Letter | Published:

Specific interaction between genotype, smoking and autoimmunity to citrullinated α-enolase in the etiology of rheumatoid arthritis

Nature Genetics volume 41, pages 13191324 (2009) | Download Citation

This article has been updated

Abstract

Gene-environment associations are important in rheumatoid arthritis (RA) susceptibility, with an association existing between smoking, HLA- DRB1 'shared epitope' alleles, PTPN22 and antibodies to cyclic citrullinated peptides(CCP)1,2. Here, we test the hypothesis that a subset of the anti-CCP response, with specific autoimmunity to citrullinated α-enolase3,4, accounts for an important portion of these associations. In 1,497 individuals from three RA cohorts, antibodies to the immunodominant citrullinated α-enolase CEP-1 epitope4 were detected in 43–63% of the anti-CCP–positive individuals, and this subset was preferentially linked to HLA-DRB1*04. In a case-control analysis of 1,000 affected individuals and 872 controls, the combined effect of shared epitope, PTPN22 and smoking showed the strongest association with the anti-CEP-1–positive subset (odds ratio (OR) of 37, compared to an OR of 2 for the corresponding anti-CEP-1–negative, anti-CCP–positive subset). We conclude that citrullinated α-enolase is a specific citrullinated autoantigen that links smoking to genetic risk factors in the development of RA.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Change history

  • 26 November 2009

    In the version of this article initially published, the x–axes of Figure 4 panels A–C were incorrectly labeled. The error has been corrected in the HTML and PDF versions of the article.

References

  1. 1.

    et al. A new model for an etiology of rheumatoid arthritis: smoking may trigger HLA-DR (shared epitope)-restricted immune reactions to autoantigens modified by citrullination. Arthritis Rheum. 54, 38–46 (2006).

  2. 2.

    et al. Refining the complex rheumatoid arthritis phenotype based on specificity of the HLA-DRB1 shared epitope for antibodies to citrullinated proteins. Arthritis Rheum. 52, 3433–3438 (2005).

  3. 3.

    et al. Identification of citrullinated alpha-enolase as a candidate autoantigen in rheumatoid arthritis. Arthritis Res. Ther. 7, R1421–R1429 (2005).

  4. 4.

    et al. Antibodies to citrullinated alpha-enolase peptide 1 are specific for rheumatoid arthritis and cross-react with bacterial enolase. Arthritis Rheum. 58, 3009–3019 (2008).

  5. 5.

    , & The shared epitope hypothesis. An approach to understanding the molecular genetics of susceptibility to rheumatoid arthritis. Arthritis Rheum. 30, 1205–1213 (1987).

  6. 6.

    , & Cigarette smoking increases the risk of rheumatoid arthritis. Results from a nationwide study of disease-discordant twins. Arthritis Rheum. 39, 732–735 (1996).

  7. 7.

    et al. Smoking increases peptidylarginine deiminase 2 enzyme expression in human lungs and increases citrullination in BAL cells. Ann. Rheum Dis. 67, 1488–1492 (2008).

  8. 8.

    , , , & Immunity to citrullinated proteins in rheumatoid arthritis. Annu. Rev. Immunol. 26, 651–675 (2008).

  9. 9.

    et al. The diagnostic properties of rheumatoid arthritis antibodies recognizing a cyclic citrullinated peptide. Arthritis Rheum. 43, 155–163 (2000).

  10. 10.

    , & Anticitrullinated protein/peptide antibody and its role in the diagnosis and prognosis of early rheumatoid arthritis. Neth. J. Med. 60, 383–388 (2002).

  11. 11.

    et al. Alpha-enolase: a target of antibodies in infectious and autoimmune diseases. Autoimmun. Rev. 6, 176–182 (2007).

  12. 12.

    et al. Synovial fluid is a site of citrullination of autoantigens in inflammatory arthritis. Arthritis Rheum. 58, 2287–2295 (2008).

  13. 13.

    et al. Presence of autoantibodies to the glycolytic enzyme alpha-enolase in sera from patients with early rheumatoid arthritis. Arthritis Rheum. 46, 1196–1201 (2002).

