Analysis of Interaction between DC and HIV virions

DC和HIV病毒颗粒之间的相互作用分析

• 批准号: 6451012
• 负责人: XIAOYUN WU
• 金额: $ 25.35万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-06-01 至 2004-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6451012
• 关键词: Langerhans' cell; Macaca mulatta; binding proteins; clinical research; communicable disease transmission; dendritic cells; enzyme linked immunosorbent assay; flow cytometry; fluorescence microscopy; gastrointestinal epithelium; green fluorescent proteins; heparan sulfate; host organism interaction; human immunodeficiency virus 1; human tissue; immunofluorescence technique; laboratory mouse; lymph nodes; mucosa; simian immunodeficiency virus; tissue /cell culture; transcytosis; virion; virus infection mechanism; virus replication

Human immunodeficiency virus type 1 (HIV-1) infection continues to spread globally and predominantly by heterosexual contact. In order for HIV to be transmitted through sexual intercourse (heterosexual and homosexual) the virus must cross the epithelial barrier of the mucosa. Rational prophylactic strategies for controlling HIV transmission rely upon a detailed cellular and molecular understanding of the initial interactions that occur between the virus and host after sexual exposure. While advances have been made to identify the kinds of cell that are first infected, very little progress has been made to delineate and understand the earliest interaction(s) of HIV-1 virions with the mucosal surface. On uninjured mucosal membranes, the first cells encountered are CD4-negative epithelial cells. Studies of mucosal transmission (vaginal) in a non-human primate model demonstrated that the first cells infected (expressing viral RNA 18 hrs after infection) are Langerhans' cells, which are located within the mucosa, beneath the epithelial cells. Although evidence continues to accumulate that suggests other cellular molecules mediate attachment of virions to the surface of CD4-negative cells there remains a great void in our understanding of the molecular and cellular events that proceed infection of Langerhans' cells. This is due in great measure to the lack of experimental methods with sufficient sensitivity and specificity to directly analyze the interactions of HIV-1 virions with the mucosa. Our previous finding clearly demonstrated the ability to directly visualize infectious HIV-1 virions that have been labeled with GFP. We have also demonstrated that GFP+ virions can be quantitatively visualized on both cell and mucosal surfaces. Thus, we propose to conduct a detailed and dynamic analysis of HIV-1 mucosal transmission. The research proposed in this grant address questions regarding the molecular determinants (viral and cellular) of initial virus attachment, penetration of virus into or across the mucosa and virus-host cell infection. Our central hypothesis is that luminal epithelial cells play an important role in the earliest events of HIV-1 mucosal transmission. It follows that this work has significant clinical implications for providing the experimental basis to proceed rationally with new strategies for inhibiting HIV-1 transmission. To test our central hypothesis we propose: (1) To define the nature of the interaction between HIV-1 and primary epithelial cells - at the level of virus binding, entry and infection; (2) To define the host-cell and virus-associated molecular determinants which mediate the physical interaction between HIV-1 virions and primary epithelial cells; (3) To define the nature of the interaction between HIV-1 virions and epithelial cells using an organ culture system and human and monkey mucosal tissues (vaginal and intestinal) and (4) To analyze the earliest events of HIV-1 mucosal transmission in animal models.

人类免疫缺陷病毒1型(HIV-1)感染继续在全球传播，并主要通过异性接触传播。为了使艾滋病毒通过性交(异性恋和同性恋者)传播，病毒必须越过粘膜的上皮屏障。控制艾滋病毒传播的合理预防策略依赖于对性接触后病毒与宿主之间发生的初始相互作用的详细的细胞和分子理解。虽然在识别最初感染的细胞种类方面取得了进展，但在描绘和了解艾滋病毒-1病毒粒子与粘膜表面最早的相互作用(S)方面进展甚微。在未受损伤的粘膜上，首先遇到的细胞是CD4阴性的上皮细胞。在非人灵长类动物模型中对粘膜传播(阴道)的研究表明，第一个被感染的细胞(在感染18小时后表达病毒RNA)是朗格汉斯细胞，它们位于粘膜内， 在上皮细胞下面。尽管不断积累的证据表明，其他细胞分子介导了病毒粒子与CD4阴性细胞表面的附着，但我们对朗格汉斯细胞感染之前的分子和细胞事件的了解仍然是一个巨大的空白。这在很大程度上是由于缺乏足够灵敏度和特异度的实验方法来直接分析HIV-1的相互作用。 病毒粒子与粘膜结合。我们之前的发现清楚地证明了直接可视化标记了GFP的传染性HIV-1病毒粒子的能力。我们还证明了GFP+病毒粒子可以在细胞和粘膜表面定量显示。因此，我们建议对HIV-1的粘膜传播进行详细和动态的分析。这项拨款中提出的研究涉及初始病毒附着的分子决定因素(病毒和细胞)、病毒进入或穿过粘膜和病毒宿主细胞感染的问题。我们的中心假设是，腔上皮细胞在HIV-1粘膜传播的最早事件中发挥着重要作用。因此，这项工作具有重要的临床意义，为合理开展抑制HIV-1传播的新策略提供了实验基础。为了检验我们的中心假设，我们建议：(1)在病毒结合、进入和感染的水平上定义艾滋病毒-1与原代上皮细胞之间相互作用的性质；(2)定义宿主细胞和病毒相关的分子决定因素，它们介导艾滋病毒-1病毒粒子与原代上皮细胞之间的物理相互作用；(3)利用器官培养系统以及人和猴子的粘膜组织(阴道和肠道)来定义艾滋病毒-1病毒粒子与上皮细胞之间相互作用的性质；(4)在动物模型中分析艾滋病毒-1粘膜传播的最早事件。