title: Antibodies specific for carbamylated proteins precede the onset of clinical symptoms in mice with collagen induced arthritis.
abstract: OBJECTIVE
  The immune response to post-translationally modified antigens is a key characteristic of rheumatoid arthritis. Carbamylation is such a posttranslational modification. Recently, we demonstrated that autoantibodies recognizing carbamylated proteins are present in sera of rheumatoid arthritis. The molecular mechanisms underlying the break of tolerance and hence the induction of anti-CarP antibody responses are unknown as well as their appearance in mouse models for systemic arthritis. Therefore we analyzed their appearance in the mouse collagen-induced arthritis model.
  
  
  METHODS
  collagen induced arthritis was induced by immunization with type II collagen in complete Freund's adjuvant. Arthritis severity was monitored by clinical scoring and anti-CarP antibody levels were determined by ELISA.
  
  
  RESULTS
  Anti-CarP antibodies were detectable in mice with collagen induced arthritis. We did not detect ACPA in mice with collagen induced arthritis. The specificity of the antibodies for carbamylated proteins was confirmed by inhibition assays and immunoblotting. Injection with complete Freund's adjuvant without type II collagen could also induce anti-CarP antibodies, however, in mice with arthritis, the anti-CarP antibody response was stronger and developed more rapidly. The onset of collagen induced arthritis was preceded by an increase of anti-CarP IgG2a levels in the serum.
  
  
  CONCLUSIONS
  In mice with collagen induced arthritis we did not observe an immune response against citrullinated antigens, but we did observe an immune response against carbamylated antigens. This anti-CarP response already appeared before disease onset, indicating that collagen induced arthritis can be used as an in vivo model to study anti-CarP antibodies. Our data also indicate that the tolerance to carbamylated proteins, in contrast to the response to citrullinated proteins, is easily broken and that arthritis boosts the immune response against these proteins. The anti-CarP response in mice with CIA can be used as a model for immune responses to post-translationally modified proteins.
authors.affiliation: Department of Rheumatology, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Jeroen N
authors.initials: JN
authors.lastname: Stoop
authors.affiliation: Department of Rheumatology, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Bi-Sheng
authors.initials: BS
authors.lastname: Liu
authors.affiliation: Department of Rheumatology, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Jing
authors.initials: J
authors.lastname: Shi
authors.affiliation: Department of Rheumatology, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Diahann T S L
authors.initials: DT
authors.lastname: Jansen
authors.affiliation: Immunoscience Research Unit, Pfizer Worldwide Research and Development, Cambridge, Massachusetts, United States of America.
authors.forename: Martin
authors.initials: M
authors.lastname: Hegen
authors.affiliation: Department of Rheumatology, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Tom W J
authors.initials: TW
authors.lastname: Huizinga
authors.affiliation: Department of Rheumatology, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Leendert A
authors.initials: LA
authors.lastname: Trouw
authors.affiliation: Department of Rheumatology, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: René E M
authors.initials: RE
authors.lastname: Toes
mesh_terms: D000818:Animals; D000918:Antibody Specificity / Q000276:immunology*; D001169:Arthritis, Experimental / Q000276:immunology*; D001323:Autoantibodies / Q000097:blood / Q000276:immunology*; D002219:Carbamates / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D007132:Immunoglobulin Isotypes / Q000097:blood / Q000276:immunology; D051379:Mice; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000276:immunology* / Q000378:metabolism*
chemical_list: D001323:Autoantibodies; D002219:Carbamates; D007132:Immunoglobulin Isotypes; D011506:Proteins