Frequency, distribution and clonality of chromosome damage in human lymphocytes by multi-color FISH

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Abstract
Whole chromosome painting was used to determine whether the use of different sets of paints would influence results obtained from the analysis of human peripheral blood lymphocytes from 27 healthy unexposed subjects. Painting was also used to determine if aberration frequencies are proportional to the size of selected chromosomes in human lymphocytes irradiated in vitro. The in vitro results showed that the frequencies of radiation-induced stable aberrations (i.e. translocations and insertions) in chromosomes 3,5 and 6 painted in unique colors are proportional to chromosome size. Aberration frequencies in the normal subjects were measured using two different sets of paints, one set for chromosomes 3,5 and 6 where each chromosome was labeled in a unique color and one set where chromosomes 1,2 and 4 were painted in a single color. The frequency of aberrations among chromosomes 1-6 in the population as a whole was also found to be proportional to chromosome size. However, some individual subjects had a distribution of damage that was not proportional to chromosome size due to the presence of clones of abnormal cells. Aberration frequencies measured in chromosomes 3, 5 and 6 as a set were highly correlated with those observed in chromosomes 1,2 and 4 as a set, after adjusting for the different amounts of the genome that were painted. We also determined whether differences exist in the aberration frequencies measured by two scoring systems: the classical method, where reciprocal exchanges are scored as single events, and PAINT, where each break junction is scored as a single event The two scoring systems gave highly correlated results for translocations and differed by a constant value (PAINT X 0.58 = classical method). Approximately 27% of translocations were observed to be nonreciprocal due to a failure to detect exchanges involving small amounts of material or to a non-reciprocal exchange mechanism. Our results support the hypothesis that cytogenetic evaluations for biodosimetry can be performed with any one or more of the chromosomes studied here and indicate that the aberration frequency measurements are independent of the scoring system selected for the evaluation. We also present a simple statistical method for identifying subjects that may possess clonal aberrations

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