Use this URL to cite or link to this record in EThOS: https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.572403
Title: Investigating interleukin-1β² processing, secretion and targeting in a zebrafish model
Author: Ogryzko, Nikolay
ISNI:       0000 0004 2737 9145
Awarding Body: University of Sheffield
Current Institution: University of Sheffield
Date of Award: 2013
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Abstract:
Uncontrolled inflammation contributes to a number of autoinflammatory pathological conditions, and is a significant factor in a number of other areas such as atherosclerosis the leading cause of death in the developed world. The inflammatory response is very tightly regulated, and controlled at its apex by Interleukin-1β, a key proinflammatory cytokine. This key initiator provides an ideal target for therapeutic intervention, yet many aspects of its biology are poorly understood, including its novel release mechanism. Mammalian in vitro studies have demonstrated IL-1β is processed by a protein complex called the inflammasome and that this processing is linked to release via membrane-derived microvesicles from the cell surface. However, it has not yet been possible to show the functional release of IL-1β microvesicles in vitro, determine the mechanism of release and how it is linked to processing and the biological function of these vesicles upon downstream cells. We hypothesise that components of the ESCRT complex are responsible for IL-1β secretion; however, the tools to test this hypothesis in vitro have not yet been developed. IL-1β structure is highly conserved between zebrafish and humans based on structure prediction software, which shows zebrafish IL-1β shares the same secondary and tertiary structural elements. Zebrafish IL-1β mRNA is induced in response to injury and the inflammatory response is downregulated in response to treatment with IL-1β pathway inhibitors. To investigate IL-1β processing in vivo, I have tested a FRET based reporter for caspase-3 activation and characterised the use of such a reporter in the zebrafish as a proof of concept. I have also designed a version of this reporter specific to caspase-1 activation. Alongside the work on an inflammasome reporter, I also show the first in vivo evidence of IL-1β secretion from inflammatory cells in response to injury using a macrophage specific IL-1β fusion reporter. In this thesis I demonstrate the utility of the zebrafish as a model of IL-1β biology with regards to induction of IL-1β signaling and the susceptibility of the zebrafish innate immue response to treatment with IL-1β pathway inhibitors, and I present the first evidence of putative IL-1β containing microvesicle release from macrophages in vivo.
Supervisor: Wilson, H. L. Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID: uk.bl.ethos.572403  DOI: Not available
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