August 1994
Volume 35, Issue 9
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Articles  |   August 1994
Increased basal levels of free plasminogen activator activity found in human aqueous humor.
Author Affiliations
  • Y Wang
    Department of Pathology, University of Connecticut Health Center, Farmington 06030-3105.
  • D M Taylor
    Department of Pathology, University of Connecticut Health Center, Farmington 06030-3105.
  • D M Smalley
    Department of Pathology, University of Connecticut Health Center, Farmington 06030-3105.
  • R E Cone
    Department of Pathology, University of Connecticut Health Center, Farmington 06030-3105.
  • J O'Rourke
    Department of Pathology, University of Connecticut Health Center, Farmington 06030-3105.
Investigative Ophthalmology & Visual Science August 1994, Vol.35, 3561-3566. doi:
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    • Get Citation

      Y Wang, D M Taylor, D M Smalley, R E Cone, J O'Rourke; Increased basal levels of free plasminogen activator activity found in human aqueous humor.. Invest. Ophthalmol. Vis. Sci. 1994;35(9):3561-3566.

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Abstract

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.

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