, Volume 94, Issue 2, pp 132–138 | Cite as

Scl 70 autoantibodies from scleroderma patients recognize a 95 kDa protein identified as DNA topoisomerase I

  • Hans-Herbert Guldner
  • Carin Szostecki
  • Hans-Peter Vosberg
  • Heinz-Jürgen Lakomek
  • Edward Penner
  • Friedlinde A. Bautz


Sera of patients suffering from the autoimmune disease progressive systemic sclerosis (PSS) are known to contain autoantibodies which have been reported to recognize a 70 kDa antigenic protein, designated the Scl 70 antigen. By immunoblotting of nuclear extracts from HeLa cells with sera from scleroderma patients we observed that the size of the antigen present in such cells depends on the conditions of antigen isolation. When protease inhibitors were included in the extraction buffer, a 95 kDa protein was identified instead of a 70 kDa protein. When protease inhibitors were omitted, a number of polypeptides in the size range 66 to 95 kDa was found. Furthermore, antibodies which had been affinity purified on the 95 kDa antigen, crossreacted with the 66 to 95 kDa polypeptides. These results suggest that the smaller proteins were degradation products of the 95 kDa antigen. Immunofluorescence studies on PtK-2 cells with the antibody specific for the 95 kDa protein gave staining of nuclei, nucleoli and of chromosomes and the nucleolar organizer region in mitotic cells. Since this distribution of antigens within the nucleus was reminiscent of the intranuclear distribution of DNA topoisomerase I found by others we probed purified DNA topoisomerase I from calf thymus directly with the autoantibodies from PSS patients, and also the 95 kDa antigens of HeLa cell nuclei with antibodies raised against the bovine DNA topoisomerase I. From the crossreaction pattern observed with the different antigens and antibodies we conclude that DNA topoisomerase I is one of the antigenic components against which autoantibodies are formed in scleroderma patients.


HeLa Cell Scleroderma Antigenic Protein Nucleolar Organizer Region Nucleolar Organizer 
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Copyright information

© Springer-Verlag 1986

Authors and Affiliations

  • Hans-Herbert Guldner
    • 1
  • Carin Szostecki
    • 1
  • Hans-Peter Vosberg
    • 2
  • Heinz-Jürgen Lakomek
    • 3
  • Edward Penner
    • 4
  • Friedlinde A. Bautz
    • 1
  1. 1.Institut für Molekulare GenetikUniversität HeidelbergHeidelbergFederal Republic of Germany
  2. 2.Abteilung Molekulare BiologieMax-Planck-Institut für Medizinische ForschungHeidelbergFederal Republic of Germany
  3. 3.Medizinische Klinik C, Endokrinologie und RheumatologieUniversität DüsseldorfFederal Republic of Germany
  4. 4.Erste Universitätsklinik für Gastroenterologie und Hepatologie, Universität WienAustria

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