Development of a dynamic analysis method for phagocytic activity utilizing flow cytometry

建立了利用流式细胞术(FCM)分析吞噬活性的新方法。我们观察了细胞吞噬微球分数的变化，认为微球在V79细胞系中的吞噬作用可用一个两参数的实验方程来描述。研究结果显示在以下12个部分。1)给出了流式细胞仪测定吞噬功能的典型例子。2)将FCM测量结果与显微镜测量结果进行了比较，证实了FCM测量的可靠性。3)说明了吞噬微球与贴壁微球的区别。4)考察了适合FCM测量的微球尺寸。5)考察了细胞周围微球的平衡性。6)荧光直方图中峰的变异系数的增大与微球在细胞内的定位有关。7)比较巨噬细胞和V79细胞的吞噬活性。8)证明了一个描述吞噬作用的实验方程。9)考察了微球浓度对吞噬参数k和n的影响。10)给出了分析吞噬活性的计算机程序。11)讨论了吞噬活性对细胞周期的依赖关系。12)探讨了羟基脲对巨噬细胞吞噬功能的影响。

A new method to analyze the phagoctic activity utilizing flow cytometry (FCM) was developed. We observed the changes in the fraction of cell phagocytizing microspheres and concluded that the phagocytosis of microspheres in V79 cultured cell line could be described with an experimental equation with two parameters. The results were shown in the following twelve sections. 1) Representative examples of FCM measurements of phagocytosis were shown. 2) Results form FCM and microscopic measurements were compared and the reliability of FCM measurements was confirmed. 3) How to distinguish phagocytized microspheres from attached those was explained. 4) Microsphere size suitable to the FCM measurements was examined. 5) Equilibrium of microspheres surrounding cells was examined. 6) Increase in coefficient of variation of peaks in fluorescence histograms was explained by the localization of microspheres in cells. 7) Phagocytic activity was compared between macrophages and V79 cells. 8) An experimental equation describing pliagocytosis was demonstrated. 9) Effects of microsphere concentration on the phagocytosis parameters k and n was examined. 10) A computer program to analyze the phagocytic activity was shown. 11) Cell cycle dependency of phagocytic activity was discussed. 12) Effects of hydroxyurea on the phagoeytosis were discussed.