Evaluation of a dual-color flow cytometry immunophenotyping panel in a multicenter quality assurance program

Authors

  • Eve Levine Schenker,

    1. Department of Medicine and Jonsson Comprehensive Cancer Center, UCLA School of Medicine, Los Angeles, California 90024
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  • Lance E. Hultin,

    1. Department of Medicine and Jonsson Comprehensive Cancer Center, UCLA School of Medicine, Los Angeles, California 90024
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  • Kenneth D. Bauer,

    1. Department of Pathology, Northwestern University Medical School, Chicago, Illinois 60611
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  • John Ferbas,

    1. Department of Infectious Diseases and Microbiology, University of Pittsburgh Graduate School of Public Health, Pittsburgh, Pennsylvania 15261
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  • Joseph B. Margolick,

    1. Department of Environmental Health Sciences, Johns Hopkins University School of Hygiene and Public Health, Baltimore, Maryland 21205
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  • Janis V. Giorgi Ph.D.

    Corresponding author
    1. Department of Medicine and Jonsson Comprehensive Cancer Center, UCLA School of Medicine, Los Angeles, California 90024
    • UCLA School of Medicine, Department of Medicine/Cellular Immunology and Cytometry, 12-240 Factor Building, Los Angeles, CA 90024-1745
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  • This work was supported by National Institutes Of Health Awards AI-72631, AI-72632, AI-72634, AI-72676, AI-32535, and CA-16042.

  • From the Multicenter AIDS Cohort Study Group Centers and Investigators: Baltimore: Johns Hopkins University School of Hygiene and Public Health: A.J. Saah, H. Armenian, H. Farzadegan, N. Graham, J. Mcarthur, J. Margolick, J. Palenicek. Chicago: Howard Brown Memorial Clinic—Northwestern University Medical School: J.P. Phair, K. Bauer, J.S. Chimel, B. Cohen, D. Variakojis, J. Wesch, S.M. Wolinsky. Los Angeles: Ucla Schools of Medicine and Public Health, and the Jonsson Comprehensive Cancer Center: R. Detels, B.R. Visscher, I. Chen, J. Dudley, J.L. Fahey, J.V. Giorgi, D.T. Imagawa, O. Martinez-Maza, P. Nishanian, J. Taylor, J. Zack. Pittsburgh: University of Pittsburgh School of Public Health: C.R. Rinaldo, Jr., J.T. Becker, J. Ferbas, P. Gupta, M. Ho, L. Kingsley. Data Coordinating Center: Johns Hopkins University School of Hygiene and Public Health: A. Muñoz, D. Hoover, L.P. Jacobson, K.-Y. Liang, C. Meinert, L. Park, S. Su, S. Zeger. NIH/National Institute of Allergy and Infectious Disease: L. Schrager, R.A. Kaslow, M.J. Vanraden, NIH/Na-tional Cancer Institute: D. Seminara.

Abstract

A basic immunophenotyping panel that employed dual-color combinations of fluorescein isothiocyanate (FITC) and phycoerythrin (PE) conjugated monoclonal antibodies (mAb; FITC-CD45/PE-CD14, FITC-IgG1/PE-IgG2, FITC-CD3/PE-CD8, FITC-CD3/PE-CD4, FITC-CD3/PE-CD16+PE-CD56, and PE-CD19) was utilized in a quality assurance program to determine whether the 4 laboratories participating in a multicenter AIDS study obtained similar lymphocyte subset percentage values for T cells, B cells, NK cells, and CD4+ and CD8+ T cells. Over a 1½ year period, 78 shared peripheral blood specimens Were prepared and analyzed in each laboratory. The CD45brightCD14 percentage for each specimen was used to correct that individual's lymphocyte subset values. Interlaboratory coefficients of variation (CV) for the human immunodeficiency virus type I (HIV) seronegative (n = 38) and HIV-seropositive (n = 40) specimens using this panel were <3% for total T cells; <5% for CD4+ T cells and CD8+ T cells; ≤17% for B and NK cells; and <8% for CD4T/CD8T ratios. The 6-tube basic immunophenotyping panel has several notable features: (a) for clinical studies, it permits comprehensive evaluation of an individual's major lymphocyte subsets, i.e., T, B, NK, and CD4+ and CD8+ T cells; (b) for interlaboratory proficiency testing programs, it allows the detection of differences among laboratories in measurements of several functionally distinct cell populations; and (c) for within-sample quality assurance, it provides several quality control checks, including the lym-phosum, i.e., the sum of an individual's corrected T + B + NK values, a sum that was generally 100 ± 5% on the HIV-seronegative specimens analyzed in this study. © 1993 Wiley-Liss, INc.

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