A recombinant topoisomerase I used for autoantibody detection in sera from patients with systemic sclerosis

• Published in: Clinical and experimental immunology Vol. 80; no. 1; pp. 38 - 43
• Main Authors: VERHEIJEN, R., HOOGEN, F., BEIJER, R., RICHTER, A., PENNER, E., HABETS, W. J., VENROOIJ, W. J.
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Publishing Ltd 01.04.1990; Blackwell
• Subjects: Amino Acid Sequence; Autoantibodies - analysis; Base Sequence; Biological and medical sciences; Cloning, Molecular; DNA Topoisomerases, Type I - immunology; DNA, Recombinant; epitope regions; Epitopes - analysis; fusion proteins; Humans; Medical sciences; Molecular Sequence Data; Recombinant Fusion Proteins - immunology; Sarcoidosis. Granulomatous diseases of unproved etiology. Connective tissue diseases. Elastic tissue diseases. Vasculitis; Scleroderma, Systemic - immunology; systemic sclerosis; topoisomerase I; Connective tissue disease; Skin disease; Antigenic determinant; DNA topoisomerase; Fusion protéine; Autoantibody; Gene expression; Complementary DNA; Systemic disease; Immunoblotting assay; Recombinant protein; Detection; Scleroderma
• ISSN: 0009-9104; 1365-2249
• DOI: 10.1111/j.1365-2249.1990.tb06438.x

SUMMARY We report the expression of a cDNA clone encoding 695 carboxyl‐terminal amino acids of human DNA topoisomerase I (topoI) in Escherichia coli. More than 96% of the anti‐HeLa topoI‐positive sera from patients with a connective tissue disease displayed also an immunoreactivity with this recombinant protein (the HTopoA protein). Sera from patients with a definite diagnosis systemic sclerosis and reacting with HeLa topoI, all reacted with the HTopoA protein as well. Sera from patients with systemic sclerosis that did not contain anti‐topoI antibodies (about 30% of the systemic sclerosis sera), as concluded from HeLa immunoblot, displayed also no immunoreactivity with our recombinant antigen. By expressing different fragments of HTopoA, we were able to assign at least three different autoimmune epitope regions on the HTopoA protein and we show that over a period of 5 years the amount of anti‐topoI antibodies against these regions may fluctutate.