Kawai Y, Ujiie M, Amano H, Sonoda A, Watanabe K
Department of Laboratory Medicine, Keio University, School of Medicine, Tokyo.
Rinsho Byori. 1995 Sep;43(9):931-9.
Determination of the human lymphocyte subpopulation selected by immunofluorescence and the phenotypic analysis of hematological malignant cells by laser flow cytometry have become popular and useful tests in various laboratories. However, several lines of evidence have questioned the accuracy and reproducibility of these analysis. We examined the problems of laser flow cytometric analysis to measure the lymphocyte subpopulation and determine the phenotypic expression of hematological malignancies. In lymphocyte subset analysis, no survey has been applied to reveal the accuracy and reproducibility of these tests. We compared the accuracy of gating events and the ratio of lymphocytes using leuco GATE/simul SET analysis to those by manual gate method analysis. We found that there were some patients with SLE in which the accurate lymphocyte subpopulation was difficult to calculate due to the gating of lymphocytes by either method. Furthermore, apparent differences in the lymphocyte population were observed between these methods. In the phenotypic analysis of hematological malignancies, there have been several problems over 30% of the total cells had to be abnormal cells. Second, the malignant cells were difficult to gate unless the information of the size, shape and cellular density were obvious. Third, the phenotype of malignant cells were often different from that of the normal matured cells in the some lineage. However, flow cytometric analysis was useful to determine the cell lineage of peroxidase-negative cells and to diagnose the hybrid leukemia. In summary, the phenotypic analysis using flow cytometry and various monoclonal antibodies are clinically useful tests to diagnose the immunological disorders and hematological malignancies. However, there remain several problems to be solved in the near future.
通过免疫荧光法测定人淋巴细胞亚群以及利用激光流式细胞术对血液系统恶性细胞进行表型分析，已成为各实验室常用且实用的检测方法。然而，有几条证据对这些分析的准确性和可重复性提出了质疑。我们研究了激光流式细胞术分析在测量淋巴细胞亚群及确定血液系统恶性肿瘤表型表达方面的问题。在淋巴细胞亚群分析中，尚未有研究用于揭示这些检测的准确性和可重复性。我们将使用白细胞门控/模拟设置分析的门控事件准确性和淋巴细胞比例与手动门控方法分析的结果进行了比较。我们发现，有一些系统性红斑狼疮患者，无论采用哪种方法对淋巴细胞进行门控，都难以准确计算淋巴细胞亚群。此外，这两种方法在淋巴细胞群体上观察到明显差异。在血液系统恶性肿瘤的表型分析中，存在几个问题：首先，超过30%的总细胞必须为异常细胞；其次，除非细胞大小、形状和细胞密度信息明显，否则恶性细胞难以进行门控；第三，在某些谱系中，恶性细胞的表型往往与正常成熟细胞不同。然而，流式细胞术分析对于确定过氧化物酶阴性细胞的细胞谱系以及诊断混合性白血病很有用。总之，使用流式细胞术和各种单克隆抗体进行表型分析是诊断免疫紊乱和血液系统恶性肿瘤的临床有用检测方法。然而，在不久的将来仍有几个问题有待解决。
