PATHOGENESIS OF LYME NEUROBORRELIOSIS: STUDIES EX VIVO & IN VIVO

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。 莱姆氏疏螺旋体病是由伯氏螺旋体引起的，通常表现为神经认知缺陷。作为莱姆氏疏螺旋体病的一个可能机制，我们假设伯氏伯氏杆菌在中枢神经系统诱导炎症介质的产生，并伴随着神经元和/或神经胶质细胞的凋亡。为了验证我们的假设，我们建立了一个由新鲜采集的L猕猴大脑皮层切片组成的体外模型，并允许活的伯氏杆菌穿透组织。通过DNA微阵列分析，大量调控炎症以及少突胶质细胞和神经元凋亡的基因的转录受到显著干扰。实时定量聚合酶链式反应阵列分析也检测到细胞因子肿瘤坏死因子-α和趋化因子IL-8的转录倍数分别增加了7.43倍(p=0.005)和2.31倍(p=0.016)。免疫荧光染色和共聚焦显微镜观察免疫介质IL-6、IL-8、IL-1β、COX-2和CXCL13在胶质细胞中的原位表达。随之而来的是，在螺旋体刺激的组织中，发生凋亡的少突胶质细胞和神经元的比例都很大。在接受活体伯氏杆菌脑实质内立体定向接种的恒河猴中，也检测到星形胶质细胞产生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上的证明。包括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.