Modes of cell death of feline T lymphocytes by ionizing radiation

Funayama, Tomoo; Kakizaki, Takehiko; Wada, Seiichi; Hamada, Nobuyuki*; Sakashita, Tetsuya; Hodatsu, Tsutomu*; Yamada, Naoaki*; Sano, Tadashi*; Natsuhori, Masahiro*; Kobayashi, Yasuhiko; Ito, Nobuhiko*

Responses of two lines of feline T lymphocytes, FeT-J and FL-4, to ionizing radiation were examined in the present study. FL-4 cells, but not FeT-J, were persistently infected with feline immunodeficiency virus. Cells were irradiated with Co- rays at 2 Gy/min. Surviving fractions were evaluated with clonogenic assay. Apoptosis was detected using TdT-mediated dUTP-biotin nick end labeling (TUNEL) method. Changes in nuclear morphology of viable cells were evaluated by double staining with Hoechst 33342 and ethidium bromide. Mean surviving fractions of FL-4 cells were slightly higher than FeT-J for each radiation dose given. However, dose required to reduce to 63 % of surviving fraction was 1.9 Gy in both cell lines. TUNEL assay, however, revealed the maximum frequency of apoptosis in FL-4 cells (20 %) was lower than in FeT-J cells (40 %). The time of frequency peak of TUNEL-positive cells in FL-4 was shorter than that of FeT-J. FL-4 cells reached peak level within 24 hours after irradiation, but FeT-J cells needed later than 48 hours after irradiation. Exposure of FL-4 cells to rays resulted in the giant- and the multi- nucleus formation. Nuclear swelling occurred to less extent in FeT-J than FL-4 cells. In conclusion, we found the difference in cellular responses to radiation in two lines of feline T lymphocytes. Modes of cell death of FL-4 cells were non-apoptotic and more research would discover new mechanism associated with apoptosis.