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Fox, Jane (2011) Utility of the precision cut lung slice model to investigate airway smooth muscle contraction. PhD thesis, University of Nottingham.

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Abstract

Asthma is characterised by airway remodelling and an increase in airway resistance. A greater understanding of the mechanisms involved in airway inflammation and airway hyper-responsiveness (AHR) may highlight therapeutic opportunities for asthma. This study initially aimed to optimise the preparation of precision cut lung slices (PCLS) in mouse and pig to investigate the influence of calcium (Ca2+) homeostasis on airway smooth muscle (ASM) contraction as a prelude to human studies. The PCLS technique was then applied to a murine model of allergic airway disease to explore the inflammatory process and pathogenesis of airway hyper-reactivity in sensitised mice.

Initial experiments using murine and porcine airways validated the PCLS model and demonstrated the significance of release and refilling of Ca2+ from internal stores to induce and maintain an airway contraction. Results also highlight interesting species differences in agonist sensitivity, with the porcine system sharing similar pharmacology to human airways.

Using a murine model of allergic airway disease, agonist induced contractile responses in peripheral airways were measured in vitro using the PCLS technique. BALB/c mice underwent initial sensitisation by intraperitoneal administration of ovalbumin, receiving a 3 day challenge with aerosolised OVA l% (vlv), for varying periods of up to 3 weeks for acute, mid-chronic and chronic sensitisation protocols. To investigate the influence of the inflammatory environment, naive murine lung slices were incubated with selected inflammatory mediators. OVA sensitisation led to progressive structural remodelling and AHR to methacholine (MCh) challenge. However, this hyperresponsiveness was decreased 48 hours post lung removal. Of the inflammatory mediators selected for lung slice incubation, IL-33 significantly increased AHR to MCh. IL-33 is a proinflammatory cytokine with transcriptional repressor properties, playing a role in initiating the TH2 inflammatory response. In lung slices prepared from IL-33 receptor (ST2) KO mice IL-33 was unable to sensitise the contractile response. These data suggest the inflammatory environment promotes AHR and disassociates this airway sensitivity from structural remodelling. These data suggest a key role for IL-33 in mediating AHR in this murine model.

Investigation of the mechanisms involved in airway hyper-reactivity revealed mRNA expression of IL-33 and the IL-33 receptor (ST2) in soluble and membrane bound forms were significantly increased in the mid-chronic and chronic ovalbumin sensitised murine lung tissue. Further quantitative analysis in human lung showed expression of IL-33 in epithelial and ASM cells. The human ST2 receptor (also known as IL-IRL-l) was expressed in mast cells. Together these results suggest IL-33 is a sensor of tissue damage; indirectly inducing AHR through further inflammatory cell activation to target ASM. This study demonstrates IL-33's role as an inflammatory marker of asthma and suggests a novel therapeutic intervention by targeting of the ST2 receptor.

Item Type:	Thesis (University of Nottingham only) (PhD)
Supervisors:	Hall, I.P. Sayers, I.
Keywords:	Respiratory muscles, Smooth muscle, Inflammatory process, Airway hyper-responsiveness
Subjects:	W Medicine and related subjects (NLM Classification) > WF Respiratory system
Faculties/Schools:	UK Campuses > Faculty of Medicine and Health Sciences > School of Clinical Sciences
Item ID:	30380
Depositing User:	Blore, Mrs Kathryn
Date Deposited:	05 Oct 2015 09:45
Last Modified:	15 Oct 2017 21:48
URI:	https://eprints.nottingham.ac.uk/id/eprint/30380

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