Immunization with Porphyromonas gingivalis enolase induces autoimmunity to mammalian α-enolase and arthritis in DR4-IE-transgenic mice

Arthritis Rheum. 2011 Dec;63(12):3818-23. doi: 10.1002/art.30639.

Abstract

Objective: To examine the hypothesis that the subset of rheumatoid arthritis (RA) characterized by antibodies to citrullinated α-enolase is mediated by Porphyromonas gingivalis enolase in the context of DR4 alleles.

Methods: Recombinant human α-enolase and P gingivalis enolase, either citrullinated or uncitrullinated, were used to immunize DR4-IE-transgenic mice and control mice (class II major histocompatibility complex-deficient [class II MHC(-/-)] and C57BL/6 wild-type mice). Arthritis was quantified by measurement of ankle swelling in the hind paws and histologic examination. Serum IgG reactivity with α-enolase and citrullinated α-enolase was assayed by Western blotting and enzyme-linked immunosorbent assay (ELISA). Antibodies to peptide 1 of citrullinated α-enolase (CEP-1) and its arginine-bearing control peptide, REP-1, were also assessed by ELISA.

Results: Significant hind-ankle swelling (≥0.3 mm) occurred in DR4-IE-transgenic mice immunized with citrullinated human α-enolase (9 of 12 mice), uncitrullinated human α-enolase (9 of 12 mice), citrullinated P gingivalis enolase (6 of 6 mice), and uncitrullinated P gingivalis enolase (6 of 6 mice). Swelling peaked on day 24. None of the control groups developed arthritis. The arthritic joints showed synovial hyperplasia and erosions, but there was a paucity of leukocyte infiltration. Antibodies to human α-enolase, both citrullinated and unmodified, and to CEP-1 and REP-1 were detectable in all immunized mice except the class II MHC(-/-) control mice.

Conclusion: This is the first animal model that links an immune response to P gingivalis enolase to an important subset of RA, defined by antibodies to citrullinated α-enolase in the context of DR4. The fact that arthritis and anti-CEP-1 antibodies were induced independent of citrullination of the immunizing antigen suggests that the unmodified form of α-enolase may be important in initiating the corresponding subset of human RA.

MeSH terms

  • Animals
  • Arthritis, Experimental / blood
  • Arthritis, Experimental / chemically induced
  • Arthritis, Experimental / immunology*
  • Autoimmunity / drug effects*
  • Autoimmunity / immunology
  • Disease Models, Animal
  • HLA-DR4 Antigen / genetics*
  • Histocompatibility Antigens Class II / genetics
  • Humans
  • Immunization*
  • Immunoglobulin G / blood
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Mice, Transgenic
  • Phosphopyruvate Hydratase / adverse effects
  • Phosphopyruvate Hydratase / immunology
  • Phosphopyruvate Hydratase / pharmacology*
  • Porphyromonas gingivalis / enzymology*
  • Recombinant Proteins / pharmacology

Substances

  • HLA-DR4 Antigen
  • Histocompatibility Antigens Class II
  • Immunoglobulin G
  • Recombinant Proteins
  • Phosphopyruvate Hydratase