Probing molecular changes induced in DNA by reactive oxygen species with monoclonal antibodies.
Antibodies reactive with native double stranded DNA are characteristic of the chronic inflammatory disease systemic lupus erythematosus. Native DNA is however, a poor immunogen and the mechanism of anti-DNA antibody production is incompletely understood. Modification of DNA can increase its immunogenicity and in inflammatory disease states reactive oxygen species produced from phagocytic cells have been shown to thus modify DNA. In this study, monoclonal antibodies produced spontaneously by two mice strains with lupus-like disease were used in a competition ELISA to monitor changes to DNA induced by reactive oxygen species. Different procedures for reactive oxygen species generation were found to cause distinct and characteristic changes to DNA involving modifications of base residues, the sugar-phosphate backbone and the gross conformational structure of double-stranded DNA. In view of this, it may be possible to use these antibodies further to probe DNA and infer the source and nature of the reactive oxygen species it has been exposed to, particularly in vivo.
Blount
S
S
Molecular Toxicity Group, Glenfield Hospital, Leicester, UK.
Griffiths
H R
HR
Staines
N A
NA
Lunec
J
J
D000974:Antibodies, Antinuclear / Q000494:pharmacology*; D000911:Antibodies, Monoclonal; D004247:DNA / Q000528:radiation effects; D004249:DNA Damage / Q000187:drug effects; D003676:Deferoxamine / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay; D000431:Ethanol / Q000494:pharmacology; D005720:Gamma Rays; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D066298:In Vitro Techniques; D008353:Mannitol / Q000494:pharmacology; D015394:Molecular Structure; D017382:Reactive Oxygen Species / Q000009:adverse effects*; D013482:Superoxide Dismutase / Q000494:pharmacology; D013890:Thiourea / Q000494:pharmacology; D014466:Ultraviolet Rays
D000974:Antibodies, Antinuclear; D000911:Antibodies, Monoclonal; D017382:Reactive Oxygen Species; D000431:Ethanol; D008353:Mannitol; D004247:DNA; D006861:Hydrogen Peroxide; D013482:Superoxide Dismutase; D013890:Thiourea; D003676:Deferoxamine