LDL-associated phospholipids (PLs) may be transferred into platelets. In this work, we characterized the role of VLDLs as PL donors. VLDL transferred radiolabeled PLs to platelets in a temperature-and concentration-dependent manner. LPL stimulated this process through its action on VLDL lipolysis, because it was abolished by tetrahydrolipstatin. LPL also stimulated the platelet production of thromboxane B 2 (TXB 2 ). Both LPL actions were inhibited in the presence of fatty acid-free albumin, suggesting that they were attributable to fatty acids generated during VLDL lipolysis. To study the relationship between PL transfers and platelet activation, we performed incubations in the presence of HDL, a physiological acceptor of PL released from VLDL. HDL antagonized the transfer of PL from VLDL to platelets but had no effect on the production of TXB 2 , suggesting that PL transfers were driven by platelet activation. Confirming this idea, thrombin stimulated both the production of TXB 2 and the transfers of PL. In conclusion, VLDL can transfer PL to platelets. These transfers are stimulated by LPL and thrombin through their action on platelet activation. They might be enhanced in pathologies characterized by increased VLDL concentrations.-Ibrahim, S., A. Djimet-Baboun, V. Pruneta-Deloche, C. Calzada, M. Lagarde, and G. Ponsin. Transfer of very low density lipoprotein-associated phospholipids to activated human platelets. J. Lipid Res. 2006. 47: 341-348.