Chemokine receptor antagonists in inflammatory disease

描述(由申请人提供)： 在动脉粥样硬化和类风湿关节炎中观察到的炎症反应有许多相似之处。趋化因子及其受体在这两种疾病中都很重要。该应用的长期目标是研究C-C趋化因子受体2(CCR2)和CCR1/CCR5拮抗剂在啮齿类关节炎中的临床和生物学效应。此外，我们计划在CCR5/CCR2拮抗剂存在的情况下，确定关键的促炎细胞因子(TNF-α和IL-1β)和Th1促进细胞因子(IL-12和IL-18)对2D61L-12 T细胞和内皮细胞中CCR5和CCR2受体后信号转导事件的影响。在体内炎症性疾病(RA)模型中使用CCR2/CCR5拮抗剂，再加上从体外研究致炎细胞因子对CCR2/CCR5信号通路影响的机制所获得的知识，将有助于我们理解和设计更有效的在RA和动脉粥样硬化的体内研究。一些研究已经使用拮抗剂、结合蛋白和反义序列来靶向致炎细胞因子，包括肿瘤坏死因子-α(拮抗剂；依那西普)、IL-18(结合蛋白)和趋化因子受体CCR2/CCR5(拮抗剂、抗体和基因敲除)，以期减轻动脉粥样硬化和类风湿关节炎的炎症反应。为了研究IL-12和IL-18对CCR5的影响以及肿瘤坏死因子-α、IL-1β和IL-8对CCR2下游信号通路的影响，我们在细胞因子处理的细胞中免疫沉淀CC趋化因子受体，并通过Western印迹分析检测相关通路。使用CCR拮抗剂的目的是验证细胞因子是否通过这些受体发挥作用。此外，我们将在大鼠佐剂性关节炎(AIA)的体内炎症模型中使用CCR2/CCR5拮抗剂，以检测它们降低病情严重程度和延缓疾病发病的能力。为此，我们将确定炎症指标的进展情况，如关节炎指数、关节周长、足爪体积、关节计数、关节炎严重程度、细胞类型募集和骨破坏标志。实现这些目标可能为我们提供有关动脉粥样硬化和类风湿关节炎分子炎症机制的有价值的信息。

DESCRIPTION (provided by applicant): There are many similarities in inflammatory responses observed in atherosclerosis and rheumatoid arthritis (RA). Chemokines and their receptors are important in both diseases. The long-term objectives of this application are to study the clinical and biological effects of C-C chemokine receptor 2 (CCR2) and CCR1/CCR5 antagonists in rodent arthritis. Furthermore we plan to determine the effect of key proinflammatory (TNF-alpha and IL-1beta) and Th1 promoting (IL-12 and IL-18) cytokines on CCR5 and CCR2 post-receptor signaling events in the 2D61L-12 T cell line and endothelial cells in the presence of CCR5/CCR2 antagonists. Using CCR2/CCR5 antagonists in a model of inflammatory disease (RA) in vivo in addition to the knowledge acquired from studying the mechanism of proinflammatory cytokines effects on CCR2/CCR5 signaling pathways in vitro will help us understand and design more efficient in vivo studies in both RA and atherosclerosis. Several studies have used antagonists, binding proteins and antisense sequences to target proinflammatory cytokines, including TNF-alpha (antagonist; Etanercept), IL-18 (binding protein) and chemokine receptors CCR2/CCR5 (antagonists, antibodies and gene knockouts) in hope of mitigating the inflammatory reaction in atherosclerosis and RA. In order to investigate the effect of IL-12 and IL-18 on CCR5 and the effect of TNF-alpha, IL-1beta and IL-8 on CCR2 downstream signaling pathways, we will immunoprecipitate CC chemokine receptors in cytokine treated cells and detect associated pathways by Western blot analysis. The purpose of using CCR antagonists is to validate that the cytokines exert their effect through these receptors. Additionally we will use CCR2/CCR5 antagonists in an in vivo inflammatory model of RA (rat adjuvant induced arthritis (AIA)) to examine their ability to decrease the severity and delay the onset of the disease. For this purpose we will determine the progression of indicators of inflammation, such as arthritis index, joint circumference, paw volume, joint count, arthritis severity, cell type recruitment and markers of bone destruction. Achieving these goals may give us valuable information in regard to molecular inflammatory mechanisms involved in atherosclerosis and RA.