XENOGENEIC ENDOTHELIAL CELLS ACTIVATE HUMAN PROTHROMBIN1,2

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Abstract

Background

Delayed xenograft rejection is characterized by platelet activation and fibrin deposition and is thought to occur independently of complement activation. We have therefore investigated the potential for xenogeneic endothelial cells (EC) to regulate the conversion of prothrombin to thrombin, a central component of the final common pathway of coagulation and an important platelet agonist.

Methods and Results

Quiescent porcine aortic EC (PAEC) were found to convert high levels of human prothrombin to thrombin (0.234±0.019 IU/ml) when compared with human aortic EC (0.017±0 IU/ml, 30-min time point, chromogenic assay; P<0.001). PAEC activation by human complement resulted in comparable levels of thrombin generation. Prothrombin conversion by PAEC as determined by generation of F1+2 (1.909±0.119 nmol/L) and formation of thrombin-antithrombin III complexes (125.611±6.373 μg/L) was significantly greater than the matched human aortic EC values (F1+2: 1.539±0.03 nmol/L, P<0.001; thrombin-antithrombin III: 1.833±0.104 μg/L, P<0.001). Sequential analysis of prothrombin activation by PAEC indicated generation of the intermediate meizothrombin followed by autolytically accelerated thrombin formation. Subsequent experiments established important cross-species' incompatibilities with respect to porcine thrombomodulin interaction with human thrombin and protein C in that PAEC had a reduced capacity to generate activated human protein C in vitro.

Conclusion

These observations indicate a potentially important molecular barrier involving blood coagulation that may impact on the planned clinical application of porcine transgenic organs.

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