title: A romatic DNA adduct levels in human peripheral blood lymphocytes and total white blood cells by 32P postlabelling: need for validation.
abstract: Long lived lymphocytes tend to have higher 32P postlabelling measured levels of adducts than short lived granulocytes in environmental and life style associated i.e. smoking exposures. With the aim of investigating this issue for occupational exposure to PAH and contributing to further validation of some technical aspects of the 32P postlabelling assay, two Italian laboratories analysed PAH-DNA adducts from lymphocytes and total white blood cells WBC. Seventy seven blood samples from coke oven workers employed at a steel plant located in Taranto, Southern Italy, and 14 samples from control subjects were collected. At the University of Padua, DNA was purified from peripheral blood lymphocytes PBL. Two years later, at the University of Bari, white blood cells WBC were isolated from replicate blood samples stored at- 80 C and DNA purified by the same method. In both cases, the nuclease P1 modified postlabelling assay was used to determine aromatic DNA adduct levels. The mean adduct levels were 5.13 3.37 Padua and 2.48 1.27 Bari per 108 nucleotides. Both laboratories observed large inter individual variations of adduct levels ranging from 0.09 to 18.93 per 108 nucleotides. Both the correlation and the agreement of the two sets of data were assessed. Slight correlation r = 0.39; p 0.01 and a poor level of agreement were found, the intra class correlation coefficient being equal to 0.05. Better correlation coefficient r = 0.54, p 0.01 and intra class correlation coefficient r = 0.50 were observed comparing only the adduct levels determined on the diagonal zone DRZ. Our findings seem to confirm the same divergence reported in the literature on DNA adduct levels between lymphocytes and granulocytes.
authors.lastname: Giuseppina Brescia Vito Foa Cristina Viezzer Lucia Celotti Giorgio Assennato