A new role for FcgammaRIIA in the potentiation of human platelet activation induced by weak stimulation.
The low affinity receptor for immunoglobulin G, FcgammaRIIA, is expressed in human platelets, mediates heparin-induced thrombocytopenia and participates to platelet activation induced by von Willebrand factor. In this work, we found that stimulation of platelets with agonists acting on G-protein-coupled receptors resulted in the tyrosine phosphorylation of FcgammaRIIA, through a mechanism involving a Src kinase. Treatment of platelets with the blocking monoclonal antibody IV.3 against FcgammaRIIA, but not with control IgG, inhibited platelet aggregation induced by TRAP1, TRAP4, the thromboxane analogue U46619, and low concentrations of thrombin. By contrast, platelet aggregation induced by high doses of thrombin was unaffected by blockade of FcgammaRIIA. We also found that the anti-FcgammaRIIA monoclonal antibody IV.3 inhibited pleckstrin phosphorylation and calcium mobilization induced by low, but not high, concentrations of thrombin. In addition, thrombin- or U46619-induced tyrosine phosphorylation of several substrates typically involved in FcgammaRIIA-mediated signalling, such as Syk and PLCgamma2, was clearly reduced by incubation with anti-FcgammaRIIA antibody IV.3. Upon stimulation with thrombin, FcgammaRIIA relocated in lipid rafts, and thrombin-induced tyrosine phosphorylation of FcgammaRIIA occurred within these membrane domains. Controlled disruption of lipid rafts by depleting membrane cholesterol prevented tyrosine phosphorylation of FcgammaRIIA and impaired platelet aggregation induced by U46619 or by low, but not high, concentrations of thrombin. These results indicate that FcgammaRIIA can be activated in human platelets downstream G-protein-coupled receptors and suggest a novel general mechanism for the reinforcement of platelet activation induced by low concentrations of agonists.
Canobbio
Ilaria
I
Centre of Excellence for Applied Biology, Department of Biochemistry, University of Pavia, via Bassi 21, 27100 Pavia, Italy.
Stefanini
Lucia
L
Guidetti
Gianni F
GF
Balduini
Cesare
C
Torti
Mauro
M
D019796:15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid / Q000494:pharmacology*; D015703:Antigens, CD / Q000378:metabolism / Q000502:physiology*; D001792:Blood Platelets / Q000378:metabolism*; D002478:Cells, Cultured; D006801:Humans; D021962:Membrane Microdomains / Q000737:chemistry / Q000378:metabolism; D015539:Platelet Activation; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism; D017452:Receptors, IgG / Q000378:metabolism / Q000502:physiology*; D015398:Signal Transduction; D013917:Thrombin / Q000494:pharmacology*
D015703:Antigens, CD; C080568:Fc gamma receptor IIA; D043562:Receptors, G-Protein-Coupled; D017452:Receptors, IgG; D019796:15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid; D013917:Thrombin