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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; Teresa, Ana; Sachetto, 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, Risom; Laakmann, Katrin; Langer, Florian; Latysheva, Nadezhda; Lucien, Fabrice; Marques De Menezes, Erika; 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; Ata, Sachetto; Hansen, Guldenpfennig M; Sr, Kristensen; Langer, Laakmann K; Menezes, De; J...

Authors

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

Dorothée Faille

Sophie Featherby

Corentin Franco

Maite Guldenpfennig

John-Bjarne Hansen

Coralie Judicone

Yohan Kim

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

Sachetto Ata

Guldenpfennig M Hansen

Kristensen Sr

Laakmann K Langer

De Menezes

Paramo Ja



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. The ISTH SSC Subcommittee on Vascular Biology initiated a multicenter study to compare the sensitivity, specificity and reproducibility of these assays.
Materials and 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 LPS stimulation. Twenty-one laboratories measured EV-TF activity and antigen in the prepared samples using their own assays representing 19 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 to antigen assays. In addition, there was a large intra-assay and inter-assay 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 to 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 30 presence of an anti-TF antibody.

Citation

Bonifay, A., Mackman, N., Hisada, Y., Teresa, A., Sachetto, 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., …Ja, P. (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