Regulation of fibrinolysis by platelet-released plasminogen activator inhibitor 1: light scattering and ultrastructural examination of lysis of a model platelet-fibrin thrombus.
We have investigated the role of plasminogen activator inhibitor 1 (PAI-1) in the regulation of fibrinolysis using a model thrombus composed of thrombin-stimulated platelets, fibrin(ogen), plasminogen, and recombinant tissue-type plasminogen activator. Laser light scattering kinetic measurements showed that clot lysis was significantly delayed both by thrombin-stimulated platelets and their cell-free releasate. This delay in lysis was almost fully reversed by the addition of a PAI-1-specific monoclonal antibody that blocks the ability of PAI-1 to inhibit plasminogen activators. Lysis half-times exhibited a linear dependence on the concentration of PAI-1 antigen present, as determined by enzyme-linked immunosorbent assay (ELISA). Sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) followed by immunoblotting confirmed the presence of PAI-1 antigen in the platelet releasates. Scanning electron micrographs of the model thrombus components sampled late in lysis showed considerable unproteolyzed fibrin still attached to platelets. Immunogold cytochemistry detected large amounts of PAI-1 antigen in the partially lysed platelet-fibrin thrombi. This PAI-1 appeared to be bound to the fibrin network rather than to the platelet surface itself. We conclude that the residual clots observed late in lysis represent platelet-associated fibrin to which platelet-released PAI-1 has bound, rendering it less susceptible to degradation.
Braaten
J V
JV
Department of Biochemistry, Bowman Gray School of Medicine, Wake-Forest University, Winston-Salem, NC 27157-1016.
Handt
S
S
Jerome
W G
WG
Kirkpatrick
J
J
Lewis
J C
JC
Hantgan
R R
RR
D000328:Adult; D000911:Antibodies, Monoclonal / Q000276:immunology; D001792:Blood Platelets / Q000502:physiology*; D005337:Fibrin / Q000378:metabolism*; D005342:Fibrinolysis; D006801:Humans; D008027:Light; D008854:Microscopy, Electron; D016253:Microscopy, Immunoelectron; D017395:Plasminogen Activator Inhibitor 1 / Q000032:analysis / Q000502:physiology*; D012542:Scattering, Radiation; D013917:Thrombin / Q000494:pharmacology; D013927:Thrombosis / Q000209:etiology* / Q000473:pathology
D000911:Antibodies, Monoclonal; D017395:Plasminogen Activator Inhibitor 1; D005337:Fibrin; D013917:Thrombin