Immunoperoxidase detection of 8-hydroxydeoxyguanosine in aflatoxin B1-treated rat liver and human oral mucosal cells

Abstract

An immunoperoxidase method using a monoclonal antiserum that recognizes 8-hydroxydeoxyguanosine has been developed for detection and quantitation of oxidative damage in single cells. The method was initially applied to cultured cells treated with H2O2 or aflatoxin B1 and then to cryostat liver sections of rats treated with aflatoxin B1. To demonstrate that the method has sufficient sensitivity for detection of damage in human samples, oral mucosal cells from a total of 12 pairs of smokers and nonsmokers were analyzed. Mean staining intensity of oral cells of smokers was 1.6-fold higher than in nonsmokers. The immunoperoxidase method, requiring a small number of cells and eliminating the need for isolation of DNA, will be useful for evaluation of oxidative damage in a wide range of biological samples.