[Generation of monoclonal anti-dermatan sulfate antibodies cross-reacting to calf thymus DNA. Analysis of "anti-DNA antibody" directly binding to dermatan sulfate].
Monoclonal anti-dermatan sulfate antibodies were produced by immunizing BALB/c mouse with dermatan sulfate (DS) derived from porcine skin. These antibodies were tested for cross-reactivity to double stranded calf thymus DNA(ds.DNA) and heat-denatured DNA (ss.DNA). Five clones (including 2H8 and 3C4 clones) had binding activities to DS, ds.DNA and ss.DNA in solid phase enzyme linked immunosorbent assay, but no clones binding to DS alone were produced. 2H8 and 3C4 clones could activate mouse complement when they bound to the DS and ss.DNA. The abilities of DS, ds.DNA and ss.DNA to inhibit binding of 2H8 and 3C4 clones to solid phased DS, ds.DNA and ss.DNA were examined. Their binding activities were inhibited by any of DS, ds.DNA and ss.DNA. DNase I treatment abolished the binding activities of 2H8 and 3C4 clones to ds.DNA and ss.DNA completely, but had no effect on the binding activities to Ds. On the other hand, chondroitinase ABC treatment of DS augmented the binding activities of 2H8 and 3C4 to DS. DS digested completely with chondroitinase ABC inhibited the binding activities to DS of 2H8 and 3C4 clones, but not any of delta Di-4S and delta Di-diSB produced from DS digested with this enzyme inhibited the binding activities. Therefore, it was suggested that the epitope recognized by 2H8 and 3C4 clones could not be in delta Di-4S or delta Di-diSB but in the linkage regions of DS.
Ohnishi
K
K
Second Department of Medicine, Hokkaido University School of Medicine, Sapporo, Japan.
D000818:Animals; D000974:Antibodies, Antinuclear / Q000032:analysis*; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D002417:Cattle; D002807:Chondroitin / Q000031:analogs & derivatives*; D002999:Clone Cells; D003429:Cross Reactions; D004247:DNA / Q000276:immunology*; D003871:Dermatan Sulfate / Q000276:immunology*; D006863:Hydrogen-Ion Concentration; D051379:Mice; D008807:Mice, Inbred BALB C
D000974:Antibodies, Antinuclear; D000911:Antibodies, Monoclonal; D003871:Dermatan Sulfate; D002807:Chondroitin; D004247:DNA