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

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 未经治疗的莱姆病患者中，有10%-15%表现为莱姆病患者患有莱姆病。我们推测莱姆病螺旋体伯氏疏螺旋体(BB)诱导胶质细胞中的炎症介质，从而导致神经胶质细胞和神经元的损伤，从而导致LNB的发病。在这里，我们通过使用抗炎药物地塞米松来评估炎症在介导脑组织少突胶质细胞(OLGs)凋亡中的作用。少突胶质细胞是脑内产生髓鞘的胶质细胞，由BB诱导。我们通过多重酶联免疫吸附试验检测培养上清液中免疫介质的水平，评价活体BB在人MO3.13 OLG体外分化培养中诱导炎性介质的能力。用原位末端标记法对这些培养物中伴随的细胞凋亡进行定量。在不同浓度抗炎药地塞米松存在和不存在的情况下，用BB刺激48h后对上述现象进行评估。BB诱导的细胞因子IL-6、趋化因子IL-8和CCL2(MCP-1)水平显著高于基础水平。与培养液对照组相比，这些BB诱导的培养物的细胞凋亡率也显著升高。BB以剂量依赖方式诱导炎性介质和细胞凋亡。地塞米松在较低浓度(5微摩尔和15微摩尔)时显著降低BB诱导的免疫介质水平和细胞凋亡。地塞米松对莱姆病螺旋体BB诱导的OLG炎症介质水平及伴随的细胞凋亡具有调节作用，提示BB诱导的炎症反应参与了OLG的细胞凋亡。由于OLGs对神经元的功能和生存至关重要，BB诱导的OLGs的炎症和随后的凋亡可能参与了LNB的发病。

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.