Direct red blood cell effect on thrombosis is dependent on the interaction of tissue factor and calcium with membrane phosphatidylserine

Adam D Price; Ryan C Chae; Taylor E Wallen; Ellen R Becker; Matthew R Baucom; Rebecca Schuster; Lisa England; Timothy A Pritts; Michael D Goodman

doi:10.1097/TA.0000000000004340

Direct red blood cell effect on thrombosis is dependent on the interaction of tissue factor and calcium with membrane phosphatidylserine

J Trauma Acute Care Surg. 2024 Apr 12. doi: 10.1097/TA.0000000000004340. Online ahead of print.

Authors

Adam D Price¹, Ryan C Chae, Taylor E Wallen, Ellen R Becker, Matthew R Baucom, Rebecca Schuster, Lisa England, Timothy A Pritts, Michael D Goodman

Affiliation

¹ Department of Surgery, University of Cincinnati; Cincinnati, OH.

PMID: 38605437
DOI: 10.1097/TA.0000000000004340

Abstract

Background: Prior literature has implicated Red Blood Cells (RBCs) in the initiation of thrombosis and suggests that post-transfusion hypercoagulability may occur secondary to the effects of RBCs. Elevated serum tissue factor is a known sequelae of acute trauma. Phosphatidylserine is a pro-thrombotic phospholipid present within the RBC cell membrane. We hypothesized that RBC aggregation is dependent on the interaction between RBC membrane bound (exposed) PS, extracellular calcium, and tissue factor.

Methods: Human whole blood (WB) was separated into components including red blood cells (RBC) and platelet-rich plasma (PRP). WB, PRP, and RBCs underwent impedance aggregometry utilizing arachidonic acid (AA), ADP, collagen, calcium, and tissue factor (TF)-based agonists. RBCs then underwent impedance aggregometry utilizing combined calcium and TF agonists. RBCs were pre-treated with Annexin V, a known PS blocking agent, and underwent impedance aggregometry with combined calcium and TF agonists to determine if the mechanism of calcium/TF-induced RBC aggregability is dependent on PS. RBCs treated with calcium, TF, calcium+TF, and pre-treated with Annexin V followed by calcium+TF were perfused through an in vitro model of pulmonary microcirculatory flow.

Results: RBC aggregation was significantly higher than that of WB and PRP when utilizing a TF agonist, an effect unique to TF. The combination of calcium and TF demonstrated significantly higher RBC aggregation than either agonist alone. Pre-treatment with Annexin V resulted in a significantly reduced aggregability of RBC following treatment with TF + calcium. RBCs aged to 42 days did not exhibit significant change in aggregation. Exposure to calcium and TF significantly reduced time to thrombosis of RBCs perfused through a pulmonary microcirculatory model.

Conclusion: Treatment with both TF and calcium synergistically induces RBC aggregation. PS appears to play an integral role in the TF/calcium-based, age-independent RBC aggregation response. RBCs treated with TF + calcium exhibit more rapid thrombus formation in an in vitro model of pulmonary microcirculatory perfusion.Study Type: human sample-based study.

Level of evidence: basic science paper.