Binding of circulating antibodies to reactive oxygen species modified-DNA and detecting DNA damage by a monoclonal...

Circulating antibodies in the sera of normal, healthy humans of different age groups to native DNA (nDNA) and reactive oxygen species modified DNA (ROS-DNA) was studied by competition ELISA. Sera from the young population (< 50 years of age) showed negligible levels of anti-DNA antibodies. In contrast, anti-DNA antibodies were found in three sera from the moderately aged group (50-59 years) with a high binding to ROS-DNA (43-47%). In the aged group (> 60 years), four sera showed higher recognition of ROS-DNA (> 60% inhibition) over nDNA (55-60%). Oxidative lesions in human genomic DNA were immunochemically detected using the monoclonal anti-ROS-DNA antibody as a probe. The antibody has a high specificity for ROS-DNA and preferentially recognizes ROS-modified epitopes on nucleic acids. The study indicates low recognition of DNA isolated from the younger population, while in the age group 50-59 years, one DNA isolate showed a high inhibition (57%) in monoclonal antibody binding. Four DNA isolates from the aged group showed substantial inhibition in antibody activity to the extent of 49, 53, 64 and 69%. The results demonstrate an age-related increase in the levels of anti-DNA antibodies, with a higher recognition and binding to ROS-DNA. A high reactivity of DNA isolated from aged individuals by the monoclonal antibody indicates increased oxidative stress leading to DNA damage. It is suggested that free radical damage to DNA in vivo, particularly in the aged, alters its antigenicity and stimulates an immune response against modified DNA. These antibodies are cross-reactive to nDNA.