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。 我们最近发现，小鼠J774巨噬细胞与IL-10和伯氏疏螺旋体孵育后，其SOCS3的表达水平增加。这与IL-10抑制几种促炎细胞因子的表达有关。我们推测，伯氏杆菌和IL-10诱导的巨噬细胞中SOCS3的表达在莱姆病的炎症控制中具有重要的功能，SOCS3可能是巨噬细胞中IL-10抗炎活性的关键介质。为了解决这一假设，我们使用SOCS3小干扰RNA沉默了J774细胞中SOCS3基因的表达，以确定沉默对伯氏杆菌诱导的促炎介质IL-10抑制的影响。用IL-10和伯氏杆菌共刺激转基因细胞；收集RNA样本，用小鼠寡核苷酸芯片进行全基因组转录。这些研究表明，在IL-10和伯氏杆菌刺激的细胞中，SOCS3基因上调(~40倍)。在SOCS3存在的情况下，IL-10可以下调伯氏杆菌刺激的巨噬细胞中的几种促炎介质。然而，这些巨噬细胞中SOCS3基因的沉默显著逆转了IL-10对许多炎症介质的下调作用，这些炎症介质包括细胞因子/受体、趋化因子、干扰素诱导基因、凋亡基因和肿瘤坏死因子配体/受体基因。本研究表明，SOCS3部分介导了伯氏杆菌刺激的巨噬细胞中IL-10的抗炎作用。

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. We recently showed that mouse J774 macrophages incubated with IL-10 and Borrelia burgdorferi spirochetes augmented their SOCS3 expression levels. This correlated with the IL-10 inhibition of expression of several pro-inflammatory cytokines. We hypothesized that SOCS3 expression induced by B. burgdorferi and IL-10 in macrophages is functionally important in the control of inflammation in Lyme disease, with SOCS3 potentially acting as a key mediator of the IL-10 anti-inflammatory activity in macrophages. To begin to address this hypothesis, we silenced socs3 gene expression in J774 cells using SOCS3 small interfering RNA to determine the effect of silencing on IL-10 inhibition of pro-inflammatory mediators induced by B. burgdorferi. Transfected cells were co-stimulated with IL-10 and B. burgdorferi spirochetes; RNA samples were collected and subjected to genome-wide transcriptomics using mouse oligonucleotide microarrays. These studies revealed that the socs3 gene is up-regulated (~40-fold) in cells stimulated with IL-10 and B. burgdorferi. In the presence of SOCS3, IL-10 could down-modulate several pro-inflammatory mediators in B. burgdorferi-stimulated macrophages. However, silencing of the socs3 gene in these macrophages resulted in a marked reversal of the down-modulatory effect exerted by IL-10 on many of these inflammatory mediators including cytokines/receptors, chemokines, IFN-inducible, apoptotic and TNF ligand/receptor genes. This study demonstrates that SOCS3 in part mediates the IL-10 anti-inflammatory effect in macrophages stimulated with B. burgdorferi spirochetes.