Localization of human tissue factor antigen by immunostaining with monospecific, polyclonal anti-human tissue factor antibody

Abstract

Tissue factor, the cofactor for factor VIIa-catalyzed activation of factors IX and X, plays an important role in the initiation of hemostasis. However, the distribution of tissue factor in the body has not been defined until recently. In the present study frozen sections of non-malignant human tissues were immunostained using polyclonal, monospecific rabbit anti-human tissue factor antibodies. Specificity of the anti-tissue factor antibody was established by Western blotting. Sensitivity of the immunostaining technique for tissue factor antigen was confirmed by correlating staining of non-perturbed and perturbed cultured human umbilical vein endothelial cells with their surface membrane tissue factor coagulant activity. Brain, lung and placenta, all known to possess large amounts of tissue factor procoagulant activity, stained strongly for tissue factor, as did peripheral nerves and autonomic ganglia. Epithelium of skin, mucosa, and glomeruli also stained; however, epithelium lining excretory ducts failed to stain. Skeletal muscle did not stain, but cardiac muscle stained faintly. Smooth muscle also did not stain except for the muscularis mucosa of the esophagus, which stained brightly. Fibroblasts varied in stainability; those found in the adventitia of vessels stained strongly. The endothelium, tunica intima and tunica media of blood vessels consistently failed to stain. The distribution of tissue factor antigen as demonstrated by immunostaining supports the hypothesis that maintenance of a physical barrier between tissue factor activity and blood is key to the normal regulation of hemostasis.