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

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。 背景：许多神经退行性疾病，包括阿尔茨海默病、脑缺血和艾滋病-痴呆复合体，被认为是中枢神经系统(CNS)炎症的结果。莱姆氏疏螺旋体病同样被认为是中枢神经系统炎症的结果。伯氏疏螺旋体可引起莱姆病，并可诱导中枢神经系统神经胶质细胞产生炎症介质。我们假设，神经细胞，凭借接近，可能会在这种环境中受到损害，最终导致神经认知症状，在神经疏螺旋体病中看到。方法：为了确定伯氏疏螺旋体感染后神经细胞和神经胶质细胞的反应，我们设计了BB与原代恒河猴皮质细胞单独或与SH-SY5Y神经母细胞瘤细胞共同培养的体外模型。结果：夹心ELISA法观察到BB刺激的大脑皮层细胞表达和释放炎性细胞因子/趋化因子IL-6和IL-8，并诱导产生肿瘤坏死因子-β。当同样的BB刺激原代胶质细胞与神经细胞系SH-SY5Y或HCN-1结合时，细胞凋亡率持续增加。用疏螺旋体独立刺激的神经细胞系表达相对微不足道的炎性细胞因子，在这种环境下它们对凋亡具有抵抗力。相反，当神经元与混合胶质细胞或纯小胶质细胞共同培养时，通过TUNEL试验评估，神经元因凋亡而死亡：这些发现表明：1)伯氏杆菌不会损害神经元本身；2)小胶质细胞，无论是单独的还是与其他类型的胶质细胞联合培养，都能够在存在伯氏杆菌的情况下对神经元产生有害影响。

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. Background: Many neurodegenerative disorders including Alzheimer's disease, cerebral ischemia and the AIDS-dementia complex are thought to result from inflammation in the central nervous system (CNS). Lyme neuroborreliosis is likewise considered to be a consequence of inflammation elicited in the CNS. The bacterial spirochete Borrelia burgdorferi (Bb) causes Lyme disease and is known to induce the production of inflammatory mediators in glial cells of the CNS. We hypothesized that neuronal cells, by virtue of proximity, may become impaired in this environment, eventually leading to the neurocognitive symptoms seen in neuroborreliosis. Methods: In order to determine the neuronal and glial cell responses to Borrelia burgdorferi infection we have designed an in-vitro model where Bb is co-cultivated with cells from primary rhesus cortex either alone or in combination with SH-SY5Y neuroblastoma cells. Results: Using sandwich ELISA we observed robust expression and release of the inflammatory cytokines / chemokines IL-6 and 8, and induction of TNF-¿, in the cortex cells stimulated with Bb. When these same stimulations of primary glial cells with Bb are combined with the neuronal cell lines SH-SY5Y or HCN-1, increases in cellular apoptosis consistently occur. The neuronal cell lines independently stimulated with Borrelia express comparatively negligible amounts of inflammatory cytokine and they are resistant to apoptosis in this environment. In contrast, when neurons are co-cultivated either with mixed glial cells or with pure microglia, the neurons die by apoptosis, as assessed via the TUNEL assay: These are findings to suggest that 1) B. burgdorferi does not damage neurons per se; 2) microglia, either alone or in concert with other glial cell types, are able to generate a deleterious effect on neurons in the presence of B. burgdorferi.