Opioids, LPS and HIV

描述(申请人提供)：虽然已知阿片类药物对免疫调节活性有深刻影响，并在艾滋病毒-1疾病的免疫发病机制中起辅助作用，但其作用机制(S)仍有待确定。关于阿片类药物的使用与血浆脂多糖水平升高之间的关联的现有信息很少，脂多糖是革兰氏阴性细菌细胞壁的主要成分，也是艾滋病毒感染中全身免疫激活的可能原因[1]。特别是，在脂多糖存在的情况下使用阿片类药物对HIV-1靶向免疫细胞(如CD4T细胞、单核细胞和小胶质细胞)的细胞microRNA和基因表达的影响尚不清楚。因此，确定阿片类药物对CD4T细胞、单核细胞和小胶质细胞感染的miRNAs和相关基因的变化是人们感兴趣的领域，这可能有助于识别先前未知的参与HIV疾病发病机制。这一建议的理论基础是基于最近的报告，表明血液中血浆内毒素水平的增加是来自泄漏肠道的细菌产物移位的结果，这可能是慢性艾滋病毒感染中全身免疫激活的原因[1]，药物滥用与血浆高内毒素[2]有关。此外，最近研究证明，CD4T细胞和单核/巨噬细胞中的细胞内抗HIV细胞因子在抑制HIV感染/复制方面起着关键作用[3，4]。因此，我们假设海洛因等药物的使用与血浆内毒素水平升高有关，阿片类药物(海洛因或吗啡)抑制免疫细胞和中枢神经系统细胞内的抗HIV先天免疫，促进HIV感染。我们提出了三个具体的目标来解决以前未知的影响和机制，阿片类药物和/或脂多糖通过这些影响和机制损害宿主细胞的天然免疫，增加免疫和中枢神经系统细胞对HIV-1感染和损伤的易感性。在目标1中，我们将在体内检测海洛因使用对CD4T细胞和单核细胞微易位标志物表达以及miRNA/基因谱的影响，重点研究JAK/STAT通路。在目标2中，我们将在体外检测吗啡和/或内毒素对miRNA表达的影响，重点是新发现的在CD4T细胞、单核细胞和巨噬细胞中抗HIV的miRNAs。我们将确定吗啡/内毒素介导的细胞内天然免疫和HIV感染的分子机制。在目标3中，我们将确定阿片类药物和/或内毒素对星形胶质细胞和小胶质细胞中抗HIV miRNAs表达的影响。我们推测，吗啡和/或内毒素通过抑制中枢神经系统细胞(原代星形胶质细胞、小胶质细胞和神经元)的天然免疫，增加小胶质细胞和星形胶质细胞对HIV感染的易感性。这些研究具有重要的意义和创新性，因为我们将解决以下新问题：在中国海洛因依赖者中，阿片类药物的使用是否与血浆内毒素升高呈正相关，以及阿片类药物和/或内毒素是否会抑制免疫细胞和中枢神经系统细胞内的抗HIV天然免疫，从而促进HIV感染。 公共卫生相关性：该项目提出了体外和体内研究，以解决先前未知的影响和机制，阿片类药物和/或脂多糖通过这些影响和机制破坏宿主细胞内的天然免疫，并增强免疫和中枢神经系统细胞对HIV-1感染和损伤的易感性。

DESCRIPTION (provided by applicant): While it is known that opioids exert a profound influence on immunomodulating activity and have a cofactor role in the immunopathogenesis of HIV-1 disease, the mechanism(s) of their action remains to be determined. There is little information available about the association of opioid use with elevated plasma levels of lipopolysaccharide (LPS), a major component of gram-negative bacteria cell walls and a likely cause of systemic immune activation in HIV infection [1]. Particularly, it is unknown about the effect of opioid use in the presence of LPS on cellular microRNA and gene expression in HIV-1-targeted immune cells such as CD4+ T cells, monocytes, and microglia. Thus, to determine changes of miRNAs and genes related HIV infection of CD4+ T cells, monocytes, and microglia exposed to opioids is the area of interest, which may help to identify previously unidentified mechanisms involved in pathogenesis of HIV disease. The rationale of this proposal is based on the recent reports, showing that increased levels of plasma LPS in the blood is a consequence of translocation of bacterial products from a leaky gut, which is likely responsible for the systemic immune activation in chronic HIV infection [1] and that substance abuse is associated with high plasma LPS [2]. In addition, it has been recently documented that intracellular anti-HIV cellular factors in CD4+ T cells and monocytes/macrophages have a critical role in suppression of HIV infection/replication [3, 4]. Therefore, we hypothesize that drugs of use such as heroin is associated with elevated plasma levels of LPS and that opioids (heroin or morphine) suppress intracellular anti- HIV innate immunity in the immune cells and CNS cells, promoting HIV infection. We propose three specific aims to address previously unrecognized impact and mechanisms by which opioids and /or LPS compromise host cell innate immunity, increasing the susceptibility of immune and the CNS cells to HIV-1 infection and injury. In aim 1, we will examine in vivo impact of heroin use on the expression of markers of microtranslocation as well as on miRNA/gene profile of CD4+ T cells and monocytes with focus on the JAK/STAT pathways. In aim 2, we will examine in vitro impact of morphine and/or LPS on the expression of miRNA profiles with focus on newly identified anti-HIV miRNAs in CD4+ T cells, monocytes and macrophages. We will determine the molecular mechanisms responsible for morphine/LPS-mediated actions on intracellular innate immunity and HIV infection. In aim 3, we will determine the impact of opioids and/or LPS on the expression of anti-HIV miRNAs in astrocytes and microglia. We hypothesize that morphine and/or LPS, through the suppression of innate immunity in the CNS cells (primary astrocytes, microgila, and neurons), increase the susceptibility of microglia and astrocytes to HIV infection. The proposed studies are highly significant and innovative, as we will address the novel questions of whether opioid use is positively associated elevated plasma LPS in Chinese heroin dependent subjects, and whether opioids and/or LPS suppress intracellular anti-HIV innate immunity in the immune and CNS cells, promoting HIV infection. PUBLIC HEALTH RELEVANCE: This project proposes in vitro and in vivo studies to address previously unrecognized impact and mechanisms by which opioids and /or LPS compromise host intracellular innate immunity and enhance the susceptibility of immune and the CNS cells to HIV-1 infection and injury.