Projects per year
Methods: Blood neutrophils were isolated from CF patients, CF pigs and appropriate controls. Neutrophils were also obtained from CF patients before and after commencing Ivacaftor. Apoptosis was assessed by morphology and flow cytometry. NET formation was determined by fluorescent microscopy and DNA release assays. NET interaction with macrophages was examined by measuring by cytokine generation by ELISA and qRT-PCR.
Results: CF neutrophils live longer due to decreased apoptosis. This was observed in both CFTR null piglets and CF patients, and furthermore was reversed by ivacaftor (CFTR potentiator) in patients with gating (G551D) mutations. CF neutrophils formed more NETs and this was reversed by cyclin-dependent kinase inhibitor (CDKi) exposure. NETs provided a pro-inflammatory stimulus to macrophages, which was enhanced in CF.
Conclusions: CF neutrophils have a pro-survival phenotype that is associated with an absence of CFTR function and allows increased NET production, which can in turn induce inflammation. Augmenting neutrophil apoptosis in CF may allow more appropriate neutrophil disposal, decreasing NET formation and thus inflammation.
1/04/11 → 31/03/17
1/04/11 → 31/03/14