ANALYSIS OF RAT LYMPHOCYTE-ACTIVATION OF BENZO[A]PYRENE, 2-ACETYLAMINOFLUORENE, AND SEVERAL OF THEIR METABOLITES TO MUTAGENIC AND DNA-DAMAGING SPECIES INVITRO

Rat lymphocytes are a potentially useful and convenient cell system for monitoring the genotoxic effects of chemicals in vivo, but little is known about the ability of these cells to metabolize promutagens to genotoxic species. In this study, Fischer 344 rat lymphocytes were treated in vitro with benzo[a]pyrene (BaP), 2-acetylaminofluorene (2-AAF), and several of their metabolites, and DNA damage was measured using nucleoid sedimentation analysis. Of the BaP derivatives, BaP 4,5-oxide and BaP 7,8-diol-9,10-epoxide decreased lymphocyte nucleoid sedimentation, whereas BaP and BaP 7,8-dihydrodiol had little effect. Among the 2-AAF derivatives, N-acetoxy-2-AAF, N-hydroxy-2-AAF, and N-hydroxy-2-aminofluorene damaged rat lymphocyte nucleoids, whereas 2-AAF, 2-aminofluorene, and fluorene produced little detectable damage. The decrease in nucleoid sedimentation produced by N-hydroxy-2-AAF was not inhibited by paraoxon, salicylamide, or 2-aminofluorene, whereas paraoxon inhibited damage produced by N-acetoxy-2-AAF. In co-culture assays, rat lymphocytes increased the mutagenicity of N-hydroxy-2-AAF in Salmonella typhimurium strain TA98, but mediated little or no mutagenic response with BaP, BaP 7,8-dihydrodiol, and 2-AAF. Also, human lymphocytes, but not rat lymphocytes, mediated a positive mutagenic response with BaP 7,8-dihydrodiol in Chinese hamster ovary UV5 cells. Although rat lymphocytes may metabolize certain proximal genotoxic chemicals to DNA-damaging species (e.g., N-hydroxy-2-AAF), these data suggest that in vivo lymphocyte DNA damage is more likely to result from lymphocytes encountering reactive chemical derivatives produced by other cells. It is also clear that differences exist between the ability of human and rat lymphocytes to activate promutagens, and this may impact on the use of the rat model to predict the genotoxicity of chemicals in humans.