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。 我们最近证实，IL-10与活体螺旋体或其他刺激物(包括螺旋体脂蛋白和声学螺旋体)一起体外刺激小鼠J774巨噬细胞，可额外增强这些细胞中细胞因子信号转导抑制物(SOCS)3的表达。SOCS3的表达与IL-10介导的抑制几种伴随而来的促炎细胞因子的表达有关。此外，活体和超声处理的螺旋体以及脂蛋白单独诱导SOCS3表达的事实表明，螺旋体也可能通过一种独立于IL-10的机制来控制炎症。我们的假设是，伯氏杆菌单独诱导巨噬细胞表达SOCS3在莱姆病病程中的炎症控制中具有重要的功能。为了解决这一假设，用声波刺激的伯氏杆菌刺激J774巨噬细胞，并在刺激后24小时收集RNA样本用于实时聚合酶链式反应阵列分析。聚合酶链式反应阵列数据显示了几种细胞因子和趋化因子的表达上调和下调。为了确定SOCS3在伯氏杆菌诱导的炎症反应中的中介作用，我们沉默了声波螺旋体刺激的巨噬细胞中的SOCS3基因。在24小时收集RNA样本，并使用小鼠寡核苷酸微阵列进行全基因组转录。抑制SOCS3基因可逆转多种细胞因子(IL-1β、IL-16、IL 15ra、TGFbeta1)、趋化因子(CCL4、CCL5、CCL6)和其他炎症介质(CD40lg、TOLLIP TNFrsf1b)的表达。细胞因子ELISA法也证实了SOCS3在蛋白水平上介导了IL-1β的产生。这项研究表明，伯氏杆菌单独诱导的SOCS3在莱姆病的炎症控制中具有重要的功能。

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 demonstrated that in vitro stimulation of mouse J774 macrophages with IL-10 together with either live spirochetes or other stimulants, including spirochetal lipoproteins and sonicated spirochetes, additively augmented the expression of the suppressor of cytokine signaling (SOCS) 3 in these cells. The expression of SOCS3 correlated with IL-10-mediated inhibition of the expression of several concomitantly elicited pro-inflammatory cytokines. Also, the fact that live and sonicated spirochetes as well as lipoproteins alone induced SOCS3 expression indicates that the spirochete may also control inflammation via a mechanism independent from that involving IL-10. Our hypothesis is that the expression of SOCS3 induced by B. burgdorferi alone in macrophages is functionally important in the control of inflammation during the course of Lyme disease. To begin to address this hypothesis J774 macrophages were stimulated with sonicated B. burgdorferi spirochetes and RNA samples were collected at 24 hr post-stimulation for real time PCR array analyses. The PCR array data revealed the up and down-regulation of the expression of several cytokines and chemokines. To determine the role of SOCS3 as a mediator of the inflammatory response induced by B. burgdorferi, we silenced the SOCS3 gene in macrophages stimulated with sonicated spirochetes. RNA samples were collected at 24 hr and subjected to genome-wide transcriptomics using mouse oligonucleotide microarrays. Silencing of the SOCS3 gene in macrophages stimulated with sonicated spirochetes resulted in a reversal of the expression of several cytokines (IL-1beta, IL-16, IL15ra, TGFbeta1) chemokines (CCL4, CCL5, CCL6) and other inflammatory mediators (CD40lg, TOLLIP TNFrsf1b). Cytokine ELISA also verified that SOCS3 mediates the production of IL-1beta at the protein level. This study demonstrates that SOCS3 induced by B. burgdorferi alone is functionally important in the control of inflammation in Lyme disease.