IMMUNOPHENOTYPING OF ACUTE-LEUKEMIA BY FLOW CYTOMETRIC ANALYSIS - USE OF CD45 AND RIGHT-ANGLE LIGHT SCATTER TO GATE ON LEUKEMIC BLASTS IN 3-COLOR ANALYSIS
IMMUNOPHENOTYPING OF ACUTE-LEUKEMIA BY FLOW CYTOMETRIC ANALYSIS - USE OF CD45 AND RIGHT-ANGLE LIGHT SCATTER TO GATE ON LEUKEMIC BLASTS IN 3-COLOR ANALYSIS
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DOI：
10.1093/ajcp/100.5.534
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发表时间：
1993-11-01
影响因子：
3.5
通讯作者：
STELZER, GT
中科院分区：
文献类型：
--
作者：
BOROWITZ, MJ;GUENTHER, KL;STELZER, GT
本文介绍了一种常规三色流式细胞术分析急性白血病的方法。该技术结合CD45强度和直角光散射（RALS）来区分白血病细胞与正常淋巴细胞、单核细胞、中性粒细胞、嗜酸性粒细胞和有核红细胞。在此显示中，白血病细胞占据一个独特的母细胞区，其特征是中等CD45密度和低RALS，在正常骨髓中，该区域占细胞总数的5%以下。该方法应用于39例急性白血病和8例骨髓增生异常或骨髓增生性疾病。流式细胞术分析估计的细胞数量与白血病细胞形态学计数在很大范围内高度相关。此外，CD45-RALS显示的模式对于不同的法国-美国-英国白血病亚型是不同的，这表明这种模式可能对分类有用。当CD45-peridin叶绿素α蛋白与其他对异硫氰酸荧光素和藻红蛋白偶联试剂结合时，可以在白血病原细胞上设置分析窗口，并显示关于白血病群体中两种附加标记表达的双参数（绿色荧光与红色荧光）数据。这种门控策略优于传统的前角和RALS显示，因为它在分析分离白血病细胞方面做得更好。
This article describes a procedure for performing routine three-color flow cytometric analysis for acute leukemia on lysed whole bone marrow preparations. This technique uses the combination of CD45 intensity and right-angle light scatter (RALS) to distinguish leukemic cells from normal lymphocytes, monocytes, neutrophils, eosinophils, and nucleated red blood cells. On this display, leukemic cells occupy a unique blast region characterized by intermediate CD45 density and low RALS, which, in normal marrows, contains less than 5% of the total cells. This approach was applied to 39 cases of acute leukemia and 8 cases of myelodysplasia or myeloproliferative disorders. The estimate of blasts by flow cytometric analysis was correlated highly with morphologic leukemic cell counts over a wide range. Moreover, the pattern seen on the CD45-RALS display was different for different French-American-British subtypes of leukemia, suggesting that this pattern might be useful for categorization. When CD45-peridin chlorophyll alpha protein was combined with other pairs of fluorescein isothiocyanate- and phycoerythrin-conjugated reagents, it was possible to set an analysis window on the leukemic blasts and display dual-parameter tie, green vs. red fluorescence) data regarding expression of two additional markers on the leukemic population. This gating strategy was superior to traditional forward-angle versus RALS displays in that it did a better job of isolating the leukemic cells analytically.