Screening for autoantibodies to the nucleolar U3- and Th(7-2) ribonucleoproteins in patients' sera using antisense riboprobes.
In this study we report the detection of autoantibodies to the nucleolar U3- and Th(7-2) ribonucleoprotein (RNP) particles in sera from patients with connective tissue diseases. The method described employs radioactively labelled antisense U3- and Th RNA which are hybridized to immunoprecipitated U3- or Th RNA from a HeLa cell extract. Of the 66 sera that were screened with this method seven sera (11%) precipitated only Th RNP, 16 sera (24%) precipitated only U3 RNP and 4 sera (6%) precipitated both U3- and Th RNP. Both anti-U3 RNP and anti-Th RNP activity appeared to be mostly associated with scleroderma or scleroderma-associated diseases. Using this method we also showed that some of the Th RNP particles in a cell extract are associated with the La autoantigen. We conclude that for the identification of immunoprecipitated RNAs this method is very sensitive and provides unambiguous data.
Verheijen
R
R
Department of Biochemistry, University of Nijmegen, Netherlands.
Wiik
A
A
De Jong
B A
BA
Høier-Madsen
M
M
Ullman
S
S
Halberg
P
P
Van Venrooij
W J
WJ
D000974:Antibodies, Antinuclear / Q000032:analysis*; D001324:Autoantigens / Q000276:immunology; D001483:Base Sequence; D015152:Blotting, Northern; D004722:Endoribonucleases / Q000235:genetics / Q000276:immunology*; D006367:HeLa Cells; D006801:Humans; D008180:Lupus Erythematosus, Systemic / Q000276:immunology; D008969:Molecular Sequence Data; D009697:Nucleolus Organizer Region / Q000276:immunology; D011233:Precipitin Tests; D012313:RNA / Q000276:immunology; D016372:RNA, Antisense / Q000276:immunology*; D017411:Ribonucleoproteins, Small Nuclear / Q000235:genetics / Q000276:immunology*; D012595:Scleroderma, Systemic / Q000276:immunology*
D000974:Antibodies, Antinuclear; D001324:Autoantigens; D016372:RNA, Antisense; D017411:Ribonucleoproteins, Small Nuclear; D012313:RNA; D004722:Endoribonucleases; C058330:mitochondrial RNA-processing endoribonuclease