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Blood, 15 November 2003, Vol. 102, No. 10, pp. 3637-3645.
Prepublished online as a Blood First Edition Paper on July 31, 2003; DOI 10.1182/blood-2003-02-0496.


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HEMOSTASIS, THROMBOSIS, AND VASCULAR BIOLOGY

Physical proximity and functional interplay of PECAM-1 with the Fc receptor Fc{gamma}RIIa on the platelet plasma membrane

Le M. Thai, Leonie K. Ashman, Stacey N. Harbour, P. Mark Hogarth, and Denise E. Jackson

From the Austin Research Institute, Austin Hospital, Heidelberg, Victoria, Australia; University of Newcastle, Callaghan, New South Wales, Australia.

We and others have recently defined that Platelet Endothelial Cell Adhesion Molecule-1 (PECAM-1/CD31) functions as a negative regulator of platelet-collagen interactions involving the glycoprotein VI/Fc receptor gamma chain (GPVI/FcR-{gamma} chain) signaling pathway.1,2 In this study, we hypothesized that PECAM-1 may be physically and functionally associated with Fc{gamma}RIIa on the platelet membrane. The functional relationship between PECAM-1 and Fc{gamma}RIIa was assessed by determining the effect of anti-PECAM-1 monoclonal antibody Fab fragments on Fc{gamma}RIIa-mediated platelet aggregation and heparin-induced thrombocytopenia (HITS)-mediated platelet aggregation. Preincubation of washed platelets with monoclonal antibody fragments of 2BD4 directed against PECAM-1 and IV.3 directed against Fc{gamma}RIIa completely blocked Fc{gamma}RIIa-mediated platelet aggregation and HITS-mediated platelet aggregation, whereas anti-CD151 antibody had no blocking effect. Coengagement of Fc{gamma}RIIa and PECAM-1 resulted in negative regulation of Fc{gamma}RIIa-mediated phospholipase C{gamma}2 activation, calcium mobilization, and phosphoinositide 3-kinase-dependent signaling pathways. In addition, the physical proximity of Fc{gamma}RIIa and PECAM-1 was confirmed by using fluorescence resonance energy transfer and coimmunoprecipitation studies. These results indicate that PECAM-1 and Fc{gamma}RIIa are colocalized on the platelet membrane and PECAM-1 down-regulates Fc{gamma}RIIa-mediated platelet responses. (Blood. 2003;102:3637-3645)


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