The active metabolite of leflunomide, A77 1726, increases the production of IL-1 receptor antagonist in human synovial fibroblasts and articular chondrocytes

Arthritis Research & Therapy

Gaby Palmer¹  Danielle Burger²  Françoise Mezin¹  David Magne¹  Cem Gabay¹  Jean-Michel Dayer²  Pierre-André Guerne¹ 

[1] Division of Rheumatology, University Hospital, and Department of Pathology, University of Geneva School of Medicine, Geneva, Switzerland

[2] Division of Immunology and Allergy, Department of Internal Medicine, University Hospital, Geneva, Switzerland

关键词: synovium;    leflunomide;    IL-1 receptor antagonist;    articular cartilage;   

Others  :  1101282
DOI  :  10.1186/ar1157

 received in 2003-09-12, accepted in 2004-02-04,  发布年份 2004

【 摘 要 】

Leflunomide is an immunomodulatory agent used for the treatment of rheumatoid arthritis. In this study, we investigated the effect of A77 1726 – the active metabolite of leflunomide – on the production of IL-1 receptor antagonist (IL-1Ra) by human synovial fibroblasts and articular chondrocytes. Cells were incubated with A77 1726 alone or in combination with proinflammatory cytokines. IL-1Ra production was determined by ELISA. A77 1726 alone had no effect, but in the presence of IL-1β or tumour necrosis factor-α it markedly enhanced the secretion of IL-1Ra in synovial fibroblasts and chondrocytes. The effect of A77 1726 was greatest at 100 μmol/l. In synovial fibroblasts and de-differentiated chondrocytes, A77 1726 also increased IL-1β-induced IL-1Ra production in cell lysates. Freshly isolated chondrocytes contained no significant amounts of intracellular IL-1Ra. A77 1726 is a known inhibitor of pyrimidine synthesis and cyclo-oxygenase (COX)-2 activity. Addition of exogenous uridine did not significantly modify the effect of A77 1726 on IL-1Ra production, suggesting that it was not mediated by inhibition of pyrimidine synthesis. Indomethacin increased IL-1β-induced IL-1Ra secretion in synovial fibroblasts and de-differentiated chondrocytes, suggesting that inhibition of COX-2 may indeed enhance IL-1β-induced IL-1Ra production. However, the stimulatory effect of indomethacin was consistently less effective than that of A77 1726. A77 1726 increases IL-1Ra production by synovial fibroblasts and chondrocytes in the presence of proinflammatory cytokines, and thus it may possess chondroprotective effects. The effect of A77 1726 may be partially mediated by inhibition of COX-2, but other mechanisms likely concur to stimulate IL-1Ra production.

【 授权许可】

   
2004 Palmer et al., licensee BioMed Central Ltd. This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the article's original URL.

