Analysis of in vivo Effects of HIV, LPS and Buprenorphine on the Brain Proteome

HIV感染的小胶质细胞和星形胶质细胞产生的炎症介质和病毒产物扰乱了 邻近未感染的神经细胞和神经胶质细胞的功能和活性，并在发病机制中起作用 艾滋病毒相关神经认知障碍(手)。我们假设感染艾滋病毒的小胶质细胞和 星形胶质细胞对血清内毒素水平升高的促炎效应表现出更高的敏感性 在艾滋病毒感染者身上检测到可能有助于手部发育的病毒。我们测试了这个 用我们的JR-CSF/Hu-CycT1小鼠模型进行的假说是转基因的全长整合 在CD4启动子的控制下，感染HIV-1前病毒和人类周期蛋白T1基因，以及 证明在体内注射脂多糖能更有效地激活Jr-CSF/Hu-CycT1小鼠小胶质细胞 星形胶质细胞与对照组小鼠小胶质细胞和星形胶质细胞比较。不断扩大的作用 丁丙诺啡治疗HIV感染的静脉吸毒者阿片成瘾 增加了确定丁丙诺啡与艾滋病毒诱导的协同作用能力的重要性 促进艾滋病毒神经认知功能下降的途径。来描述我们发现的关于 HIV和内毒素对小胶质细胞和星形胶质细胞激活的协同作用及其可能的解决方法 丁丙诺啡的贡献影响，我们建议在定义的生物学背景下分析(艾滋病毒， 炎性刺激(内毒素，丁丙诺啡)HIV、炎性刺激和丁丙诺啡对血管内皮细胞的影响 大脑蛋白质组图谱。定量脑蛋白质组学将使用稳定的同位素标记 饲喂N15或天然标签饲料的动物(Silam)。高pH碳酸盐汽提的靶向分析 从大脑中提取的膜将被执行。这将提供对表情或 HIV感染诱导的受体和转运体的修饰，HIV相关的炎症和 丁丙诺啡。用固定化的抗磷酸化酪氨酸研究酪氨酸磷酸化的变化 分离酪氨酸磷酸化蛋白和多肽。样品将被存档，以供未来分析： 脑区胞浆、神经肽、丝氨酸/苏氨酸磷酸化的多肽、血清/血浆和 其他纸巾。总体而言，这些数据将提供对感染艾滋病毒的大脑的蛋白质组的详细研究 对LPPS和丁丙诺啡的反应在人类受试者中是不可行的。这些数据将提供 对神经认知改变的生物标志物以及丁丙诺啡使用的相关见解，并可能建议 艾滋病毒和阿片剂在手部发育过程中相互作用的新途径。

inflammatory mediators and viral products produced by HIV-infected microglia and astrocytes perturb the function and viability of adjacent uninfected neuronal and glial cells and contribute to the pathogenesis of HIV-associated neurocognitive disorders (HAND). We hypothesized that HIV-infected microglia and astrocytes display increased sensitivity to the proinflammatory effects of the increased levels of serum LPS detected in HIV-infected individuals which may contribute to the development of HAND. We tested this hypothesis using our JR-CSF/hu-cycT1 mouse model that is transgenic for an integrated full-length nfectious HIV-1 provirus and the human cyclin T1 gene under the control of a CD4 promoter, and demonstrated that in vivo administered LPS more potently activated JR-CSF/hu-cycT1 mouse microglia and astrocytes as compared to control littermate mouse microglia and astrocytes. The expanding role of buprenorphine for the treatment of opiate addiction in the population of HIV-infected intravenous drug users increases the importance of determining the capacity of buprenorphine to synergize with HIV-induced pathways to promote HIV neurocognitive decline. To delineate the mechanistic basis of our finding about the synergistic effects of HIV and LPS on microglial and astrocyte activation and to address the possible contributory impact of buprenorphine, we propose to analyze in a defined biological context (¿ HIV, ¿ inflammatory stimulus (LPS), ¿ buprenorphine) the effects of HIV, inflammatory stimuli and buprenorphine on the brain proteome profile. Quantitative brain proteomics will be carried out using stable isotope labeling of the animals (SILAM) fed with N15 or natural labeled chow. Targeted analysis of high pH carbonate stripped membranes from the brain will be performed. This will provide insights into the changes in expression or modification of receptors and transporters induced by HIV infection, HIV-associated inflammation and buprenorphine. Changes in tyrosine phosphorylation will be studied using immobilized anti-phosphotyrosine to isolate tyrosine phosphorylated proteins and peptides. Samples will be archived for future analysis of: brain region cytosol, neuropeptides, peptides that are serine/threonine phosphorylated, serum/plasma, and other tissues. Overall, these data will provide a detailed study of the proteome of the HIV-infected brain in response to LPPS and buprenorphine which is not feasible in human subjects. These data will provide correlative insights for biomarkers of neurocognitive changes as well as buprenorphine use and may suggest new pathways by which HIV and opiates interact in the development of HAND.