PATHOGENESIS OF LYME NEUROBORRELIOSIS IN THE RHESUS MONKEY: STUDIES IN VITRO

恒河猴莱姆病神经疏螺旋体病的发病机制：体外研究

• 批准号: 8172979
• 负责人: MARIO TOMAS PHILIPP
• 金额: $ 6.18万
• 依托单位: TULANE UNIVERSITY OF LOUISIANA
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8172979
• 关键词: Apoptosis; Borrelia burgdorferi; Bystander Effect; CCL2 gene; CCL3 gene; CCL4 gene; Cell Culture Techniques; Cell Survival; Cells; Coculture Techniques; Computer Retrieval of Information on Scientific Projects Database; Confocal Microscopy; Distress; Exhibits; Funding; Genes; Grant; IL8 gene; Image; Impairment; In Vitro; Inflammation Mediators; Inflammatory; Inflammatory Response; Institution; Interleukin-6; Lyme Neuroborreliosis; Macaca mulatta; Mediating; Microarray Analysis; Microglia; Myeloid Cells; Neuroblastoma; Neurocognitive; Neurons; Pathogenesis; Pathway interactions; Preparation; Production; RANTES; Research; Research Personnel; Resistance; Resources; Signal Transduction; Source; Staining method; Stains; Symptoms; TP53 gene; Toll-Like Receptor 2; Transcript; United States National Institutes of Health; Up-Regulation; cell type; cytokine; neuron apoptosis; receptor; research study

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. Borrelia burgdorferi is known to induce the production of inflammatory mediators in a variety of cells. We hypothesized that in this inflammatory milieu neuronal apoptosis may occur, leading to neuroborreliosis. In experiments where B. burgdorferi was co-cultured in vitro with primary microglia, we observed robust release of IL-6 and IL-8, CCL2 (MCP-1), CCL3 (MIP-1¿), CCL4 (MIP-1¿) and CCL5 (RANTES), but we detected no induction of microglial apoptosis. In contrast, SH-SY5Y (SY) neuroblastoma cells co-cultured with B. burgdorferi expressed negligible amounts of inflammatory mediators but also remained resistant to apoptosis. When SY cells were co-cultured with microglia and B. burgdorferi, neuronal apoptosis consistently occurred. Confocal microscopy imaging of these cell cultures stained for apoptosis and with cell type-specific markers confirmed that it was predominantly the SY cells that were dying. Microarray analysis further demonstrated an intense microglia mediated inflammatory response to B. burgdorferi including up-regulation in gene transcripts for proinflammatory cytokines, Toll-like receptor 2 (TLR-2) and NF¿¿. Interestingly, the pathway that exhibited the most profound changes in regard to inflammatory signaling was triggering receptor expressed on myeloid cells-1 (TREM1). Significant transcript alterations in essential p53 pathway genes also occurred in neuronal cells cultured in the presence of microglia and B. burgdorferi, which indicated a shift from cell survival to preparation for apoptosis when compared to SY cells cultured in the presence of B. burgdorferi alone. These findings indicate that B. burgdorferi is not directly toxic to SY cells; rather, these cells become distressed and die in the inflammatory surroundings generated by microglia through a bystander effect. Such a neuronal impairment may eventually contribute to the neurocognitive symptoms seen in neuroborreliosis.

该副本是使用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这是调查员的机构。 众所周知，Borrelia burgdorferi会诱导各种细胞中炎症介质的产生。我们假设在这种炎症性环境中可能会发生神经凋亡，从而导致神经红细胞增多。在B. burgdorferi与原发性小胶质细胞共培养的实验中，我们观察到IL-6和IL-8和IL-8，CCL2（MCP-1），CCL3（MIP-1¿），CCL4（MIP-1¿）和CCL5（rantes）的稳健释放，但我们检测到了微胶质细胞的诱导。相比之下，与B. burgdorferi共同培养的SH-SY5Y（SY）神经母细胞瘤细胞表达了可忽略不计的炎症介质数量，但仍然对凋亡具有抗性。当SY细胞与小胶质细胞和爆发芽孢杆菌共培养时，神经元凋亡始终发生。这些细胞培养物和细胞类型特异性标记物染色的细胞培养物的共聚焦显微镜成像证实，主要是SY细胞死亡的主要是。微阵列分析进一步表明，对爆发芽孢杆菌的炎症反应有强烈的炎症反应，包括促炎细胞因子的基因转录物上的上调，Toll样受体2（TLR-2）和NFS.有趣的途径，这种途径在炎症信号传播中触发了最深刻的途径。基本p53途径基因的重大转录改变也发生在存在于小胶质细胞和B. burgdorferi的神经元细胞中，这表明与单独存在B. burgdorferi存在的Sy细胞相比，与Sy细胞相比，从细胞存活到准备凋亡的准备。研究结果表明，伯格多菲尔河芽孢杆菌对SY细胞没有直接毒性。相反，这些细胞在小胶质细胞通过旁观者效应产生的炎症环境中变得困扰并死亡。这种神经元障碍最终可能有助于神经性化的神经认知符号。