Multisite comparison of methods for the quantitation of the surface expression of CD38 on CD8+ T lymphocytes

John L. Schmitz, Mary Ann Czerniewski, Mark Edinger, Susan Plaeger, Rebecca Gelman, Cynthia L. Wilkening, Jerome A. Zawadzki, Susan B. Wormsley, D. Asthana, Y. Bauer, P. Blair, B. Blais, T. Brennan, P. Bucy, D. Campbell, S. Chiu, M. Czerniewski, T. Denny, M. Edinger, P. FranksR. Gelman, C. Gonzaga, K. Helm, J. Kagan, A. Landay, E. Larson, D. Livnat, K. Luzuriaga, T. McCloskey, B. McFarland, J. Nichols, D. Nickishcer, M. Nokta, S. Pahwa, A. Patki, S. Perfetto, S. Plaeger, T. Quinn, K. Reimann, N. Roberts, H. Rosenblatt, D. Sabath, J. Schmitz, A. Sevin, C. Spina, J. Thomas, F. Valentine, D. Weng, B. Zhang

Research output: Contribution to journalArticle

8 Scopus citations

Abstract

We evaluated the effect of specimen processing variations and quantitation methods on quantitative determination of CD38 expression on CD8 T lymphocytes. Neither lysing reagent (ammonium chloride versus BD FACSlyse), fixation (paraformaldehyde versus no final fixation step), nor acquisition delay (acquisition within 6 h after fixation versus 24 h after fixation) had a significant effect on CD38 relative fluorescent intensity or CD38 quantitative estimates (RFI or antibodies bound per cell). The only significant difference in fluorescent intensity and CD38 antibodies bound per cell (ABC) was encountered when whole blood was held for 24 h prior to staining and fixation and then acquired after another 24-h hold. However, for all sample processing methods above, the CD4 biologic calibrator and QuantiBRITE bead methods gave significantly different estimates of CD38 intensity. In many cases, however, these differences are relatively small and were more pronounced in certain laboratories. We conclude that there is some flexibility in sample processing methods for quantitative CD38 determination; however, it is preferable for a laboratory to employ one method of fluorescence quantitation calculation consistently because small differences are detected between different methods. (C) 2000 Wiley-Liss, Inc.

Original languageEnglish (US)
Pages (from-to)174-179
Number of pages6
JournalCommunications in Clinical Cytometry
Volume42
Issue number3
DOIs
StatePublished - Jun 15 2000

Keywords

  • CD38
  • CD4
  • Comparison
  • QuantiBRITE
  • Quantitation

ASJC Scopus subject areas

  • Hematology
  • Cell Biology
  • Pathology and Forensic Medicine
  • Biophysics
  • Endocrinology

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    Schmitz, J. L., Czerniewski, M. A., Edinger, M., Plaeger, S., Gelman, R., Wilkening, C. L., Zawadzki, J. A., Wormsley, S. B., Asthana, D., Bauer, Y., Blair, P., Blais, B., Brennan, T., Bucy, P., Campbell, D., Chiu, S., Czerniewski, M., Denny, T., Edinger, M., ... Zhang, B. (2000). Multisite comparison of methods for the quantitation of the surface expression of CD38 on CD8+ T lymphocytes. Communications in Clinical Cytometry, 42(3), 174-179. https://doi.org/10.1002/1097-0320(20000615)42:3<174::AID-CYTO3>3.0.CO;2-2