Human neutrophil-derived extracellular vesicles induce renal endothelial inflammation in critical illness: an ex vivo investigation
File(s)
Accepted version
Author(s)
Type
Journal Article
Abstract
Background
Circulating neutrophil-derived extracellular vesicles (NEVs) may contribute to the pathophysiology of acute kidney injury by causing glomerular endothelial inflammation.
Methods
NEVs were first isolated from ex vivo, lipopolysaccharide stimulation of whole blood taken from healthy volunteers (median age [interquartile range {IQR}]: 32 [26–42] yr; 47% female), and also from plasma of COVID-19 patients with acute respiratory distress syndrome (median age [IQR]: 59 [52–66] yr; 45% female). NEVs were incubated for 4 h in a co-culture of peripheral blood mononuclear cells and either human umbilical vein endothelial cells or renal glomerular endothelial cells. Enzyme-linked immunoassays (tumour necrosis factor-alpha [TNF]) and flow cytometry (median fluorescence intensity [MFI]) were used to quantify cell-specific markers of inflammation/activation, in the presence/absence of pharmacological inhibitors.
Results
NEVs were internalised by monocytes, leading to their activation via the p38 mitogen-activated protein kinase pathway and increased release of TNF (median [IQR]: 676 [474–1731] pg ml−1 after NEV internalisation, compared with controls (27 [18–29] pg ml−1, P=0.003). This proinflammatory response increased cell adhesion molecule expression (E-selectin) on human umbilical vein endothelial cells (median MFI [IQR]; NEVs: 4120 [3671–4858] vs untreated: 1438 [1252–1708], P=0.008) and human renal glomerular endothelial cells (median MFI [IQR]; NEVs: 2960 [2471–4991] vs untreated: 931 [881–1181], P=0.003). NEVs contained substantial amounts of matrix metalloproteinase-8 and -9, inhibitors of which (doxycycline, MMP-8 inhibitor or MMP-9 inhibitor) prevented endothelial cell inflammation.
Conclusions
Circulating NEVs may contribute to acute kidney injury through renal endothelial inflammation in a monocyte-dependent fashion in patients with acute respiratory distress syndrome.
Circulating neutrophil-derived extracellular vesicles (NEVs) may contribute to the pathophysiology of acute kidney injury by causing glomerular endothelial inflammation.
Methods
NEVs were first isolated from ex vivo, lipopolysaccharide stimulation of whole blood taken from healthy volunteers (median age [interquartile range {IQR}]: 32 [26–42] yr; 47% female), and also from plasma of COVID-19 patients with acute respiratory distress syndrome (median age [IQR]: 59 [52–66] yr; 45% female). NEVs were incubated for 4 h in a co-culture of peripheral blood mononuclear cells and either human umbilical vein endothelial cells or renal glomerular endothelial cells. Enzyme-linked immunoassays (tumour necrosis factor-alpha [TNF]) and flow cytometry (median fluorescence intensity [MFI]) were used to quantify cell-specific markers of inflammation/activation, in the presence/absence of pharmacological inhibitors.
Results
NEVs were internalised by monocytes, leading to their activation via the p38 mitogen-activated protein kinase pathway and increased release of TNF (median [IQR]: 676 [474–1731] pg ml−1 after NEV internalisation, compared with controls (27 [18–29] pg ml−1, P=0.003). This proinflammatory response increased cell adhesion molecule expression (E-selectin) on human umbilical vein endothelial cells (median MFI [IQR]; NEVs: 4120 [3671–4858] vs untreated: 1438 [1252–1708], P=0.008) and human renal glomerular endothelial cells (median MFI [IQR]; NEVs: 2960 [2471–4991] vs untreated: 931 [881–1181], P=0.003). NEVs contained substantial amounts of matrix metalloproteinase-8 and -9, inhibitors of which (doxycycline, MMP-8 inhibitor or MMP-9 inhibitor) prevented endothelial cell inflammation.
Conclusions
Circulating NEVs may contribute to acute kidney injury through renal endothelial inflammation in a monocyte-dependent fashion in patients with acute respiratory distress syndrome.
Date Issued
2025-10-01
Date Acceptance
2025-06-07
Citation
British Journal of Anaesthesia, 2025, 135 (4), pp.920-929
ISSN
0007-0912
Publisher
Elsevier
Start Page
920
End Page
929
Journal / Book Title
British Journal of Anaesthesia
Volume
135
Issue
4
Copyright Statement
Copyright © 2025 Published by Elsevier Ltd on behalf of British Journal of Anaesthesia. This is the author’s accepted manuscript made available under a CC-BY licence in accordance with Imperial’s Research Publications Open Access policy (www.imperial.ac.uk/oa-policy)
License URL
Identifier
https://www.ncbi.nlm.nih.gov/pubmed/40877108
Subjects
COVID-19
critical illness
extracellular vesicle
matrix metalloproteinase (MMP)
renal glomerular inflammation
Publication Status
Published
Coverage Spatial
England
Date Publish Online
2025-08-27