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Preferential Induction of Prodestructive Matrix Metalloproteinase-1 and Proinflammatory Interleukin 6 and Prostaglandin E2 in Rheumatoid Arthritis Synovial Fibroblasts via Tumor Necrosis Factor Receptor-55
SAIFEDDIN ALSALAMEH, RAYYA J. AMIN, ELKE KUNISCH, HUGO E. JASIN, and RAIMUND W. KINNE
ABSTRACT.
Methods. Seventh to 9th passage RA SFB and OA SFB were analyzed for TNF-R expression by FACS. The SFB were then stimulated with TNF-a (1–10 ng/ml) or agonistic anti-TNF-R55 (HTR-9) and anti-TNF-R75 (UTR-1) monoclonal antibodies (1–25 µg/ml each). Matrix metalloproteinase-1 (MMP-1), tissue inhibitor of matrix metalloproteinase-1 (TIMP-1), interleukin 6 (IL-6), and prostaglandin E2 (PGE2) in culture supernatants were quantified by ELISA, and DNA fragmentation by TUNEL assay. Results. RA SFB variably expressed TNF-R55 (7.2 ± 2.2% positive cells, mean ± SEM) and TNF-R75 (0.6 ± 0.3%), similarly to OA SFB (6.8 ± 2.1% and 1.6 ± 0.8%, respectively). RA SFB constitutively secreted large amounts of TIMP-1 (1700 ng/ml), but only small amounts of MMP-1 (23.7 ng/ml), IL-6 (4.4 ng/ml), and PGE2 (0.34 ng/ml). OA SFB secreted comparable amounts of TIMP-1 (2470 ng/ml), MMP-1 (37 ng/ml), and IL-6 (5.0 ng/ml), but significantly higher amounts of PGE2 (0.58 ng/ml; p ³ 0.05). TNF-a stimulation induced IL-6 secretion by RA SFB (3-fold) and OA SFB (4-fold), as well as MMP-1 secretion (RA, 85-fold; OA, 29-fold), with significant differences between RA and OA. This was exclusively mediated by separate stimulation with agonistic anti-TNF-R55 Mab. Strikingly, RA SFB were completely unresponsive to TIMP-1 mRNA and protein induction by TNF-a, whereas TIMP-1 mRNA and/or protein in OA SFB was significantly upregulated by TNF-a (2-fold; p ³ 0.05, OA > RA) and by separate stimulation of both TNF receptors. TNF-a-induced PGE2 release by RA SFB (92-fold) and OA SFB (56-fold) was mediated by both TNF receptors; however, predominantly by TNF-R55. DNA fragmentation was induced exclusively by high concentrations of anti-TNF-R55 Mab and only in RA SFB. Conclusion. These results indicate preferential induction of prodestructive and proinflammatory mediators in RA SFB by the TNF-R55, with potential implications for understanding the pathogenesis of RA and the development of more specific therapeutic strategies. (J Rheumatol 2003;30:1680-90) Key Indexing Terms:
TUMOR NECROSIS FACTOR-a
From the Department of Medicine, Division of Rheumatology and Clinical Immunology, University of Arkansas for Medical Sciences and Veterans Administration Hospital, Little Rock, Arkansas, USA; and Experimental Rheumatology Unit, Friedrich Schiller University, Jena, Germany. Supported in part by grants from the Trinity Foundation and from the German Federal Ministry of Education and Research (FKZ 01ZZ9602). S. Alsalameh, MD, Associate Professor of Internal Medicine and Rheumatology, Research Instructor of Internal Medicine, Division of Rheumatology and Clinical Immunology, Department of Medicine, University of Arkansas for Medical Sciences (Present address: Outpatient Clinic for Rheumatic Diseases, Bahnhofstr. 30, D-35037 Marburg, Germany); R.J. Amin, MD, MSc, Postdoctoral Fellow, Division of Rheumatology and Clinical Immunology, Department of Medicine, University of Arkansas for Medical Sciences (Present address: Goethestr. 5, D-35043 Marburg, Germany); H.E. Jasin, MD, Professor of Medicine and Rheumatology, Head, Division of Rheumatology and Clinical Immunology, Department of Medicine, University of Arkansas for Medical Sciences; E. Kunisch, PhD; R.W. Kinne, MD, Associate Professor of Experimental Rheumatology, Head, Experimental Rheumatology Unit, Friedrich Schiller University. Address reprint requests to Dr. R.W. Kinne, Experimental Rheumatology Unit, Friedrich Schiller University Jena, Bachstrasse 18, D-07740 Jena, Germany. E-mail: Raimund.W.Kinne@rz.uni-jena.de Submitted April 3, 2002; revision accepted January 22, 2003. |