title: Type I interferons might form the link between Toll-like receptor (TLR) 3/7 and TLR4-mediated synovial inflammation in rheumatoid arthritis (RA).
abstract: BACKGROUND
  Rheumatoid arthritis (RA) has been associated with an increased risk of infections, but the underlying pathways have not yet been identified. Toll-like receptors (TLR) probably play a role in synovial inflammation and may also contribute to the understanding of the role of infections in RA.
  
  
  OBJECTIVE
  To investigate if the synovial expression of TLR3 and TLR7 in RA correlates with that of inflammatory cytokines, and to assess whether this has functional consequences for local cytokine production and to study potential links between the TLR3/7 axis and TLR4 in RA synovium.
  
  
  METHODS
  Immunohistochemistry was used to study the expression of TLR3, TLR7, interferon alpha (IFNalpha), tumour necrosis factor alpha (TNFalpha) and interleukins IL1beta, IL12, IL17 and IL18 in RA synovium obtained by arthroscopy from 34 patients with RA. Monocytes, monocyte-derived dendritic cells (MoDCs) and RA synovial fibroblasts were stimulated via TLR3 (poly-IC) and TLR7 (loxorubin), after which IL1beta, IL6 and TNFalpha were measured by Luminex bead array technology. Following preincubation with IFNalpha, IL1beta and IL18, TLR3 and TLR7 mRNA expression was assessed using real-time PCR. Cytokine production after preincubation with IFNalpha and subsequent TLR stimulation was measured.
  
  
  RESULTS
  Synovial TLR3/7 expression was co-expressed with IFNalpha, IL1beta and IL18, but not with TNFalpha, IL12 and IL17. Stimulation of TLR3/TLR7 on monocytes, MoDCs or synovial fibroblasts led to secretion of type I IFN but no biologically active IL1beta or IL18 could be detected. Type I IFNalpha increased TLR3/7 mRNA expression whereas IL1beta and IL18 did not. In spite of the fact that the mRNA level of TLR4 remained unchanged, IFNalpha enhanced the response to TLR4 agonists, a phenomenon that was clearly more marked in patients with RA.
  
  
  CONCLUSIONS
  Type I interferons are highly co-expressed with TLR3/TLR7 in RA synovium. They enhance TLR3/TLR7-mediated cytokine production and also TLR4-mediated responses.
authors.affiliation: Department of Rheumatology, Radboud University Nijmegen Medical Center, Nijmegen 6500 HB, The Netherlands.
authors.forename: M F
authors.initials: MF
authors.lastname: Roelofs
authors.forename: M H
authors.initials: MH
authors.lastname: Wenink
authors.forename: F
authors.initials: F
authors.lastname: Brentano
authors.forename: S
authors.initials: S
authors.lastname: Abdollahi-Roodsaz
authors.forename: B
authors.initials: B
authors.lastname: Oppers-Walgreen
authors.forename: P
authors.initials: P
authors.lastname: Barrera
authors.forename: P L C M
authors.initials: PL
authors.lastname: van Riel
authors.forename: L A B
authors.initials: LA
authors.lastname: Joosten
authors.forename: D
authors.initials: D
authors.lastname: Kyburz
authors.forename: W B
authors.initials: WB
authors.lastname: van den Berg
authors.forename: T R D J
authors.initials: TR
authors.lastname: Radstake