Rossmann Eva D, Lenkei Rodica, Lundin Jeanette, Mellstedt Håkan, Osterborg Anders
Department of Oncology (Radiumhemmet), Karolinska University Hospital, Stockholm, Sweden.
Cytometry B Clin Cytom. 2007 Nov;72(6):450-7. doi: 10.1002/cyto.b.20359.
The fluorescence intensities of CD3, CD4 on T cells and CD20, CD22 molecules on B cells were quantitatively measured on lymphocytes from chronic lymphocytic leukemia (CLL) patients and healthy donors.
The performance of three different types of microbeads was compared, i.e. Quantum molecules of equivalent soluble fluorochrome (Q-MESF), Quantum simply cellular (QSC), and QuantiBRITE (QB). As all PE-conjugates had a F/P ratio of 1:1, the MESF units represented also the antibody binding capacity (ABC).
The ABCs of CD4 and CD20 antigens estimated with QSC (ABC(QSC)) were higher than those assigned with QB (ABC(QB)) with an average difference 49%. Higher numbers of antigenic sites were obtained with Q-MESF than with QSC for CD20 antigen. On the contrary, CD4 antigenic sites numbers estimated with QSC were higher than those estimated with Q-MESF. ABC values estimated with Quantum MESF PE (ABC(Q-MESF)) were approximately 15% higher than ABC(QSC), whereas ABC(Q-MESF) was approximately 49% higher than ABC(QB). Statistically significant correlations were found between the values obtained using various standards. The present study is the first to report down-regulation of CD3 antigen on T cells from patients with CLL.
This study emphasizes the relevance of quantitative measurement of fluorescence intensity by flow cytometry as a standardized approach to measure and interpret the expression of some CLL markers and reduce variability of results obtained at different sites in multi-center clinical studies.
对慢性淋巴细胞白血病（CLL）患者及健康供体的淋巴细胞进行定量检测T细胞上CD3、CD4以及B细胞上CD20、CD22分子的荧光强度。
比较了三种不同类型微珠的性能，即等效可溶性荧光染料量子分子（Q-MESF）、量子单细胞（QSC）和定量荧光（QB）。由于所有PE偶联物的F/P比均为1:1，MESF单位也代表抗体结合能力（ABC）。
用QSC估算的CD4和CD20抗原的ABC（ABC(QSC)）高于用QB估算的值（ABC(QB)），平均差异为49%。对于CD20抗原，用Q-MESF获得的抗原位点数量比用QSC多。相反，用QSC估算的CD4抗原位点数量高于用Q-MESF估算的值。用量子MESF PE估算的ABC值（ABC(Q-MESF)）比ABC(QSC)高约15%，而ABC(Q-MESF)比ABC(QB)高约49%。在使用各种标准获得的值之间发现了具有统计学意义的相关性。本研究首次报道了CLL患者T细胞上CD3抗原的下调。
本研究强调了通过流式细胞术对荧光强度进行定量测量作为一种标准化方法来测量和解释某些CLL标志物表达并减少多中心临床研究中不同地点获得结果的变异性的相关性。