  14. 14.

    et al. Antibodies towards several citrullinated antigens are enriched in the joints of RA patients. Arthritis Rheum. (2009)(in the press).

  15. 15.

    et al. Quantification of the influence of cigarette smoking on rheumatoid arthritis: results from a population based case-control study, using incident cases. Ann. Rheum. Dis. 62, 835–841 (2003).

  16. 16.

    , & The Arthritis and Rheumatism Council's national family material repository. Br. J. Rheumatol. 32, 443–444 (1993).

  17. 17.

    & The Norfolk Arthritis Register (NOAR). Clin. Exp. Rheumatol. 21, S94–S99 (2003).

  18. 18.

    et al. The HLA-DRB1 shared epitope alleles are primarily a risk factor for anti-cyclic citrullinated peptide antibodies and are not an independent risk factor for development of rheumatoid arthritis. Arthritis Rheum. 54, 1117–1121 (2006).

  19. 19.

    , & HLA class II association with rheumatoid arthritis: facts and interpretations. Hum. Immunol. 61, 1254–1261 (2000).

  20. 20.

    et al. Multiple antibody reactivities to citrullinated antigens in sera from rheumatoid arthritis patients-association with HLA-DRB1 alleles. Ann. Rheum. Dis. 68, 736–743 (2008).

  21. 21.

    , & Concepts of interaction. Am. J. Epidemiol. 112, 467–470 (1980).

  22. 22.

    & Confidence interval estimation of interaction. Epidemiology 3, 452–456 (1992).

  23. 23.

    et al. A missense single-nucleotide polymorphism in a gene encoding a protein tyrosine phosphatase (PTPN22) is associated with rheumatoid arthritis. Am. J. Hum. Genet. 75, 330–337 (2004).

  24. 24.

    et al. Gene-gene and gene-environment interactions involving HLA-DRB1, PTPN22, and smoking in two subsets of rheumatoid arthritis. Am. J. Hum. Genet. 80, 867–875 (2007).

  25. 25.

    et al. Different patterns of associations with anti-citrullinated protein antibody-positive and anti-citrullinated protein antibody-negative rheumatoid arthritis in the extended major histocompatibility complex region. Arthritis Rheum. 60, 30–38 (2009).

  26. 26.

    et al. Brd2 is a TBP-associated protein and recruits TBP into E2F–1 transcriptional complex in response to serum stimulation. Mol. Cell. Biochem. 294, 45–54 (2007).

  27. 27.

    , , & Arthritis provoked by linked T and B cell recognition of a glycolytic enzyme. Science 286, 1732–1735 (1999).

  28. 28.

    et al. How antibodies to a ubiquitous cytoplasmic enzyme may provoke joint-specific autoimmune disease. Nat. Immunol. 3, 360–365 (2002).

  29. 29.

    et al. The American Rheumatism Association 1987 revised criteria for the classification of rheumatoid arthritis. Arthritis Rheum. 31, 315–324 (1988).

  30. 30.

    et al. The influence of HLA-DRB1 alleles and rheumatoid factor on disease outcome in an inception cohort of patients with early inflammatory arthritis. Arthritis Rheum. 42, 2174–2183 (1999).

  31. 31.

    et al. Replication of putative candidate-gene associations with rheumatoid arthritis in >4,000 samples from North America and Sweden: association of susceptibility with PTPN22, CTLA4, and PADI4. Am. J. Hum. Genet. 77, 1044–1060 (2005).

  32. 32.

    , , , & Calculating measures of biological interaction. Eur. J. Epidemiol. 20, 575–579 (2005).

  33. 33.

    , & Calculating measures of biological interaction using R. Eur. J. Epidemiol. 21, 571–573 (2006).

  34. 34.

    Estimability and estimation in case-referent studies. Am. J. Epidemiol. 103, 226–235 (1976).

