Analysis of DNA damage recovery processes in the adaptive response to ionizing radiation in human lymphocytes

It has been frequently suggested that the adaptive response to ionizing radiation involves the induction of a chromosomal repair mechanism. Although several lines of evidence favour this assumption, direct proof is lacking. We have chosen to study this question with the help of the comet assay. Lymphocytes from three human donors were given an adapting dose of 0.05 Gy 16 h after mitogenic stimulation and a challenging dose of 2 Gy 5 h thereafter. While a portion of the cells was removed from the cultures for the comet assay, remaining cells were harvested at 52 h culture time and screened for chromosomal aberrations. In some experiments an analysis of cell proliferation was additionally carried out by flow cytometry. In the comet assay a reduced level of initial damage and an increased repair capacity was observed in the adapted + challenged cells; however, this did not result in a reduction of the aberration frequencies. No effect of the adapting dose on cell proliferation was detectable. The analysis of comet distributions revealed that the observed enhanced repair capacity was due to the presence of a subpopulation of slowly repairing cells in the challenged lymphocytes and the lack of such a subpopulation in the adapted + challenged cells. We assume that the slowly repairing cells were quiescent G0 lymphocytes which were removed from the adapted + challenged cell population, probably by apoptotic-like processes.