SUPPRESSORS OF CYTOKINE SIGNALING AND IL-10 INHIBITORY EFFECT IN LYME DISEASE

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 莱姆病是由伯氏疏螺旋体引起的一种炎症性疾病。由螺旋体引起的炎症在疾病的发病机制中起着重要作用。抗炎细胞因子IL-10通过控制各种促炎细胞因子的产生和功能，被证明是莱姆病炎症反应的关键调节因子。最近，我们观察到伯氏杆菌和IL-10共同诱导巨噬细胞表达细胞因子信号转导抑制因子(SOCS)1和3。SOCS1/SOCS3的表达与IL-10介导的对几种促炎细胞因子的抑制有关。我们推测，伯氏杆菌和IL-10诱导的巨噬细胞SOCs的表达在IL-10介导的巨噬细胞炎症控制中具有重要的功能。我们使用RNAi沉默了活的伯氏杆菌(BB)或脂化的外表面蛋白A(L-OspA)单独刺激的小鼠J774巨噬细胞中的SOCS3基因，或者两种刺激剂与IL-10一起刺激。使用SOCS3特异性的4个siRNA分子池，我们实现了SOCS3 siRNA细胞中SOCS3基因表达的抑制，与非靶向对照siRNA细胞相比，实时荧光RT-PCR检测到了高达79%的下调。刺激剂联合IL-10和SOCS3 siRNA后，培养上清液中IL-6的产量较对照siRNA明显增加。综上所述，这些数据表明SOCS3可能参与了IL-10对巨噬细胞炎症的控制。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Lyme disease, caused by the spirochete Borrelia burgdorferi, is an inflammatory disease. Inflammation, as induced by the spirochete, plays a major role in disease pathogenesis. The anti-inflammatory cytokine IL-10 has been shown to be a key regulator of inflammatory responses in Lyme disease, by controlling the production and function of various pro-inflammatory cytokines. Recently we observed that B. burgdorferi together with IL-10 additively induced the expression of the suppressor of cytokine signaling (SOCS)1 and 3 in macrophages. SOCS1/SOCS3 expression correlated with the IL-10-mediated inhibition of several pro-inflammatory cytokines. We hypothesized that the expression of SOCS induced by B. burgdorferi and IL-10 in macrophages is functionally important in the IL-10-mediated control of inflammation in macrophages. We used RNAi to silence the SOCS3 gene in mouse J774 macrophages stimulated with either live B. burgdorferi (Bb) or lipidated outer surface protein A (L-OspA) alone, or each stimulant together with IL-10. Using a SOCS3 specific pool of 4 siRNA molecules we achieved up to 79% knockdown of SOCS3 gene expression in cells transfected with the SOCS3 siRNA as compared to cells transfected with a nontargeting control siRNA, as assessed by real-time RT-PCR. IL-6 production was increased in culture supernatants with stimulants combined with IL-10 and SOCS3 siRNA as compared to those containing control siRNA. Taken together, these data suggest that SOCS3 may be involved in the IL-10 control of inflammation in macrophages.