title: Phosphotidylserine exposure and neutrophil extracellular traps enhance procoagulant activity in patients with inflammatory bowel disease.
abstract: 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.
authors.forename: Zhangxiu
authors.initials: Z
authors.lastname: He
authors.forename: Yu
authors.initials: Y
authors.lastname: Si
authors.forename: Tao
authors.initials: T
authors.lastname: Jiang
authors.forename: Ruishuang
authors.initials: R
authors.lastname: Ma
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhang
authors.forename: Muhua
authors.initials: M
authors.lastname: Cao
authors.forename: Tao
authors.initials: T
authors.lastname: Li
authors.forename: Zhipeng
authors.initials: Z
authors.lastname: Yao
authors.forename: Lu
authors.initials: L
authors.lastname: Zhao
authors.forename: Shaohong
authors.initials: S
authors.lastname: Fang
authors.forename: Bo
authors.initials: B
authors.lastname: Yu
authors.forename: Zengxiang
authors.initials: Z
authors.lastname: Dong
authors.forename: Hemant S
authors.initials: HS
authors.lastname: Thatte
authors.forename: Yayan
authors.initials: Y
authors.lastname: Bi
authors.forename: Junjie
authors.initials: J
authors.lastname: Kou
authors.forename: Shufen
authors.initials: S
authors.lastname: Yang
authors.forename: Daxun
authors.initials: D
authors.lastname: Piao
authors.forename: Lirong
authors.initials: L
authors.lastname: Hao
authors.forename: Jin
authors.initials: J
authors.lastname: Zhou
authors.affiliation: Jialan Shi, MD, PhD, or Jin Zhou, MD, PhD, or Lirong Hao, MD, PhD, Department of Medicine, or Daxun Piao, MD, PhD, Department of General Surgery, First Hospital, Harbin Medical University, 23 Youzheng Street, Nangang District, Harbin 150001, China, E-mail: jialan_shi@hms.harvard.edu.
authors.forename: Jialan
authors.initials: J
authors.lastname: Shi
mesh_terms: D000328:Adult; D000368:Aged; D000954:Antigens, Surface / Q000494:pharmacology; D001777:Blood Coagulation / Q000187:drug effects*; D055252:Cell-Derived Microparticles / Q000187:drug effects* / Q000502:physiology; D002478:Cells, Cultured; D003850:Deoxyribonuclease I / Q000494:pharmacology*; D065206:Extracellular Traps / Q000187:drug effects* / Q000378:metabolism*; D015943:Factor Va / Q000378:metabolism; D015951:Factor Xa / Q000378:metabolism; D005260:Female; D005337:Fibrin / Q000378:metabolism; D006801:Humans; D015212:Inflammatory Bowel Diseases / Q000097:blood*; D008297:Male; D008875:Middle Aged; D008894:Milk Proteins / Q000494:pharmacology; D010718:Phosphatidylserines / Q000494:pharmacology*; D011485:Protein Binding / Q000187:drug effects; D019851:Thrombophilia / Q000097:blood*
chemical_list: D000954:Antigens, Surface; C098942:MFGE8 protein, human; D008894:Milk Proteins; D010718:Phosphatidylserines; D015943:Factor Va; D005337:Fibrin; D003850:Deoxyribonuclease I; D015951:Factor Xa
keywords: IBD hypercoagulable disorders; Neutrophil Extracellular Traps; phosphatidylserine; procoagulant activity