How to cite this article: Wang L, Abbasi F, Jasper GA, Kreitman RJ, Liewehr DJ, Marti GE, Stetler-Stevenson M. Variables in the Quantification of CD4 in Normals and Hairy Cell Leukemia Patients. Cytometry Part B 2011; 80B: 51–56.
Article first published online: 4 AUG 2010
Published 2010 Wiley-Liss, Inc.
Cytometry Part B: Clinical Cytometry
Volume 80B, Issue 1, pages 51–56, January 2011
How to Cite
Wang, L., Abbasi, F., Jasper, G. A., Kreitman, R. J., Liewehr, D. J., Marti, G. E. and Stetler-Stevenson, M. (2011), Variables in the quantification of CD4 in normals and hairy cell leukemia patients. Cytometry, 80B: 51–56. doi: 10.1002/cyto.b.20541
Disclaimer: Certain commercial equipment, instruments, and materials are identified in this paper to specify adequately the experimental procedure. In no case does such identification imply recommendation or endorsement by the National Institute of Standards and Technology, nor does it imply that the materials or equipment are necessarily the best available for the purpose.
This article is a US government work and, as such, is in the public domain in the United States of America.
- Issue published online: 22 DEC 2010
- Article first published online: 4 AUG 2010
- Manuscript Accepted: 17 MAY 2010
- Manuscript Revised: 5 MAY 2010
- Manuscript Received: 28 JAN 2010
- quantitative flow cytometry;
- CD4 quantification;
- anti-CD4 PE unimolar conjugate;
- sample preparation method
Quantitative flow cytometry (QFCM) is being applied in the clinical flow cytometry laboratory. Quantitative normal T-cell CD4 expression represents a biologic standard and quality control agent. However, low levels of CD4 expression were detected in normal T-cells in Hairy Cell Leukemia (HCL) samples.
The QuantiBrite System® was used to determine the level of CD4 expression (mean antibody bound per cell, ABC) in fresh and shipped HCL blood and fresh normal donor blood (NDB). The effects of shipping, lysing reagent, cell preparation method, and antibody lot were evaluated.
Shipped HCL specimens (n = 69) had a significantly lower mean CD4 ABC of 38,788 (CV = 9.1%) compared to fresh specimens (n = 105) CD4 value of 40,330 (CV = 8.4%) (P < 0.05). In NDB, significant differences were seen for fresh versus shipped specimens using the stain/lyse method but not for lyse/stain method. Consistent differences in CD4 ABC based upon antibody lot were observed in fresh HCL and NDB samples. Stain/lyse and lyse/stain methods using NH4Cl lyse were compared in NDB using identical samples and antibodies. The NDB CD4 ABC values obtained with the lyse (NH4Cl)/stain method (45,562, 3.7% CV) were lower than those obtained with the stain/lyse (NH4Cl) method (49,955, 3.3% CV) with P < 0.001.
CD4 expression in HCL patient samples is not inherently different from that observed in NDB and therefore may serve as a biological control in clinical QFCM. Technical variables impact significantly on QFCM of CD4. Published 2010 Wiley-Liss, Inc.