Procoagulant Activity of T Lymphoblastoid Cells in Extrinsic and Intrinsic Coagulation Systems

Summary

We have measured the procoagulant activity (PCA) of four T lymphoblastoid cell lines (Jurkat, CEM, HSB-2 and Molt 4) as well as normal peripheral blood T lymphocytes, before and after stimulation with phytohaemagglutinin (PHA), using clotting and amidolytic methods.

Of the four cell lines only one, Jurkat, gave enhanced PCA after stimulation with PH A. This activity was shown to be tissue factor-like by its dependence on factor VII in plasma and in an amidolytic assay with purified factors VII and X. Jurkat was also the only one of the four cell lines to secrete interleukin-2. All four cell lines promoted the generation of large amounts of thrombin in platelet-free plasma in glass tubes. This activity was dependent on the presence of plasma factor VIII, and was probably due to phospholipids in the cell membranes. Normal T lymphocytes gave intrinsic PCA in the thrombin generation test which was only 15% of that of the lymphoma cells.

These results show that some T lymphocytes can develop PCA in both intrinsic and extrinsic systems and this should be taken into account in studies of the PCA of mixed leukocyte populations.

Keywords

Leukocytes - Tissue factor - Procoagulant activity

• References

• 1 Addison W. On the colourless corpuscles, and on the molecules and cytoblasts in the blood. London Med Gaz 1842; 30: 144-146
• 2 Lisiewicz J, Astaldi G, Okulski J. On the influence of normal and leukemic human leukocytes on blood coagulation. Acta Haematol 1970; 44: 332-340
• 3 Garg SK, Niemetz J. Tissue factor activity of normal and leukaemic cells. Blood 1973; 42: 729-735
• 4 Levy GA, Edgington TS. Lymphocyte co-operation is required for amplification of macrophage procoagulant activity. J Exp Med 1980; 154: 1232-1244
• 5 Hogg N. Human monocytes are associated with the formation of fibrin. J Exp Med 1983; 157: 473-485
• 6 Rickies FR, Hardin JA, Pitlick FA, Hoyer LW, Conrad ME. Tissue factor activity in lymphocyte cultures from normal individuals and patients with hemophilia A. J Clin Invest 1973; 52: 1427-1434
• 7 Edwards RL, Rickies FR. Macrophage procoagulants. Prog Haemo-stas Thromb 1984; 7: 183-209
• 8 Prydz HP B. (ed.). The cell biology of triggers in coagulation. Haemostasis 1984; 14: 373-444
• 9 Rivers RP A, Hathaway WE, Weston WL. The endotoxin-induced coagulant activity of human monocytes. Br J Haematol 1975; 30: 311-316
• 10 Prydz H, Lyberg T, Deteix P, Allison AC. In vitro stimulation of tissue thromboplastin (factor III) activity in human monocytes by immune complexes and lectins. Thromb Res 1979; 15: 465-474
• 11 Dean RT, Prydz H. Inflammatory particles stimulate thromboplastin production by human monocytes. Thromb Res 1983; 30: 357-367
• 12 Semeraro N, Biondi A, Lorenzet R, Locati D, Mantovani A, Donati MB. Direct induction of tissue factor synthesis by endotoxin in human macrophages from diverse sites. Immunology 1983; 50: 529
• 13 Edwards RL, Rickies FR, Ewan VA. Monocyte procoagulant activity: recognition by monoclonal anti-tissue factor antibody and evidence for in vivo regulation. Thromb Haemostas 1983; 50: 176 (abstract).
• 14 Edwards RL, Rickies FR. The role of human T cells (and T cell products) for monocyte tissue factor generation. J Immunol 1980; 125: 606-609
• 15 Helin HJ, Fox RI, Edgington TS. The instructor cell for the human procoagulant monocyte response to bacterial lipolysaccharide is a Leu-3a T cell by fluorescence-activated cell sorting. J Immunol 1983; 131: 749-752
• 16 Rickies FR, Bobrove AM. Tissue factor in mononuclear cell cultures. Blood 1975; 46: 92 (abstract).
• 17 Schwarz BS, Edgington TS. Lymphocyte collaboration is required for induction of murine monocyte procoagulant activity by immune complexes. J Immunol 1981; 127: 438-443
• 18 Barrowcliffe TW, Marshall DA, Trickett LP, Hubbard AR, Thorpe R. Procoagulant activity of human T lymphocytes in intrinsic and extrinsic systems. Thromb Haemostas 1987; 58: 102 (abstract).
• 19 Gearing AJ H, Johnstone AP, Thorpe R. Production and assay of the interleukins. J Immunol Meth 1985; 83: 01-27
• 20 Henry C. Cell separations. In: Selected Methods in Cellular Immunology. Mishell BB, Shiigi SM. (eds.). W H Freeman; San Francisco, CA: 1980: 182-185
• 21 Bird C, Callus M, Trickett L, Thorpe R. Immunochemical characterization of a new platelet specific monoclonal antibody and its use to demonstrate the cytoskeletal association of the platelet glycoprotein Ilb-IIIa complex. Biosci Rep 1985; 6: 323-333
• 22 Austen DE G, Rhymes IL. A Laboratory Manual of Blood Coagulation Blackwell Scientific Publications; Oxford: 1975
• 23 Barrowcliffe TW, Stocks J, Gray E. Preparation of a stable phospholipid reagent for coagulation assays. Haemostasis 1982; 11: 096-101
• 24 Giddings JC. Hereditary coagulation disorders: laboratory techniques. In: Blood Coagulation and Haemostasis. Thomson JM. (ed.). Churchill Livingstone; Edinburgh: 1980: 117-157
• 25 Hubbard AR, Jennings CA. Inhibition of the tissue factor-Factor VII complex: involvement of Factor Xa and lipoproteins. Thromb Res 1987; 46: 527-537
• 26 Modi GJ, Blajchman MA, Ofosu FA. The isolation of prothrombin, Factor IX and Factor X from human Factor IX concentrates. Thromb Res 1984; 36: 537-547
• 27 Zwaal RF A, Hemker C. Blood cell membranes and haemostasis. Haemostasis 1982; 11: 12-39
• 28 Zwaal RF A, Rosing J, Tans G, Bevers EM, Hemker HC. Topological and kinetic aspects of phospholipids in blood coagulation. In: The Regulation of Coagulation. Mann Tylor. (ed.). Elsevier/North-Holland; Amsterdam: 1980: 095-115
• 29 Lyberg T. Clinical significance of increased thromboplastin activity on the monocyte surface-a brief review. Haemostasis 1984; 14: 430-439
• 30 Geczy CL, Meyer PA. Leukocyte procoagulant activity in man, as in vitro correlate of delayed-type hypersensitivity. J Immunol 1982; 128: 331-336
• 31 Zur M, Nemerson Y. Tissue factor pathways of blood coagulation. In: Haemostasis and Thrombosis. Bloom AL, Thomas DP. (eds.). 2nd ed. Churchill Livingstone; Edinburgh: 1987: 148-164
• 32 Osterud B, Rapaport SI. Activation of factor IX by the reaction product of tissue factor and factor VII: additional pathway for initiating blood coagulation. Proc Natl Acad Sci USA 1977; 74: 5250-5264
• 33 Marlar RA, Kleiss AJ, Griffin JH. An alternative extrinsic pathway of human blood coagulation. Blood 1982; 60: 1353-1358