title: Synovial fibroblasts infected with Salmonella enterica serovar Typhimurium mediate osteoclast differentiation and activation.
abstract: The mechanisms whereby arthritogenic organisms may induce cartilage and bone erosions in infection-triggered arthritis remain unknown. In this study, we asked whether an arthritogenic organism could contribute to osteoclast differentiation and activation through regulation of the receptor activator of NF-kappaB ligand (RANKL) in synovial fibroblasts. Rat synovial fibroblasts were infected in vitro with Salmonella enterica serovar Typhimurium and monitored over time. The expression of RANKL in resting and infected synovial fibroblasts was quantified by reverse transcription-PCR and Western blotting. Osteoclast progenitors, isolated from femurs of 8-week-old rats and cultured in the presence of macrophage colony-stimulating factor, were cocultured with either infected or noninfected synovial fibroblasts for 2 to 4 days. Differentiation and maturation of osteoclasts were determined by morphology and tartrate-resistant acid phosphatase (TRAP) staining and by a bone resorption bioassay. RANKL expression was undetectable in resting synovial fibroblasts but was dose-dependently upregulated in cells after Salmonella infection. Osteoprotegerin was constitutively expressed by synovial fibroblasts and was not upregulated by infection. Further, we observed the formation of multinucleated TRAP-positive cells and formation of bone resorption pits in cocultures of bone marrow-derived osteoclast precursors with synovial fibroblasts infected with Salmonella but not with heat-killed Salmonella or noninfected cells. Arthritogenic bacteria may alter bone structure via synovial fibroblast intermediaries, since infected synovial fibroblasts (i) upregulate RANKL expression and (ii) enhance osteoclast precursor maturation into multinucleated, TRAP-positive, bone-resorbing, osteoclast-like cells. These data provide a link between infection and osteoclastogenesis. A better understanding of infection-mediated osteoclast differentiation and activation may provide new therapeutic strategies for inflammatory joint disease.
authors.affiliation: Division of Rheumatology, Toronto Western Hospital, ECW8-005, 399 Bathurst St., Toronto, Ontario M5T 2S8, Canada.
authors.forename: Xiang
authors.initials: X
authors.lastname: Zhang
authors.forename: Jane E
authors.initials: JE
authors.lastname: Aubin
authors.forename: Tae-Hwan
authors.initials: TH
authors.lastname: Kim
authors.forename: Ursula
authors.initials: U
authors.lastname: Payne
authors.forename: Basil
authors.initials: B
authors.lastname: Chiu
authors.forename: Robert D
authors.initials: RD
authors.lastname: Inman
mesh_terms: D000818:Animals; D001862:Bone Resorption / Q000209:etiology; D002352:Carrier Proteins / Q000235:genetics; D002454:Cell Differentiation; D005347:Fibroblasts / Q000382:microbiology / Q000502:physiology*; D006023:Glycoproteins / Q000235:genetics; D008562:Membrane Glycoproteins / Q000235:genetics; D010010:Osteoclasts / Q000166:cytology / Q000502:physiology*; D053244:Osteoprotegerin; D053245:RANK Ligand; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D011917:Rats, Inbred Lew; D018160:Receptors, Cytoplasmic and Nuclear / Q000235:genetics; D018124:Receptors, Tumor Necrosis Factor; D012486:Salmonella typhimurium / Q000472:pathogenicity*; D013583:Synovial Membrane / Q000166:cytology / Q000382:microbiology*
chemical_list: D002352:Carrier Proteins; D006023:Glycoproteins; D008562:Membrane Glycoproteins; D053244:Osteoprotegerin; D053245:RANK Ligand; D012333:RNA, Messenger; D018160:Receptors, Cytoplasmic and Nuclear; D018124:Receptors, Tumor Necrosis Factor; C506032:Tnfrsf11b protein, rat