Elsevier

Thrombosis Research

Volume 96, Issue 1, 1 October 1999, Pages 51-56
Thrombosis Research

Regular article
The Flow Cytometer Model Markedly Affects Measurement of ex Vivo Whole Blood Platelet-Bound P-Selectin Expression in Patients with Chest Pain: Are We Comparing Apples with Oranges?

https://doi.org/10.1016/S0049-3848(99)00063-8Get rights and content

Abstract

Surface expression of P-selectin is known to be a marker of platelet activation in patients with acute coronary syndromes. However, direct comparisons of flow cytometer data may be obscured by differences in methodology, artifactual platelet activation during washing procedures, choice of antibodies, absence of control measurements, and possible observer bias due to unblinded data collection. We sought to test the hypothesis that the model of flow cytometer represents another variable affecting P-selectin measurements. Platelet P-selectin in whole blood was measured by FACScan (Becton Dickinson, Inc., San Diego, CA, USA) or EPICS XL (Coulter Corporation, Hialeah, FL, USA) flow cytometry in 338 patients presenting with chest pain to the emergency departments of three community hospitals as part of a multicenter diagnostic trial. Platelet expression of P-selectin (% of cell positivity) was consistently higher for each discharge diagnosis when measured with FACScan flow cytometer (13.2±4.1 for myocardial infarction, 10.0± 3.6 for unstable angina, 9.9±3.5 for heart failure, 4.7±0.1 for gastrointestinal illness, and 6.3±0.7 for patients with noncardiac chest pain) when compared with results obtained from the EPICS XL instrument 2.4±0.2, 2.5±0.2, 2.5±0.1, 1.8±0.1, and 2.3±0.1 respectively, p=0.0001 for all groups. This study reveals marked discrepancies in the level of platelet P-selectin measurement based exclusively on the model of flow cytometer used. If P-selectin is to become a diagnostic tool for differentiating an etiology of chest pain, standardized measurements must be defined for each model of flow cytometer.

Section snippets

Patients

Three hundred thirty-eight consecutive patients ad- mitted to the Chest Pain Center of the emergency department of The Christ Hospital (Cincinnati, OH; n=143), St. Agnes Hospital (Baltimore, MD; n=98), and Sinai Hospital (Baltimore, MD; n=97) between December 1997 and February 1998 and meeting enrollment criteria were included in this study. Patients who were at least 21 years old with chest pain occurring at rest and lasting at least 5 minutes and with an initial onset occurring within 24

Results

Although the patterns of P-selectin changes were overall similar between the sites, platelet expression of P-selectin was consistently three times higher or greater when measured with the FACScan flow cytometer when compared to the results obtained from the EPICS XL instrument. Table 1 summarizes the data by discharge diagnosis in patients with chest pain. Individual patient data are presented at Figure 1.

Discussion

This is the first large prospective multicenter study to evaluate the diagnostic utility of two different models of flow cytometer in measuring whole blood platelet-bound P-selectin in patients with chest pain. These data demonstrate that despite similar demographics of the chest pain population evaluated, identical sampling, quality control assurance, and staining procedures between sites, the level of platelet P-selectin expression is overwhelmingly dependent on the model of flow cytometer

Acknowledgements

This study was supported in part by Centocor Diagnostics of Pennsylvania (Malvern, PA, USA). We are indebted to the staff of emergency department of the Christ Hospital (Cincinnati, Ohio, USA), St. Agnes and Sinai Hospitals (Baltimore, Maryland, USA).

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