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Increased basal levels of free plasminogen activator activity found in human aqueous humor
Y Wang1, D M Taylor, D M Smalley
1Department of Pathology, University of Connecticut Health Center, Farmington 06030-3105.
Purpose:
To quantify the basal free t-PA activity in human aqueous humor.
Methods:
Human aqueous humor obtained by simplified pipette paracentesis at cataract surgery was tested for free t-PA activity in a revised 15-hour amidolytic assay using a t-PA standard curve (n = 15). Total antigenic levels of PAI-1, the principal PA inhibitor, were determined using an ELISA kit. The available PAI-1 activity was tested indirectly using anti-human PAI-1 antibody blocking before t-PA activity assay (n = 11). Plasminogen activator type was determined by anti-human t-PA and urokinase (u-PA) antibody blocking before activity assay (n = 7).
Results:
Free PA activity ranged widely (0.072 to 0.47 IU/ml; mean, 0.20 +/- 0.10 IU/ml) and was almost completed inactivated (> or = 89%) by antibody against human t-PA but not by the u-PA antibody. PAI-1 total antigen also ranged widely between (0.25 to 8.0 ng/ml; mean, 2.25 +/- 2.54 ng/ml). However, pretreatment of samples with PAI-1 antibody or by acidification (pH 3.2) to inactivate inhibitors did not increase t-PA activity levels.
Conclusions:
A basal-free t-PA activity, which is predominantly t-PA, is present in human aqueous humor at approximately 20 times higher levels than previously found with an earlier assay. This activity is estimated to represent roughly 10% of total released t-PA antigen. PAI-1 is present in aqueous humor at levels considerably lower than reported values for plasma in a predominantly PA-complexed or inactive form.