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【 参考文献 】

• [1]Strand V, Cohen S, Schiff M, Weaver A, Fleischmann R, Cannon G, Fox R, Moreland L, Olsen N, Furst D, Caldwell J, Kaine J, Sharp J, Hurley F, Loew-Friedrich I: Treatment of active rheumatoid arthritis with leflunomide compared with placebo and methotrexate. Leflunomide Rheumatoid Arthritis Investigators Group. Arch Intern Med 1999, 159:2542-2550.
• [2]Smolen JS, Kalden JR, Scott DL, Rozman B, Kvien TK, Larsen A, Loew-Friedrich I, Oed C, Rosenburg R: Efficacy and safety of leflunomide compared with placebo and sulphasalazine in active rheumatoid arthritis: a double-blind, randomised, multi-centre trial. European Leflunomide Study Group. Lancet 1999, 353:259-266.
• [3]Emery P, Breedveld FC, Lemmel EM, Kaltwasser JP, Dawes PT, Gomor B, Van Den Bosch F, Nordstrom D, Bjorneboe O, Dahl R, Horslev-Petersen K, Rodriguez De La Serna A, Molloy M, Tikly M, Oed C, Rosenburg R, Loew-Friedrich I: A comparison of the efficacy and safety of leflunomide and methotrexate for the treatment of rheumatoid arthritis. Rheumatology (Oxford) 2000, 39:655-665.
• [4]Breedveld FC, Dayer JM: Leflunomide: mode of action in the treatment of rheumatoid arthritis. Ann Rheum Dis 2000, 59:841-849.
• [5]Herrmann ML, Schleyerbach R, Kirschbaum BJ: Leflunomide: an immunomodulatory drug for the treatment of rheumatoid arthritis and other autoimmune diseases. Immunopharmacology 2000, 47:273-289.
• [6]Williams JW, Mital D, Chong A, Kottayil A, Millis M, Longstreth J, Huang W, Brady L, Jensik S: Experiences with leflunomide in solid organ transplantation. Transplantation 2002, 73:358-366.
• [7]Cherwinski HM, Byars N, Ballaron SJ, Nakano GM, Young JM, Ransom JT: Leflunomide interferes with pyrimidine nucleotide biosynthesis. Inflamm Res 1995, 44:317-322.
• [8]Williamson RA, Yea CM, Robson PA, Curnock AP, Gadher S, Hambleton AB, Woodward K, Bruneau JM, Hambleton P, Spinella-Jaegle S, Morand P, Courtin O, Sautes C, Westwood R, Hercend T, Kuo EA, Ruuth E: Dihydroorotate dehydrogenase is a target for the biological effects of leflunomide. Transplant Proc 1996, 28:3088-3091.
• [9]Manna SK, Aggarwal BB: Immunosuppressive leflunomide metabolite (A77 1726) blocks TNF-dependent nuclear factor-kappa B activation and gene expression. J Immunol 1999, 162:2095-2102.
• [10]Manna SK, Mukhopadhyay A, Aggarwal BB: Leflunomide suppresses TNF-induced cellular responses: effects on NF-kappa B, activator protein-1, c-Jun N-terminal protein kinase, and apoptosis. J Immunol 2000, 165:5962-5969.
• [11]Hamilton LC, Vojnovic I, Warner TD: A77 the active metabolite of leflunomide, directly inhibits the activity of cyclo-oxygenase-2 in vitro and in vivo in a substrate-sensitive manner. Br J Pharmacol 1999, 127:1589-1596.
• [12]Burger D, Begue-Pastor N, Benavent S, Gruaz L, Kaufmann MT, Chicheportiche R, Dayer JM: The active metabolite of leflunomide, A77 inhibits the production of prostaglandin E(2), matrix metalloproteinase 1 and interleukin 6 in human fibroblast-like synoviocytes. Rheumatology (Oxford) 2003, 42:89-96.
• [13]Mattar T, Kochhar K, Bartlett R, Bremer EG, Finnegan A: Inhibition of the epidermal growth factor receptor tyrosine kinase activity by leflunomide. FEBS Lett 1993, 334:161-164.
• [14]Xu X, Williams JW, Gong H, Finnegan A, Chong AS: Two activities of the immunosuppressive metabolite of leflunomide, A77 1726. Inhibition of pyrimidine nucleotide synthesis and protein tyrosine phosphorylation. Biochem Pharmacol 1996, 52:527-534.
• [15]Elder RT, Xu X, Williams JW, Gong H, Finnegan A, Chong AS: The immunosuppressive metabolite of leflunomide, A77 affects murine T cells through two biochemical mechanisms. J Immunol 1997, 159:22-27.
• [16]Deage V, Burger D, Dayer JM: Exposure of T lymphocytes to leflunomide but not to dexamethasone favors the production by monocytic cells of interleukin-1 receptor antagonist and the tissue-inhibitor of metalloproteinases-1 over that of interleukin-1beta and metalloproteinases. Eur Cytokine Netw 1998, 9:663-668.
• [17]Cutolo M, Sulli A, Ghiorzo P, Pizzorni C, Craviotto C, Villaggio B: Anti-inflammatory effects of leflunomide on cultured synovial macrophages from patients with rheumatoid arthritis. Ann Rheum Dis 2003, 62:297-302.
• [18]Elkayam O, Yaron I, Shirazi I, Judovitch R, Caspi D, Yaron M: Active leflunomide metabolite inhibits interleukin 1beta, tumour necrosis factor alpha, nitric oxide, and metalloproteinase-3 production in activated human synovial tissue cultures. Ann Rheum Dis 2003, 62:440-443.
• [19]Seitz M, Zwicker M, Loetscher P: Effects of methotrexate on differentiation of monocytes and production of cytokine inhibitors by monocytes. Arthritis Rheum 1998, 41:2032-2038.
