Screening for autoantibodies to the nucleolar U3- and Th(7-2) ribonucleoproteins in patients' sera using antisense riboprobes

J Immunol Methods. 1994 Mar 10;169(2):173-82. doi: 10.1016/0022-1759(94)90261-5.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Antinuclear / analysis*
  • Autoantigens / immunology
  • Base Sequence
  • Blotting, Northern
  • Endoribonucleases / genetics
  • Endoribonucleases / immunology*
  • HeLa Cells
  • Humans
  • Lupus Erythematosus, Systemic / immunology
  • Molecular Sequence Data
  • Nucleolus Organizer Region / immunology
  • Precipitin Tests
  • RNA / immunology
  • RNA, Antisense / immunology*
  • Ribonucleoproteins, Small Nuclear / genetics
  • Ribonucleoproteins, Small Nuclear / immunology*
  • Scleroderma, Systemic / immunology*

Substances

  • Antibodies, Antinuclear
  • Autoantigens
  • RNA, Antisense
  • Ribonucleoproteins, Small Nuclear
  • RNA
  • Endoribonucleases
  • mitochondrial RNA-processing endoribonuclease