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Monocyte-derived tissue factor contributes to stent thrombosis in an in vitro system

Tullio Palmerini, MD^*,*, Barry S. Coller, MD, Vittorio Cervi, BSc^*, Luciana Tomasi, BSc^*, Antonio Marzocchi, MD^*, Cinzia Marrozzini, MD^*, Ornella Leone, MD, Milena Piccioli, BSC and Angelo Branzi, MD^*

^* Istituto di Cardiologia
Istituto di Anatomia Patologica, Policlinico S. Orsola, University of Bologna, Bologna, Italy
Laboratory of Blood and Vascular Biology, Rockefeller University, New York, New York

View larger version (159K): [in a new window]   Figure 1 Thrombus formation after 120 min perfusion on stent surface exposed to blood pretreated with either saline (control) or with a monoclonal antibody against tissue factor (cH36). Note the striking reduction of thrombus formation on the stent in which blood was pretreated with cH36.

View larger version (124K): [in a new window]   Figure 2 Histologic assessment of thrombi formed on a stent surface showing intense tissue factor staining (red). Monocytes (arrows) found in the thrombus displayed strong tissue factor staining, while granulocytes displayed weak and irregular staining. Original magnification 400x.

View larger version (37K): [in a new window]   Figure 3 Tissue factor (TF) expression on monocyte membranes at baseline (A) and at the end of the perfusion (B). In two experiments monocytes were subjected to two-color analysis using TF-FITC as the abscissa and CD42b-PE as the ordinate: at baseline (C) there are some platelet-monocyte aggregates that do not display TF staining. At the end of the perfusion (D) only monocytes with attached platelets displayed TF staining.

View larger version (30K): [in a new window]   Figure 4 Whole blood before (A) and after (B) anti-CD14 bead addition. Note in B the dramatic reduction of monocyte count. In controls that were subjected to all of the procedures used to deplete monocytes except for adding the anti-CD14-coupled beads, no difference was noticed between the baseline sample (C) and the sample after the procedures (D).

View larger version (98K): [in a new window]   Figure 5 Immunohistochemistry of thrombi formed on stents perfused with whole blood (A) or monocyte-depleted blood (B). In B a strong reduction in tissue factor staining (red) was observed compared with A. Original magnification 200x.