Rheumatoid Arthritis: Klein MJ

Mindrescu C, Dias AA, Olszewski RJ, Klein MJ, Reis LF, Wisniewski HG. · New York University School of Medicine, New York, New York 10016, USA. · Arthritis Rheum. · Pubmed #12355494 links to  free full text

Abstract: OBJECTIVE: Expression of TSG-6 (tumor necrosis factor-stimulated gene 6) is induced by proinflammatory cytokines. This study was undertaken to examine the effects of local expression of TSG-6 in arthritic joints of TSG-6 transgenic mice, in the collagen-induced arthritis (CIA) model. METHODS: We generated transgenic mice that harbored the TSG-6 gene under the control of the T cell-specific lck promoter. Arthritis was induced by immunization with bovine type II collagen (CII), and its progression was monitored based on the incidence of arthritis, the arthritis index, and footpad swelling. Anti-CII antibodies and cytokine production were determined by enzyme-linked immunosorbent assay. Gene expression arrays were used to compare gene expression profiles of transgenic and control mice at various stages of CIA. RESULTS: TSG-6 was expressed in limbs of transgenic mice after immunization with CII, while its expression in nontransgenic animals was insignificant. The incidence of CIA was reduced in TSG-6 transgenic animals, its onset delayed, and all parameters of clinical arthritis significantly reduced. However, the immune response against CII was not significantly inhibited in TSG-6 transgenic mice. CONCLUSION: TSG-6 expression has been demonstrated in patients with rheumatoid and other forms of arthritis. Our data show that local expression of TSG-6 at sites of inflammation results in potent inhibition of inflammation and joint destruction in a model of autoimmune arthritis in mice. Therefore, it is likely that TSG-6 plays a similar modulatory role in human rheumatoid arthritis and related diseases and may have potential for the treatment of autoimmune arthritis in humans.

Hou WS, Li W, Keyszer G, Weber E, Levy R, Klein MJ, Gravallese EM, Goldring SR, Brömme D. · Mount Sinai School of Medicine, New York, New York 10029, USA. · Arthritis Rheum. · Pubmed #11920402 links to  free full text

Abstract: OBJECTIVE: To determine and compare the expression of cathepsins K and S proteins in joints with rheumatoid arthritis (RA) and osteoarthritis (OA) and to determine the effect of interleukin-1 beta (IL-1 beta) and tumor necrosis factor alpha (TNF alpha) on the expression of cathepsin K in fibroblast-like synoviocytes. METHOD: Expression and localization of cathepsins K and S were determined by immunohistochemistry in the synovium of 10 RA- and 8 OA-affected joints. Northern and Western blot analyses were performed to analyze cathepsin K and S expression in primary fibroblast-like synoviocyte cultures from RA and OA patients. The effect of IL-1 beta and TNF alpha on the expression and secretion of cathepsin K in primary cultures of synoviocytes was determined by real-time polymerase chain reaction and Western blot analysis. Staining of in situ activity was used to identify active cathepsin K enzyme in primary synovial fibroblast cultures. RESULTS: Cathepsin K and S protein expression was identified in the synovium from patients with RA and OA. Cathepsin K protein was localized in synovial fibroblasts, stromal multinucleated giant cells, and, to a lesser degree, in CD68+ macrophage-like synoviocytes. Of note is the expression of cathepsin K in synovial fibroblasts and mononuclear macrophage-like cells at sites of cartilage erosion in RA and in interdigitating cells of lymphocyte-rich areas. In contrast, cathepsin S expression was restricted to CD68+ macrophage-like synoviocytes, interdigitating cells, and endothelial cells of blood vessels. Cathepsin K protein expression in the interstitial areas and perivascular regions of RA-derived synovial specimens was 2-5 times higher than in OA samples (P < 0.001), whereas the expression of cathepsin S did not significantly differ in these diseases. Cathepsin K expression levels in normal synovium were low and restricted to fibroblast-like cells. Of note, cathepsin K also was expressed in repairing fibrocartilage in 1 OA specimen. Primary cell cultures of RA- and OA-derived synovial fibroblasts expressed comparable amounts of cathepsin K at the transcript and protein levels. Both cell cultures secreted mature cathepsin K as well as procathepsin K, and expressed active cathepsin K in cytosolic vesicles. In contrast, neither RA- nor OA-derived fibroblasts expressed detectable levels of cathepsin S. IL-1 beta and TNF alpha stimulated the transcript (7-8-fold) and protein expression (2-fold) of cathepsin K (P < 0.05) in primary synovial fibroblast cultures, without differences in expression between RA- and OA-derived synovial fibroblasts. CONCLUSION: The presence of cathepsin K polypeptide in synovial fibroblasts and macrophage-like cells in normal, OA, and RA synovia suggests a constitutive expression of this protease and a role in synovial remodeling. The comparable increase in cathepsin K expression after stimulation of RA- and OA-derived synovial fibroblasts with IL-1 beta and TNF alpha further suggests that the expression of cathepsin K is independent of cellular alterations leading to the invasive phenotype of RA-synovial fibroblasts. However, the overexpression of cathepsin K in RA synovia due to an increase in the number of cathepsin K-expressing cells identifies this enzyme as a candidate protease for the pathologic degradation of articular cartilage. Cathepsin S expression in macrophage-like synoviocytes suggests dual activity in antigen presentation and matrix degradation in the inflamed synovia.

