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来源获得了主要资金， 因此可在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 我们已经报道了体外实验，表明在对活的伯氏疏螺旋体（Bb）及其纯化的外表面脂蛋白A（L-OspA）的应答中，IL-10抑制小鼠J774巨噬细胞中的原型IL-6、IL-12、TNF-α和IL-1 β炎症应答。我们还表明，与IL-10一起孵育的小鼠巨噬细胞与活的Bb或L-OspA相加增强了这些细胞中细胞因子信号传导抑制因子（SOCS）3的表达。在该研究中，我们首先使用趋化因子/细胞因子多重方法来确定响应于活Bb和L-OspA的巨噬细胞中的炎症介质的IL-10介导的抑制程度。然后使用RNA干扰来沉默巨噬细胞中的socs 3基因表达，以确定沉默对响应于活Bb和L-OspA的炎性介质的IL-10抑制的影响。多重分析显示，IL-10显著（P0.05）下调了用活Bb或L-OspA刺激的巨噬细胞中几种细胞因子（TNF-α、IL-12、IL-6、IL-1 α、IL-1 β、IL-9）和趋化因子（G-CSF、CXCL 1、CXCL 5、CXCL 10和CCL 5）的产生。在所有情况下，活Bb诱导的细胞因子和趋化因子水平高于L-OspA。在巨噬细胞中由两种刺激物诱导的所有介质以类似的方式被IL-10下调，并且与这些细胞中增强的SOCS 3表达相关。沉默巨噬细胞中的SOCS 3表达消除了响应于活Bb和L-OspA的IL-10抗炎作用。该研究表明，SOCS 3部分介导用活B刺激的巨噬细胞中的IL-10抗炎作用。伯氏螺旋体和脂蛋白。在SOCS 3沉默之前和之后的巨噬细胞全基因组转录组学数据的比较将最终鉴定响应于B的SOCS 3介导的抗IL-10抑制作用的程度。burgdorferi。

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 have reported on experiments in vitro showing that in response to live Borrelia burgdorferi (Bb) and its purified outer surface lipoprotein A (L-OspA), IL-10 dampens the prototypic IL-6, IL-12, TNF-alpha and IL-1beta inflammatory responses in mouse J774 macrophages. We also showed that mouse macrophages incubated with IL-10 together with either live Bb or L-OspA additively augmented the expression of the suppressor of cytokine signaling (SOCS) 3 in these cells. In this study we first used a chemokine/cytokine multiplex approach to determine the extent of the IL-10-mediated inhibition of inflammatory mediators in macrophages in response to live Bb and L- OspA. RNA interference was then used to silence socs3 gene expression in macrophages to determine the effect of silencing on IL-10 inhibition of inflammatory mediators in response to live Bb and L-OspA. Multiplex analysis revealed that IL-10 significantly (P0.05) down-regulated the production of several cytokines (TNF-alpha, IL-12, IL-6, IL-1alpha, IL-1beta, IL-9) and chemokines (G-CSF, CXCL1, CXCL5, CXCL10 and CCL5) in macrophages stimulated with live Bb or L-OspA. In all instances live Bb induced higher levels of cytokines and chemokines than did L-OspA. All mediators induced by both stimulants in macrophages were down-modulated by IL-10 in a similar fashion and correlated with enhanced SOCS3 expression in these cells. Silencing of SOCS3 expression in macrophages abrogated the IL-10-anti-inflammatory actions against IL-6 production in response to live Bb and L-OspA. This study demonstrates that SOCS3 in part mediates the IL-10 anti-inflammatory effect in macrophages stimulated with live B. burgdorferi spirochetes and lipoproteins. Comparisons of macrophage genome-wide transcriptomics data before and after the silencing of SOCS3 will ultimately identify the extent of the SOCS3-mediated anti-IL-10 inhibitory effect in response to B. burgdorferi.