title: Evidence for a rapid inhibitor to tissue plasminogen activator in plasma.
abstract: An improved assay method for tissue plasminogen activator (t-PA) in plasma samples is described, which allows the plasma t-PA activity to be determined within 3 hours. The method minimizes the action of possible inhibitors by immediate acidification of blood samples prior to separation of the blood cells. In almost all plasma samples variable amounts of a rapid t-PA inhibitor could be demonstrated. The inhibitor was present in plasma and seemed not to be associated with the blood cells. A quantitative method for determination of the inhibitor content in plasma was developed. In this investigation 1 unit of inhibitor is defined as the amount that neutralized 1 unit (0.06 pmol) of t-PA in 10 minutes. The content of t-PA inhibitor in plasma samples from healthy individuals was determined as 0.6 U/ml +/- 0.5 U/ml (range 0-3.2, median 0.4). In a group of patients (n = 75) with suspected disturbance of the haemostatic system, elevated levels were found (3.8 +/- 2.7, U/ml median 3.6 U/ml, range 0 - 15 U/ml). Kinetic analysis with plasma from three patients with elevated inhibitor content showed very similar rate of t-PA inhibition and assuming the formation of a 1:1 t-PA-inhibitor complex a second order rate constant of 10(7) M(-1) s(-1) was calculated. A regulatory function may be anticipated for this inhibitor, but its exact patophysiological role has still to be established.
authors.affiliation: Dept of Clinical Chemistry and Blood Coagulation, Karolinska Hospital, S-10401 Stockholm, Sweden.
authors.forename: J
authors.initials: J
authors.lastname: Chmielewska
authors.forename: M
authors.initials: M
authors.lastname: Rånby
authors.forename: B
authors.initials: B
authors.lastname: Wiman
mesh_terms: D000328:Adult; D001681:Biological Assay; D001800:Blood Specimen Collection; D016022:Case-Control Studies; D005260:Female; D006487:Hemostasis; D020141:Hemostatic Disorders / Q000097:blood*; D006801:Humans; D007700:Kinetics; D008297:Male; D008875:Middle Aged; D015849:Plasminogen Inactivators / Q000097:blood*; D010959:Tissue Plasminogen Activator / Q000037:antagonists & inhibitors* / Q000097:blood; D055815:Young Adult
chemical_list: D015849:Plasminogen Inactivators; D010959:Tissue Plasminogen Activator