Characterization of the cell-surface procoagulant activity of T-lymphoblastoid cell lines

W. Pickering, E. Gray, A. H. Goodall, S. Ran, P. E. Thorpe, T. W. Barrowcliffe

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

The procoagulant activity (PCA) of four T-lymphoblastoid cell lines (CEM-CCRF, Jurkat, Molt-4 and A3.01) at different stages of differentiation has been characterized and compared with that of a monocytoid cell line (THP-1). Four assay systems were employed; the activated partial thromboplastin time (APTT); prothrombin time/tissue factor (TF) activity; a purified factor (F)Xa generation system and cancer procoagulant. High levels of TF activity were seen only with the monocytic cells. However the more differentiated of the T-lymphoblastoid cells (Molt-4 and A3.01) were more active than monocytic cells in supporting FXa generation. This pattern was not repeated for the APTT assay, which was related to cell-surface TF activity, since it was partially inhibited by antiTF antibody. Annexin V totally inhibited the activity observed in all three assay systems, indicating that the PCA of T-lymphoblastoid cells is primarily due to expression of negatively charged phospholipids. However, antiphosphatidyl-serine antibody even at a high concentration gave only partial inhibition of the activity observed in the APTT and FXa generation systems for the cells compared with almost total inhibition for the phospholipid standard, suggesting either that cellular phosphatidylserine (PS) is less accessible to the antibody, or that PS is not the sole negatively charged phospholipid responsible for this activity. Flow cytometry studies using propidium iodide and annexin V showed that the PCA, although linked to PS exposure, was not the result of apoptosis.

Original languageEnglish (US)
Pages (from-to)459-467
Number of pages9
JournalJournal of Thrombosis and Haemostasis
Volume2
Issue number3
DOIs
StatePublished - Mar 2004

Fingerprint

Cell Line
Partial Thromboplastin Time
Phosphatidylserines
Thromboplastin
Phospholipids
Annexin A5
cancer procoagulant
Antibodies
Factor Xa
Propidium
Prothrombin Time
Serine
Flow Cytometry
Apoptosis

Keywords

  • Coagulation
  • Phospholipid
  • Procoagulant activity
  • T-cells
  • Tissue factor

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Characterization of the cell-surface procoagulant activity of T-lymphoblastoid cell lines. / Pickering, W.; Gray, E.; Goodall, A. H.; Ran, S.; Thorpe, P. E.; Barrowcliffe, T. W.

In: Journal of Thrombosis and Haemostasis, Vol. 2, No. 3, 03.2004, p. 459-467.

Research output: Contribution to journalArticle

Pickering, W. ; Gray, E. ; Goodall, A. H. ; Ran, S. ; Thorpe, P. E. ; Barrowcliffe, T. W. / Characterization of the cell-surface procoagulant activity of T-lymphoblastoid cell lines. In: Journal of Thrombosis and Haemostasis. 2004 ; Vol. 2, No. 3. pp. 459-467.
@article{d8b0cd81ca504c48bf3de389b8b0bb21,
title = "Characterization of the cell-surface procoagulant activity of T-lymphoblastoid cell lines",
abstract = "The procoagulant activity (PCA) of four T-lymphoblastoid cell lines (CEM-CCRF, Jurkat, Molt-4 and A3.01) at different stages of differentiation has been characterized and compared with that of a monocytoid cell line (THP-1). Four assay systems were employed; the activated partial thromboplastin time (APTT); prothrombin time/tissue factor (TF) activity; a purified factor (F)Xa generation system and cancer procoagulant. High levels of TF activity were seen only with the monocytic cells. However the more differentiated of the T-lymphoblastoid cells (Molt-4 and A3.01) were more active than monocytic cells in supporting FXa generation. This pattern was not repeated for the APTT assay, which was related to cell-surface TF activity, since it was partially inhibited by antiTF antibody. Annexin V totally inhibited the activity observed in all three assay systems, indicating that the PCA of T-lymphoblastoid cells is primarily due to expression of negatively charged phospholipids. However, antiphosphatidyl-serine antibody even at a high concentration gave only partial inhibition of the activity observed in the APTT and FXa generation systems for the cells compared with almost total inhibition for the phospholipid standard, suggesting either that cellular phosphatidylserine (PS) is less accessible to the antibody, or that PS is not the sole negatively charged phospholipid responsible for this activity. Flow cytometry studies using propidium iodide and annexin V showed that the PCA, although linked to PS exposure, was not the result of apoptosis.",
keywords = "Coagulation, Phospholipid, Procoagulant activity, T-cells, Tissue factor",
author = "W. Pickering and E. Gray and Goodall, {A. H.} and S. Ran and Thorpe, {P. E.} and Barrowcliffe, {T. W.}",
year = "2004",
month = "3",
doi = "10.1111/j.1538-7836.2004.00607.x",
language = "English (US)",
volume = "2",
pages = "459--467",
journal = "Journal of Thrombosis and Haemostasis",
issn = "1538-7933",
publisher = "Wiley-Blackwell",
number = "3",

