on July 12, 2004
Published online before print July 12, 2004, doi: 10.1161/01.CIR.0000134963.77201.55
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Submitted on November 21, 2003
From Divisione IV Clinica Medica (P.P., D.F., F.V.) and Dipartimento di Medicina Sperimentale e Patologia (P.P., L.L., F.V.), Università di Roma "La Sapienza," and Dipartimento di Medicina Interna (V.S.) and Dipartimento di Medicina Sperimentale (A.F., P.R.), Università di Roma Tor Vergata, Rome, Italy. * To whom correspondence should be addressed. E-mail: Francesco.Violi{at}uniroma1.it.
Background--CD40 ligand (CD40L) expression on platelets is mediated by agonists, but the underlying mechanism is still unclear. Methods and Results--CD40L expression was measured in platelets from healthy subjects both with and without the addition of antioxidants or a phospholipase A2 (PLA2) inhibitor and in platelets from 2 patients with an inherited deficiency of gp91phox. Immunoprecipitation analysis was also performed to determine whether normal platelets showed gp91phox expression. Unlike catalase and mannitol, superoxide dismutase inhibited agonist-induced platelet CD40L expression in healthy subjects. Immunoprecipitation analysis also showed that platelets from healthy subjects expressed gp91phox. In 2 male patients with inherited gp91phox deficiency, collagen-, thrombin-, and arachidonic acid-stimulated platelets showed an almost complete absence of superoxide anion (O2-) and CD40L expression. Incubation of platelets from healthy subjects with a PLA2 inhibitor almost completely prevented agonist-induced O2- and CD40L expression. Conclusions--These data provide the first evidence that platelet CD40L expression occurs via arachidonic acid-mediated gp91phox activation.
Revised on May 20, 2004
Accepted on May 20, 2004
gp91phox-Dependent Expression of Platelet CD40 Ligand
P. Pignatelli MD,
Key words: CD40 ligand • NADPH oxidase • oxidative stress • platelets
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