Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Correspondence
  • Published:

Autoantibodies to GPI and creatine kinase in RA

Nature Immunology volume 3, page 411 (2002)Cite this article

Rheumatoid arthritis (RA) is a systemic chronic inflammatory disease that primarily affects the joints. The trigger that activates autoreactive T and B cells in RA patients is still unknown1. The glycolytic enzyme glucose-6-phosphate-isomerase (GPI) has been identified as the antigen recognized by arthritogenic antibodies in a spontaneously developing murine model of arthritis that has several of the features of human RA2. It is, therefore, of interest to determine whether GPI is also relevant to human arthritides, such as RA. Thus, we expressed recombinant human GPI fused to a histidine tag in Escherichia coli. The expressed protein had the expected enzymatic activity (data not shown) and was used to examine serial dilutions of sera from patients with RA, other rheumatic diseases (including ankylosing spondylitis, vasculitis and connective tissue disease) or healthy controls for the presence of antibodies to GPI. At a dilution of 1:50, only 2/61 sera from RA patients bound recombinant human GPI (Web Fig. 1 online). This contrasts with an article by Schaller et al. published in Nature Immunology in which 64% of sera (diluted 1:50) from RA patients had detectable IgG to a commercial GPI preparation, which was isolated from rabbit muscle3. When we used the commercial GPI preparation for ELISA analysis, only a few samples contained detectable anti-GPI at dilutions >1:50 (data not shown). At the 1:50 dilution, we detected increased antibody titers in 15/61 RA serum samples, 9/21 serum samples from patients with other rheumatic diseases and 1/32 serum samples from healthy donors (Web Fig. 2 online).

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on SpringerLink
  • Instant access to the full article PDF.

USD 39.95

Prices may be subject to local taxes which are calculated during checkout

Figure 1: CK-M is contained in the commercial GPI preparation and recognized by sera from RA patients.

References

  1. Kamradt, T. & Mitchison, N. A. N. Engl. J. Med. 344, 655–664 (2001).

    Article  CAS  Google Scholar 

  2. Matsumoto, I., Staub, A., Benoist, C. & Mathis, D. Science 286, 1732–1735 (1999).

    Article  CAS  Google Scholar 

  3. Schaller, M., Burton, D. R. & Ditzel, H. J. Nature Immunol. 2, 746–753 (2001).

    Article  CAS  Google Scholar 

  4. Lamer, S. & Jungblut, P. R. J. Chromatogr. B. Biomed. Sci. Appl. 752, 311–322 (2001).

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Deutsches Rheumaforschungszentrum, Berlin, Germany

    David Schubert, Dietmar Zaiss & Thomas Kamradt

  2. Max-Planck-Institut für Infektionsbiologie, Berlin, Germany

    Monika Schmidt & Peter R. Jungblut

  3. Universitätsklinikum Charité, Berlin, Germany

    Thomas Kamradt

Authors
  1. David Schubert
    View author publications

    Search author on:PubMed Google Scholar

  2. Monika Schmidt
    View author publications

    Search author on:PubMed Google Scholar

  3. Dietmar Zaiss
    View author publications

    Search author on:PubMed Google Scholar

  4. Peter R. Jungblut
    View author publications

    Search author on:PubMed Google Scholar

  5. Thomas Kamradt
    View author publications

    Search author on:PubMed Google Scholar

Additional information

Note: Supplementary information is available on the Nature Immunology website.

Supplementary information

Web Figure 1.

Sera from RA patients or controls rarely recognize recombinant human GPI. ELISAs were performed on sera (diluted 1:50) from 61 patients with RA, other rheumatic diseases (e.g. ankylosing spondylitis, vasculitis, connective tissue disease; n=21), or healthy donors (n=32) to detect antibodies against recombinant human GPI (produced in E. coli in our laboratory). The cutoff for a positive value was defined as 2 standard deviations above the mean optical density of the 32 samples from healthy donors (dotted line). (GIF 6 kb)

Web Figure 2.

Recognition of a commercial GPI-preparation, purified from rabbit muscle, by sera from RA patients or controls. ELISAs were performed on sera (diluted 1:50) from 61 patients with RA, other rheumatic diseases (e.g. ankylosing spondylitis, vasculitis, connective tissue disease; n=21), or healthy donors (n=32) to detect antibodies against GPI purified from rabbit muscle (Sigma, St. Louis, MO). The cutoff for a positive value was defined as 2 standard deviations above the mean optical density of the 32 samples from healthy donors (dotted line). (GIF 16 kb)

Web Figure 3.

MALDI-MS identifies one of the proteins recognized by patient sera as creatine kinase. The ≈40 kD band was excised from the gel and after tryptic digestion the peptides were analyzed by MALDI-MS. 21/24 peaks with a sequence coverage of 64% (insert) are identical with the M-chain of rabbit creatine kinase. (GIF 29 kb)

Web Figure 4.

Recognition of CK-M by sera from RA patients or controls. ELISAs were performed on sera (diluted 1:50) from 61 patients with RA, other rheumatic diseases (n=21), or healthy donors (n=32) to detect antibodies against CK-M purified from rabbit muscle (Sigma, St. Louis, MO). The cutoff for a positive value was defined as 2 standard deviations above the mean optical density of the 32 samples from healthy donors (dotted line). (GIF 7 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Schubert, D., Schmidt, M., Zaiss, D. et al. Autoantibodies to GPI and creatine kinase in RA. Nat Immunol 3, 411 (2002). https://doi.org/10.1038/ni0502-411a

Download citation

  • Issue date:

  • DOI: https://doi.org/10.1038/ni0502-411a

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing