REGULATION OF SOCS EXPRESSION IN MACROPHAGES IN RESPONSE TO BORRELIA AND IL-10

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。 炎症是由伯氏疏螺旋体在多个器官中引起的，在疾病的发病机制中起主要作用，但其调节机制尚不清楚。我们最近发现，小鼠J774巨噬细胞与IL-10孵育后，加入伯氏疏螺旋体(冻融、活或超声)或脂化外表面蛋白A(L-OspA)可增强其细胞因子信号转导抑制物(SOCS)1和SOCS3的表达，其中以SOCS3的表达更为丰富。这与IL-10介导的对几种促炎细胞因子表达的抑制在时间上相关。我们假设，与单独使用刺激剂刺激相比，通过共刺激增强SOCs的表达要么导致共同的信号通路的加强，要么导致不同信号通路的汇聚以获得最佳的SOCS表达。为了解决这一假说，我们使用小鼠基因芯片来研究巨噬细胞中SOCS诱导的介体。用IL-10和伯氏杆菌共同刺激J774巨噬细胞，分别在刺激后2、4和24小时采集RNA样本，进行基因芯片分析。根据3的折叠变化截断标准，我们观察到500多个在巨噬细胞中上调的基因，其中最主要的是SOCS3基因(5-6的log2倍变化)。基因表达数据揭示了在所有时间点上的独特和重叠的基因集。进一步的基因本体论分析显示，在改变了微阵列图谱的基因中，来自TLR、JAK/STAT和p38MAPK信号通路的编码介体的基因过度表达。已知这些途径参与了SOCS的诱导，因此提示，在IL-10和声学螺旋体共同刺激的巨噬细胞中，可能有不止一条途径参与了SOCS的诱导。

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. Inflammation, as induced by the spirochete Borrelia burgdorferi in multiple organs, plays a major role in disease pathogenesis, but its regulation is not understood We recently showed that mouse J774 macrophages incubated with IL-10 and added B. burgdorferi spirochetes (freeze-thawed, live or sonicated) or lipidated outer surface protein A (L-OspA) augmented their suppressor of cytokine signaling (SOCS)1 and SOCS3 expression, with SOCS3 being the more abundant. This temporally correlated with the IL-10-mediated inhibition of expression of several pro-inflammatory cytokines. We hypothesized that augmentation of SOCS expression by co-stimulation compared to stimulation with individual stimulants led either to reinforcement of a common signaling pathway or convergence of different signaling pathways for optimal SOCS expression. To begin to address this hypothesis, we used murine cDNA microarray to investigate mediators of SOCS induction in macrophages. J774 macrophages were co-stimulated with IL-10 and sonicated B. burgdorferi spirochetes and RNA samples were collected at 2, 4 and 24 hr post-stimulation for microarray analyses. Based on a fold-change cut-off criterion of 3, we observed over 500 up-regulated genes in macrophages, chief amongst which was the SOCS3 gene (log2 fold change of 5-6). The gene expression data revealed both unique and overlapping sets of genes at all time points. Further Gene Ontology analysis revealed amongst genes with altered microarray profiles, the over-representation of genes encoding mediators from TLR, JAK/STAT and p38 MAPK signaling pathways. These pathways are known to be involved in the induction of SOCS, thus suggesting that more than one pathway maybe involved in SOCS induction in macrophages co-stimulated with IL-10 and sonicated spirochetes.