CD45作门的三色流式细胞术在白血病免疫分型中的应用

目的 介绍一种直接裂解骨髓样本进行急慢性白血病分型的三色流式细胞术。方法 将这种方法应用于115例急性白血病和10例慢性白血病的免疫分型。利用CD45和侧向散射将白血病细胞从正常的淋巴细胞，单核细胞，粒细胞和有核红细胞中区分出来后，用双参数流式细胞术进行分析。结果 流式细胞术测定得到的幼稚细胞比例在很大程度上与形态学计数相关，不同的FAB亚型白血病在CD45/SS图中的形式也不完全相同，当PC5标记的CD45与异硫氰酸荧光黄和藻红蛋白标记的其他抗体联合应用时，可以在对白血病细胞作门的基础上，同时进行白血病细胞表型表达的双参数分析。结论 在白血病免疫分型时，这种作门技术明显优于传统的前向散射对侧向散射作门方法。

Application of Three-Color Flow Cytometry Gated with CD45 in Leukemia Immunophenotyping

Objective To describe the procedure to perform three - color flow cytometric analysis on lysed whole bone marrow preparations for acute and chronic leukemia. Methods This approach was used to 115 patients with acute leukemia and 10 patients of chronic leukemia. Leukemia cells were analyzed by dual -parameter flow cytometry to distinguish normal lymphocytes, monocytes, granulocytes and nucleated red blood cells with combination of CD45 intensity and side scatter. Results The estimate of blasts by flow cytometric analysis was correlated highly with morphologic leukemia cell counts over a wide range. The pattern on the CD45/SS display varied in different FAB subtypes of leukemia. When CD45 - PCS was combined with FITC - and PE - conjugated antibodies, it was possible to set a gate on leukemia blasts and demonstate the dual -parameter analysis of the expressions of two additional markers in the leukemia population. Furthermore, this method also showed pattern display on the specimens of remission satisfactorily . Conclusion This gating strategy was superior to traditional FS versus SS display in immunophenotyping of leukemia.