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

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 recombi...

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Published in:Clinical and experimental immunology 1990-04, Vol.80 (1), p.38-43
Main Authors: VERHEIJEN, R., HOOGEN, F., BEIJER, R., RICHTER, A., PENNER, E., HABETS, W. J., VENROOIJ, W. J.
Format: Article
Language:English
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
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Summary: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.
ISSN:0009-9104
1365-2249
DOI:10.1111/j.1365-2249.1990.tb06438.x