Methamphetamine and Immune Cells in NeuroAIDS

神经艾滋病中的甲基苯丙胺和免疫细胞

• 批准号: 8067738
• 负责人: Maria Cecilia Garibaldi Marcondes
• 金额: $ 28.43万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2013-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8067738
• 关键词: AIDS neuropathy; Address; Affect; Animals; Antigen Presentation Pathway; Antiviral Agents; Behavior; Binding; Biological Assay; Blood - brain barrier anatomy; Brain; CCR5 gene; Cell Cycle; Cell Line; Cells; Characteristics; Comorbidity; Complication; Control Groups; Data; Detection; Drug abuse; Environment; Epigenetic Process; Exposure to; Freezing; Future; Gene Expression; Gene Proteins; Genes; HIV; HIV Infections; HIV-1; Hepatitis C virus; Human; Human Cell Line; Imagery; Immune; Immune response; Immunohistochemistry; In Vitro; Incidence; Individual; Infection; Inflammatory; Lead; Liver; Lymphocyte; Macaca; Macaca mulatta; Maps; Mediating; Methamphetamine; Microglia; Minor; Mitochondria; Modeling; Molecular; Neuroglia; Neurologic; Neurons; Oxidative Stress; Paraffin Embedding; Pathology; Pathway interactions; Pattern; Performance; Peripheral; Pharmaceutical Preparations; Phenotype; Post-Translational Protein Processing; Process; Proteins; Quantitative Reverse Transcriptase PCR; Reporting; Research; Risk; SIV; Severities; Signal Pathway; Signal Transduction; Spleen; Stimulus; Surface; Suspension substance; Suspensions; Systems Biology; Therapeutic; Therapeutic Intervention; Tissues; Validation; Viral; Viral Load result; Virus; Virus Diseases; Western Blotting; base; brain tissue; chemokine receptor; cytokine; in vivo; macrophage; methamphetamine abuse; neuropathology; novel; outcome forecast; prevent; protein expression

DESCRIPTION (provided by applicant): Methamphetamine (Meth) abuse leads to behaviors that increase the risk of exposure to HIV1-3. NeuroAIDS, which affects >40% of HIV+ individuals, is a particularly severe complication of the infection causing neurological deficits, not decreased by antiviral drugs that do not cross the blood-brain-barrier efficiently 4. NeuroAIDS and peripheral pathologies associated to this co-morbidity have been reported to be aggravated by Meth5 6. Thus, the effects of Meth are predominant, but not restricted to the CNS. We have found that Meth modulates the phenotype of brain and peripheral innate immune cells in the SIV/Rhesus macaque model, increasing cytokine secretion and enriching virus targets CCR5+ cells7, and in vitro. We will investigate the hypothesis of direct action of Meth on innate immune cells, causing alterations that may enhance virus-triggered tissue-pathologies in HIV+ Meth abusers. In the present proposal, we will focus on innate immune cells existing in the brain and in the periphery, especially macrophages and microglia cells. Given that, we hypothesize that Meth can interfere with the anti-viral host immune response, by directly affecting innate immune cells of the macrophage/ microglia lineage. This can be an important component of increased neuropathology, particularly in neuroAIDS, since HIV brain targets are macrophages and microglia. We will determine if Meth modulates gene expression in CNS-derived immune cells in SIV-infected macaques, and changes sets of genes triggered by SIV in the brain by comparing gene expression in spleen and brain- cryopreserved immune cells (depleted of neurons and astro/oligoglia) in healthy and SIV-infected macaques, treated or not with Meth. Expression data will be compared using an integrative systems biology approach and validated. We will prioritize high score pathways, but favor molecules involved in cell cycle, oxidative stress, and inflammatory phenotype, in which the effects of Meth have been observed. Validation will be performed using qRTPCR and protein detection (western blot on brain and spleen frozen tissue fragments, FACS of frozen cell suspensions, and immunohistochemistry on paraffin-embedded tissue - already processed and available for all animals). In parallel, we will assess the ability of Meth to directly modulate macrophage/ microglia gene/protein expression and functional characteristics, using macrophage cell lines and human microglia primary cultures. Gene expression and functional assays assess the effects of Meth on cell cycle, oxidative stress, chemokine receptor and inflammatory molecule expression, mitochondrial functions and antigen processing/ presentation. This will allow the observation of direct effects on innate immune cells that get infected and predominate in the brain. These studies will help understand mechanisms regulating the severity of brain and other tissues pathology in HIV+ Meth users and reveal direct Meth targets on innate immune cells. PUBLIC HEALTH RELEVANCE: Among Methamphetamine (Meth) users, there is a large overlap with HIV infection, being NeuroAIDS an important complication reported to be aggravated by this drug, although its effects on pathology are not restricted to the CNS. Given that the mechanisms by which the drug aggravates pathology and operates systemically are not well understood, there are no medications for treating the pathological consequences of Meth use in HIV or in other viral infections. To help understand mechanisms regulating the severity of brain and other tissue pathologies in HIV+ Meth, the present exploratory study will investigate direct effects on innate immune cells, which are predominantly infected in the brain and present also in other inflammatory conditions, aiming a molecular basis for therapeutic intervention, and for a new paradigm of immune-mediated pathology in drug abuse.

