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Title: Functional characterisation of novel WFDC proteins
Author: Glasgow, Arlene Mina Ann
Awarding Body: Queen's University Belfast
Current Institution: Queen's University Belfast
Date of Award: 2013
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SLPI and elafin are members of the whey acidic protein (WAP) four disulphide core (WFDC} family of proteins and have multiple contributions to innate host defence including inhibition of neutrophil serine proteases, antimicrobial properties and inhibition of the inflammatory response to LPS. This research aimed to investigate novel functions of WAP2 and eppin, two lesser characterised WFDC proteins that are found expressed in the lung. Recombinant expression and purification of WAP2 were first optimised in E. coli for use in a range of functional assays. Recombinant WAP2 inhibited cathepsin G protease activity. Monocytic cells pre-treated with recombinant WAP2 before LPS stimulation produced significantly lower levels of IL-8 and MCP-l compared to cells stimulated with LPS alone. Recombinant WAP2 became conjugated to fibronectin in a transglutaminase-mediated reaction and retained antiprotease activity. WAP2 was detected at variable levels in bronchoalveolar lavage fluid from cystic fibrosis patients. WAP2 and eppin were both shown by immunohistochemistry to be expressed in epithelial and immune cells of the lung. Eppin exerted potent antibacterial effects against the lung-colonising bacteria P. aeruginosa and S. aureus, in contrast to WAP2 which was ineffective. Together these results suggest a role for both WAP2 and eppin in the innate host response. Further characterisation is required to fully understand the range and implications of their physiological functions.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available