Variables in the quantification of CD4 in normals and hairy cell leukemia patients
| Autor: | Robert J. Kreitman, Gregory A. Jasper, Gerald E. Marti, Lili Wang, Maryalice Stetler-Stevenson, David J. Liewehr, Fatima Abbasi |
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| Rok vydání: | 2010 |
| Předmět: |
CD4-Positive T-Lymphocytes
Leukemia Hairy Cell Histology Lysis medicine.diagnostic_test Cell preparation Cell Biology Biology medicine.disease Molecular biology Stain Article Immunophenotyping Pathology and Forensic Medicine Flow cytometry Reference Values Case-Control Studies CD4 Antigens medicine biology.protein Humans Hairy cell leukemia Antibody Stain method |
| Zdroj: | Cytometry Part B: Clinical Cytometry. :51-56 |
| ISSN: | 1552-4949 |
| Popis: | Background: 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. Methods: 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. Results: 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. Conclusions: 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. |
| Databáze: | OpenAIRE |
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