INDUCTION OF TISSUE FACTOR EXPRESSION IN HUMAN MONOCYTE/ENDOTHELIUM COCULTURES

Induction of tissue factor (TF) expression on monocyctes and endothelial cells is central to the development of septic coagulopathy, Serum concentrations of endotoxin in septic patients who develop disseminated intravascular coagulation (DIG) do not, however, reach the levels that would directly stimulate TF expression on either monocytes or endothelium. We show, using an in vitro coculture system, that the interaction of monocytes with endothelium induces the expression of significant levels of TF, Unstimulated cocultures of monocytes (2 x 10(4)/well) and endothelial cells (2 x 10(4)/well) produced 35.3+/-8.5 mU of PCA/well, representing a 5-fold increase over the combined PCA of each cell type cultured alone (7.1+/-1.5 mU, n = 6, P < 0.001), Significant enhancement was also found in the presence of low concentrations of LPS, Induction of TF protein was confirmed by Western blotting, Fixation of monocytes with paraformaldehyde completely abolished TF induction in cocultures, whereas fixation of endothelium had no effect, suggesting that TF induction occurred in monocytes rather than endothelial cells. Induction of TF in cocultures could be further augmented by preincubating the endothelial cells with IFN-gamma. When endothelium was prestimulated with 500 U/ml IFN-gamma there was 142+/-11% increase over unstimulated cocultures (n = 5, P < 0.01). TF induction was inhibited by 32+/-6% in the presence of anti-ICAM-1 mAb (n = 5, P < 0.01). Our results suggest that monocyte interactions with vascular endothelium, regulated by inflammatory cytokines, and mediated by adhesive ligand binding, leads to the induction of functional monocyte TF protein, which may be responsible for the initiation of DIC in sepsis.