描述（由申请人提供）：甲基苯丙胺（甲基苯丙胺）滥用导致行为增加了暴露于HIV1-3的风险。影响> 40％的HIV+个体的神经剂是引起神经系统缺陷的感染特别严重的并发症，抗病毒药不会降低，这些药物没有有效地越过血脑障碍。 CNS。我们发现，在SIV/Rhesus猕猴模型中，METH调节大脑和外周先天性免疫细胞的表型，从而增加细胞因子分泌，并富集病毒靶标CCR5+细胞7和体外。我们将研究METH对先天免疫细胞的直接作用的假设，从而导致可能增强病毒触发的HIV+ METH滥用者的组织 - 组织病理。在本提案中，我们将专注于大脑中存在的先天免疫细胞，尤其是巨噬细胞和小胶质细胞。鉴于这一点，我们假设METH可以直接影响巨噬细胞/小胶质细胞谱系的先天免疫细胞来干扰抗病毒宿主免疫反应。这可能是神经病理增加的重要组成部分，尤其是在神经辅酶中，因为HIV脑靶标是巨噬细胞和小胶质细胞。我们将确定METH是否在CNS衍生的猕猴中的CNS衍生的免疫细胞中调节基因表达，以及通过比较Spleen和脑冷冻保存的免疫细胞中SIV触发的基因的变化集（由脾脏和脑冷​​冻保存的免疫细胞中的基因表达（由健康和SATRO/ASTRO/ASTORIA）在健康和siv-Infected tefected macaques中，用seftero蛋白耗尽）。表达数据将使用综合系统生物学方法进行比较并经过验证。我们将优先考虑高分途径，但有利于在细胞周期，氧化应激和炎症表型中涉及的分子，其中已经观察到了甲基甲基苯酚的作用。将使用QRTPCR和蛋白质检测（在脑和脾脏冷冻的组织片段，冷冻细胞悬浮液的FAC上进行蛋白质印迹，以及在石蜡包裹的组织上的免疫组织化学 - 已经对所有动物进行处理并使用）。同时，我们将使用巨噬细胞系和人类小胶质细胞原代培养物评估甲基直接调节巨噬细胞/小胶质细胞基因/蛋白表达和功能特征的能力。基因表达和功能测定评估甲基对细胞周期，氧化应激，趋化因子受体和炎症分子表达，线粒体功能和抗原加工/表现的影响。这将允许观察到对先天免疫细胞感染和占主导地位的直接影响。这些研究将有助于理解调节艾滋病毒+甲基甲基使用者中大脑和其他组织病理严重程度的机制，并揭示对先天性免疫细胞的直接甲基苯甲酸酯靶标。 公共卫生相关性：在甲基苯丙胺（甲基苯丙胺）使用者中，与艾滋病毒感染存在很大的重叠，是神经助理的一种重要的并发症，据报道，这种药物对病理学的影响不仅限于CNS。鉴于该药物加剧病理并系统地作战的机制尚未得到充分了解，因此没有药物可以治疗HIV或其他病毒感染中使用甲基甲基苯丙胺的病理后果。为了帮助了解调节HIV+ METH中大脑和其他组织病理严重程度的机制，本探索性研究将调查对先天免疫细胞的直接影响，这主要是在大脑中感染的，并且在其他炎症状态下也呈现，旨在用于治疗干预的分子基础，以进行治疗疗法，并针对药物滥用的新范围。