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Tumor necrosis factor alpha-induced endothelial tissue factor is located on
the cell surface rather than in the subendothelial matrix
AB Mulder, KS Hegge-Paping, CP Magielse, NR Blom, JW Smit, J van der Meer, MR Hallie and VJ Bom
Division of Haemostasis, University Hospital, Groningen, The Netherlands.
Because there is no consensus regarding the precise distribution of induced
endothelial tissue factor (TF), we studied TF activity in and on tumor
necrosis factor alpha-stimulated cultured human umbilical vein endothelial
cells (ECs) and their underlying matrix. TF was mainly expressed on the
cell surface. Only small traces were found on the apical surface suggesting
that TF is predominantly located on the basolateral side of the cell
membrane. The presence of TF on the cell surface was confirmed by flow
cytometry. Subendothelial TF activity appeared to be dependent upon the
procedure used to remove the stimulated EC monolayer. Whereas ammonium
hydroxide or hypotonic lysis resulted in relatively high levels of
matrix-associated TF, virtually no TF was found on the matrix after mild
enzymatic detachment of stimulated ECs. Cell removal with EDTA resulted in
intermediate levels of matrix-associated TF. Neither the enzymatic
treatment nor EDTA degraded or removed this TF activity. Similar patterns
were observed for matrix-associated TF antigen and EC surface markers.
Electron microscopic analysis showed cell fragments on the matrix after
monolayer lysis. The findings strongly suggest that induced endothelial TF
associated with the subendothelial matrix actually represents TF on EC
remnants.
Volume 84,
Issue 5,
pp. 1559-1566,
09/01/1994
Copyright © 1994 by The American Society of Hematology
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