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Leflunomide induces immunosuppression in collagen-induced arthritis rats by upregulating CD4+CD25+ regulatory T cells

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This study was to investigate the effect of leflunomide on the immunosuppressive CD4+CD25+ regulatory T cells (CD4+CD25+ Tregs) in collagen-induced arthritis (CIA) rats. CIA was induced by collagen type II in Wistar rats. Immunofluorescence flow cytometry and RT-PCR were used to determine the proportion of CD4+CD25+ Tregs and the expression of Foxp3 mRNA, respectively. Proliferation of T lymphocytes was assayed with MTT reagent, and the level of transforming growth factor 1 (TGF-1) in the supernatant of concanavalin A (Con A)-induced T lymphocytes was determined by ELISA kit. Our investigations demonstrated that inhibition of arthritis by leflunomide was related to changes in CD4+CD25+ Tregs. In addition, A771726, which is the active metabolite of leflunomide, promoted the differentiation of spleen lymphocytes into CD4+CD25+ Tregs, increased antiinflammatory cytokine TGF-1 secretion, and adjusted the activity of Con A-induced lymphocytes in vitro.

La présente étude a examiné l’effet du léflunomide sur les cellules T régulatrices CD4+ et CD25+ (CD4+CD5+ Treg) immunosuppressives chez des rats souffrant d’arthrite induite au collagène (AIC). On a induit l’AIC chez des rats Wistar au moyen de collagène de type II. On a utilisé la cytométrie en flux couplée à l’immunofluorescence et la technique RT-PCR pour déterminer la proportion de CD4+CD5+ Treg et l’expression de l’ARNm de Foxp3, respectivement. On a évalué la prolifération de lymphocytes T par la méthode du MTT. On a déterminé, à l’aide d’une trousse Elisa, le taux de TGF-1 dans le surnageant des lymphocytes T induits par Con A. Nos examens ont démontré que l’inhibition de l’arthrite par le léflunomide était associée à des modifications dans les cellules CD4+CD5+ Treg. De plus, le métabolite actif du léflunomide, A771726, a facilité la différenciation des lymphocytes spléniques dans les CD4+CD5+ Treg, augmenté la sécrétion des cytokines anti-inflammatoires TGF-1 et ajusté l’activité des lymphocytes induits par CON A.

Document Type: Research Article

Publication date: January 1, 2010

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