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

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

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish (US)
Pages (from-to)2910-2921
Number of pages12
JournalJournal of Thrombosis and Haemostasis
Volume22
Issue number10
DOIs
StatePublished - Oct 2024

Keywords

  • blood coagulation
  • extracellular vesicles
  • flow cytometry
  • functional assays
  • tissue factor

ASJC Scopus subject areas

  • Hematology

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