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An effective analysis of individual cells using flow cytometry and its clinical applicatjion.

使用流式细胞仪对单个细胞进行有效分析及其临床应用。

基本信息

批准号：
13470516
负责人：
NAKAHARA Kazuhiko
金额：
$ 5.7万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2003
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13470516/
关键词：
flow cytometry cell surface antigen antigen molecule density cell size FF index quantification of cell surface molecule hematological malignancy clinical application 抗原密度解析ソフトリストモード急性白血病伝染性単核球症 list mode date 標準ビーズ

项目摘要

The purpose of this study is to explore an effective application of flow cytometry (FCM) to clinical medicine by using the information of individual cells. Until now, cytometric immunophenotyping using FCM has focused mainly on the quantification of the proportion of cells bearing certain combinations of molecules and the change of fluorescence intensity (FI) dose not reflect the antigen density directly. For the qualification of cell surface molecule density, the forward light scatter (FSC) and FI, obtained by FCM on an individual cell basis were plotted in a scattergram where x-and y-axis show FSC and FI respectively. Where a proportional relationship between FI and FSC can been seen, the slope of the line of regression (or its absolute value) may be used as an indicator of the molecule density, which is referred to as the FF index. In this way, the FI vs FSC scattergram enables the evaluation of the relationship between the size of the cell and the amount of antigen expressed. Additionally we explored the computer software which enables to practice automatically these analysis by using the listmode of FCM. We analyzed normal peripheral lymphocytes, hematological malignant cells and lymphocytes of hemodyalisis patients and found the potent usefulness of more precise clinical analysis using this software. In future, more clinical studies, the evaluation and standardization of the developed software and actual quantification of cell surface antigens are needed.

本研究旨在利用单个细胞的信息，探索流式细胞术(FCM)在临床医学中的有效应用。到目前为止，流式细胞术的免疫表型主要集中在定量细胞携带某种分子组合的比例，而荧光强度的变化并不能直接反映抗原的密度。为了确定细胞表面分子密度，将FCM在单个细胞基础上获得的前向光散射(FSC)和FI绘制在散射图中，其中x轴和y轴分别表示FSC和FI。在FI和FSC之间可以看到成比例的关系时，可以使用回归直线的斜率(或其绝对值)作为分子密度的指示器，这被称为FF指数。通过这种方式，FI VS FSC散点图能够评估细胞大小和所表达的抗原量之间的关系。此外，我们还开发了能够利用FCM的列表模式自动练习这些分析的计算机软件。我们分析了正常外周淋巴细胞、血液系统恶性肿瘤细胞和止血患者的淋巴细胞，发现使用该软件进行更精确的临床分析具有潜在的实用价值。在未来，还需要更多的临床研究，所开发的软件的评估和标准化，以及细胞表面抗原的实际定量。