Thrombocytopenia and thrombosis following treatment with the integrin Ī±IIbĪ²3 antagonist eptifibatide are rare complications caused by patient antibodies specific for ligand-occupied Ī±IIbĪ²3. Whether such antibodies induce platelet clearance by simple opsonization, by inducing mild platelet activation, or both is poorly understood. To gain insight into the mechanism by which eptifibatide-dependent antibodies initiate platelet clearance, we incubated normal human platelets with patient serum containing an Ī±IIbĪ²3-specific, eptifibatide-dependent antibody. We observed that in the presence of eptifibatide, patient IgG induced platelet secretion and aggregation as well as tyrosine phosphorylation of the integrin Ī²3 cytoplasmic domain, the platelet FcĪ³RIIa Fc receptor, the protein-tyrosine kinase Syk, and phospholipase CĪ³2. Each activation event was inhibited by preincubation of the platelets with Fab fragments of the FcĪ³RIIa-specific mAb IV.3 or with the Src family kinase inhibitor PP2. Patient serum plus eptifibatide did not, however, activate platelets from a patient with a variant form of Glanzmann thrombasthenia that expressed normal levels of FcĪ³RIIa and the Ī±IIbĪ²3 complex but lacked most of the Ī²3 cytoplasmic domain. Taken together, these data suggest a novel mechanism whereby eptifibatide-dependent antibodies engage the integrin Ī²3 subunit such that FcĪ³RIIa and its downstream signaling components become activated, resulting in thrombocytopenia and a predisposition to thrombosis.