title: Flow cytometric quantification of mutant T cells with altered expression of the T-cell receptor: detecting somatic mutants in humans and mice.
abstract: Spontaneously generated mutant T cells defective in T-cell receptor (TCR) gene expression are detectable at the frequency of 2×10(-4) in vivo, and the mutant fractions are dose dependently increased by exposure to genotoxic agents such as ionizing radiation. Mutant cells with altered expression of TCRα 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 incomplete TCRαβ/CD3 complexes 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.
authors.affiliation: Laboratory of Immunology, Department of Radiobiology, Radiation Effects Research Foundation, 5-2 Hijiyama Park, Minami Ward, Hiroshima, 732-0815, Japan, kyoizumi@rerf.or.jp.
authors.forename: Seishi
authors.initials: S
authors.lastname: Kyoizumi
authors.forename: Yoichiro
authors.initials: Y
authors.lastname: Kusunoki
authors.forename: Tomonori
authors.initials: T
authors.lastname: Hayashi
mesh_terms: D000818:Animals; D015496:CD4-Positive T-Lymphocytes / Q000378:metabolism*; D002469:Cell Separation; D005434:Flow Cytometry; D015870:Gene Expression; D006801:Humans; D051379:Mice; D009154:Mutation; D011948:Receptors, Antigen, T-Cell / Q000235:genetics / Q000378:metabolism*
chemical_list: D011948:Receptors, Antigen, T-Cell