Detection of benzo[a]pyrene-DNA adducts in cultured cells treated with benzo[a]pyrene diol-epoxide by quantitative immunofluorescence microscopy and 32P-postlabelling; immunofluorescence analysis of benzo[a]pyrene-DNA adducts in bronchial cells from smoking individuals.
Monoclonal antibodies were raised against the reaction product of benzo[a]pyrene diol-epoxide (BPDE) and deoxyguanosine-5'-monophosphate. The antibodies were used for detection of DNA adducts in situ in BPDE-treated cultured human fibroblasts by immunofluorescence microscopy. Analogue-digital conversion of the fluorescence signal and further image processing allowed measurement of the immunospecific fluorescence in the nuclei of these cells. The results are compared with the adduct levels measured in isolated DNA by 32P-postlabelling. Preliminary results are shown of the application of the immunofluorescence method to the analysis of DNA adducts in bronchial cells obtained from smoking individuals.
D015123:7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide / Q000032:analysis* / Q000276:immunology; D001980:Bronchi / Q000032:analysis*; D002478:Cells, Cultured; D004247:DNA / Q000032:analysis* / Q000276:immunology; D018736:DNA Adducts; D004249:DNA Damage; D004101:Dihydroxydihydrobenzopyrenes / Q000032:analysis*; D005455:Fluorescent Antibody Technique; D006801:Humans; D010761:Phosphorus Radioisotopes; D012907:Smoking / Q000378:metabolism*
D018736:DNA Adducts; D004101:Dihydroxydihydrobenzopyrenes; D010761:Phosphorus Radioisotopes; C043931:benzo(a)pyrene-7,8-dihydrodiol-9,10-epoxide-DNA; D015123:7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide; D004247:DNA