Impact of illicit drugs, HIV, and ART on neuroinflammation and BBB disruption

我们建议在人类免疫缺陷病毒存在的情况下研究HIV介导的炎症的分子机制 抗逆转录病毒疗法(ART)和滥用药物。物质滥用往往会加剧神经信息传递。我们 将使用甲基苯丙胺(冰毒)，因为它已被证明会增强艾滋病毒中枢神经系统疾病，而多巴胺作为一种 药物滥用的模型，因为所有滥用的物质都会增加中枢神经系统细胞外的多巴胺。人类免疫缺陷病毒感染 中枢神经系统会导致慢性炎症，导致超过50%的感染者出现认知障碍。这 ART不能减轻炎症和随后的中枢神经系统损害。因此，引发炎症是一个关键 艾滋病毒疾病的进程和限制免疫激活和神经炎症的治疗必须发展到 提高艾滋病毒感染者的生活质量。HIV在外周感染后不久进入中枢神经系统并 尽管有抗逆转录病毒药物，但它仍然存在于受感染的细胞内。HIV进入大脑的媒介，至少部分是通过感染 单核细胞跨血脑屏障(BBB)迁移。一个成熟的单核细胞亚群，表达 CD14和CD16是人类免疫缺陷病毒中枢神经系统疾病的关键介质，外周血中CD14和CD16数量增加 艾滋病病毒感染者。这些单核细胞能够高效地感染艾滋病毒，并为跨越血脑屏障做好了准备。 在中枢神经系统内，感染艾滋病毒的单核细胞可能分化为巨噬细胞，可持续数年。这 导致中枢神经系统细胞感染/激活，导致慢性神经炎和病毒的产生 和/或病毒蛋白、细胞因子和趋化因子。趋化因子，特别是CCL2，增加了 外周血单核细胞，持续的神经炎症。因此，慢性炎症被认为是 在很大比例的感染者中，神经元损伤的机制尚不清楚。我们会 描述冰毒、HIV、TAT和ART对单核细胞进入中枢神经系统和随后的影响的特征 神经炎。我们将测试潜在的治疗方法来限制炎症并指导ART的疗效。我们 冰毒与HIV感染合并使用可加重神经炎症并损害血脑屏障的假设 完整性，增加未感染和感染艾滋病毒的单核细胞向大脑的迁移，导致 认知障碍。根据我们的新的初步数据，替诺福韦增加了细胞上的连接蛋白 BBB，我们还假设某些抗逆转录病毒疗法，特别是在冰毒滥用者中，可能会产生负面影响 通过与冰毒协同作用增加血脑屏障通透性和神经炎症的认知功能。我们会 描述冰毒、HIV、TAT和ART对细胞因子、趋化因子和黏附分子的影响 由CD14+CD16+单核细胞阐述和表达，促进进入中枢神经系统，并在导致 血脑屏障通透性和CD14+CD16+单核细胞移行致炎作用及其影响 艺术。我们将使用小鼠模型来评估冰毒、HIV及其蛋白和ART对血脑屏障的体内影响。 炎症细胞进入大脑的渗透性和迁移性。我们还将检查冰毒或 多巴胺、HIV TAT和ART对中枢神经系统细胞释放细胞因子的影响，这些细胞因子介导了炎症。

We propose is to examine molecular mechanisms of HIV-mediated inflammation in the presence of antiretroviral therapy (ART) and drugs of abuse. Substance abuse often exacerbates neuroinlammation. We will use methamphetamine (meth) as it has been shown to enhance HIV CNS disease, and dopamine as a model for drug abuse as all substances of abuse increase CNS extracellular dopamine. HIV infection of the CNS results in chronic inflammation that leads to cognitive deficits in more than 50% of infected people. This inflammation and subsequent CNS damage is not mitigated with ART. Thus, triggering inflammation is a key process in HIV disease and therapies to limit immune activation and neuroinflammation must be developed to improve the quality of life of HIV infected people. HIV enters the CNS soon after peripheral infection and despite ART, persists within infected cells. HIV entry into the brain is mediated, at least in part, by infected monocyte transmigration across the blood brain barrier (BBB). A mature subset of monocytes that expresses CD14 and CD16 is a key mediator of HIV CNS disease and is increased in number in the peripheral blood of HIV infected people. These monocytes are productively infected with HIV and are primed to cross the BBB. Within the CNS, HIV infected monocytes may differentiate into macrophages that can persist for years. This leads to infection/activation of CNS cells, resulting in chronic neuroinflammation with the production of virus and/or viral proteins, cytokines and chemokines. Chemokines, in particular CCL2, increase transmigration of peripheral blood monocytes, continuing neuroinflammation. Thus, chronic inflammation is thought to mediate neuronal damage in a large percentage of infected individuals by mechanisms not well understood. We will characterize the effects of meth, HIV and tat, and ART on monocyte entry into the CNS and on subsequent neuroinflammation. We will test potential therapeutics to limit inflammation and guide efficacy of ART. We hypothesize that meth use combines with HIV infection to exacerbate neuroinflammation and compromise BBB integrity, increasing transmigration of uninfected and HIV-infected monocytes into the brain, leading to cognitive impairment. Based on our new preliminary data that tenofovir increases junctional proteins on the BBB, we also hypothesize that certain ART regimens, especially in meth abusers, may negatively impact cognitive function by synergizing with meth to increase BBB permeability and neuroinflammation. We will characterize effects of meth, HIV and tat, and ART on cytokines, chemokines, and adhesion molecules elaborated by and expressed on CD14+CD16+ monocytes that facilitate entry into the CNS, and in causing BBB permeability and transmigration of CD14+CD16+ monocytes resulting in inflammation, and the impact of ART. We will use a murine model to evaluate the in vivo impact of meth, HIV and its proteins, and ART on BBB permeability and migration of inflammatory cells into the brain. We will also examine effects of meth or dopamine, HIV tat, and ART on cytokine release from CNS cells that mediate inflammation.