IgM, IgG, and IgA anti-DNA topoisomerase I antibodies in systemic sclerosis.
Abstract
Anti-DNA topoisomerase I (anti-topo I) antibodies have been broadly studied in systemic sclerosis (SSc). The use of different native and molecularly cloned antigens has shown a predominant IgG response, and a variable frequency of positive IgM and IgA tests. We report herein the serological findings of SSc using a recombinant topo I obtained through a standard bacterial system. Anti-topo I antibodies were studied in 45 SSc patients and 85 healthy controls through ELISA and western blot. Escherichia coli XL1-blue strain and pT7-7 vector were used to amplify a DNA topo I cDNA clone, and to obtain the recombinant polypeptide. The latter was purified by affinity chromatography, and the enzymatic and antigenic properties were evaluated through specific tests. A native antigen was included for comparison. The SSc group disclosed positive IgM (20%), IgG (86.6%), and IgA (26.6%) anti-topo I tests with the recombinant polypeptide, and a purified calf thymus antigen yielded similar results. IgG autoantibodies were frequently associated with skin involvement, esophageal dysfunction, and restrictive lung disease. The recombinant protein showed a molecular weight of 86.6 kDa, a positive topo I activity using a supercoiled pBR322 DNA relaxat...Continue Reading
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