The Journal of Immunology, Vol 155, Issue 9 4427-4436, Copyright © 1995 by American Association of Immunologists

ARTICLES

Fc gamma II receptor-mediated platelet activation induced by anti-CD9 monoclonal antibody opens Ca2+ channels which are distinct from those associated with Ca2+ store depletion

K Kuroda, Y Ozaki, R Qi, N Asazuma, Y Yatomi, K Satoh, S Nomura, M Suzuki and S Kume
Department of Clinical and Laboratory Medicine, Yamanashi Medical University, Japan.

Anti-human platelet CD9 mAb, NNKY1-19, induced platelet activation in a Fc gamma RII-dependent manner in terms of aggregation and secretion of intracellular granule contents. These responses were considerably suppressed by aspirin. [Ca2+]i elevation in the absence of extracellular Ca2+ ([Ca2+]e), which represents the amount of Ca2+ released from intracellular Ca2+ ([Ca2+]i) stores, was also greatly reduced, whereas Ca2+ influx was sustained at similar levels. We thus investigated the mechanism that leads to the opening of Ca2+ channels in platelets incubated with aspirin. IP3 production and Ca2+ efflux were below detectable levels. 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'- tetraacetic acid loading of platelets to chelate [Ca2+]i did not reduce Ca2+ influx, as assessed by 45Ca2+ measurement. These findings suggested that NNKY1-19 induces Ca2+ channels to open without [Ca2+]i mobilization or by depleting the [Ca2+]i stores. The magnitude of Ca2+ influx was evaluated by adding [Ca2+]e to a platelet suspension activated by various agonists in the absence of [Ca2+]e. The dose dependence of the Ca2+ influx on [Ca2+]e concentrations differed according to the mode of activation. The ED50 value of Ca2+ after thrombin or thapsigargin stimulation was 0.6 mM, whereas that of NNKY1- 19 activation was about 3 mM. The addition of anti-Fc gamma RII mAb, IV.3, even 10 min after the initiation of platelet activation induced by NNKY1-19, inhibited the Ca2+ influx. These findings suggest that the Fc gamma RII-dependent activation of platelets induced by NNKY1-19 directly opens Ca2+ channels, which are distinct from those opened by thrombin or thapsigargin.

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