PATHOGENESIS OF LYME NEUROBORRELIOSIS: STUDIES EX VIVO & IN VIVO

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。 莱姆神经疏螺旋体病是由伯氏疏螺旋体螺旋体引起的，通常表现为神经认知缺陷。作为莱姆神经疏螺旋体病的可能机制，我们假设伯氏疏螺旋体诱导中枢神经系统产生炎症介质，并伴随神经元和/或神经胶质细胞凋亡。为了检验我们的假设，我们建立了一个离体模型，该模型由从恒河猴大脑皮层新鲜采集的切片组成，并允许活伯氏疏螺旋体穿透组织。通过 DNA 微阵列分析评估，调节炎症以及少突胶质细胞和神经元凋亡的许多基因转录本受到显着干扰。实时 PCR 阵列分析还检测到细胞因子 TNF-α 的转录倍数增加了 7.43 (p = 0.005)，趋化因子 IL-8 的转录倍数增加了 2.31 (p = 0.016)。通过免疫荧光染色和共聚焦显微镜在胶质细胞中原位观察免疫介质 IL-6、IL-8、IL-1beta、COX-2 和 CXCL13。与此同时，螺旋体刺激的组织中存在显着比例的经历凋亡的少突胶质细胞和神经元。在接受了活伯氏疏螺旋体体内实质内立体定位接种的恒河猴中，也检测到星形胶质细胞产生的 IL-6 和少突胶质细胞凋亡，尽管水平较低。这些结果为我们的假设提供了概念证明。包含这些结果的手稿发表在《美国病理学杂志》上。

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. Lyme neuroborreliosis is caused by the spirochete B. burgdorferi, and is often manifest by neurocognitive deficits. As a possible mechanism for Lyme neuroborreliosis we hypothesized that B. burgdorferi induces in the central nervous system the production of inflammatory mediators, with concomitant neuronal and/or glial apoptosis. To test our hypothesis we set up an ex vivo model consisting of freshly collected slices from brain cortex of l rhesus macaques, and allowed live B. burgdorferi to penetrate the tissue. Numerous transcripts of genes that regulate inflammation as well as both oligodendrocyte and neuronal apoptosis were significantly perturbed, as assessed by DNA microarray analysis. Transcriptional fold-increases of 7.43 (p = 0.005) for the cytokine TNF-alpha and 2.31 (p = 0.016) for the chemokine IL-8 were also detected by real-time-PCR array analysis. The immune mediators IL-6, IL-8, IL-1beta, COX-2, and CXCL13 were visualized in glial cells in situ by immunofluorescence staining and confocal microscopy. Concomitantly, significant proportions of both oligodendrocytes and neurons undergoing apoptosis were present in spirochete-stimulated tissues. IL-6 production by astrocytes, and oligodendrocyte apoptosis were also detected, albeit at lower levels, in rhesus macaques that had received in vivo intraparenchymal stereotaxic inoculations of live B. burgdorferi. These results provide proof of concept for our hypothesis. A manuscript including these results was published in the American Journal of Pathology.