Download references

Acknowledgements

The authors wish to acknowledge M. Andersson, M. Mullazhi and M. Widhe for their input regarding the development of the anti-CEP-1 ELISA. We would also like to thank investigators and study participants from the EIRA, the NR and the NOAR studies for their contributions. This work was supported by grants from the Arthritis Research Campaign, the Swedish Research Council, the Swedish Council for Working Life and Social Research, the Arbetsmarknadens Försäkrings Aktibebolag insurance foundation, the US National Institutes of Health (grant P60 AR047782), FAMRI (Flight Attendants Medical Research Institute), the Swedish Rheumatism Association, King Gustav V's 80-year foundation and the Swedish Fund for Research Without Animal Experiments. This study is part of the EU funded research project AutoCure, within the sixth framework program.

Author information

Author notes

    • Hiba Mahdi
    •  & Benjamin A Fisher

    These authors contributed equally to this work.

Affiliations

  1. Rheumatology Unit, Karolinska Institutet, Center for Molecular Medicine, Stockholm, Sweden.

    • Hiba Mahdi
    • , Vivianne Malmström
    • , Leonid Padyukov
    •  & Lars Klareskog
  2. Kennedy Institute of Rheumatology, Imperial College London, London, UK.

    • Benjamin A Fisher
    • , Peter Charles
    • , Patrick J Venables
    •  & Karin Lundberg
  3. Institute for Environmental Medicine, Karolinska Institutet, Stockholm, Sweden.

    • Henrik Källberg
    • , Bo Ding
    •  & Lars Alfredsson
  4. Arthritis Research Campaign Epidemiology Unit, University of Manchester, Manchester, UK.

    • Darren Plant
    •  & Deborah P M Symmons
  5. Unit of Clinical Immunology, Rudbeck Laboratory, Uppsala University, Uppsala, Sweden.

    • Johan Rönnelid

Authors

  1. Search for Hiba Mahdi in:

  2. Search for Benjamin A Fisher in:

  3. Search for Henrik Källberg in:

  4. Search for Darren Plant in:

  5. Search for Vivianne Malmström in:

  6. Search for Johan Rönnelid in:

  7. Search for Peter Charles in:

  8. Search for Bo Ding in:

  9. Search for Lars Alfredsson in:

  10. Search for Leonid Padyukov in:

  11. Search for Deborah P M Symmons in:

  12. Search for Patrick J Venables in:

  13. Search for Lars Klareskog in:

  14. Search for Karin Lundberg in:

Contributions

H.M. performed ELISA assays on EIRA, performed statistical analyses and produced the figures. B.F. performed ELISA assays on NR and NOAR, performed statistical analyses and contributed to writing of the paper. H.K. performed statistical analyses on EIRA, produced the tables and contributed to the writing of the Supplementary Note and Methods sections. D.P. performed statistical analyses and HLA-DRB1 genotyping on NR and NOAR. V.M. supervised laboratory experiments and supervised statistical analysis on EIRA. J.R. developed the CEP-1 ELISA. P.C. supervised laboratory experiments on NR and NOAR. B.D. performed the SNP analyses. L.A. supervised statistical analyses on EIRA and wrote the Supplementary Note and Methods section on interaction analyses. L.P. performed the HLA-DRB1 and PTPN22 genotyping on EIRA. D.P.M.S. is responsible for the NOAR study and supervised statistical analyses on NR and NOAR. P.J.V. designed the study and supervised laboratory experiments on NR and NOAR. L.K. is responsible for the EIRA study and the design of the study. K.L. performed ELISA assays on NR and NOAR, designed the study and wrote the paper. All authors contributed to the final paper.

Competing interests

A patent for the diagnostic use of the CEP-1 peptide (patent application number: WO0890360, published on 31 July 2008) is jointly held by two of the authors (P.J.V. and K.L.) and funded by Imperial Innovations, Imperial College London.

Corresponding author

Correspondence to Karin Lundberg.

Supplementary information

PDF files

  1. 1.

    Supplementary Text and Figures

    Supplementary Tables 1–3 and Supplementary Note

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/ng.480

Further reading