}

TY - JOUR

T1 - Characterization of the cell-surface procoagulant activity of T-lymphoblastoid cell lines

AU - Pickering, W.

AU - Gray, E.

AU - Goodall, A. H.

AU - Ran, S.

AU - Thorpe, P. E.

AU - Barrowcliffe, T. W.

PY - 2004/3

Y1 - 2004/3

N2 - The procoagulant activity (PCA) of four T-lymphoblastoid cell lines (CEM-CCRF, Jurkat, Molt-4 and A3.01) at different stages of differentiation has been characterized and compared with that of a monocytoid cell line (THP-1). Four assay systems were employed; the activated partial thromboplastin time (APTT); prothrombin time/tissue factor (TF) activity; a purified factor (F)Xa generation system and cancer procoagulant. High levels of TF activity were seen only with the monocytic cells. However the more differentiated of the T-lymphoblastoid cells (Molt-4 and A3.01) were more active than monocytic cells in supporting FXa generation. This pattern was not repeated for the APTT assay, which was related to cell-surface TF activity, since it was partially inhibited by antiTF antibody. Annexin V totally inhibited the activity observed in all three assay systems, indicating that the PCA of T-lymphoblastoid cells is primarily due to expression of negatively charged phospholipids. However, antiphosphatidyl-serine antibody even at a high concentration gave only partial inhibition of the activity observed in the APTT and FXa generation systems for the cells compared with almost total inhibition for the phospholipid standard, suggesting either that cellular phosphatidylserine (PS) is less accessible to the antibody, or that PS is not the sole negatively charged phospholipid responsible for this activity. Flow cytometry studies using propidium iodide and annexin V showed that the PCA, although linked to PS exposure, was not the result of apoptosis.

AB - The procoagulant activity (PCA) of four T-lymphoblastoid cell lines (CEM-CCRF, Jurkat, Molt-4 and A3.01) at different stages of differentiation has been characterized and compared with that of a monocytoid cell line (THP-1). Four assay systems were employed; the activated partial thromboplastin time (APTT); prothrombin time/tissue factor (TF) activity; a purified factor (F)Xa generation system and cancer procoagulant. High levels of TF activity were seen only with the monocytic cells. However the more differentiated of the T-lymphoblastoid cells (Molt-4 and A3.01) were more active than monocytic cells in supporting FXa generation. This pattern was not repeated for the APTT assay, which was related to cell-surface TF activity, since it was partially inhibited by antiTF antibody. Annexin V totally inhibited the activity observed in all three assay systems, indicating that the PCA of T-lymphoblastoid cells is primarily due to expression of negatively charged phospholipids. However, antiphosphatidyl-serine antibody even at a high concentration gave only partial inhibition of the activity observed in the APTT and FXa generation systems for the cells compared with almost total inhibition for the phospholipid standard, suggesting either that cellular phosphatidylserine (PS) is less accessible to the antibody, or that PS is not the sole negatively charged phospholipid responsible for this activity. Flow cytometry studies using propidium iodide and annexin V showed that the PCA, although linked to PS exposure, was not the result of apoptosis.

KW - Coagulation

KW - Phospholipid

KW - Procoagulant activity

KW - T-cells

KW - Tissue factor

UR - http://www.scopus.com/inward/record.url?scp=6944222765&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=6944222765&partnerID=8YFLogxK

U2 - 10.1111/j.1538-7836.2004.00607.x

DO - 10.1111/j.1538-7836.2004.00607.x

M3 - Article

VL - 2

SP - 459

EP - 467

JO - Journal of Thrombosis and Haemostasis

JF - Journal of Thrombosis and Haemostasis

SN - 1538-7933

IS - 3

ER -