Measurement of spontaneous and X-irradiation-induced 6-thioguanine-resistant human blood lymphocytes using a T-cell cloning technique

Abstract

Lymphocytes separated from human peripheral blood were cultured in vitro, in the presence of 6-thioguanine (TG), to select and clone rare TG-resistant (TGr) cells present in the circulation in vivo. The incidence of such TGr cells ranged from 0.83 X 10(-5) to 2.53 X 10(-5) (mean 1.48 X 10(-5) ) in healthy individuals aged between 19 and 79 years; did not differ between males and females; but increased significantly with age at a rate of 2.4 cells/10(7) lymphocytes/year. Exposure of lymphocytes (G0) in vitro to X-ray doses of upto 200 rad resulted in a dose-dependent increase in TGr cell frequencies. The rates of increase were approximately in proportion to the square of the dose and these rates were closely similar to those obtained in cultured skin fibroblasts and suggest that the bulk of these mutations are a consequence of chromosome structural aberrations. The cloned TGr cells are considered to be HPRT- mutants and the mutation frequencies in lymphocytes determined using this cloning technique were compared with the variant frequencies obtained in earlier experiments utilising an autoradiographic technique to detect azaguanine-resistant (AGr) variant cells. Mutation frequencies with the cloning technique were 10-20-fold lower than variant frequencies with the autoradiographic method.