Spontaneous and induced genetic damage in T lymphocyte subsets evaluated by the Comet assay.

High inter- and intra-individual variability was reported in the level of DNA damage, both spontaneous and induced, when peripheral blood mononuclear leukocytes were used to perform the Comet assay. In order to find out the underlying causes for such variability, different subsets of T lymphocytes were isolated by immunomagnetic cell sorting. The level of DNA damage was evaluated with the alkaline version of the Comet assay by using three different parameters: tail moment, tail length and amount of DNA in the tail (%). Helper T cells (CD4+), cytotoxic T cells (CD8+), their negative fraction and the mixed cell population were evaluated both in untreated cells and after 10 and 20 microM H(2)O(2) treatments. Differences between cell subsets were only observed after H(2)O(2) treatment. The results indicate that, although CD4+ is the fraction with the highest induced level of genetic damage, this value is not high enough to explain the large inter- and intra-individual variability found.