Hou WS, Li Z, Gordon RE, Chan K, Klein MJ, Levy R, Keysser M, Keyszer G, Brömme D. · Department of Human Genetics, Mount Sinai School of Medicine, New York, New York 10029-6574, USA. · Am J Pathol. · Pubmed #11733367 links to  free full text

Abstract: Synovial fibroblasts (SFs) play a critical role in the pathogenesis of rheumatoid arthritis (RA) and are directly involved in joint destruction. Both SF-resident matrix metalloproteases and cathepsins have been implicated in cartilage degradation although their identities and individual contributions remain unclear. The aims of this study were to investigate the expression of cathepsin K in SFs, the correlation between cathepsin K expression and disease severity, and the contribution of cathepsin K to fibroblast-mediated collagen degradation. Immunostaining of joint specimens of 21 patients revealed high expression of cathepsin K in SFs in the synovial lining and the stroma of synovial villi, and to a lesser extent in CD68-positive cells of the synovial lining. Cathepsin K-positive SFs were consistently observed at sites of cartilage and bone degradation. Expression levels of cathepsin K in the sublining and vascularized areas of inflamed synovia showed a highly significant negative correlation with results derived from the Hannover Functional Capacity Questionnaire (r = 0.78, P = 0.003; and r = 0.70, P = 0.012, respectively) as a measure of the severity of RA in individual patients. For comparison, there was no correlation between Hannover Functional Capacity Questionnaire and cathepsin S whose expression is limited to CD-68-positive macrophage-like synoviocytes. The expression of cathepsin K was also demonstrated in primary cell cultures of RA-SFs. Co-cultures of SFs on cartilage disks revealed the ability of fibroblast-like cells to phagocytose collagen fibrils whose intralysosomal hydrolysis was prevented in the presence of a potent cathepsin K inhibitor but not by an inhibitor effective against cathepsins L, B, and S. The selective and critical role of cathepsin K in articular cartilage and subchondral bone erosion was further corroborated by the finding that cathepsin K has a potent aggrecan-degrading activity and that cathepsin K-generated aggrecan cleavage products specifically potentiate the collagenolytic activity of cathepsin K toward type I and II collagens. This study demonstrates for the first time a critical role of cathepsin K in cartilage degradation by SFs in RA that is comparable to its well-known activity in osteoclasts.

Mindrescu C, Thorbecke GJ, Klein MJ, Vilcek J, Wisniewski HG. · New York University School of Medicine, New York 10016, USA. · Arthritis Rheum. · Pubmed #11145024 links to  free full text

Abstract: OBJECTIVE: To examine the effect of recombinant TSG-6 on collagen-induced arthritis (CIA) in DBA/1J mice. TSG-6 is a tumor necrosis factor (TNF)/ interleukin-1 (IL-1)-inducible hyaluronan-binding protein produced by synovial cells and chondrocytes that is present in synovial fluids of patients with rheumatoid arthritis. METHODS: To determine the effect of TSG-6 on chronic inflammatory joint disease, we induced CIA in DBA/1J mice by immunization with bovine type II collagen. Animals were treated with 12 intraperitoneal doses of 200 microg of recombinant TSG-6, beginning 3 days before the expected onset of disease symptoms. Progression of arthritis was monitored by determining the disease incidence, arthritis index, and footpad swelling. Levels of IgG1, IgG2a, and IgG2b antibodies against bovine and murine type II collagen and serum concentrations of IL-6 were determined at various time points. Histologic examination of affected joints was performed approximately 20 days after the onset of arthritis. RESULTS: Treatment with recombinant TSG-6 protein had a potent ameliorative effect, manifested by decreases in the disease incidence, arthritis index, and footpad swelling. Histologic examination of affected joints in TSG-6-treated animals revealed little pannus formation and cartilage erosion, features which were conspicuous in control mice. Animals treated with recombinant TSG-6 developed significantly reduced levels of IgG1, IgG2a, and IgG2b antibodies against bovine and murine type II collagen. CONCLUSION: The antiinflammatory effect of the TNF/IL-1-inducible TSG-6 protein in murine CIA suggests a role for this protein as an endogenous regulator of the inflammatory process.