Quantitation of anti-RNP and anti-Sm antibodies in MCTD and SLE patients by immunoblotting

A quantitative immunoblotting assay (QIBA) for the determination of specific antibody titres in human autoimmune sera is described. In this assay, a total HeLa nuclear protein fraction, immobilized on nitrocellulose blot strips, was used as source of antigens and immunoreactive species of autoantibo...

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Published inClinical and experimental immunology Vol. 59; no. 2; pp. 457 - 466
Main Authors HABETS, W. J, DE ROOIJ, D. J, HOET, M. H, VAN DE PUTTE, L. B, VAN VENROOIJ, W. J
Format Journal Article
LanguageEnglish
Published Oxford Blackwell 01.02.1985
Subjects
Adult
Aged
Antibodies, Antinuclear - analysis
Antibody Specificity
Antigens - immunology
Autoantigens
Biological and medical sciences
Collodion
Dose-Response Relationship, Immunologic
Female
Follow-Up Studies
Humans
Immunologic Techniques
Lupus Erythematosus, Systemic - immunology
Medical sciences
Mixed Connective Tissue Disease - immunology
Ribonucleoproteins - immunology
Ribonucleoproteins, Small Nuclear
Sarcoidosis. Granulomatous diseases of unproved etiology. Connective tissue diseases. Elastic tissue diseases. Vasculitis
snRNP Core Proteins
Human
Immunopathology
Connective tissue disease
Skin disease
Ribonucleoprotein
Antibody
Lupus erythematosus
Sharp syndrome
Disseminated
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Summary:A quantitative immunoblotting assay (QIBA) for the determination of specific antibody titres in human autoimmune sera is described. In this assay, a total HeLa nuclear protein fraction, immobilized on nitrocellulose blot strips, was used as source of antigens and immunoreactive species of autoantibodies were quantitated by an enzyme linked second antibody procedure. Besides being more discriminative, QIBA appeared to be up to 500 times more sensitive than immunodiffusion or immunoelectrophoresis. In this study we used 21 sera from patients with SLE or MCTD for a quantitative analysis of their specific autoantibody content. Within this group, a very diverse spectrum of antibody populations was observed; anti-RNP sera appeared to contain, among others, high tired antibody versus 70K and 31K polypeptides while all (n = 6) anti-Sm sera recognized a 25kD protein doublet. In a follow-up study of two MCTD patients significant flares in specific antibody content could be observed.
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ISSN:0009-9104
1365-2249