Comparison of assays measuring extracellular vesicle tissue factor in plasma samples: communication from the ISTH SSC Subcommittee on Vascular Biology

Bonifay, Amandine; Mackman, Nigel; Hisada, Yohei; Sachetto, Ana Teresa Azevedo; Hau, Chi; Gray, Elaine; Hogwood, John; Aharon, Anat; Badimon, Lina; Barile, Lucio; Baudar, Justine; Beckmann, Lennart; Benedikter, Birke; Bolis, Sara; Bouriche, Tarik; Brambilla, Marta; Burrello, Jacopo; Camera, Marina; Campello, Elena; Ettelaie, Camille; Faille, Dorothée; Featherby, Sophie; Franco, Corentin; Guldenpfennig, Maite; Hansen, John Bjarne; Judicone, Coralie; Kim, Yohan; Kristensen, Soren Risom; Laakmann, Katrin; Langer, Florian; Latysheva, Nadezhda; Lucien, Fabrice; de Menezes, Erika Marques; Mullier, François; Norris, Philip; Nybo, Jette; Orbe, Josune; Osterud, Bjarne; Paramo, Jose A; Radu, Claudia M; Roncal, Carmen; Samadi, Nazanin; Snir, Omri; Suades, Rosa; Wahlund, Casper; Chareyre, Corinne; Abdili, Evelyne; Martinod, Kimberly; Thaler, Johannes; Dignat-George, Françoise; Nieuwland, Rienk; Lacroix, Romaric

Abstract

Background: Scientific and clinical interest in extracellular vesicles (EVs) is growing. EVs that expose tissue factor (TF) bind factor VII/VIIa and can trigger coagulation. Highly procoagulant TF-exposing EVs are detectable in the circulation in various diseases, such as sepsis, COVID-19, or cancer. Many in-house and commercially available assays have been developed to measure EV-TF activity and antigen, but only a few studies have compared some of these assays. Objectives: The International Society on Thrombosis and Haemostasis Scientific and Standardization Committee Subcommittee on Vascular Biology initiated a multicenter study to compare the sensitivity, specificity, and reproducibility of these assays. Methods: Platelet-depleted plasma samples were prepared from blood of healthy donors. The plasma samples were spiked either with EVs from human milk or EVs from TF-positive and TF-negative cell lines. Plasma was also prepared from whole human blood with or without lipopolysaccharide stimulation. Twenty-one laboratories measured EV-TF activity and antigen in the prepared samples using their own assays representing 18 functional and 9 antigenic assays. Results: There was a large variability in the absolute values for the different EV-TF activity and antigen assays. Activity assays had higher specificity and sensitivity compared with antigen assays. In addition, there was a large intra-assay and interassay variability. Functional assays that used a blocking anti-TF antibody or immunocapture were the most specific and sensitive. Activity assays that used immunocapture had a lower coefficient of variation compared with assays that isolated EVs by high-speed centrifugation. Conclusion: Based on this multicenter study, we recommend measuring EV-TF using a functional assay in the presence of an anti-TF antibody.

Citation

Bonifay, A., Mackman, N., Hisada, Y., Sachetto, A. T. A., Hau, C., Gray, E., Hogwood, J., Aharon, A., Badimon, L., Barile, L., Baudar, J., Beckmann, L., Benedikter, B., Bolis, S., Bouriche, T., Brambilla, M., Burrello, J., Camera, M., Campello, E., Ettelaie, C., …Lacroix, R. (2024). Comparison of assays measuring extracellular vesicle tissue factor in plasma samples: communication from the ISTH SSC Subcommittee on Vascular Biology. Journal of Thrombosis and Haemostasis, https://doi.org/10.1016/j.jtha.2024.05.037

• Journal Article Type: Article
• Acceptance Date: May 23, 2024
• Online Publication Date: Jun 25, 2024
• Publication Date: 2024
• Deposit Date: Jun 28, 2024
• Publicly Available Date: Jun 26, 2025
• Journal: Journal of Thrombosis and Haemostasis
• Print ISSN: 1538-7933
• Electronic ISSN: 1538-7836
• Publisher: Elsevier
• Peer Reviewed: Peer Reviewed
• DOI: https://doi.org/10.1016/j.jtha.2024.05.037
• Keywords: Blood coagulation; Extracellular vesicles; Flow cytometry; Functional assays; Tissue factor
• Public URL: https://hull-repository.worktribe.com/output/4721018
• Publisher URL: https://www.sciencedirect.com/science/article/pii/S1538783624003672