Reduction of variation in T-cell subset enumeration among 55 laboratories using single-platform, three or four-color flow cytometry based on CD45 and SSC-based gating of lymphocytes
Article first published online: 17 APR 2002
Copyright © 2002 Wiley-Liss, Inc.
Special Issue: CD4: 20 Years and Counting
Volume 50, Issue 2, pages 92–101, 15 April 2002
How to Cite
Gratama, J. W., Kraan, J., Keeney, M., Granger, V. and Barnett, D. (2002), Reduction of variation in T-cell subset enumeration among 55 laboratories using single-platform, three or four-color flow cytometry based on CD45 and SSC-based gating of lymphocytes. Cytometry, 50: 92–101. doi: 10.1002/cyto.10084
- Issue published online: 26 DEC 2002
- Article first published online: 17 APR 2002
- Manuscript Accepted: 4 FEB 2002
- Manuscript Received: 14 JAN 2002
- Directorate General XII of the European Commission (Brussels, Belgium). Grant Number: QLRI-2000-00436
- flow cytometry;
- lymphocyte immunophenotyping;
- multicenter study;
Enumeration of CD4+ and CD8+ T-cell subsets provides relevant information for diagnosis and monitoring of patients with cellular immunodeficiencies. As a result, an external quality assurance scheme was implemented in Belgium, The Netherlands, and Luxemburg in 1995. A workshop was held to train the participants in state-of-the art technology for assessment of absolute T-cell subset counts (i.e., a three or four-color, single-platform assay with lymphocyte gating based on CD45 and sideward light scatter) with the aim to achieve between-site coefficients of variation (CVs) <10% and within-site CVs <5% for ≥75% of the participants. Methods: Three send-outs of stabilized blood from a healthy donor were distributed to 55 laboratories, each with the request to perform the standard assay on three occasions. For comparison, each laboratory performed its local technique in parallel. Results: With the standard technique, between-site CVs of ∼8% (CD3+ T cells), ∼9% (CD4+ T cells), and ∼10% (CD8+ T cells) were achieved. Within-site CVs were <5% for 82% (CD3+ T cells) and ∼70% (CD4+ and CD8+ subsets) of the participants. Local techniques yielded between-site CVs of 13%–17% for CD3+, CD4+, and CD8+ T cells. Conclusions: The state-of-the-art technology for T-cell subset enumeration was implemented successfully among 55 Belgian-Dutch laboratories and resulted in significant reductions of between-site variation of absolute CD3+, CD4+, and CD8+ T-cell counts. Cytometry (Clin. Cytometry) 50:92–101, 2002. © 2002 Wiley-Liss, Inc.