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的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得主要资金， 因此可以在其他CRISP条目中表示。所列机构为 中心，不一定是研究者的机构。 我们最近发现，小鼠J774巨噬细胞与IL-10和伯氏疏螺旋体螺旋体孵育增加了它们的SOCS 3表达水平。这与IL-10抑制几种促炎细胞因子的表达有关。我们推测B.巨噬细胞中的伯氏螺旋体和IL-10在莱姆病的炎症控制中是功能上重要的，其中SOCS 3潜在地充当巨噬细胞中IL-10抗炎活性的关键介体。为了开始解决这一假设，我们使用SOCS 3小干扰RNA沉默J774细胞中的SOCS 3基因表达，以确定沉默对由B诱导的促炎介质的IL-10抑制的影响。burgdorferi。用IL-10和B共刺激转染的细胞。收集RNA样品并使用小鼠寡核苷酸微阵列进行全基因组转录组学。这些研究表明，socs 3基因在用IL-10和B刺激的细胞中上调（~40倍）。burgdorferi。在SOCS 3存在的情况下，IL-10可下调B中的几种促炎介质。burgdorferi刺激的巨噬细胞。然而，这些巨噬细胞中socs 3基因的沉默导致IL-10对许多这些炎性介质（包括细胞因子/受体、趋化因子、IFN诱导型、凋亡和TNF配体/受体基因）施加的下调作用的显著逆转。该研究表明，SOCS 3部分介导了用B刺激的巨噬细胞中的IL-10抗炎作用。伯氏螺旋体

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.