Brain-derived gangliosides prime human platelet aggregation and induce platelet-leukocyte aggregate formation

Abstract

Background: Platelet activation and interaction with leukocytes are crucial in inflammation. Gangliosides, sialic acid-containing glycosphingolipids, have been linked to different inflammatory conditions related to cardio- and neurodegenerative disorders. The role of gangliosides in platelet and leukocyte function, although reported, still needs further investigation.

Objectives: We aimed to study the role of gangliosides in platelet activation and platelet-leukocyte interaction in vitro.

Methods: Platelet activation was studied through aggregometry in platelet-rich plasma from apparently healthy human volunteers. Signaling protein phosphorylation was analyzed by immunoblotting. Platelet P-selectin expression and platelet-leukocyte aggregate formation were measured by flow cytometry.

Results: The gangliosides monosialoganglioside GM1, disialoganglioside GD1a, and trisialoganglioside GT1b did not induce by themselves any platelet aggregation. Conversely, when preincubated with platelets, they potentiate platelet aggregation induced by submaximal adenosine diphosphate and collagen concentrations and increased P-selectin expression. Incubation of platelets with free sialic acid and the soluble part of monosialoganglioside GM1 induced a similar potentiating effect on platelet aggregation but not on platelet P-selectin expression. Consistently, analyzing the signaling protein phosphorylation, only the entire gangliosides activated extracellular stimuli-responsive kinase 1/2 suggesting that a complete ganglioside is crucial for its action on platelets. Both the priming effect on platelet aggregation and ERK1/2 activation were prevented by aspirin. Moreover, incubation of citrated whole blood with gangliosides induced platelet-leukocyte aggregate formation accompanied by increased expression of granulocyte and monocyte CD11b compared with untreated blood, suggesting a primary leukocyte activation.

Conclusion: Gangliosides may act in vitro both on platelet and leukocyte activation and on their interaction. The observed effects might contribute to inflammatory processes in clinical conditions.

Keywords: ganglioside; platelet aggregation; platelet-leukocyte aggregate; thromboinflammation.