Neurokinin-1R Antagonists- Anti-HIV Mechanisms

项目1。该项目的总体目标是了解神经激肽-1受体（NK-1 R），P物质（SP）偏好受体，拮抗剂介导的抗HIV活性在人类免疫细胞中的机制。我们早期的研究表明，NK-1 R拮抗剂（CP-96，345）至少部分通过下调HIV进入巨噬细胞的主要辅助受体CCR 5来抑制巨噬细胞的HIV感染。此外，CP-96，345抑制免疫细胞的内源性SP产生。为了支持这些体外研究结果，我们已经证明，在HIV感染者中， 与未感染的受试者相比，HIV感染会诱导人免疫细胞中SP的表达。这些重要的体外和体内数据表明，有必要进一步研究使用NK-1 R拮抗剂作为HIV疾病的治疗方法的潜力。在本提案中，我们将通过测试其他NK-1 R拮抗剂在感染不同HIV毒株的CD 4 + T细胞和巨噬细胞中的抗HIV能力来扩展我们的研究范围。我们将研究NK-1 R拮抗剂抗HIV活性的机制。我们的总体假设是NK-1 R拮抗剂是具有抗病毒和免疫调节作用的强效抑制剂。我们还将研究假设， NK-1受体拮抗剂可降低HIV感染及其蛋白（gp 120或达特）对人神经细胞的神经毒性作用。我们提出了新的体外研究，以确定NK-1 R拮抗剂在正常和HIV感染受试者的PBMC中的抗HIV活性（目的1和2）。NK-1 R拮抗剂对免疫细胞耐药HIV感染的影响，以及它们与常用抗逆转录病毒药物的协同作用，也将是这项拟议研究的新焦点（目的1）。结合项目2，我们将研究NK-1 R拮抗剂抗HIV活性的机制（目的3）。在目的4中，我们将确定NK-1 R拮抗剂是否在小胶质细胞（CNS中的主要HIV靶细胞）中具有抗HIV活性，以及NK-1 R拮抗剂是否对人类神经元细胞具有神经保护作用。这些研究代表了一系列独特的实验，旨在阐明NK-1 R拮抗剂的抗HIV能力和机制，并将提供 使用NK-1 R拮抗剂作为治疗HIV疾病的潜在治疗剂的进一步科学证据。

Project 1. The overall goal of this project is to understand the mechanisms involved in the neurokinin-1 receptor (NK-1R), the substance P (SP) preferring receptor, antagonist-mediated anti-HIV activity in human immune cells. Our earlier studies have shown that the NK-1R antagonist (CP-96,345) inhibits HIV infection of macrophages, at least in part, through down-regulation of CCR5, a principal co-receptor for HIV entry into macrophages. In addition, CP-96,345 inhibits endogenous SP production by the immune cells. In support of these in vitro findings, we have documented that there are elevated SP levels in HIV-infected subjects in comparison to uninfected subjects, and that HIV infection induces SP expression in human immune cells. These important in vitro and in vivo data indicate the necessity of further investigation of the potential of using NK-1R antagonists as a therapeutic approach for HIV disease. In this proposal, we will extend the scope of our investigations by testing the anti-HIV ability of other NK-1R antagonists in both CD4+ T cells and macrophages infected with different strains of HIV. We will examine the mechanisms responsible for the anti-HIV activity of the NK-1R antagonists. Our overarching hypothesis is that NK-1R antagonists are potent inhibitors with both antiviral and immunomodulatory effects. We will also examine the hypothesis that the NK-1R antagonists reduce the neurotoxic effects of HIV infection and its proteins (gp120 or Tat) on the human neuronal cells. We propose new in vitro studies to determine the anti-HIV activity of NK-1R antagonists in PBMC from both normal and HIV-infected subjects (Aims 1 and 2). The impact of the NK-1R antagonists on drug-resistant HIV infection of immune cells, as well as their synergistic effect with the commonly used antiretrovirals, also will be a new focus of this proposed study (Aim 1). In conjunction with the Project 2, we will examine the mechanism(s) responsible for the anti-HIV activity of the NK-1R antagonists (Aim 3). In Aim 4, we will determine whether the NK-1R antagonists have the anti-HIV activity in microglia, the primary HIV target cells in the CNS, and whether the NK-1R antagonists have the neuroprotective effect on human neuronal cells. These studies represent a unique series of experiments directed toward elucidating the anti-HIV ability and mechanisms of the NK-1R antagonists and will provide further scientific evidence for using the NK-1R antagonists as a potential therapeutic agent for treatment of HIV disease.