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A novel flow cytometry procoagulant assay for diagnosis of vaccine-induced immune thrombotic thrombocytopenia

Vaccine-induced immune thrombotic thrombocytopenia (VITT) is a severe prothrombotic complication of adenoviral vaccines, including the ChAdOx1 nCoV-19 (Vaxzevria) vaccine. The putative mechanism involves formation of pathological anti–platelet factor 4 (PF4) antibodies that activate platelets via th...

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Autores principales: Lee, Christine S.M., Liang, Hai Po Helena, Connor, David E., Dey, Agnibesh, Tohidi-Esfahani, Ibrahim, Campbell, Heather, Whittaker, Shane, Capraro, David, Favaloro, Emmanuel J., Donikian, Dea, Kondo, Mayuko, Hicks, Sarah M., Choi, Philip Y.-I., Gardiner, Elizabeth E., Clarke, Lisa Joanne, Tran, Huyen, Passam, Freda H., Brighton, Timothy Andrew, Chen, Vivien M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society of Hematology 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9198924/
https://www.ncbi.nlm.nih.gov/pubmed/35359002
http://dx.doi.org/10.1182/bloodadvances.2021006698
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author Lee, Christine S.M.
Liang, Hai Po Helena
Connor, David E.
Dey, Agnibesh
Tohidi-Esfahani, Ibrahim
Campbell, Heather
Whittaker, Shane
Capraro, David
Favaloro, Emmanuel J.
Donikian, Dea
Kondo, Mayuko
Hicks, Sarah M.
Choi, Philip Y.-I.
Gardiner, Elizabeth E.
Clarke, Lisa Joanne
Tran, Huyen
Passam, Freda H.
Brighton, Timothy Andrew
Chen, Vivien M.
author_facet Lee, Christine S.M.
Liang, Hai Po Helena
Connor, David E.
Dey, Agnibesh
Tohidi-Esfahani, Ibrahim
Campbell, Heather
Whittaker, Shane
Capraro, David
Favaloro, Emmanuel J.
Donikian, Dea
Kondo, Mayuko
Hicks, Sarah M.
Choi, Philip Y.-I.
Gardiner, Elizabeth E.
Clarke, Lisa Joanne
Tran, Huyen
Passam, Freda H.
Brighton, Timothy Andrew
Chen, Vivien M.
author_sort Lee, Christine S.M.
collection PubMed
description Vaccine-induced immune thrombotic thrombocytopenia (VITT) is a severe prothrombotic complication of adenoviral vaccines, including the ChAdOx1 nCoV-19 (Vaxzevria) vaccine. The putative mechanism involves formation of pathological anti–platelet factor 4 (PF4) antibodies that activate platelets via the low-affinity immunoglobulin G receptor FcγRIIa to drive thrombosis and thrombocytopenia. Functional assays are important for VITT diagnosis, as not all detectable anti-PF4 antibodies are pathogenic, and immunoassays have varying sensitivity. Combination of ligand binding of G protein–coupled receptors (protease-activated receptor-1) and immunoreceptor tyrosine–based activation motif–linked receptors (FcγRIIa) synergistically induce procoagulant platelet formation, which supports thrombin generation. Here, we describe a flow cytometry–based procoagulant platelet assay using cell death marker GSAO and P-selectin to diagnose VITT by exposing donor whole blood to patient plasma in the presence of a protease-activated receptor-1 agonist. Consecutive patients triaged for confirmatory functional VITT testing after screening using PF4/heparin ELISA were evaluated. In a development cohort of 47 patients with suspected VITT, plasma from ELISA-positive patients (n = 23), but not healthy donors (n = 32) or individuals exposed to the ChAdOx1 nCov-19 vaccine without VITT (n = 24), significantly increased the procoagulant platelet response. In a validation cohort of 99 VITT patients identified according to clinicopathologic adjudication, procoagulant flow cytometry identified 93% of VITT cases, including ELISA-negative and serotonin release assay–negative patients. The in vitro effect of intravenous immunoglobulin (IVIg) and fondaparinux trended with the clinical response seen in patients. Induction of FcγRIIa-dependent procoagulant response by patient plasma, suppressible by heparin and IVIg, is highly indicative of VITT, resulting in a sensitive and specific assay that has been adopted as part of a national diagnostic algorithm to identify vaccinated patients with platelet-activating antibodies.
