Philip P, Sartiaux C
Laboratoire d'Immunologie, Faculté de Médecine de Nancy, Vandoeuvre les Nancy, France.
Leuk Lymphoma. 1994;13 Suppl 1:45-8. doi: 10.3109/10428199409052673.
The use of flow cytometry as a common tool has opened the field of quantimetric immunophenotyping. This could help refine the definition of leukemic proliferations, and perhaps provide new prognostic criteria. We investigated whether this approach could be applied to multicentric immunophenotyping. The reproducibility of quantitative measurements on different types of flow cytometers was tested using a single batch of calibrating and "blind" fluorescent beads. Less than 7% CV was obtained between 20 different laboratories. We then explored the variations noted with control or normal lymphocytes, using various CD4 monoclonal antibodies, which were found to be lower than 10%. We finally applied this strategy to acute lymphoblastic leukemias (ALL), performing quantitative measurements of CD10 using one single aliquoted batch of FITC-CD10 in comparison with locally used reagents. PreB-ALL could indeed be further stratified depending on the density of CD10 expression. Use of the same monoclonal antibody is however recommended, since PreB-ALL also seem to display epitopic variations of the CD10 molecule.
流式细胞术作为一种常用工具的应用开启了定量免疫表型分析领域。这有助于完善白血病增殖的定义,并可能提供新的预后标准。我们研究了这种方法是否可应用于多中心免疫表型分析。使用一批校准和“盲法”荧光微球测试了不同类型流式细胞仪上定量测量的可重复性。在20个不同实验室之间获得的变异系数(CV)小于7%。然后,我们使用各种CD4单克隆抗体探索了对照或正常淋巴细胞的变异情况,发现其低于10%。我们最终将该策略应用于急性淋巴细胞白血病(ALL),使用一批单一分装的FITC-CD10与当地使用的试剂相比,对CD10进行定量测量。前B-ALL确实可以根据CD10表达密度进一步分层。然而,由于前B-ALL似乎也显示出CD10分子的表位变异,因此建议使用相同的单克隆抗体。