Increased levels of peptidylarginine deiminase 2 in synovial fluid from anti-CCP-positive rheumatoid arthritis patients: Association with disease activity and inflammatory markers

Rheumatology (Oxford). 2016 May;55(5):918-27. doi: 10.1093/rheumatology/kev440. Epub 2016 Feb 15.

Abstract

Objective: To quantify peptidylarginine deiminase 2 (PAD2) in SF of RA patients and OA patients, and to determine the association between PAD2 levels, disease activity and inflammatory markers in RA.

Methods: Blood and SF samples were obtained from 39 RA patients and 40 patients with OA. PAD2 content and PAD activity were measured by means of in-house assays. TNF-α, IL-1β, IL-6, IL-8, IL-10 and IL-12 were measured using flow cytometry.

Results: PAD2 levels and PAD activity were higher in SF from RA than OA patients (P < 0.0001 and P = 0.03, respectively), as were all cytokine levels (P < 0.0001-0.05). SF PAD2 levels were higher among anti-CCP-positive patients than among anti-CCP-negative patients (P = 0.005). PAD2 levels in SF from RA patients correlated with disease activity, as assessed by DAS28 (P < 0.005). Moreover, SF PAD2 levels correlated with circulating CRP and anti-CCP levels (P < 0.0006), as well as with leucocyte count, IL-6, IL-8 and IL-10 levels in SF (P < 0.0001-0.02). PAD activity in SF was higher in RA patients than in OA patients, and correlated with PAD2 concentration.

Conclusion: Extracellular PAD2 levels in SF correlate with disease activity in RA patients. Anti-CCP-positive RA patients have higher PAD2 levels in SF than anti-CCP-negative RA patients and OA patients. Since we could demonstrate enzymatically active PADs in SF, we propose that free, extracellular PAD is of pathogenic relevance.

Keywords: citrullination; cytokines; peptidylarginine deiminase; rheumatoid arthritis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Arthritis, Rheumatoid / enzymology*
  • Arthritis, Rheumatoid / immunology
  • Autoantibodies / blood*
  • Biomarkers / metabolism
  • C-Reactive Protein / metabolism
  • Cytokines / metabolism
  • Enzyme-Linked Immunosorbent Assay / methods
  • Female
  • Humans
  • Hydrolases / metabolism*
  • Inflammation Mediators / metabolism
  • Leukocyte Count
  • Male
  • Middle Aged
  • Osteoarthritis / enzymology
  • Osteoarthritis / immunology
  • Peptides, Cyclic / immunology*
  • Protein-Arginine Deiminase Type 2
  • Protein-Arginine Deiminases
  • Synovial Fluid / metabolism*

Substances

  • Autoantibodies
  • Biomarkers
  • Cytokines
  • Inflammation Mediators
  • Peptides, Cyclic
  • cyclic citrullinated peptide
  • C-Reactive Protein
  • Hydrolases
  • PADI2 protein, human
  • Protein-Arginine Deiminase Type 2
  • Protein-Arginine Deiminases