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spelling pubmed-91989242022-06-15 A novel flow cytometry procoagulant assay for diagnosis of vaccine-induced immune thrombotic thrombocytopenia Lee, Christine S.M. Liang, Hai Po Helena Connor, David E. Dey, Agnibesh Tohidi-Esfahani, Ibrahim Campbell, Heather Whittaker, Shane Capraro, David Favaloro, Emmanuel J. Donikian, Dea Kondo, Mayuko Hicks, Sarah M. Choi, Philip Y.-I. Gardiner, Elizabeth E. Clarke, Lisa Joanne Tran, Huyen Passam, Freda H. Brighton, Timothy Andrew Chen, Vivien M. Blood Adv Thrombosis and Hemostasis Vaccine-induced immune thrombotic thrombocytopenia (VITT) is a severe prothrombotic complication of adenoviral vaccines, including the ChAdOx1 nCoV-19 (Vaxzevria) vaccine. The putative mechanism involves formation of pathological anti–platelet factor 4 (PF4) antibodies that activate platelets via the low-affinity immunoglobulin G receptor FcγRIIa to drive thrombosis and thrombocytopenia. Functional assays are important for VITT diagnosis, as not all detectable anti-PF4 antibodies are pathogenic, and immunoassays have varying sensitivity. Combination of ligand binding of G protein–coupled receptors (protease-activated receptor-1) and immunoreceptor tyrosine–based activation motif–linked receptors (FcγRIIa) synergistically induce procoagulant platelet formation, which supports thrombin generation. Here, we describe a flow cytometry–based procoagulant platelet assay using cell death marker GSAO and P-selectin to diagnose VITT by exposing donor whole blood to patient plasma in the presence of a protease-activated receptor-1 agonist. Consecutive patients triaged for confirmatory functional VITT testing after screening using PF4/heparin ELISA were evaluated. In a development cohort of 47 patients with suspected VITT, plasma from ELISA-positive patients (n = 23), but not healthy donors (n = 32) or individuals exposed to the ChAdOx1 nCov-19 vaccine without VITT (n = 24), significantly increased the procoagulant platelet response. In a validation cohort of 99 VITT patients identified according to clinicopathologic adjudication, procoagulant flow cytometry identified 93% of VITT cases, including ELISA-negative and serotonin release assay–negative patients. The in vitro effect of intravenous immunoglobulin (IVIg) and fondaparinux trended with the clinical response seen in patients. Induction of FcγRIIa-dependent procoagulant response by patient plasma, suppressible by heparin and IVIg, is highly indicative of VITT, resulting in a sensitive and specific assay that has been adopted as part of a national diagnostic algorithm to identify vaccinated patients with platelet-activating antibodies. American Society of Hematology 2022-06-10 /pmc/articles/PMC9198924/ /pubmed/35359002 http://dx.doi.org/10.1182/bloodadvances.2021006698 Text en © 2022 by The American Society of Hematology. Licensed under Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0), permitting only noncommercial, nonderivative use with attribution. All other rights reserved. https://www.ncbi.nlm.nih.gov/pmc/pmcdoc/tagging-guidelines/article/tags.html#el-licenseThis article is made available via the PMC Open Access Subset for unrestricted reuse and analyses in any form or by any means with acknowledgment of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic or until permissions are revoked in writing. Upon expiration of these permissions, PMC is granted a perpetual license to make this article available via PMC and Europe PMC, consistent with existing copyright protections.
spellingShingle Thrombosis and Hemostasis
Lee, Christine S.M.
Liang, Hai Po Helena
Connor, David E.
Dey, Agnibesh
Tohidi-Esfahani, Ibrahim
Campbell, Heather
Whittaker, Shane
Capraro, David
Favaloro, Emmanuel J.
Donikian, Dea
Kondo, Mayuko
Hicks, Sarah M.
Choi, Philip Y.-I.
Gardiner, Elizabeth E.
Clarke, Lisa Joanne
Tran, Huyen
Passam, Freda H.
Brighton, Timothy Andrew
Chen, Vivien M.
A novel flow cytometry procoagulant assay for diagnosis of vaccine-induced immune thrombotic thrombocytopenia
title A novel flow cytometry procoagulant assay for diagnosis of vaccine-induced immune thrombotic thrombocytopenia
title_full A novel flow cytometry procoagulant assay for diagnosis of vaccine-induced immune thrombotic thrombocytopenia
title_fullStr A novel flow cytometry procoagulant assay for diagnosis of vaccine-induced immune thrombotic thrombocytopenia
title_full_unstemmed A novel flow cytometry procoagulant assay for diagnosis of vaccine-induced immune thrombotic thrombocytopenia
title_short A novel flow cytometry procoagulant assay for diagnosis of vaccine-induced immune thrombotic thrombocytopenia
title_sort novel flow cytometry procoagulant assay for diagnosis of vaccine-induced immune thrombotic thrombocytopenia
topic Thrombosis and Hemostasis
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9198924/
https://www.ncbi.nlm.nih.gov/pubmed/35359002
http://dx.doi.org/10.1182/bloodadvances.2021006698
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