Expression and Localization of Tissue Factor-Based Procoagulant Activity (PCA) in Pigeon Monocyte-Derived Macrophages

Madhu Gupta; George J Doellgas; Therese Cheng; Jon C Lewis

doi:10.1055/s-0038-1649708

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 1993; 70(06): 0963-0969
DOI: 10.1055/s-0038-1649708

Original Article

Coagulation

Schattauer GmbH Stuttgart

Expression and Localization of Tissue Factor-Based Procoagulant Activity (PCA) in Pigeon Monocyte-Derived Macrophages

Madhu Gupta

¹The Department of Pathology, Bowman Gray School of Medicine of Wake Forest University, Winston-Salem, North Carolina, USA

,

George J Doellgas

²The Department of Biochemistry, Bowman Gray School of Medicine of Wake Forest University, Winston-Salem, North Carolina, USA

³Elcatech Inc., Winston-Salem, North Carolina, USA

,

Therese Cheng

³Elcatech Inc., Winston-Salem, North Carolina, USA

,

Jon C Lewis

¹The Department of Pathology, Bowman Gray School of Medicine of Wake Forest University, Winston-Salem, North Carolina, USA

› Author Affiliations

Abstract

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Summary

Monocyte-derived macrophages, focal to initiation and progression of atherosclerosis, have been implicated in thrombotic complication of this disease. In the present study we demonstrated tissue factor based procoagulant activity in cultured macrophages from the White Carneau pigeon following endotoxin (1-2 μg/ml) stimulation. This macrophage procoagulant activity paralleled activity obtained with pigeon brain homogenate. We used Enzyme-Linked Coagulation Assay (ELCA), an ultrasensitive microtiter plate assay, to measure procoagulant activity in these cells. Through the use of clotting factors purified from pigeon plasma, procoagulant activity could be detected with as few as 1-3 cells. Tissue factor antigen, detected through the use of immunogold labelling in conjunction with a polyclonal antibody which was highly specific to human tissue factor, was distributed uniformly over the plasma membrane of the endotoxin-stimulated cells. These studies suggest that this procoagulant activity might play an important role in the pathobiology of atherosclerosis in White Carneau pigeons by initiating fibrin polymerization and thus leading to thrombotic complications of the disease.

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References

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