Gerlag D M, Ransone L, Tak P P, Han Z, Palanki M, Barbosa M S, Boyle D, Manning A M, Firestein G S
Division of Rheumatology, Allergy and Immunology, University of California, San Diego School of Medicine, La Jolla, CA 92093, USA.
J Immunol. 2000 Aug 1;165(3):1652-8. doi: 10.4049/jimmunol.165.3.1652.
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.
核因子-κB在炎症性疾病的细胞因子产生中起关键作用。在培养的Jurkat细胞和小鼠胶原诱导性关节炎（CIA）中评估了一种新型T细胞特异性核因子-κB抑制剂SP100030的作用。对化学文库进行核因子-κB抑制活性筛选。在此过程中鉴定出的化合物SP100030，在浓度为1μM时通过电泳迁移率变动分析（EMSA）抑制佛波酯/植物血凝素（PMA/PHA）激活的Jurkat细胞中的核因子-κB活化。用核因子-κB启动子/荧光素酶构建体转染Jurkat细胞和单核细胞系THP-1并进行激活。SP100030抑制Jurkat细胞中的荧光素酶产生（半数抑制浓度[IC50]=30 nM）。酶联免疫吸附测定（ELISA）和逆转录-聚合酶链反应（RT-PCR）证实，SP100030抑制激活的Jurkat细胞和其他T细胞系产生白细胞介素-2（IL-2）、白细胞介素-8（IL-8）和肿瘤坏死因子-α（TNF-α）。然而，该化合物在THP-1细胞、成纤维细胞、内皮细胞或上皮细胞中未阻断细胞因子表达。随后，用II型胶原免疫DBA/1J小鼠。用SP100030治疗（从第21天开始腹腔注射，剂量为10 mg/kg/天）从临床症状出现到研究结束的第34天显著降低了关节炎严重程度（关节炎评分，SP100030组为5.6±1.7，对照组为9.8±1.5；p<0.001）。组织学评估显示SP100030治疗的动物有改善趋势。CIA中关节炎小鼠踝关节的EMSA显示，SP100030治疗的小鼠滑膜核因子-κB结合受到抑制。SP100030抑制T细胞中的核因子-κB活化，导致核因子-κB调节的基因表达减少并减轻CIA。其对T细胞的选择性可提供比其他核因子-κB抑制剂更强的免疫抑制作用且毒性更小。
