Responses to the proinflammatory cytokines interleukin-1 and tumor necrosis factor alpha in cells derived from rheumatoid synovium and other joint tissues involve nuclear factor kappaB-mediated induction of the Ets transcription factor ESE-1
Department of Medicine, Division of Rheumatology
Arthritis, Rheumatoid; Blotting, Northern; Cell Line; Chondrocytes; DNA Primers; *DNA-Binding Proteins; Fibroblasts; Humans; Immunoenzyme Techniques; Interleukin-1; Lipopolysaccharides; Macrophages; NF-kappa B; Oligonucleotide Probes; Osteoarthritis; Osteoblasts; *Proto-Oncogene Proteins; Proto-Oncogene Proteins c-ets; RNA, Messenger; Reverse Transcriptase Polymerase Chain Reaction; Synovial Membrane; Trans-Activators; *Transcription Factors; Tumor Necrosis Factor-alpha
Musculoskeletal Diseases | Rheumatology | Skin and Connective Tissue Diseases
OBJECTIVE: To investigate the expression of the novel Ets transcription factor ESE-1 in rheumatoid synovium and in cells derived from joint tissues, and to analyze the role of nuclear factor kappaB (NF-kappaB) as one of the central downstream targets in mediating the induction of ESE-1 by proinflammatory cytokines.
METHODS: ESE-1 protein expression was analyzed by immunohistochemistry using antibodies in synovial tissues from patients with rheumatoid arthritis (RA) and osteoarthritis (OA). ESE-1 messenger RNA (mRNA) levels were analyzed by reverse transcriptase-polymerase chain reaction or Northern blotting in human chondrocytes, synovial fibroblasts, osteoblasts, and macrophages, before and after exposure to interleukin-1beta (IL-1beta), tumor necrosis factor alpha (TNFalpha), or lipopolysaccharide (LPS) with or without prior infection with an adenovirus encoding the inhibitor of nuclear factor kappaB (IkappaB). The wild-type ESE-1 promoter and the ESE-1 promoter mutated in the NF-kappaB site were cloned into a luciferase reporter vector and analyzed in transient transfections. Electrophoretic mobility shift assays (EMSAs) and supershift assays with antibodies against members of the NF-kappaB family were conducted using the NF-kappaB site from the ESE-1 promoter as a probe.
RESULTS: Immunohistochemical analysis showed specific expression of ESE-1 in cells of the synovial lining layer and in some mononuclear and endothelial cells in RA and OA synovial tissues. ESE-1 mRNA expression could be induced by IL-1beta and TNFalpha in cells such as synovial fibroblasts, chondrocytes, osteoblasts, and monocytes. Transient transfection experiments and EMSAs showed that induction of ESE-1 gene expression by IL-1beta requires activation of NF-kappaB and binding of p50 and p65 family members to the NF-kappaB site in the ESE-1 promoter. Overexpression of IkappaB using an adenoviral vector blocked IL-1beta-induced ESE-1 mRNA expression. Chromatin immunoprecipitation further confirmed that NF-kappaB binds to the ESE-1 promoter in vivo.
CONCLUSION: ESE-1 is expressed in synovial tissues in RA and, to a variable extent, in OA, and is specifically induced in synovial fibroblasts, chondrocytes, osteoblasts, and monocyte/macrophages by IL-1beta, TNFalpha, or LPS. This induction relies on the translocation of the NF-kappaB family members p50 and p65 to the nucleus and transactivation of the ESE-1 promoter via a high-affinity NF-kappaB binding site. ESE-1 may play a role in mediating some effects of proinflammatory stimuli in cells at sites of inflammation.
DOI of Published Version
Arthritis Rheum. 2003 May;48(5):1249-60. Link to article on publisher's site
Arthritis and rheumatism
Grall, Franck; Gu, Xuesong; Tan, Lujian; Cho, Je-Yoel; Inan, Mehmet Sait.; Pettit, Allison R.; Thamrongsak, Usanee; Choy, Bob K.; Manning, Cathy; Akbarali, Yasmin; Zerbini, Luiz; Rudders, Susan; Goldring, Steven R.; Gravallese, Ellen M.; Oettgen, Peter; Goldring, Mary B.; and Libermann, Towia A., "Responses to the proinflammatory cytokines interleukin-1 and tumor necrosis factor alpha in cells derived from rheumatoid synovium and other joint tissues involve nuclear factor kappaB-mediated induction of the Ets transcription factor ESE-1" (2003). Rheumatology Publications and Presentations. 38.