Flow cytometric measurement of mutant T cells with altered expression of TCR: detecting somatic mutations in humans and mice.
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.
Kyoizumi
Seishi
S
Laboratory of Immunology, Department of Radiobiology/Molecular Epidemiology, Radiation Effects Research Foundation, Hiroshima, Japan.
Kusunoki
Yoichiro
Y
Hayashi
Tomonori
T
D000818:Animals; D015496:CD4-Positive T-Lymphocytes / Q000737:chemistry / Q000378:metabolism; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D051379:Mice; D009152:Mutagenicity Tests / Q000379:methods*; D009154:Mutation / Q000235:genetics; D011948:Receptors, Antigen, T-Cell / Q000096:biosynthesis / Q000235:genetics*
D011948:Receptors, Antigen, T-Cell