Anti-β2glycoprotein I-induced neutrophil extracellular traps cause endothelial activation.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 40238197.
- Also identified by DOI 10.1093/rheumatology/keaf204 and PMC identifier 12316379.
- Licence recorded as CC BY-NC.
- Because redistribution is not established, this page shows the abstract only. Follow the links below for the full text.
Abstract
Neutrophil extracellular traps (NETs) involvement in antiphospholipid syndrome (APS) pathogenesis is known, but the role of anti-β2glycoprotein I (aβ2GPI) antibodies-induced NETs in triggering a procoagulant and proinflammatory phenotype in endothelial cells (ECs) remains to be evaluated. This study investigated whether NET-aβ2GPI can activate ECs and whether NET-aβ2GPI and NET-phorbol myristate acetate (PMA) have different proteomic profiles. Healthy donor (HD) neutrophils were stimulated with APS-aβ2GPI, normal human IgG or PMA. NETs were stained with anti-neutrophil elastase and 4'6-diamidino-2-phenylindole (DAPI), and the ability of aβ2GPI to bind NETs and inhibit DNA degradation was investigated. Following aβ2GPI, NET-aβ2GPI and NET-PMA stimuli, we evaluated EC activation investigating intra-cellular adhesion molecule (ICAM), vascular cell adhesion molecule (VCAM) and tissue factor (TF) expression using flow cytometry and reverse transcription-quantitative polymerase chain reaction (RT-qPCR); and EC dysfunction analysing extracellular microvesicles (EMVs) release via flow cytometry and NanoSight analysis. Mass spectrometry-based proteomics was performed on NET-aβ2GPI and NET-PMA. Unlike normal IgG, aβ2GPI induced NET formation and bound to NETs by colocalizing with the neutrophil elastase signal at 93.6% without preventing NET degradation. Compared with unstimulated EC, NET-aβ2GPI triggered higher mRNA and a robust expression of TF, VCAM and ICAM in EC with a change-fold median fluorescence intensity (MFI) of 6, 4.2 and 2.3. aβ2GPI induced a significant increase in EMVs compared with untreated samples and those treated with NETs. Fifty-six proteins were identified, seven resulted upregulated in NET-aβ2GPI and downregulated in PMA-induced ones. GO enrichment analysis revealed that proteins upregulated in NET-aβ2GPI were enriched for ubiquitin protein ligase binding and SLC2A4 translocation to the plasma membrane. aβ2GPI-induced NETs can cause EC activation and TF expression.
Medical subject headings
- Extracellular Traps
- beta 2-Glycoprotein I
- Neutrophils
- Antiphospholipid Syndrome
- Endothelial Cells