Measurement of the CD3− 4+ Variant T Cell Frequency by Flow Cytometry after X‐Irradiation on Mice

Abstract

Measurement of the CD3− 4+ Variant T Cell Frequency by Flow Cytometry after X‐ Irradiation on Mice: Naoki Kunugita, et al. Department of Radiation Biology and Health, School of Medicine, University of Occupational and Environmental Health, Japan—Variant cell frequency of mice T‐lymphocytes defective in the T‐cell receptor (TCR) gene expression was measured by flow cytometry. Splenic T‐lymphocytes were obtained from mice after irradiation and stained with fluorescein‐labeled anti‐L 3 T 4 (CD 4) and phycoerythrin‐ labeled anti‐CD3‐£ antibodies. They were analyzed with a flow cytometer to detect variant T cells lacking surface expression of TCR/CD 3 complexes which showed CD3− 4+ T cells. Variant T cells could be observed 3 days after 3 Gy irradiation. Frequency of variant T cells increased to the maximum level of 94×10−4 between 15 and 21 days after 3 Gy irradiation on BALB/c mice and gradually decreased with a half‐life of approximately 17 days. We examined the strain difference in radiosensitivity of splenic T‐lymphocytes in BALB/c, C57BL/6, C3H/ He and B6C3F1 mouse strains by this assay method. We observed that BALB/c showed the most radiosensitivity. This assay method is quick and easy in comparison with other methods and is considered useful for the mutagenicity test.