Flow Cytometric Measurement of Mutant T Cells With Altered Expression of TCR
Flow Cytometric Measurement of Mutant T Cells With Altered Expression of TCR
复制标题
TCR 表达改变的突变 T 细胞的流式细胞术测量
DOI：
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发表时间：
2005
期刊：
影响因子：
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通讯作者：
Tomonori Hayashi
中科院分区：
文献类型：
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作者：
S. Kyoizumi;Y. Kusunoki;Tomonori Hayashi
自发产生的T细胞受体（TCR）基因表达缺陷的突变T细胞在体内可检测到10（-4）的频率，并且突变部分通过暴露于遗传毒性物质如电离辐射而剂量依赖性地增加。在CD 4 + T细胞中，TCR-α或-β表达改变的突变细胞可通过双色流式细胞术使用不同荧光染料标记的抗CD 3和抗CD 4单克隆抗体检测为CD 3-/CD 4+细胞，因为不完整的TCR α/CD 3复合物不能转运至细胞膜。该流式细胞术突变检测方法可应用于人外周血和小鼠脾脏的CD 4 + T细胞。描述了用于制备靶细胞和检测突变细胞的方法。
Spontaneously generated mutant T cells defective in T-cell receptor (TCR) gene expression are detectable at the frequency of 10(-4) in vivo, and the mutant fractions are dose-dependently increased by exposure to genotoxic substances such as ionizing radiation. Mutant cells with altered expression of TCR-alpha or -beta among CD4+ T cells can be detected as CD3-/CD4+ cells by two-color flow cytometry using anti-CD3 and anti-CD4 monoclonal antibodies labeled with different fluorescent dyes, because an incomplete TCRalphabeta/CD3 complex cannot be transported to the cellular membrane. This flow cytometric mutation assay can be applied to CD4+ T cells from human peripheral blood and mouse spleen. Methods for both preparation of target cells and detection of the mutant cells are described.