Elevated levels of platelet- and red cell-derived extracellular vesicles in transfusion-dependent ß-thalassemia/HbE patients with pulmonary arterial hypertension.

ABSTRACT

Pulmonary arterial hypertension (PAH) is a serious complication in ß-thalassemia. The mechanism of PAH development is believed to be through chronic platelet activation and red cell (RBC) membrane abnormality contributing to a hypercoagulable state and thrombosis, which consequently leads to the development of PAH. Extracellular vesicles (EVs) shed from the plasma membrane of platelets and RBCs are found to be associated with thrombotic risk. This study aimed to investigate the involvement of phosphatidylserine (PS)-bearing cells and EVs in accelerating the progression of the hypercoagulable state in transfusion-dependent thalassemia (TDT) patients. Fresh whole blood samples from splenectomized TDT-ß-thalassemia/HbE patients (11 with PAH and 14 without PAH) and 15 normal subjects were analyzed for platelet activation by measuring P-selectin expression using flow cytometry and the number of dense granular using an electron microscope. The amounts of PS-bearing RBCs, large RBC-EVs, platelets, and medium EVs were determined by flow cytometry. Platelet activation in PAH patients was not significantly different from other groups; however, the amounts of PS-bearing large RBC-EVs, platelets, and medium platelet-derived EVs were significantly increased in PAH patients as compared to normal subjects, but they were not different from patients without PAH. This could be affected by antiplatelet therapy that reduced the levels of platelet activation and the amount of PS-bearing cells, including EVs, in PAH patients as well as in patients without PAH.