The new method investigation for more effective utilization of flow cytometry

更有效利用流式细胞术的新方法研究

• 批准号: 06454619
• 负责人: NAKAHARA Kazuhiko
• 金额: $ 3.07万
• 依托单位: UNIVERSITY OF TOKYO (1995-1996)Kyorin University (1994)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-06454619/
• 关键词: fow cytometry; cell surface antigen; new index; quntitative analysis of antigens; クローサイトメトリー; 細胞内pH; 細胞周期

The purpose of this project is to find the methods to utilize flow cytometry (FCM) more effectively. FCM has been used most frequently for cell-surface antigen analysis. Hitherto the analysis is usually utilized only to calculate relative percentages of positive cells and the quantitative analysis of antigens has not been vetermined. In this project we have devised the new index for cell-surface antigens-quantitative analysis to study more detail.The forward scatter (FSC) pattern of cells stained with flurescent dye is divided into 5 pieces, and mean values of FSC and fluorescence intensity (FI) are calculated on each piece. The mean values of FSC are plotted on the X-axis, those of FI are on the Y-axis and an inclination of the recurrent line is named FF-coefficient as the new index. When FF-coefficient is constant, it indicates that cells' fluorscence intensity must change constantly according to change of the cell size and antigen values per unit area on the cell surface are constan … More t. We have examined and got the following results with this method.1. Lineal positive relationship between FSC and FI is recognized on CD4 antigens of CD4 positive T-lymphocytes. Its FF-coefficient is 0.40(]SY.+-。[)0.04 in healthy adult male, 0.36(]SY.+-。[)0.04 in healthy adult female and significant difference is recognized between them.2. The FF-coefficient of IL-2alpha antigens on T-lymphocytes increases (1.60(]SY.+-。[)0.40, control : 0.06(]SY.+-。[)0.16) after stimulating lymphocytes by ConA.To the contrary the FF-coefficient of CD4 dose not change (0.19(]SY.+-。[)0.04, control : 0.20(]SY.+-。[)0.05).3. The FF-coefficient of CD4 antigens on memory cells (CD45RO strong positive and CD45RA negative cells) is larger than it on naive cells (CD45RA strong positive and CD45RO negative cells). Almost same values of the FF-coefficient of CD8 antigens are counted.We conclude that our proposed new index, FF-coefficient, must be useful for detail examination of diseases with the method of quntitative analysis of cell surface antigens. Less

本项目的目的是寻找更有效地利用流式细胞术(FCM)的方法。流式细胞术是最常用的细胞表面抗原分析方法。到目前为止，这种分析通常只用于计算阳性细胞的相对百分比，而对抗原的定量分析尚未确定。在本项目中，我们设计了一种新的细胞表面抗原指标-定量分析来进行更详细的研究，将荧光染料染色的细胞的前向散射(FSC)图案分成5块，并计算每一块的FSC平均值和荧光强度(FI)。FSC的平均值在X轴上，FI的平均值在Y轴上，回归线的倾斜度被称为FF系数作为新的指标。当FF系数不变时，表明细胞的荧光强度必须随着细胞大小的变化而不断变化，细胞表面单位面积的抗原值是恒定的…我们用这种方法进行了检验，得到了如下结果。FSC与FI在CD4阳性T淋巴细胞的CD4抗原上呈线性正相关。其FF系数为0.40(SY+-)。[)0.04，健康成年男性，0.36(]sy.+-。[)0.04在健康成年女性中，两者之间存在显著差异。T淋巴细胞表面IL-2α抗原的Ff系数增加(1.60±0.00)。[)0.40，对照：0.06(SY+-)。[[]0.16)用刀豆蛋白A刺激淋巴细胞后，CD_4的F_F系数无明显变化(0.19±0.16)。[)0.04，对照：0.20(SY+-)。[)0.05).3.记忆细胞(CD45RO强阳性细胞，CD45RO阴性细胞)上CD4抗原的FF系数大于幼稚细胞(CD45RA强阳性细胞，CD45RO阴性细胞)。我们的结论是，我们提出的新的指标--FF-系数，对于定量分析细胞表面抗原的疾病的详细检查是有用的。较少

项目成果

渡邉 卓: "フローサイトメトリーによるリンパ球表面抗原定量的解析法の開発" 臨床病理. 43(suppl). 177 (1995)

Takashi Watanabe：“通过流式细胞术定量分析淋巴细胞表面抗原的方法的开发”临床病理学43（增刊）177（1995）。

中原 一彦: "白血球表面抗原" 日本臨床. 54・6. 1536-1541 (1996)

中原和彦：“白细胞表面抗原”日本临床杂志 54・6（1996）。

米山彰子: "血液疾患における可溶性リンパ球抗原の有用性" 臨床血液. 36. 286-292 (1995)

Akiko Yoneyama：“可溶性淋巴细胞抗原在血液疾病中的用途”《临床血液学》36. 286-292 (1995)。

中原一彦: "臨床検査ひと口メモ" モダンメディア. 40. 580-585 (1994)

中原和彦：《临床试验笔记》现代媒体 40. 580-585 (1994)。

Yoneyama,A: "Increased levels of soluble CD8 and CD4 in patients with infectious mononucleosis." Brit,J.Haematol,. 89. 47-54 (1995)

Yoneyama,A：“传染性单核细胞增多症患者的可溶性 CD8 和 CD4 水平升高。”