Visualization of O6-methylguanine in target cell nuclei of dimethylnitrosamine-treated human pancreas by a murine monoclonal antibody.
A murine monoclonal IgG2b with high affinity and specificity for O6-methylguanine was developed. This allowed visualization of this adduct in paraffin-embedded sections by indirect fluorescence or immunoperoxidase methods. The antibody was used to study and compare the formation and persistence of this adduct detected by immunohistochemistry and the presence of 14C methyl by autoradiography in sections from adult human pancreas explants exposed to [14C methyl]dimethylnitrosamine (DMN) and organ-cultured for up to 7 days. O6-methylguanine was detectable randomly and with varied intensity in parenchymal cell nuclei and cytoplasms, 6 h after exposure to DMN. Following RNase treatment, however, only 7.5% of nuclei revealed measurable staining. The positive nuclei decreased to 2.5% within 24 h and remained about the same throughout the 7 days of culture. This correlated well with the change in percentage of positive nuclei for 14C methyl autoradiographic grains which were selectively condensed over the O6-methylguanine-stained nuclei. The data demonstrate a subpopulation of repair-deficient parenchymal cells of human pancreas exposed to DMN.
D000818:Animals; D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex / Q000032:analysis; D002250:Carbon Radioisotopes; D002467:Cell Nucleus / Q000187:drug effects / Q000648:ultrastructure*; D004128:Dimethylnitrosamine / Q000633:toxicity*; D006147:Guanine / Q000031:analogs & derivatives* / Q000032:analysis; D006801:Humans; D051379:Mice; D009924:Organ Culture Techniques; D010179:Pancreas / Q000187:drug effects / Q000473:pathology* / Q000648:ultrastructure
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D002250:Carbon Radioisotopes; D006147:Guanine; C008449:O-(6)-methylguanine; D004128:Dimethylnitrosamine