1. HETE-PEs are increased in leukocytes and platelets from APS patients.
HETE-PEs are increased in leukocytes and platelets from APS patients. (A and B) 15- and 5-HETE-PEs are increased in leukocytes from antiphospholipid syndrome (APS) patients. Total leukocytes were isolated from healthy control (HC) and APS patients, left unstimulated (US) or stimulated at 4 × 106 cells/ml with 10 μM A23187 for 30 min at 37°C, and then the lipids were analyzed for HETE-PEs. HETE-PEs were quantified as individual molecular species and then combined. Data are means ± SEM of 34 (HC) or 17 (APS) subjects. (C) 12-HETE-PEs are increased in platelets from APS patients. Washed platelets from HC and APS patients were isolated and left unstimulated or stimulated with thrombin (0.2 U/ml) for 30 min at 37°C before the amounts of 12-HETE-PE were determined. Data are means ± SEM of 18 (HC) or 12 (APS) subjects. (D) Platelets from APS patients that spontaneously aggregated contain substantial amounts of 12-HETE-PEs. During the final washing step, a number of APS platelet isolates spontaneously aggregated (n = 7 patients). Lipids were extracted and analyzed for 12-HETE-PEs. (E) Urinary 11-thromboxane B2 (TXB2) is increased in APS patients. Urine from HC and APS patients was analyzed by gas chromatography–mass spectrometry (MS) to determine the amounts of TXB2. Data are means ± SEM of 32 (HC) or 9 (APS) subjects. ng/mg creatinine (Cr). (F and G) 15- and 5-HETE-PEs are basally increased in concentration in platelets from APS patients. Platelets were prepared as described in (C) and then were analyzed to determine the amounts of the indicated HETE-PEs. Data are means ± SEM of 18 (HC) or 12 (APS) subjects. (H) 12-HETE-PEs are basally increased in abundance in leukocytes from APS patients. Total leukocytes were isolated and left unstimulated or stimulated at 4 × 106 cells/ml with 10 μM A23187 for 30 min at 37°C and then analyzed for 12-HETE-PEs. Data are means ± SEM of 34 (HC) or 18 (APS) subjects. ***P ≤ 0.001, **P ≤ 0.01, *P ≤ 0.05 by Mann-Whitney U test.
Sarah N. Lauder et al., Sci. Signal. 2017;10:eaan2787
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