Journal
RHEUMATOLOGY
Volume 52, Issue 10, Pages 1775-1784Publisher
OXFORD UNIV PRESS
DOI: 10.1093/rheumatology/ket234
Keywords
antiphospholipid syndrome; antiprothrombin antibody; tissue factor; p38 MAPK; procoagulant cell activation
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Funding
- Japanese Ministry of Health, Labour and Welfare
- Japanese Ministry of Education, Culture, Sports, Science and Technology (MTX)
- Japanese Society for the Promotion of the Science (JSPS)
- Grants-in-Aid for Scientific Research [24591436] Funding Source: KAKEN
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Objective. The aim of this study was to investigate the effects of phosphatidylserine-dependent antiprothrombin antibody (aPS/PT) on the expression of tissue factor (TF) and the signal transduction pathway in procoagulant cells. Methods. Peripheral blood mononuclear cells (PBMCs) from a healthy donor, murine monocyte RAW264.7 cells and human umbilical vein endothelial cells (HUVECs) were treated with either IgG fractions obtained from APS patients who were positive for aPS/PT or a murine monoclonal aPS/PT antibody, 231D, in the presence of prothrombin. The levels of TF mRNA were measured using real-time PCR. TF function, as measured by procoagulant activity, was determined with a clotting assay. 231D binding on the surface of treated cells was determined by flow cytometric analysis. Screening for phosphorylation of intracellular signalling proteins was conducted using an array assay. Phosphorylation of p38 MAPK was quantitatively analysed with ELISA, and SB203580 was used as a specific inhibitor of p38 MAPK. Specific siRNA for p38 MAPK was used for the knockdown assay. Results. The IgG fractions from APS patients and 231D induced TF mRNA overexpression and shortening of coagulation time in cells in the presence of prothrombin. The 231D moiety induced phosphorylation of p38 MAPK after binding to the cell surface of RAW264.7 cells. SB203580 or p38 siRNA significantly hampered TF overexpression. Conclusion. Expression of TF in procoagulant cells was induced by aPS/PT via p38MAPK phosphorylation. This phenomenon may be correlated with the thrombogenicity of APS.
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