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: 8358082
• 负责人: MARIO TOMAS PHILIPP
• 金额: $ 3.72万
• 依托单位: TULANE UNIVERSITY OF LOUISIANA
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-05-01 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8358082
• 关键词: Anti-Inflammatory Agents; Anti-inflammatory; Apoptosis; Apoptotic; Biological Assay; Biological Response Modifiers; Borrelia burgdorferi; Brain; CCL2 gene; Cell Death; Dexamethasone; Dose; Enzyme-Linked Immunosorbent Assay; Funding; Grant; Hour; Human; IL8 gene; In Situ; In Vitro; Inflammation; Inflammation Mediators; Inflammatory Response; Interleukin-6; Life; Lyme Disease; Lyme Neuroborreliosis; Macaca mulatta; Measures; Mediating; Myelin; National Center for Research Resources; Neuroglia; Neurons; Oligodendroglia; Pathogenesis; Patients; Pharmaceutical Preparations; Primates; Principal Investigator; Research; Research Infrastructure; Resources; Role; Source; TdT-Mediated dUTP Nick End Labeling Assay; United States National Institutes of Health; chemokine; cost; cytokine

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Lyme neuroborreliosis (LNB) manifests in 10-15% of Lyme disease patients that go untreated. We hypothesized that the Lyme disease spirochete Borrelia burgdorferi (Bb) induces inflammatory mediators in glial cells that contribute to glial and neuronal damage, resulting in the pathogenesis of LNB. Here we evaluate the role of inflammation in mediating apoptosis of oligodendrocytes (OLGs), the myelin producing glial cells in the brain, as induced by Bb, by using the anti-inflammatory drug dexamethasone. We evaluated the potential of live Bb to elicit inflammatory mediators in in vitro differentiated cultures of human MO3.13 OLGs by measuring the levels of immune mediators in culture supernatants using Multiplex ELISA assays. Concomitant apoptosis was quantified in these cultures by the in situ TUNEL assay. The above phenomena were evaluated after 48 hours of stimulation with Bb in the presence and absence of various concentrations of the anti-inflammatory drug dexamethasone. Bb induced significantly enhanced levels of the cytokine IL-6 and the chemokines IL-8 and CCL2 (MCP-1) as compared to basal levels in OLGs. These Bb-induced cultures also showed significantly elevated levels of apoptosis as compared to medium controls. Bb induced inflammatory mediators and apoptosis in OLGs in a dose dependent manner. Dexamethasone significantly reduced both the levels of immune mediators as well as apoptosis as induced by Bb when used at lower concentrations (5 and 15 micro molar). Dexamethasone modulates the levels of inflammatory mediators and concomitant apoptosis induced by the Lyme disease spirochete Bb in OLGs, suggesting that the inflammatory response elicited in OLGs by Bb contributes to their apoptotic cell death. As OLGs are vital for the functioning and survival of neurons, the inflammation and subsequent apoptosis of OLGs induced by Bb could contribute to the pathogenesis of LNB.

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 莱姆病神经疏螺旋体病（LNB）表现在10-15%的莱姆病患者未经治疗。我们假设莱姆病螺旋体伯氏疏螺旋体（Bb）诱导神经胶质细胞中的炎症介质，导致神经胶质细胞和神经元损伤，导致LNB的发病机制。在这里，我们评估的作用，炎症介导的少突胶质细胞（OLG），髓鞘产生的神经胶质细胞在大脑中，诱导的Bb，通过使用抗炎药物地塞米松的凋亡。我们通过使用多重ELISA测定法测量培养物上清液中的免疫介质水平，评估了活Bb在人MO3.13 OLG的体外分化培养物中引发炎症介质的潜力。通过原位TUNEL测定在这些培养物中定量伴随的细胞凋亡。在存在和不存在各种浓度的抗炎药物地塞米松的情况下，用Bb刺激48小时后评价上述现象。与OLG中的基础水平相比，Bb诱导细胞因子IL-6和趋化因子IL-8和CCL 2（MCP-1）的水平显著增加。与培养基对照相比，这些BB诱导的培养物也显示出显著升高的细胞凋亡水平。Bb以剂量依赖性方式诱导OLG中的炎症介质和凋亡。地塞米松显着降低免疫介质的水平，以及诱导的细胞凋亡的Bb时，在较低的浓度（5和15微摩尔）。地塞米松调节炎症介质的水平和伴随的细胞凋亡诱导的莱姆病螺旋体Bb在OLG中，这表明炎症反应引起的OLG的Bb有助于他们的细胞凋亡。由于OLG对神经元的功能和存活至关重要，Bb诱导的OLG的炎症和随后的凋亡可能有助于LNB的发病机制。