Antibodies Against Free Radical Modified Native DNA Recognize B-Conformation

Abstract

Hydroxyl radical, a prominent entity of reactive oxygen species, is known to modify cellular DNA and has been implicated in several human diseases. In the present studies, the radical was generated by exposure of hydrogen peroxide to 254 nm light in the presence of native calf thymus DNA. Single strand breaks, decrease in Tm and modification of adenine and thymine were some of the modifications observed in nDNA. Antibodies induced in experimental animals against the modified DNA were immunogen specific. These antibodies also recognize native B-conformation. It was observed that naturally occurring anti-native DNA autoantibodies from SLE sera recognize modified DNA in direct binding and competition ELISA. Gel retardation assay reiterated the formation of immune complexes between induced antibodies and DNA fragments of around 300 bp (B-conformation). The possible significance of these findings in the etiology of SLE has been discussed.