Immunoblotting on HEp-2 cells increases the detection of antitopoisomerase 1 antibodies in patients with systemic sclerosis.
In order to improve the detection of antitopoisomerase 1 antibodies in a cohort of 111 systemic sclerosis patients, we have performed immunoblots on protein extracts of HEp-2 cells. Using indirect immunofluorescence and ELISA, 27 patients (24.3%) had antitopoisomerase 1 antibodies, 32 (28.8%) had anticentromere antibodies, 31 (27.9%) had antinuclear antibodies with none of these two antibodies and 21 (18.9%) had no antinuclear antibody. IgG from 24/27 (88.9%) patients with antitopoisomerase 1 antibodies bound to 2 protein bands of 63 and 100 kDa identified as topoisomerase 1 by N-terminal sequencing. Antitopoisomerase 1 antibodies were detected in 9 (8.1%) patients who had no antitopoisomerase 1 antibody as determined by ELISA. Patients with antitopoisomerase 1 antibodies had an almost similar phenotype without distinction between ELISA or immunoblot approaches. Our results provide evidence for the use of a combination of ELISA and immunoblot approaches for the detection of antitopoisomerase 1 antibodies.
Tamby
Mathieu C
MC
Paris-Descartes University, UPRES-EA 4058, site Cochin, Paris, France.
Servettaz
Amélie
A
Guilpain
Philippe
P
Tamas
Nicolas
N
Berezné
Alice
A
Batteux
Frédéric
F
Reinbolt
Joseph
J
Guillevin
Loïc
L
Weill
Bernard
B
Mouthon
Luc
L
D000328:Adult; D000974:Antibodies, Antinuclear / Q000097:blood / Q000276:immunology; D001323:Autoantibodies / Q000097:blood* / Q000276:immunology; D001324:Autoantigens / Q000276:immunology; D002460:Cell Line; D002503:Centromere / Q000276:immunology; D004264:DNA Topoisomerases, Type I / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D005455:Fluorescent Antibody Technique; D006801:Humans; D015151:Immunoblotting / Q000379:methods; D007074:Immunoglobulin G / Q000097:blood* / Q000276:immunology; D008875:Middle Aged; D012595:Scleroderma, Systemic / Q000097:blood* / Q000276:immunology; D012680:Sensitivity and Specificity
D000974:Antibodies, Antinuclear; D001323:Autoantibodies; D001324:Autoantigens; D007074:Immunoglobulin G; D004264:DNA Topoisomerases, Type I