Human fibroblasts, a convenient nuclear substrate for detection of anti-nuclear antibodies including anti-centromere antibodies.
Human fibroblast monolayers were applied as a nuclear substrate in indirect immunofluorescence for the detection of anti-nuclear antibodies (ANA). Comparison of the results obtained with this substrate and with conventional rat liver sections led to the conclusion that the application of human fibroblast monolayers as a substrate has the following advantages: 1) better recognition of the distinct nuclear staining patterns; 2) more convenient serum titrations, since a cryostat is not needed and twelve tests can be performed on one slide; 3) detection of significantly higher ANA titres in sera from patients with SLE and MCTD; 4) recognition of a type of ANA that was not detected on rat liver sections. These latter antibodies produced a discrete speckled pattern of fluorescence, which was completely lost after acid elution of the fibroblasts. Using HEp-2 cells in metaphase, it was shown that the antibodies were directed against the centromere regions of the chromosomes. These anti-centromere antibodies were detected in sera from patients with severe Raynaud's phenomenon. Underlying disorders were scleroderma (8 patients, 5 of them with CREST syndrome), Sjögren's syndrome (2 patients), and Raynaud's phenomenon in combination with a few symptoms of connective tissue diseases without fulfilling the criteria for a specific disease (5 patients).
D000293:Adolescent; D000328:Adult; D000368:Aged; D000818:Animals; D000974:Antibodies, Antinuclear / Q000032:analysis*; D002478:Cells, Cultured; D002503:Centromere / Q000276:immunology*; D002875:Chromosomes / Q000276:immunology*; D003095:Collagen Diseases / Q000276:immunology*; D005260:Female; D005347:Fibroblasts; D005455:Fluorescent Antibody Technique; D006801:Humans; D005779:Immunodiffusion; D008099:Liver / Q000276:immunology; D008180:Lupus Erythematosus, Systemic / Q000276:immunology; D008297:Male; D008875:Middle Aged; D008947:Mixed Connective Tissue Disease / Q000276:immunology; D051381:Rats; D011928:Raynaud Disease / Q000276:immunology; D012859:Sjogren's Syndrome / Q000276:immunology
D000974:Antibodies, Antinuclear