The effect of a T cell-specific NF-kappa B inhibitor on in vitro cytokine production and collagen-induced arthritis.

Abstract

NF-kappa B plays a key role in the production of cytokines in inflammatory diseases. The effects of a novel T cell-specific NF-kappa B inhibitor, SP100030, were evaluated in cultured Jurkat cells and in murine collagen-induced arthritis (CIA). Chemical libraries were screened for NF-kappa B-inhibitory activity. SP100030, a compound identified in this process, inhibited NF-kappa B activation in PMA/PHA-activated Jurkat cells by EMSA at a concentration of 1 microM. Jurkat cells and the monocytic cell line THP-1 were transfected with an NF-kappa B promotor/luciferase construct and activated. SP100030 inhibited luciferase production in the Jurkat cells (IC50 = 30 nM). ELISA and RT-PCR confirmed that IL-2, IL-8, and TNF-alpha production by activated Jurkat and other T cell lines were inhibited by SP100030. However, cytokine expression was not blocked by the compound in THP-1 cells, fibroblasts, endothelial cells, or epithelial cells. Subsequently, DBA/1J mice were immunized with type II collagen. Treatment with SP100030 (10 mg/kg/day i.p. beginning on day 21) significantly decreased arthritis severity from onset of clinical signs to the end of the study on day 34 (arthritis score, 5.6 +/- 1.7 for SP100030 and 9.8 +/- 1.5 for control; p < 0.001). Histologic evaluation demonstrated a trend toward improvement in SP100030-treated animals. EMSA of arthritic mouse ankles in CIA showed that synovial NF-kappa B binding was suppressed in the SP100030-treated mice. SP100030 inhibits NF-kappa B activation in T cells, resulting in reduced NF-kappa B-regulated gene expression and decreased CIA. Its selectivity for T cells could provide potent immunosuppression with less toxicity than other NF-kappa B inhibitors.

• Authors: D M Gerlag, L Ransone, P P Tak, Z Han, M Palanki, M S Barbosa, D Boyle, A M Manning, G S Firestein
• Journal: Journal of immunology (Baltimore, Md. : 1950) (J Immunol) Vol. 165 Issue 3 Pg. 1652-8 (Aug 01 2000) ISSN: 0022-1767 [Print] United States
• PMID: 10903776 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• Cytokines
• Epitopes, T-Lymphocyte
• Immunosuppressive Agents
• N-(3,5-bis(trifluoromethyl)phenyl)-2-chloro-4-(trifluoromethyl)pyrimidine-5-carboxamide
• NF-kappa B
• Organic Chemicals
• Collagen
• DNA

Topics

• Animals
• Arthritis, Experimental (immunology, metabolism, prevention & control)
• Collagen (immunology)
• Cytokines (biosynthesis, genetics)
• DNA (drug effects, metabolism)
• Dose-Response Relationship, Immunologic
• Epitopes, T-Lymphocyte (immunology)
• Gene Expression Regulation (drug effects, immunology)
• Humans
• Immunosuppressive Agents (pharmacology, therapeutic use)
• Jurkat Cells (drug effects, immunology, metabolism)
• Male
• Mice
• Mice, Inbred DBA
• NF-kappa B (antagonists & inhibitors, metabolism, physiology)
• Organic Chemicals
• Protein Binding (drug effects, immunology)
• Synovial Membrane (drug effects, metabolism)
• T-Lymphocytes (immunology, metabolism)
• Tumor Cells, Cultured