Variables in the Quantification of CD4 in Normals and Hairy Cell Leukemia Patients
Variables in the Quantification of CD4 in Normals and Hairy Cell Leukemia Patients
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DOI：
10.1002/cyto.b.20541
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发表时间：
2011-01-01
影响因子：
3.4
通讯作者：
Stetler-Stevenson, Maryalice
中科院分区：
文献类型：
--
作者：
Wang, Lili;Abbasi, Fatima;Stetler-Stevenson, Maryalice
背景：定量流式细胞术（QFCM）正在临床流式细胞术实验室中得到应用。正常t细胞CD4的定量表达是一种生物学标准和质量控制手段。然而，在毛细胞白血病（HCL）样本的正常t细胞中检测到低水平的CD4表达。方法：采用QuantiBrite系统(R)检测新鲜、运输HCL血和新鲜正常供者血（NDB）中CD4表达水平（平均抗体结合每细胞，ABC）。评估了运输、裂解试剂、细胞制备方法和抗体批次的影响。结果：运标本（n = 69） CD4平均ABC为38,788 (CV = 9.1%)，低于新鲜标本（n = 105）的40,330 (CV = 8.4%) （P < 0.05）。在NDB中，使用染色/裂解法观察到新鲜和运输标本的显著差异，但裂解/染色法没有。CD4 ABC在新鲜HCL和NDB样品中基于抗体批次观察到一致的差异。使用相同的样品和抗体，比较NDB中使用NH4Cl lyse的染色/裂解和裂解/染色方法。lyse (NH4Cl)/染色法获得的NDB CD4 ABC值（45,562,3.7% CV）低于染色/lyse （NH4Cl）法（49,955,3.3% CV）， P < 0.001。结论：HCL患者样本中的CD4表达与NDB中观察到的CD4表达并无本质差异，因此可能作为临床QFCM的生物学对照。技术变量对CD4 QFCM有显著影响。2010年Wiley-Liss出版（匕首）
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 (R) 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.(dagger)