Whole blood coagulation in an ex vivo thrombus is sufficient to induce clot neutrophils to adopt a myeloid-derived suppressor cell signature and shed soluble Lox-1.
Blood Clot
RNA
immunity
innate
myeloid-derived suppressor cells
polymorphonuclear neutrophils
sequence analysis
Journal
Journal of thrombosis and haemostasis : JTH
ISSN: 1538-7836
Titre abrégé: J Thromb Haemost
Pays: England
ID NLM: 101170508
Informations de publication
Date de publication:
20 Dec 2023
20 Dec 2023
Historique:
received:
12
07
2023
revised:
04
12
2023
accepted:
06
12
2023
medline:
23
12
2023
pubmed:
23
12
2023
entrez:
22
12
2023
Statut:
aheadofprint
Résumé
Blood clots are living tissues that release inflammatory mediators including IL-8/CXCL8 and MCP-1/CCL2. A deeper understanding of blood clots is needed to develop new therapies for pro-thrombotic disease states and regenerative medicine. Identify a common transcriptional shift in cultured blood clot leukocytes. Differential gene expression of whole blood and cultured clots (4h 37°C) was assessed by RNA sequencing (RNAseq), RT-PCR, proteomics, and histology (23 diverse healthy human donors). Cultured clot serum bioactivity was tested in endothelial barrier functional assays. All cultured clots developed a polymorphonuclear myeloid-derived suppressor cell (PMN-MDSC) signature, including upregulation of OLR1 (mRNA encoding lectin-like oxidized low density lipoprotein receptor 1, Lox-1), IL-8/CXCL8, CXCL2, CCL2, IL10, IL1A, SPP1, TREM1 and DUSP4/MKP. Lipopolysaccharide enhanced PMN-MDSC gene expression and specifically induced a Type II interferon response with IL-6 production. Lox-1 was specifically expressed by cultured clot CD15+ neutrophils. Cultured clot neutrophils, but not activated platelets, shed copious amounts of soluble Lox-1 (sLox-1) with a donor-dependent amplitude. sLox-1 shedding was enhanced by phorbol ester and suppressed by heparin and by beta-glycerol phosphate, a phosphatase inhibitor. Cultured clot serum significantly enhanced endothelial cell monolayer barrier function, consistent with a pro-resolving bioactivity. This study suggests that PMN-MDSC activation is part of the innate immune response to coagulation which may have a protective role in inflammation. The cultured blood clot is an innovative thrombus model that can be used to study both sterile and nonsterile inflammatory states and could be used as a personalized medicine tool for drug screening.
Sections du résumé
BACKGROUND
BACKGROUND
Blood clots are living tissues that release inflammatory mediators including IL-8/CXCL8 and MCP-1/CCL2. A deeper understanding of blood clots is needed to develop new therapies for pro-thrombotic disease states and regenerative medicine.
OBJECTIVES
OBJECTIVE
Identify a common transcriptional shift in cultured blood clot leukocytes.
METHODS
METHODS
Differential gene expression of whole blood and cultured clots (4h 37°C) was assessed by RNA sequencing (RNAseq), RT-PCR, proteomics, and histology (23 diverse healthy human donors). Cultured clot serum bioactivity was tested in endothelial barrier functional assays.
RESULTS
RESULTS
All cultured clots developed a polymorphonuclear myeloid-derived suppressor cell (PMN-MDSC) signature, including upregulation of OLR1 (mRNA encoding lectin-like oxidized low density lipoprotein receptor 1, Lox-1), IL-8/CXCL8, CXCL2, CCL2, IL10, IL1A, SPP1, TREM1 and DUSP4/MKP. Lipopolysaccharide enhanced PMN-MDSC gene expression and specifically induced a Type II interferon response with IL-6 production. Lox-1 was specifically expressed by cultured clot CD15+ neutrophils. Cultured clot neutrophils, but not activated platelets, shed copious amounts of soluble Lox-1 (sLox-1) with a donor-dependent amplitude. sLox-1 shedding was enhanced by phorbol ester and suppressed by heparin and by beta-glycerol phosphate, a phosphatase inhibitor. Cultured clot serum significantly enhanced endothelial cell monolayer barrier function, consistent with a pro-resolving bioactivity.
CONCLUSIONS
CONCLUSIONS
This study suggests that PMN-MDSC activation is part of the innate immune response to coagulation which may have a protective role in inflammation. The cultured blood clot is an innovative thrombus model that can be used to study both sterile and nonsterile inflammatory states and could be used as a personalized medicine tool for drug screening.
Identifiants
pubmed: 38135253
pii: S1538-7836(23)00912-1
doi: 10.1016/j.jtha.2023.12.014
pii:
doi:
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Informations de copyright
Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.