Detection and occurrence of the 60- and 52-kD Ro (SS-A) antigens and of autoantibodies against these proteins
| Autor: | W. G. M. Damen, W. J. Van Venrooij, J.W.C.M. van der Kemp, R. L. Slobbe, G.J.M. Pruijn |
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| Rok vydání: | 1991 |
| Předmět: |
Immunoprecipitation
Immunology Autoantigens Autoimmune Diseases Antigen Species Specificity Antibody Specificity RNA Small Cytoplasmic medicine Tumor Cells Cultured Immunology and Allergy Humans Ribonucleoprotein Autoantibodies Lupus erythematosus biology Autoantibody Nuclear Proteins medicine.disease Blot Molecular Weight Ribonucleoproteins biology.protein Electrophoresis Polyacrylamide Gel Antibody Anti-SSA/Ro autoantibodies Research Article |
| Zdroj: | Clinical and experimental immunology. 86(1) |
| ISSN: | 0009-9104 |
| Popis: | SUMMARY The simultaneous detection of anti-La. anti-60-kD Ro and anti-52-kD Ro antibodies by immunoblotting is greatly improved by changing the crosslinking level in the gel to an acrylamide/bisacrylamide ratio of 19:1. Using this method for the analysis of a number of systemic lupus erythematosus (SLE) and Sjögren’s syndrome patient sera it was observed that antibody to the 52-kD Ro protein without anti-60-kD Ro antibody was restricted to Sjogren’s syndrome patients (9/26), whereas antibody to the 60-kD Ro protein without contaminating anti-52-kD Ro antibody was only found in SLE patients (8/38). Moreover, in Sjogren’s sydrome patient sera anti-Ro antibody was found only in combination with anti-La antibody (20/26). whereas in SLE patient sera anti-Ro antibody could be found without detectable anti-La specificity (4/38). Double immunofluorescence microscopy revealed that the 52-kD Ro and the 60-kD Ro proteins co-localize in the cytoplasm as well as in the nucleus, whereas immunoprecipitation of [32P]-labelled HeLa cell extract with monospecific anti-52-kD Ro and anti-60-kD Ro sera showed that both proteins are associated with the Ro RNAs. These data suggest the presence of both the 52-kD and the 60-kD Ro proteins in the same ribonucleoprolein complexes. To study the evolutionary conservation of the 52-kD Ro, the 60-kD Ro and the La proteins, extracts of cell lines derived from various mammalian species were analysed on Western blols using monospecific human antibodies. In contrast to the 60-kD Ro and the La antigens which are well conserved in evolution, the 52-kD Ro antigen could be detected in primate cells only by this immunological approach. |
| Databáze: | OpenAIRE |
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