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Vincristine induces procoagulant activity of the human lymphoblastic leukemia cell line Jurkat through the release of extracellular vesicles

  • Claire PluchartEmail author
  • Gael Poitevin
  • Maud Colinart-Thomas
  • Gregory Guimard
  • Sandra Audonnet
  • Christine Terryn
  • Philippe Nguyen
Article

Abstract

Thromboembolic events are frequent and serious complications of acute lymphoblastic leukaemia treatment. The importance of chemotherapy in the pathogenesis of this increased risk is enhanced by the fact that thrombosis rarely occurs at diagnosis. Our study aims at investigating the effect of chemotherapy on pro-coagulant activity (PCA), phosphatidylserine (PS) exposure, tissue factor (TF) activity and derived extracellular vesicles (EV) of Jurkat cells. Jurkat cells were treated with two commonly used chemotherapeutics: Vincristine (VCR) or Daunorubicin (DNR), at relevant concentrations. PCA of cells and derived EV were evaluated using Thrombin generation Assay (TGA). Cells or EV were incubated with annexin V or anti TF antibodies to assess the respective contribution of TF and PS. PS exposure on cells was analysed by flow cytometry. Derived EV were evaluated in fluorescence microscopy and flow cytometry. Untreated Jurkat cells and EV support thrombin generation. Thrombin generation was abolished when PS activity was inhibited by annexin V. VCR treatment resulted in a time dependent increase of thrombin generation. After VCR exposure, TF activity increased as well as PS exposure increased on the cell surface. The increase in TF activity was abolished by annexin V indicating that PS was required. A spontaneous release of EV from Jurkat cells was observed and VCR treatment increased the number of generated EV. Our results indicate that VCR increased the PCA of Jurkat cells predominantly through PS exposure and increased EV generation. Lymphoid blasts derived EV could be biomarkers to determine high thrombotic risk ALL patients.

Keywords

Acute lymphoblastic leukemia Phosphatidylserine Extracellular vesicles Microparticles Vincristine Thrombin generation assay 

Notes

Compliance with ethical standards

Conflicts of interest

The authors have no conflict of interest to disclose.

Supplementary material

11239_2019_1894_MOESM1_ESM.pptx (1.5 mb)
Supplementary material 1 (PPTX 1542 kb) Fig. 1 Videomicroscopy of cells and derived EV

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Paediatric Oncology/HematologyAmerican Hospital, CHU de ReimsReimsFrance
  2. 2.EA3801-HERVIUniversité de Reims Champagne-ArdenneReimsFrance
  3. 3.Plateau technique de cytométrie en flux – URCACyt, Pôle SantéUniversité de Reims Champagne-ArdenneReimsFrance
  4. 4.Plateau technique en Imagerie Cellulaire et Tissulaire (PICT), Pôle SantéUniversité de Reims Champagne-ArdenneReimsFrance
  5. 5.Department of Hematology LaboratoryCHU ReimsReimsFrance

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