α2-Macroglobulin Is a Significant In Vivo Inhibitor of Activated Protein C and Low APC:α2M Levels Are Associated with Venous Thromboembolism

Laura Martos; Luis Andrés Ramón; Julia Oto; Álvaro Fernández-Pardo; Santiago Bonanad; Ana Rosa Cid; Andras Gruber; John H. Griffin; Francisco España; Silvia Navarro; Pilar Medina

doi:10.1055/s-0038-1629902

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2018; 118(04): 630-638
DOI: 10.1055/s-0038-1629902

Coagulation and Fibrinolysis

Schattauer GmbH Stuttgart

α₂-Macroglobulin Is a Significant In Vivo Inhibitor of Activated Protein C and Low APC:α₂M Levels Are Associated with Venous Thromboembolism

Laura Martos^*

¹Grupo de Investigación en Hemostasia, Trombosis, Arteriosclerosis y Biología Vascular, Instituto de Investigación Sanitaria La Fe (IIS La Fe), Hospital Universitario y Politécnico La Fe, Valencia, Spain

,

Luis Andrés Ramón^*

¹Grupo de Investigación en Hemostasia, Trombosis, Arteriosclerosis y Biología Vascular, Instituto de Investigación Sanitaria La Fe (IIS La Fe), Hospital Universitario y Politécnico La Fe, Valencia, Spain

,

Julia Oto

¹Grupo de Investigación en Hemostasia, Trombosis, Arteriosclerosis y Biología Vascular, Instituto de Investigación Sanitaria La Fe (IIS La Fe), Hospital Universitario y Politécnico La Fe, Valencia, Spain

,

Álvaro Fernández-Pardo

¹Grupo de Investigación en Hemostasia, Trombosis, Arteriosclerosis y Biología Vascular, Instituto de Investigación Sanitaria La Fe (IIS La Fe), Hospital Universitario y Politécnico La Fe, Valencia, Spain

,

Santiago Bonanad

¹Grupo de Investigación en Hemostasia, Trombosis, Arteriosclerosis y Biología Vascular, Instituto de Investigación Sanitaria La Fe (IIS La Fe), Hospital Universitario y Politécnico La Fe, Valencia, Spain

²Unidad de Trombosis y Hemostasia, Servicio de Hematología, Hospital Universitario y Politécnico La Fe, Valencia, Spain

,

Ana Rosa Cid

¹Grupo de Investigación en Hemostasia, Trombosis, Arteriosclerosis y Biología Vascular, Instituto de Investigación Sanitaria La Fe (IIS La Fe), Hospital Universitario y Politécnico La Fe, Valencia, Spain

²Unidad de Trombosis y Hemostasia, Servicio de Hematología, Hospital Universitario y Politécnico La Fe, Valencia, Spain

,

Andras Gruber

³Department of Biomedical Engineering, Oregon Health and Science University, Portland, Oregon, United States

,

John H. Griffin

⁴Department of Molecular Medicine, The Scripps Research Institute, La Jolla, California, United States

,

Francisco España

¹Grupo de Investigación en Hemostasia, Trombosis, Arteriosclerosis y Biología Vascular, Instituto de Investigación Sanitaria La Fe (IIS La Fe), Hospital Universitario y Politécnico La Fe, Valencia, Spain

,

Silvia Navarro

¹Grupo de Investigación en Hemostasia, Trombosis, Arteriosclerosis y Biología Vascular, Instituto de Investigación Sanitaria La Fe (IIS La Fe), Hospital Universitario y Politécnico La Fe, Valencia, Spain

,

Pilar Medina

¹Grupo de Investigación en Hemostasia, Trombosis, Arteriosclerosis y Biología Vascular, Instituto de Investigación Sanitaria La Fe (IIS La Fe), Hospital Universitario y Politécnico La Fe, Valencia, Spain

› Author Affiliations

Abstract

Background Activated protein C (APC) is a major regulator of thrombin formation. Two major plasma inhibitors form complexes with APC, protein C inhibitor (PCI) and α₁-antitrypsin (α₁AT), and these complexes have been quantified by specific enzyme-linked immunosorbent assays (ELISAs). Also, complexes of APC with α₂-macroglobulin (α₂M) have been observed by immunoblotting. Here, we report an ELISA for APC:α₂M complexes in plasma.

Methods Plasma samples were pre-treated with dithiothreitol and then with iodoacetamide. The detection range of the newly developed APC:α₂M assay was 0.031 to 8.0 ng/mL of complexed APC. Following infusions of APC in humans and baboons, complexes of APC with α₂M, PCI and α₁AT were quantified. These complexes as well as circulating APC were also measured in 121 patients with a history of venous thromboembolism (VTE) and 119 matched controls.

Results In all the in vivo experiments, α₂M was a significant APC inhibitor. The VTE case–control study showed that VTE patients had significantly lower APC:α₂M and APC levels than the controls (p < 0.001). Individuals in the lowest quartile of APC:α₂M or the lowest quartile of APC had approximately four times more VTE risk than those in the highest quartile of APC:α₂M or of APC. The risk increased for individuals with low levels of both parameters.

Conclusion The APC:α₂M assay reported here may be useful to help monitor the in vivo fate of APC in plasma. In addition, our results show that a low APC:α₂M level is associated with increased VTE risk.

Keywords

activated protein C - α2-macroglobulin - ELISA - baboon - complexes

Full Text

References

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