Abstract
Tissue factor (TF), the primary initiator of blood coagulation in vivo, is expressed in vitro by a variety of cells. Previous efforts to localize TF in tissue and cells have been limited principally to the use of immunological techniques. In the present study, we describe a novel functional probe for TF expression, which can be utilized to localize functional TF in situ in human cells and tissues. This probe, a biotinylated phe-pro-arg-chloro-methyl-ketone-labeled rVIIa (FPR-ck-VIIa), interacts with TF via high-affinity binding sites. The binding of FPR-ck-VIIa, therefore, can be correlated with the ability of TF to activate clotting. In the described studies, TF antigen (TF:Ag) expression was examined immunohistochemically with various TF-specific monoclonal antibodies (MAbs) and was correlated with functional TF expression using our novel TF-binding probe (eg, FPR-ck-VIIa). Initial results indicate that TF:Ag expression correlates with the expression of functional TF (TF:VIIa), and the specificity of both types of probes was confirmed. Parallel antigenic and functional TF expression in situ was demonstrated in various human tumors. We believe this to be the first demonstration of functional TF in situ in human cells and tissues. We suggest that FPR-ck-VIIa should prove a useful reagent for studying the role of TF in the pathogenesis of clotting complications of human disease.
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- Andrews B. S., Rehemtulla A., Fowler B. J., Edgington T. S., Mackman N. Conservation of tissue factor primary sequence among three mammalian species. Gene. 1991 Feb 15;98(2):265–269. doi: 10.1016/0378-1119(91)90184-d. [DOI] [PubMed] [Google Scholar]
- Bach R. R. Initiation of coagulation by tissue factor. CRC Crit Rev Biochem. 1988;23(4):339–368. doi: 10.3109/10409238809082548. [DOI] [PubMed] [Google Scholar]
- Bevilacqua M. P., Gimbrone M. A., Jr Inducible endothelial functions in inflammation and coagulation. Semin Thromb Hemost. 1987 Oct;13(4):425–433. doi: 10.1055/s-2007-1003519. [DOI] [PubMed] [Google Scholar]
- Bloem L. J., Chen L., Konigsberg W. H., Bach R. Serum stimulation of quiescent human fibroblasts induces the synthesis of tissue factor mRNA followed by the appearance of tissue factor antigen and procoagulant activity. J Cell Physiol. 1989 May;139(2):418–423. doi: 10.1002/jcp.1041390226. [DOI] [PubMed] [Google Scholar]
- Drake T. A., Cheng J., Chang A., Taylor F. B., Jr Expression of tissue factor, thrombomodulin, and E-selectin in baboons with lethal Escherichia coli sepsis. Am J Pathol. 1993 May;142(5):1458–1470. [PMC free article] [PubMed] [Google Scholar]
- Drake T. A., Morrissey J. H., Edgington T. S. Selective cellular expression of tissue factor in human tissues. Implications for disorders of hemostasis and thrombosis. Am J Pathol. 1989 May;134(5):1087–1097. [PMC free article] [PubMed] [Google Scholar]
- Edgington T. S., Mackman N., Brand K., Ruf W. The structural biology of expression and function of tissue factor. Thromb Haemost. 1991 Jul 12;66(1):67–79. [PubMed] [Google Scholar]
- Ewan V. A., Edwards R. L., Rickles F. R. Expression of procoagulant activity in a human monocyte-like cell line. J Lab Clin Med. 1983 Mar;101(3):401–410. [PubMed] [Google Scholar]
- Korn J. H., Rickles F. R., Ewan V. A., Edwards R. L., Myers T. J., Onder O. Mononuclear cell modulation of fibroblast procoagulant activity. J Lab Clin Med. 1982 May;99(5):657–668. [PubMed] [Google Scholar]
- Mann K. G., Williams E. B., Krishnaswamy S., Church W., Giles A., Tracy R. P. Active site-specific immunoassays. Blood. 1990 Aug 15;76(4):755–766. [PubMed] [Google Scholar]
- Morrissey J. H., Fair D. S., Edgington T. S. Monoclonal antibody analysis of purified and cell-associated tissue factor. Thromb Res. 1988 Nov 1;52(3):247–261. doi: 10.1016/0049-3848(88)90084-9. [DOI] [PubMed] [Google Scholar]
- Morrissey J. H., Revak D., Tejada P., Fair D. S., Edgington T. S. Resolution of monomeric and heterodimeric forms of tissue factor, the high-affinity cellular receptor for factor VII. Thromb Res. 1988 May 15;50(4):481–493. doi: 10.1016/0049-3848(88)90197-1. [DOI] [PubMed] [Google Scholar]
- Nemerson Y. Tissue factor and hemostasis. Blood. 1988 Jan;71(1):1–8. [PubMed] [Google Scholar]
- Rao L. V., Rapaport S. I. Affinity purification of human brain tissue factor utilizing factor VII bound to immobilized anti-factor VII. Anal Biochem. 1987 Sep;165(2):365–370. doi: 10.1016/0003-2697(87)90283-1. [DOI] [PubMed] [Google Scholar]
- Rickles F. R., Hardin J. A., Pitlick F. A., Hoyer L. W., Conrad M. E. Tissue factor activity in lymphocyte cultures from normal individuals and patients with hemophilia A. J Clin Invest. 1973 Jun;52(6):1427–1434. doi: 10.1172/JCI107316. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rickles F. R., Levin J., Hardin J. A., Barr C. F., Conrad M. E., Jr Tissue factor generation by human mononuclear cells: effects of endotoxin and dissociation of tissue factor generation from mitogenic response. J Lab Clin Med. 1977 Apr;89(4):792–803. [PubMed] [Google Scholar]
- Ruf W., Edgington T. S. An anti-tissue factor monoclonal antibody which inhibits TF.VIIa complex is a potent anticoagulant in plasma. Thromb Haemost. 1991 Nov 1;66(5):529–533. [PubMed] [Google Scholar]
- Ruf W., Rehemtulla A., Edgington T. S. Antibody mapping of tissue factor implicates two different exon-encoded regions in function. Biochem J. 1991 Sep 15;278(Pt 3):729–733. doi: 10.1042/bj2780729. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wilcox J. N., Smith K. M., Schwartz S. M., Gordon D. Localization of tissue factor in the normal vessel wall and in the atherosclerotic plaque. Proc Natl Acad Sci U S A. 1989 Apr;86(8):2839–2843. doi: 10.1073/pnas.86.8.2839. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Williams E. B., Krishnaswamy S., Mann K. G. Zymogen/enzyme discrimination using peptide chloromethyl ketones. J Biol Chem. 1989 May 5;264(13):7536–7545. [PubMed] [Google Scholar]
- Zacharski L. R., Dunwiddie C., Nutt E. M., Hunt J., Memoli V. A. Cellular localization of activated factor X by Xa-specific probes. Thromb Haemost. 1991 May 6;65(5):545–548. [PubMed] [Google Scholar]



