title: Macrophage migration inhibitory factor enhances osteoclastogenesis through upregulation of RANKL expression from fibroblast-like synoviocytes in patients with rheumatoid arthritis.
abstract: BACKGROUND
  Macrophage migration inhibitory factor (MIF) is one of key regulators in acute and chronic immune-inflammatory conditions including rheumatoid arthritis (RA). We examined the effect of MIF on osteoclastogenesis, which is known to play a crucial role in bone destruction in RA.
  
  
  METHODS
  The concentration of MIF and receptor activator of nuclear factor-κB ligand (RANKL) in the synovial fluid was measured by ELISA. MIF-induced RANKL expression of RA synovial fibroblasts was determined by real-time PCR and western blot. Osteoclastogenesis was analyzed in culture of human peripheral blood mononuclear cells (PBMC) with MIF. Osteoclastogenesis was also determined after co-cultures of rhMIF-stimulated RA synovial fibroblasts with human PBMC.
  
  
  RESULTS
  Synovial fluid MIF concentration in RA patients was significantly higher than in osteoarthritis (OA) patients. The concentration of RANKL correlated with that of MIF in RA synovial fluids (r = 0.6, P < 0.001). MIF stimulated the expression of RANKL mRNA and protein in RA synovial fibroblasts, which was partially reduced by blocking of interleukin (IL)-1β. Osteoclasts were differentiated from PBMC cultures with MIF and M-CSF, even without RANKL. Osteoclastogenesis was increased after co-culture of MIF-stimulated RA synovial fibroblasts with PBMC and this effect was diminished by RANKL neutralization. Blocking of PI3 kinase, p38 MAP kinase, JAK-2, NF-κB, and AP-1 also led to a marked reduction in RANKL expression and osteoclastogenesis.
  
  
  CONCLUSIONS
  The interactions among MIF, synovial fibroblasts, osteoclasts, RANKL, and IL-1β have a close connection in osteoclastogenesis and they could be a potential gateway leading to new therapeutic approaches in treating bone destruction in RA.
authors.affiliation: Division of Rheumatology, Medical Immunology Center, Department of Internal Medicine, Konkuk University School of Medicine, 1 Hwayang-dong, Kwangjin-gu, Seoul 143-729, Korea.
authors.forename: Hae-Rim
authors.initials: HR
authors.lastname: Kim
authors.forename: Kyoung-Woon
authors.initials: KW
authors.lastname: Kim
authors.forename: Hong Geun
authors.initials: HG
authors.lastname: Jung
authors.forename: Kwang-Sup
authors.initials: KS
authors.lastname: Yoon
authors.forename: Hye-Jwa
authors.initials: HJ
authors.lastname: Oh
authors.forename: Mi-La
authors.initials: ML
authors.lastname: Cho
authors.forename: Sang-Heon
authors.initials: SH
authors.lastname: Lee
mesh_terms: D000328:Adult; D000368:Aged; D001172:Arthritis, Rheumatoid / Q000378:metabolism*; D015153:Blotting, Western; D001862:Bone Resorption / Q000378:metabolism*; D018920:Coculture Techniques; D004797:Enzyme-Linked Immunosorbent Assay; D005347:Fibroblasts / Q000378:metabolism*; D015870:Gene Expression; D006801:Humans; D007150:Immunohistochemistry; D053583:Interleukin-1beta / Q000378:metabolism; D007963:Leukocytes, Mononuclear / Q000378:metabolism; D008263:Macrophage Migration-Inhibitory Factors / Q000378:metabolism*; D008875:Middle Aged; D010010:Osteoclasts / Q000378:metabolism; D053245:RANK Ligand / Q000096:biosynthesis*; D012333:RNA, Messenger / Q000032:analysis; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013583:Synovial Membrane / Q000378:metabolism; D015854:Up-Regulation
chemical_list: D053583:Interleukin-1beta; D008263:Macrophage Migration-Inhibitory Factors; D053245:RANK Ligand; D012333:RNA, Messenger; C505688:TNFSF11 protein, human