Newer lymphocyte surface antigens.
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Antibodies, Monoclonal↗
Biomedical subjects
J Schiffenbauer
Publications and source records attributed to J Schiffenbauer.
Detection of masked anti-DNA antibodies in lupus sera by a monoclonal anti-idiotype.
We report here the use of a monoclonal anti-idiotype 3I to human anti-DNA antibodies to detect in serum idiotype-positive antigen-binding antibodies lacking DNA-binding activity as measured by conventional antigen binding assays. We studied paired serum samples from 13 patients with systemic lupus obtained at two times in the course of their disease: in each patient, one serum sample has anti-DNA activity and the second serum sample has no anti-dsDNA activity detectable by Millipore filter, ELISA, or Crithidia assay. Reactivity with 3I as detected with a radioimmunoassay (RIA) was present in all 13 sera with anti-dsDNA activity. Six patients showed a decrease in 3I reactivity to normal levels in the second serum sample, in which anti-dsDNA antibodies were not detectable by conventional antigen-binding assays. The other seven patients' second serum sample continued to show elevated 3I reactivity by RIA even though no anti-dsDNA activity was apparent. When the 3I-reactive antibodies from these latter patients' sera were eluted from a 3I affinity column, they revealed DNA-binding activity. Furthermore, dsDNA binding by these sera was apparent when they were displayed on Western blots of isoelectric focusing gels run in 8 M urea and incubated with radiolabeled dsDNA. These results indicate that the 3I anti-idiotype can detect anti-DNA antibodies in some sera of SLE patients that lack anti-DNA activity by ordinary assays. These antibodies may be inhibited in binding dsDNA by excess antigen or autologous anti-idiotype, and their DNA binding activity can be unmasked by procedures promoting immune complex dissociation.
Antibodies, Anti-Idiotypic↗
Use of monoclonal antibodies to identify shared idiotypes on human antibodies to native DNA from patients with systemic lupus erythematosus.
Antinative DNA antibodies were purified from the serum of a patient with active systemic lupus erythematosus. Using hybridoma technology, we produced several mouse monoclonal antibodies directed against idiotypic determinants on these antinative DNA antibodies. One of these monoclonal anti-idiotypic antibodies, 3I, an IgG2a Kappa, was extensively characterized. 3I is believed to be directed against an idiotypic determinant because (i) its reactivity with antinative DNA antibodies is not inhibited by a large excess of pooled human serum, (ii) it reacts with F(ab')2 fragments of antinative DNA antibodies, and (iii) it reacts with antinative DNA antibodies of all IgG subclasses. 3I is not directed against the antigen binding site in that native DNA and 3I do not compete for binding to antinative DNA antibodies. Eight of nine sera containing antinative DNA activity as determined by the Millipore filter assay reacted with 3I in a solid-phase radioimmunoassay. These findings suggest that antinative DNA antibodies from nonrelated patients with systemic lupus erythematosus share a common idiotype.
Antibodies, Anti-Idiotypic↗