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Tissue factor in microvesicles shed from U87MG human glioblastoma cells
induces coagulation, platelet aggregation, and thrombogenesis
E Bastida, A Ordinas, G Escolar and GA Jamieson
Microvesicles (diameter ca 200 nm) from the cell-free supernatant of U87MG
human glioblastoma cell caused platelet aggregation and coagulation in a
manner identical with that previously shown for the intact cells. Both
activities were inhibited by dansylarginine -N-(3- ethyl-1,5-pentanediyl)
amide (DAPA), confirming the thrombin-dependent nature of both activities.
The specific activities per microgram of protein were 2-10 times greater in
the microvesicles than in the plasma membrane fraction, suggesting
localization in specific membrane domains. Sucrose density centrifugation
gave a single protein peak (density 1.14) with congruent procoagulant and
platelet aggregating activities. Both activities required the extrinsic
pathway, as shown by studies with factor-deficient plasmas, and both were
inhibited by heating (60 min/100 degrees C), by reduction and alkylation,
and by incubation of the microvesicles with rabbit anti-bovine brain tissue
factor antibody. These observations were confirmed using microvesicles from
the HL-60 human promyelocytic leukemia cells, which are known to contain
tissue factor activity. The results suggest that both procoagulant and
proaggregating activities are causally related through the presence of
tissue factor in the microvesicles. Studies with the Baumgartner perfusion
apparatus showed that U87MG microvesicles increased the size of adherent
thrombi nearly tenfold and that these thrombi were associated with
nucleated cells from the blood. The increase in adherent thrombi did not
occur if perfusion was carried out in the presence of DAPA, confirming the
role of thrombin in their formation.
Volume 64,
Issue 1,
pp. 177-184,
07/01/1984
Copyright © 1984 by The American Society of Hematology
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