Binding of tissue plasminogen activator to human monocytes and monocytoid cells

Blood. 1991 Nov 1;78(9):2318-27.

Abstract

Monocytes and monocytoid cell lines previously have been shown to express receptors for plasminogen and urokinase (u-PA). In the present study, the monocytoid cell lines, U937 and THP-1, are shown to bind tissue plasminogen activator (t-PA) in a specific, saturable, and reversible manner. These cells bound t-PA with low affinity (kd = 0.67 to 0.97 mumol/L) and high capacity (0.71 to 3.3 x 10(6) receptors/cell). Human peripheral blood monocytes bound t-PA with a kd (0.9 mumol/L) similar to that of the monocytoid cells but with a lower capacity (0.17 x 10(6) sites/cell). These binding parameters also were similar to the low-affinity interaction of t-PA with endothelial cells as measured with the cells in suspension (kd = 0.73 mumol/L and 1.1 x 10(6) sites/cell). Lysine analogues and active or diisopropylfluorophosphate-inactivated u-PA inhibited t-PA binding to monocytes, monocytoid cells, and endothelial cells with similar IC50 (concentration producing 50% inhibition) values, suggesting that the same recognition specificity mediates t-PA binding to all of these cell types. The existence of a high-affinity binding site for t-PA on monocytoid cells was also explored in detail. Unlike endothelial cells where plasminogen activator inhibitor-1 has been implicated in mediating a high-affinity interaction of t-PA with the cells, no evidence for a role of this inhibitor in ligand binding to the monocytoid cells was found. Furthermore, using both high and low 125I-t-PA concentrations, competition analyses with lysine analogues or u-PA, or treatment of the cells with carboxypeptidase B, failed to indicate the presence of distinguishable classes of t-PA binding sites. In sum, low-affinity receptors for t-PA are expressed at high density on monocytes and monocytoid cells, identifying a new element in the fibrinolytic arsenal of these cells.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Arginine / pharmacology
  • Binding Sites
  • Cell Line
  • Endothelium, Vascular / metabolism
  • Erythrocytes / metabolism
  • Humans
  • Isoflurophate / pharmacology
  • Kinetics
  • Lysine / analogs & derivatives
  • Lysine / pharmacology
  • Monocytes / metabolism*
  • Neutrophils / metabolism
  • Recombinant Proteins / metabolism
  • Tissue Plasminogen Activator / metabolism*
  • Umbilical Veins
  • Urokinase-Type Plasminogen Activator / pharmacology

Substances

  • Recombinant Proteins
  • Isoflurophate
  • Arginine
  • Tissue Plasminogen Activator
  • Urokinase-Type Plasminogen Activator
  • Lysine