Abstract
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-α or -β 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 TCRαβ/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.
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Kyoizumi, S., Kusunoki, Y., Hayashi, T. (2005). Flow Cytometric Measurement of Mutant T Cells With Altered Expression of TCR . In: Keohavong, P., Grant, S.G. (eds) Molecular Toxicology Protocols. Methods in Molecular Biology™, vol 291. Humana Press. https://doi.org/10.1385/1-59259-840-4:197
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DOI: https://doi.org/10.1385/1-59259-840-4:197
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