Variables Affecting the Quantitation of CD22 in Neoplastic B Cells
Variables Affecting the Quantitation of CD22 in Neoplastic B Cells
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DOI：
10.1002/cyto.b.20567
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发表时间：
2011-03-01
影响因子：
3.4
通讯作者：
Stetler-Stevenson, Maryalice
中科院分区：
文献类型：
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作者：
Jasper, Gregory A.;Arun, Indu;Stetler-Stevenson, Maryalice
背景资料：定量流式细胞术（QFCM）正在临床流式细胞术实验室中应用于接受抗体治疗的患者的诊断、预后和评估。研究了急性淋巴细胞白血病（ALL）、慢性淋巴细胞白血病（CLL）、套细胞淋巴瘤（MCL）、滤泡性淋巴瘤（FCL）、毛细胞白血病（HCL）和正常B细胞的ABC值以及技术变量对CD 22定量的影响。使用QuantiBrite System（R）测定恶性和正常B细胞的CD 22表达水平（每个细胞的平均抗体结合，ABC）。试验内变异性、精密度所需的细胞数量、延迟处理的影响以及外周血标本的运输（延迟处理和暴露于非受控环境），以及多聚甲醛固定对测定结果的影响。测量CD 22 ABC的QuantiBRITE方法是精确的（中位CV 1.6%，95%置信区间，1.2-2.3%），但可靠的CD 22 ABC值需要250个恶性细胞的阈值。延迟处理和隔夜装运的标本导致显着不同的ABC值，而固定长达12小时没有显着影响。ABC测量确定，CD 22的表达低于正常的ALL，CLL，FCL，MCL，但高于正常的HCL。结论：CD 22的表达是不典型的血液淋巴细胞恶性肿瘤的研究，可能有诊断价值。技术变量（如分析的细胞数量和延迟处理或过夜运输标本）对临床实验室中QFCM测量抗原表达产生显著影响。出版2010 Wiley-Liss，Inc.
Background: Quantitative flow cytometry (QFCM) is being applied in the clinical flow cytometry laboratory for diagnosis, prognosis, and assessment of patients receiving antibody-based therapy. ABC values and the, effect of technical variables on CD22 quantitation in acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL), mantle cell lymphoma (MCL), follicular lymphoma (FCL), hairy cell leukemia (HCL) and normal B cells were studied.Methods: The QuantiBrite System (R) was used to determine the level of CD22 expression (mean antibody bound per cell, ABC) by malignant and normal B cells. The intra-assay variability, number of cells required for precision, effect of delayed processing as well as shipment of peripheral blood specimens (delayed processing and exposure to noncontrolled environments), and the effect of paraformaldehyde fixation on assay results were studied.Results: The QuantiBRITE method of measuring CD22 ABC is precise (median CV 1.6%, 95% confidence interval, 1.2-2.3%) but a threshold of 250 malignant cells is required for reliable CD22 ABC values. Delayed processing and overnight shipment of specimens resulted in significantly different ABC values whereas fixation for up to 12 h had no significant effect. ABC measurements determined that CD22 expression is lower than normal in ALL, CLL, FCL, and MCL but higher than normal in HCL.Conclusions: CD22 expression was atypical in the hematolymphoid malignancies studied and may have diagnostic utility. Technical variables such as cell number analyzed and delayed processing or overnight shipment of specimens impact significantly on the measurement of antigen expression by QFCM in the clinical laboratory. Published 2010 Wiley-Liss, Inc.