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

巨噬细胞中 SOCS 表达对疏螺旋体和 IL-10 的调节

• 批准号: 7716275
• 负责人: VIDA A DENNIS
• 金额: $ 2.4万
• 依托单位: TULANE UNIVERSITY OF LOUISIANA
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-07-21 至 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7716275
• 关键词: Address; Borrelia; Borrelia burgdorferi; CCL4 gene; Cells; Computer Retrieval of Information on Scientific Projects Database; Cytokine Inducible SH2-Containing Protein; Data; Down-Regulation; Enzyme-Linked Immunosorbent Assay; Funding; Genes; Genome; Grant; In Vitro; Inflammation; Inflammatory; Inflammatory Response; Institution; Interleukin-10; Interleukin-16; Life; Lipoproteins; Lyme Disease; Mediating; Mediator of activation protein; Mus; Oligonucleotide Microarrays; Order Spirochaetales; Polymerase Chain Reaction; Production; Proteins; RANTES; RNA; Regulation; Research; Research Personnel; Resources; Role; Sampling; Source; TOLLIP gene; Time; United States National Institutes of Health; chemokine; cytokine; human TOLLIP protein; macrophage; response; transcriptomics

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.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 我们最近证明，用 IL-10 与活螺旋体或其他兴奋剂（包括螺旋体脂蛋白和超声处理螺旋体）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传导抑制因子 (SOCS) 3 的表达。 SOCS3 的表达与 IL-10 介导的几种同时引发的促炎细胞因子表达的抑制相关。此外，活的和超声处理的螺旋体以及单独的脂蛋白诱导SOCS3表达的事实表明，螺旋体还可以通过独立于IL-10的机制来控制炎症。我们的假设是，巨噬细胞中伯氏疏螺旋体单独诱导的 SOCS3 表达对于控制莱姆病过程中的炎症具有​​重要的功能。 为了开始解决这一假设，用超声处理的伯氏疏螺旋体刺激 J774 巨噬细胞，并在刺激后 24 小时收集 RNA 样本用于实时 PCR 阵列分析。 PCR阵列数据揭示了几种细胞因子和趋化因子表达的上调和下调。为了确定 SOCS3 作为伯氏疏螺旋体诱导的炎症反应介质的作用，我们沉默了用超声螺旋体刺激的巨噬细胞中的 SOCS3 基因。 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组学分析。 用超声处理的螺旋体刺激巨噬细胞中 SOCS3 基因的沉默，导致几种细胞因子（IL-1beta、IL-16、IL15ra、TGFbeta1）、趋化因子（CCL4、CCL5、CCL6）和其他炎症介质（CD40lg、TOLLIP TNFrsf1b）的表达逆转。细胞因子 ELISA 还验证了 SOCS3 在蛋白质水平介导 IL-1beta 的产生。这项研究表明，由伯氏疏螺旋体单独诱导的 SOCS3 在控制莱姆病炎症方面具有重要的功能。