Immunoregulation of Herpes Encephalitis By Microglia

小胶质细胞对疱疹性脑炎的免疫调节

• 批准号: 6699071
• 负责人: James R Lokensgard
• 金额: $ 23.41万
• 依托单位: HENNEPIN HEALTHCARE RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-02-01 至 2008-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6699071
• 关键词: Herpes simplex disease; T lymphocyte; athymic mouse; benzodiazepine receptor; cell migration; chemokine; encephalitis virus; herpes simplex virus 1; immunopathology; infectious encephalitis; inflammation; laboratory mouse; leukocyte depletion therapy; microglia; neuroimmunomodulation; neuropathology; neurotoxicology; neutralizing antibody; pathologic process; tissue /cell culture

DESCRIPTION (provided by applicant): Herpes simplex virus 1 (HSV) is an important opportunistic pathogen in HIV-1-infected patients as well as the cause of a devastating CNS infection in normal hosts. Although immune responses to HSV have been the subject of intense investigation, the role of immune-mediated pathology in HSV-related brain damage is unknown. In this proposal, the central hypothesis to be tested is that chemokines produced by microglial cells in response to HSV infection initiate a cascade of neuroimmune responses that result in the serious brain damage seen during herpes encephalitis. To test this hypothesis, chemokine production in the brains of HSV-infected mice will be compared to that in cultures of highly purified murine glial and neuronal cells, and in murine organotypic brain slices infected with HSV. This approach will allow us to differentiate microglial cell chemokine production from that of cells of the somatic immune system. Additionally, the use of organotypic brain slice cultures will enable us to specifically deplete microglial cells for "loss-of-function" experiments. We will then investigate the neurotoxic effects of microglial cell-produced immune mediators on cultured murine neurons. Microglia-driven leukocyte trafficking into the brain will be investigated by determining if neutralizing antibodies to chemokines inhibit T-cell infiltration. The neuropathogenic role of T-cell infiltration will be studied by determining if depletion of T-cells in vivo will delay encephalitis and whether adoptive transfer of HSV-specific lymphocytes restores the encephalitis phenotype. Comparing neuropathology in brain slice cultures with and without the transfer of HSV-specific CD4 + and CD8 +lymphocytes, will allow us to distinguish between injury generated by viral infection and brain damage provoked by immunopathogenic mechanisms. Downregulation of microglial cell chemokine production through peripheral benzodiazepine (BDZ) receptor-mediated cellular deactivation will then be examined. We will determine if deactivation of microglia with BDZs suppresses the production of neurotoxic factors. Finally, we will study the effects of BDZ ligands on chemokine production, T-cell trafficking, and the development of encephalitis in vivo. These in vivo, in vitro, and ex vivo models will provide us with the ability to investigate neuropathogenesis, neuroinflammation, neurotoxicity, and neuroimmune-mediated pathology occurring during herpes encephalitis. Knowledge gained from these studies will increase our understanding of the role of microglial cells and chemokine networks that regulate brain inflammation during herpes encephalitis with the ultimate goal of finding new therapy for this serious brain infection.

描述（由申请人提供）：单纯疱疹病毒1 （HSV）是hiv -1感染患者中一种重要的机会性病原体，也是正常宿主中破坏性中枢神经系统感染的原因。尽管对单纯疱疹病毒的免疫反应一直是深入研究的主题，但免疫介导的病理在单纯疱疹病毒相关脑损伤中的作用尚不清楚。在这一提议中，需要验证的中心假设是，小胶质细胞对HSV感染的反应产生的趋化因子启动了一系列神经免疫反应，导致疱疹性脑炎期间出现的严重脑损伤。为了验证这一假设，将单纯疱疹病毒感染小鼠的大脑中趋化因子的产生与高度纯化的小鼠胶质细胞和神经元细胞的培养以及感染单纯疱疹病毒的小鼠器官型脑切片中的趋化因子的产生进行比较。这种方法将使我们能够将小胶质细胞趋化因子的产生与体细胞免疫系统的细胞区分开来。此外，使用器官型脑切片培养将使我们能够专门消耗小胶质细胞进行“功能丧失”实验。然后，我们将研究小胶质细胞产生的免疫介质对培养的小鼠神经元的神经毒性作用。通过确定趋化因子的中和抗体是否抑制t细胞浸润，将研究小胶质细胞驱动的白细胞运输到大脑。通过确定体内t细胞的消耗是否会延迟脑炎，以及单纯疱疹病毒特异性淋巴细胞的过继转移是否会恢复脑炎表型，将研究t细胞浸润的神经致病作用。比较有和没有转移hsv特异性CD4 +和CD8 +淋巴细胞的脑切片培养的神经病理学，将使我们能够区分由病毒感染引起的损伤和由免疫致病机制引起的脑损伤。通过外周苯二氮卓（BDZ）受体介导的细胞失活，下调小胶质细胞趋化因子的产生将被检查。我们将确定BDZs的小胶质细胞失活是否会抑制神经毒性因子的产生。最后，我们将研究BDZ配体在体内趋化因子产生、t细胞运输和脑炎发展中的作用。这些体内、体外和离体模型将为我们提供研究疱疹脑炎期间发生的神经发病机制、神经炎症、神经毒性和神经免疫介导的病理的能力。从这些研究中获得的知识将增加我们对疱疹性脑炎期间调节脑炎症的小胶质细胞和趋化因子网络的作用的理解，最终目标是为这种严重的脑感染找到新的治疗方法。