• [20]Lotz M, Clark-Lewis I, Ganu V: HIV-1 transactivator protein Tat induces proliferation and TGF beta expression in human articular chondrocytes. J Cell Biol 1994, 124:365-371.
• [21]Klareskog L, Forsum U, Malmnas Tjernlund UK, Kabelitz D, Wigren A: Appearance of anti-HLA-DR-reactive cells in normal and rheumatoid synovial tissue. Scand J Immunol 1981, 14:183-192.
• [22]Gabay C, Smith MF, Eidlen D, Arend WP: Interleukin 1 receptor antagonist (IL-1Ra) is an acute-phase protein. J Clin Invest 1997, 99:2930-2940.
• [23]Fox RI, Herrmann ML, Frangou CG, Wahl GM, Morris RE, Strand V, Kirschbaum BJ: Mechanism of action for leflunomide in rheumatoid arthritis. Clin Immunol 1999, 93:198-208.
• [24]Seckinger P, Lowenthal JW, Williamson K, Dayer JM, MacDonald HR: A urine inhibitor of interleukin 1 activity that blocks ligand binding. J Immunol 1987, 139:1546-1549.
• [25]Arend WP, Malyak M, Guthridge CJ, Gabay C: Interleukin-1 receptor antagonist: role in biology. Annu Rev Immunol 1998, 16:27-55.
• [26]Muzio M, Polentarutti N, Facchetti F, Peri G, Doni A, Sironi M, Transidico P, Salmona M, Introna M, Mantovani A: Characterization of type II intracellular IL-1 receptor antagonist (IL-1ra3): a depot IL-1ra. Eur J Immunol 1999, 29:781-788.
• [27]Watson JM, Lofquist AK, Rinehart CA, Olsen JC, Makarov SS, Kaufman DG, Haskill JS: The intracellular IL-1 receptor antagonist alters IL-1-inducible gene expression without blocking exogenous signaling by IL-1 beta. J Immunol 1995, 155:4467-4475.
• [28]Palmer G, Mezin F, Juge-Aubry CE, Plater-Zyberk C, Gabay C, Guerne PA: Interferon-beta stimulates interleukin-1 receptor antagonist production in human articular chondrocytes and synovial fibroblasts. Ann Rheum Dis 2004, in press.
• [29]Palmer G, Guerne PA, Mezin F, Maret M, Guicheux J, Goldring MB, Gabay C: Production of interleukin-1 receptor antagonist by human articular chondrocytes. Arthritis Res 2002, 4:226-231. BioMed Central Full Text
• [30]Palmer G, Talabot-Ayer D, Szalay-Quinodoz I, Maret M, Arend WP, Gabay C: Mice transgenic for intracellular interleukin-1 receptor antagonist type 1 are protected from collagen-induced arthritis. Eur J Immunol 2003, 33:434-440.
• [31]Bresnihan B: Effects of anakinra on clinical and radiological outcomes in rheumatoid arthritis. Ann Rheum Dis 2002, Suppl 2:ii74-ii77.
• [32]Dayer JM, Bresnihan B: Targeting interleukin-1 in the treatment of rheumatoid arthritis. Arthritis Rheum 2002, 46:574-578.
• [33]Dayer JM: The pivotal role of interleukin-1 in the clinical manifestations of rheumatoid arthritis. Rheumatology (Oxford) 2003, Suppl 2:ii3-ii10.
• [34]Granowitz EV, Clark BD, Vannier E, Callahan MV, Dinarello CA: Effect of interleukin-1 (IL-1) blockade on cytokine synthesis: I. IL-1 receptor antagonist inhibits IL-1-induced cytokine synthesis and blocks the binding of IL-1 to its type II receptor on human monocytes. Blood 1992, 79:2356-2363.
• [35]Chabaud M, Page G, Miossec P: Enhancing effect of IL-1, IL-17, and TNF-alpha on macrophage inflammatory protein-3alpha production in rheumatoid arthritis: regulation by soluble receptors and Th2 cytokines. J Immunol 2001, 167:6015-6020.
• [36]Cohen S, Cannon GW, Schiff M, Weaver A, Fox R, Olsen N, Furst D, Sharp J, Moreland L, Caldwell J, Kaine J, Strand V: Two-year, blinded, randomized, controlled trial of treatment of active rheumatoid arthritis with leflunomide compared with methotrexate. Utilization of Leflunomide in the Treatment of Rheumatoid Arthritis Trial Investigator Group. Arthritis Rheum 2001, 44:1984-1992.
• [37]van de Loo FA, van den Berg WB: Gene therapy for rheumatoid arthritis. Lessons from animal models, including studies on interleukin-4, interleukin-10, and interleukin-1 receptor antagonist as potential disease modulators. Rheum Dis Clin North Am 2002, 28:127-149.
• [38]Huh YH, Kim SH, Kim SJ, Chun JS: Differentiation status-dependent regulation of cyclooxygenase-2 expression and prostaglandin E2 production by epidermal growth factor via mitogen-activated protein kinase in articular chondrocytes. J Biol Chem 2003, 278:9691-9697.
• [39]Pelletier JP, Mineau F, Ranger P, Tardif G, Martel-Pelletier J: The increased synthesis of inducible nitric oxide inhibits IL-1ra synthesis by human articular chondrocytes: possible role in osteoarthritic cartilage degradation. Osteoarthritis Cartilage 1996, 4:77-84.
• [40]Al-Ramadi BK, Welte T, Fernandez-Cabezudo MJ, Galadari S, Dittel B, Fu XY, Bothwell AL: The Src-protein tyrosine kinase Lck is required for IL-1-mediated costimulatory signaling in Th2 cells. J Immunol 2001, 167:6827-6833.
• [41]Funakoshi-Tago M, Tago K, Sonoda Y, Tominaga S, Kasahara T: TRAF6 and C-SRC induce synergistic AP-1 activation via PI3-kinase-AKT-JNK pathway. Eur J Biochem 2003, 270:1257-1268.