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Clinical Chemistry 53: 1231-1234, 2007. First published May 10, 2007; 10.1373/clinchem.2007.085332
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Right arrow Hemostasis and Thrombosis
(Clinical Chemistry. 2007;53:1231-1234.)
© 2007 American Association for Clinical Chemistry, Inc.

Hemostasis and Thrombosis

Increased Plasma Concentrations of Soluble CD40 Ligand in Acute Coronary Syndrome Depend on in Vitro Platelet Activation

Boris T. Ivandic1,a,2, Eberhard Spanuth2,2, Detlef Haase3, Heiko-Gundmar Lestin3 and Hugo A. Katus1

1 Department of Medicine III, University of Heidelberg, Heidelberg, Germany.
2 Roche Diagnostics GmbH, Mannheim, Germany.
3 Institute of Laboratory Medicine, Helios Kliniken, Schwerin, Germany.

aAddress correspondence to this author at: Innere Medizin, Abt. III, Universitaetsklinikum Heidelberg, Im Neuenheimer Feld 410, 69120 Heidelberg, Germany. Fax 49-6221-56-5235; e-mail boris.ivandic{at}med.uni-heidelberg.de.

Background: Soluble CD40 ligand (sCD40L) was suggested as a novel biomarker of cardiovascular risk. We examined the effect of preanalytical variation on the measurement of sCD40L concentration.

Methods: From healthy control individuals (n = 20) and patients with acute coronary syndrome (ACS) (n = 20) or sepsis (n = 20), we obtained blood drawn into 5 tubes containing citrate or a mixture of citrate, theophylline, adenosine, and dipyridamole (CTAD). The tubes were incubated for 30 min at room temperature or 0 °C before a single or double centrifugation (15 min, 2500g) at room temperature or 4 °C, respectively. sCD40L, ß-thromboglobulin (ßTG), and platelet factor 4 (PF4) concentrations were measured using immunoassays.

Results: Concentrations of sCD40L were very low in all CTAD and citrated samples maintained at 0 °C (median ≤0.076 µg/L). Although increased ßTG and PF4 confirmed disease-related in vivo platelet activation, sCD40L was not higher in patients than in controls. In contrast, if the samples were processed at room temperature, sCD40L was significantly higher in ACS patients than in controls (P <0.02 in CTAD and citrated plasma at room temperature). Moreover, the ßTG:PF4 ratio decreased in patient but not control CTAD samples, suggesting a greater susceptibility of patient platelets to in vitro activation.

Conclusions: Increased sCD40L concentrations resulted from in vitro platelet activation during sample preparation. Disease-related in vivo activation did not contribute to sCD40L concentrations in plasma. Therefore, published studies of sCD40L demand cautious interpretation, because their preanalytical conditions were not standardized.




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