The role of Interleukin-33 in the development of Idiopathic Pulmonary Fibrosis

Lead Research Organisation: University of Nottingham
Department Name: School of Medicine

Abstract

The student will stimulate primary lung fibroblasts with increasing concentrations of recombinant IL-33 and measure markers of fibrosis including pSmad2/3, alphaSMA, collagen gene and protein expression. They will compare the effect of IL-33 on fibroblasts from patients with and without IPF. They will stimulate human macrophages (PMA stimulated THP1 cells, blood derived monocytes and primary alveolar macrophages) with IL-33 and measure
macrophage polarization and profibrotic cytokines, including TGF-beta. The effect of IL-33 function blocking antibodies on markers of fibrosis and macrophage polarization from cells obtained from patients with IPF, and controls, will be assessed.
Aim 2: NHBE cells will be stimulated with poly I:C and recombinant GM-CSF individually, and in combination, to induce IL-33 expression in the presence of intracellular (Caspase 1, 2, and 7) and extracellular proteases (neutrophil elastase and cathepsin) known to cleave IL-33. Levels, and isoforms, of intracellular IL-33 will be assessed by quantitative PCR and immunoblot. Levels of extracellular IL-33 will be measured by ELISA. The effect of extracellular IL-33 on macrophage polarization will be assessed by culturing macrophages in the conditioned media from epithelial cells with anti-IL-33 blocking reagents. Due to the labile nature of IL-33 (Cohen et al, Submitted), co-culture experiments will be performed to confirm results. The effect of epithelial apoptosis on IL-33 expression the effect of staurosporin on IL-33 secretion will be assessed. The cellular co-localization of IL-33 and intracellular proteases in lung tissue from patients, and controls, will be assessed using paraffin sections. Subsequently the levels of IL-33 and its cleavage fragments will be measured by immunoblot from frozen tissue obtained from patients or controls.

Publications

10 25 50
 
Description Bursary to attend 16th ERS Lung Science Conference: Cell-matrix interactions in lung disease and regeneration
Amount € 600 (EUR)
Organisation European Respiratory Society (ERS) 
Sector Charity/Non Profit
Country United Kingdom
Start 03/2018 
End 03/2018
 
Description Cambridge Science Festival 2019 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Public/other audiences
Results and Impact Volunteered as part of AstraZeneca (Industrial sponsor) team at the Cambridge Science Festival 2019. The practical activity aimed to teach children about the importance of drug formulation. The activity was incredibly popular with very positive feedback. In particular, several parents reported that their child had an increased interest in science after doing the activity. Although the practical activity was aimed predominately at children the event sparked discussions with multiple parents about the drug development process. Many also asked about my PhD and the focus of my research.
Year(s) Of Engagement Activity 2019
 
Description Nottingham Festival of Science & Curiosity 2018 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact Part of the Respiratory Medicine team that volunteered at Nottingham Festival of Science & Curiosity 2018. Ran the DNA extraction part of the workshop. Feedback suggested that members of the public had learnt more about the techniques we frequently use during our research and were more interested in respiratory medicine.
Year(s) Of Engagement Activity 2018
 
Description University of Nottingham 'Wonder 2017' event 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Public/other audiences
Results and Impact Part of the team that delivered a range of Respiratory Medicine themed, interactive activities at the Wonder 2017 event at the University of Nottingham. Had many discussions with members of the public about my research and received positive feedback. Parents also reported that their children were more interested in respiratory biology after participating in our workshop/activities
Year(s) Of Engagement Activity 2017