Single Cell with One Particle Entry (SCOPE) for Study of HIV Infection

用于 HIV 感染研究的单细胞单粒子进入 (SCOPE)

• 批准号: 8146689
• 负责人: Wei Cheng
• 金额: $ 233.25万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-30 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8146689
• 关键词: Binding; CD4 Positive T Lymphocytes; Cell surface; Cells; DNA Integration; Event; Goals; HIV; HIV Infections; HIV-1; Incubated; Infection; Lead; Life; Ligands; Link; Measures; Methods; Molecular; Pathway interactions; Population; Probability; T-Lymphocyte; Techniques; Time; Viral; Virion; Virus; abstracting; base; chemokine; laser tweezer; macromolecular assembly; nanoparticle; new technology; novel; particle; public health relevance; receptor; research study; therapeutic development; uptake

DESCRIPTION (Provided by the applicant) Abstract: Through infection of CD4 positive (CD4+) T lymphocytes, the Human Immunodeficiency Virus type 1 (HIV-1) has claimed twenty-five million lives since its discovery in 1983. Although it has been well established that HIV-1 initiates a T cell infection by binding to CD4 and chemokine coreceptors on the cell surface, the early events in HIV-1 infection of CD4+ T cells remain poorly understood. A variety of different techniques have been used over the years to study mechanisms of HIV-1 entry. However, one technical limitation that is inherent in all these methods is the inability to track the fate of a single HIV-1 virion from the very beginning of viral entry to chromosomal integration. In all these experiments, a population of viruses and cells were incubated and measured. Because each entry event can lead to proviral DNA integration with a finite probability, it is therefore difficult to establish a causative link between entry pathway and productive infection. The goal of this project is to develop a set of nanoscopic and novel technologies that we can harness to define the pathway and interactions by which HIV-1 infects CD4+ T cells. We are developing a technique based on optical tweezers that can manipulate a single HIV-1 virion, deliver it to CD4+ T cell and thus allows us to determine the fate of CD4+ T cell upon entry by a single virion. This technique will allow us to unambiguously define the molecular mechanisms of HIV-1 infection. Furthermore, we propose to measure directly the interactions between a single HIV-1 virion and receptors in the context of a live T cell. Collectively, these studies will contribute to a definitive and quantitative understanding of early events in HIV infection, which may help therapeutic development that is aimed to block HIV-1 entry to CD4+ T cells. The techniques developed herein can be useful for studying cellular uptake of not only viruses but other molecules, macromolecular assemblies and nanoparticles, and are applicable to a wide-range of ligand-receptor interactions on the cell surface. Public Health Relevance: The early events in HIV infection of CD4+ T cells are poorly understood. This proposal aims to develop a set of nanoscopic and other novel techniques to study HIV infection of CD4+ T cells in real time, one virion at a time. If successful, the results from this study will establish for the first time a causative link between HIV entry pathway and the productive infection of CD4+ T cells, which will help therapeutic development that is aimed to block HIV-1 entry to CD4+ T cells.

描述（由申请人提供） 摘要：通过感染CD 4阳性（CD 4+）T淋巴细胞，人类免疫缺陷病毒1型（HIV-1）自1983年发现以来已夺去了2500万人的生命。虽然已经确定HIV-1通过与细胞表面的CD 4和趋化因子辅助受体结合来启动T细胞感染，但HIV-1感染CD 4 + T细胞的早期事件仍然知之甚少。多年来，各种不同的技术已被用于研究HIV-1进入的机制。然而，所有这些方法中固有的一个技术限制是不能从病毒进入染色体整合的最开始追踪单个HIV-1病毒粒子的命运。在所有这些实验中，孵育并测量了一群病毒和细胞。由于每个进入事件都可能以有限的概率导致前病毒DNA整合，因此很难在进入途径和生产性感染之间建立因果关系。该项目的目标是开发一套纳米级的新技术，我们可以利用这些技术来定义HIV-1感染CD 4 + T细胞的途径和相互作用。我们正在开发一种基于光镊的技术，该技术可以操纵单个HIV-1病毒体，将其递送到CD 4 + T细胞，从而使我们能够确定单个病毒体进入后CD 4 + T细胞的命运。这项技术将使我们能够明确定义HIV-1感染的分子机制。此外，我们建议直接测量单个HIV-1病毒粒子和受体在活T细胞的背景下之间的相互作用。总的来说，这些研究将有助于对HIV感染的早期事件进行明确和定量的了解，这可能有助于开发旨在阻断HIV-1进入CD 4 + T细胞的治疗方法。本文开发的技术不仅可用于研究病毒，而且可用于研究其他分子、大分子组装体和纳米颗粒的细胞摄取，并且可适用于细胞表面上的广泛的配体-受体相互作用。 公共卫生相关性：对HIV感染CD 4 + T细胞的早期事件知之甚少。该提案旨在开发一套纳米级和其他新技术，以真实的时间研究HIV感染CD 4 + T细胞，一次一个病毒体。如果成功，这项研究的结果将首次建立HIV进入途径与CD 4 + T细胞的生产性感染之间的因果关系，这将有助于旨在阻断HIV-1进入CD 4 + T细胞的治疗开发。