Open AccessA recombinant topoisomerase I used for autoantibody detection in sera from patients with systemic sclerosis
Author(s) -
VERHEIJEN R.,
HOOGEN F.,
BEIJER R.,
RICHTER A.,
PENNER E.,
HABETS W. J.,
VENROOIJ W. J.
Publication year - 1990
Publication title -
clinical & experimental immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.329
H-Index - 135
eISSN - 1365-2249
pISSN - 0009-9104
DOI - 10.1111/j.1365-2249.1990.tb06438.x
Subject(s) - autoantibody , recombinant dna , antibody , epitope , autoimmune disease , immunology , connective tissue disease , systemic disease , hela , microbiology and biotechnology , antigen , biology , complementary dna , topoisomerase , dna , immunopathology , gene , cell culture , biochemistry , genetics
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.