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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   *
1   Grupo 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   *
1   Grupo 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
1   Grupo 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
1   Grupo 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
1   Grupo 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
2   Unidad de Trombosis y Hemostasia, Servicio de Hematología, Hospital Universitario y Politécnico La Fe, Valencia, Spain
,
Ana Rosa Cid
1   Grupo 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
2   Unidad de Trombosis y Hemostasia, Servicio de Hematología, Hospital Universitario y Politécnico La Fe, Valencia, Spain
,
Andras Gruber
3   Department of Biomedical Engineering, Oregon Health and Science University, Portland, Oregon, United States
,
John H. Griffin
4   Department of Molecular Medicine, The Scripps Research Institute, La Jolla, California, United States
,
Francisco España
1   Grupo 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
1   Grupo 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
1   Grupo 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
Further Information

Publication History

22 September 2017

02 January 2018

Publication Date:
15 February 2018 (online)

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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

Authors' Contributions

S.N. performed the experiments, analysed the data and critically revised the manuscript. L.M. performed the experiments and analysed the data. L.A.R. performed the experiments. J.O. performed the experiments. A.F-P. performed the experiments. S.B.S. performed the plasma collection from VTE patients and controls and revised the manuscript. A.R.C. performed the plasma collection from VTE patients and controls and revised the manuscript. A.G. designed the experiments and revised the manuscript. J.H.G. designed the experiments and critically revised the manuscript. F.E. designed and performed the experiments, analysed the data and wrote the manuscript. P.M. designed and performed the experiments, analysed the data and wrote the manuscript.


* These authors contributed equally to this study.


 

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