-Aspergillus immune responses is not known. The studies in this thesis aimed to explore innate immune responses by CF phagocytes against A. fumigatus. Peripheral blood mononuclear cells (PBMC), monocytes (MNC) and polymorphonuclear cells (PMN) were isolated from blood samples of CF patients with at least one copy of the F508del mutation. CF phagocytes efficiently cleared A. fumigatus similar to healthy controls. Microtubuleassociated proteins 1A/1B light chain 3B (LC3) expression was found to be attenuated in CF PMN and MNC, indicating a disbalance in the autophagy or LC3-associated phagocytosis pathway. Regarding inflammatory responses, it was found that upon phagocytosis a resting A. fumigatus conidia, CF phagocytes produce up to 4-fold more reactive oxygen species (ROS) compared to controls. The excessive ROS was then shown not to be necessary for adequate killing, suggesting the increased ROS to be redundant in the antifungal response. Patient metrics obtained from the clinic showed that the excessive ROS production correlated to exacerbations and lung function. Liquid chromatography-mass spectrometry (LC-MS) analysis revealed that CF PMN only express 20-25% of the 4 haemoglobin subunits compared to healthy controls. Combining the LC-MS data suggests that CF PMN are under hypoxic stress. In conclusion, the effective clearance of A. fumigatus by CF phagocytes comes at the cost of an excessive respiratory burst which correlates to disease severity. Our data indicate that CF PMN are under hypoxic stress while circulating in the blood stream, which is likely to contribute to the hyperinflammatory phenotype observed upon interaction with A. fumigatus.
| Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:752687 |
| Date | January 2018 |
| Creators | Brunel, Franz Shan |
| Contributors | Warris, Adilia ; Devereux, Graham |
| Publisher | University of Aberdeen |
| Source Sets | Ethos UK |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Source | http://digitool.abdn.ac.uk:80/webclient/DeliveryManager?pid=237839 |
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