The influence of DNA size on the binding of antibodies to DNA in the sera of normal human subjects and patients with systemic lupus erythematosus (SLE).
To elucidate antibody recognition of DNA in normal and aberrant immunity, the binding of sera of normal human subjects (NHS) and patients with SLE was tested with mammalian and bacterial DNA varying in size. Klebsiella pneumoniae (KP) and calf thymus (CT) single-stranded DNA (ssDNA) were investigated as model antigens using the restriction enzyme HinfI to generate fragments with the size range of 800-5000 base pairs. The influence of size on activity was assessed by ELISA by both titration of serum as well as coating antigen concentration. In both assay formats, SLE sera bound equivalently to intact CT and KP DNA, but had dramatically reduced reactivity to fragments of both antigens. In contrast, NHS bound similarly to intact KP DNA and its fragments but had low reactivity to CT DNA. These results suggest that SLE and NHS anti-DNA react with different antigenic determinants on DNA, as shown by cross-reactivity as well as size dependency in solid-phase assays.
Pisetsky
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DS
Medical Research Service, Department of Veterans Affairs Medical Center, Division of Rheumatology, Allergy and Clinical Immunology, Duke University Medical Center, Durham, NC, USA. dpiset@acpub.duke.edu
Gonzalez
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TC
D000974:Antibodies, Antinuclear / Q000276:immunology*; D004247:DNA / Q000276:immunology*; D004269:DNA, Bacterial / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007711:Klebsiella pneumoniae / Q000235:genetics; D008180:Lupus Erythematosus, Systemic / Q000276:immunology*
D000974:Antibodies, Antinuclear; D004269:DNA, Bacterial; D004247:DNA