Phosphotidylserine exposure and neutrophil extracellular traps enhance procoagulant activity in patients with inflammatory bowel disease

Zhangxiu He; Yu Si; Tao Jiang; Ruishuang Ma; Yan Zhang; Muhua Cao; Tao Li; Zhipeng Yao; Lu Zhao; Shaohong Fang; Bo Yu; Zengxiang Dong; Hemant S. Thatte; Yayan Bi; Junjie Kou; Shufen Yang; Daxun Piao; Lirong Hao; Jin Zhou; Jialan Shi

doi:10.1160/TH15-09-0710

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2016; 115(04): 738-751
DOI: 10.1160/TH15-09-0710

Coagulation and Fibrinolysis

Schattauer GmbH

Phosphotidylserine exposure and neutrophil extracellular traps enhance procoagulant activity in patients with inflammatory bowel disease

Zhangxiu He^*

¹Department of Hematology, First Hospital, Harbin Medical University, Harbin, China

²Department of Nephrology, First Hospital, Harbin Medical University, Harbin, China

⁶The Key Laboratory of Myocardial Ischemia, Ministry of Education, Heilongjiang Province, Harbin, China

,

Yu Si^*

¹Department of Hematology, First Hospital, Harbin Medical University, Harbin, China

⁶The Key Laboratory of Myocardial Ischemia, Ministry of Education, Heilongjiang Province, Harbin, China

,

Tao Jiang^*

³Department of General Surgery, First Hospital, Harbin Medical University, Harbin, China

,

Ruishuang Ma

¹Department of Hematology, First Hospital, Harbin Medical University, Harbin, China

⁶The Key Laboratory of Myocardial Ischemia, Ministry of Education, Heilongjiang Province, Harbin, China

,

Yan Zhang

¹Department of Hematology, First Hospital, Harbin Medical University, Harbin, China

,

Muhua Cao

¹Department of Hematology, First Hospital, Harbin Medical University, Harbin, China

⁶The Key Laboratory of Myocardial Ischemia, Ministry of Education, Heilongjiang Province, Harbin, China

,

Tao Li

¹Department of Hematology, First Hospital, Harbin Medical University, Harbin, China

,

Zhipeng Yao

¹Department of Hematology, First Hospital, Harbin Medical University, Harbin, China

,

Lu Zhao

¹Department of Hematology, First Hospital, Harbin Medical University, Harbin, China

,

Shaohong Fang

⁶The Key Laboratory of Myocardial Ischemia, Ministry of Education, Heilongjiang Province, Harbin, China

,

Bo Yu

⁵Department of Cardiology of the Second Hospital, Harbin Medical University, Harbin, China

⁶The Key Laboratory of Myocardial Ischemia, Ministry of Education, Heilongjiang Province, Harbin, China

,

Zengxiang Dong

⁴Department of Cardiology, First Hospital, Harbin Medical University, Harbin, China

,

Hemant S. Thatte

⁷Department of Surgery, Brigham and Women’s Hospital, VA Boston Healthcare System, Harvard Medical School, Boston, Massachusetts, USA

,

Yayan Bi

⁴Department of Cardiology, First Hospital, Harbin Medical University, Harbin, China

,

Junjie Kou

⁵Department of Cardiology of the Second Hospital, Harbin Medical University, Harbin, China

,

Shufen Yang

⁶The Key Laboratory of Myocardial Ischemia, Ministry of Education, Heilongjiang Province, Harbin, China

,

Daxun Piao

³Department of General Surgery, First Hospital, Harbin Medical University, Harbin, China

,

Lirong Hao

²Department of Nephrology, First Hospital, Harbin Medical University, Harbin, China

,

Jin Zhou

¹Department of Hematology, First Hospital, Harbin Medical University, Harbin, China

,

Jialan Shi

¹Department of Hematology, First Hospital, Harbin Medical University, Harbin, China

⁷Department of Surgery, Brigham and Women’s Hospital, VA Boston Healthcare System, Harvard Medical School, Boston, Massachusetts, USA

› Author Affiliations

Abstract

Summary

Inflammatory bowel disease (IBD)-associated thromboembolic event often lacks precise aetiology. The aim of this study was to investigate the contribution of phosphatidylserine (PS) exposure and neutrophil extracellular traps (NETs) towards the hypercoagulable state in IBD. We demonstrated that the levels of PS exposed MPs and the sources of MP-origin, platelets, erythrocytes, leukocytes and cultured endothelial cells (ECs) were higher in IBD groups than in healthy controls using flow cytometry and confocal microscopy. Wright-Giemsa and immunofluorescence staining demonstrated that the elevated NETs were released by activated IBD neutrophils or by control neutrophils treated with IBD sera obtained from patients with the active disease. MPs and MP-origin cells in IBD groups, especially in active stage, markedly shortened coagulation time and had increased levels of fibrin, thrombin and FXa production as assessed by coagulation function assays. Importantly, we found that on stimulated ECs, PS rich membranes provided binding sites for FXa and FVa, promoting fibrin formation while TNF blockage or IgG depletion attenuated this effect. Treatment of control neutrophils with TNF and isolated IgG from PR3-ANCA-positive active IBD patients also resulted in the release of NETs. Blockade of PS with lactadherin prolonged coagulation time, decreased fibrin formation to control levels, and inhibited the procoagulant enzymes production in the MPs and MP-origin cells. NET cleavage by DNase I partly decreased PCA in IBD or stimulated neutrophils. Our study reveals a previously unrecognised link between hypercoagulable state and PS exposure or NETs, and may further explain the epidemiological association of thrombosis within IBD patients.

Keywords

IBD hypercoagulable disorders - phosphatidylserine - Neutrophil Extracellular Traps - procoagulant activity

Full Text

References

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