Role of cell-to-cell HIV-1 spread in vivo during intravenous transmission

HIV-1 静脉内传播过程中体内细胞间传播的作用

• 批准号: 8602035
• 负责人: KENNETH Marvin LAW
• 金额: $ 3.67万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-01 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8602035
• 关键词: Acute; Address; Adherens Junction; Animal Model; Area; Biological Assay; CCR5 gene; CD4 Positive T Lymphocytes; Cells; Data; Development; Disease Progression; Environment; Event; Evolution; Exposure to; Flow Cytometry; HIV; HIV-1; Human; Image Cytometry; Imagery; Imaging Techniques; Immune; In Vitro; Infection; Intravenous; Label; Laboratories; Lead; Life; Lung; Lymphocyte; Lymphoid; Mediating; Methods; Microscopy; Modeling; Mus; Organ; Patients; Pattern; Proviruses; Role; SIV; Sequence Analysis; Site; Spleen; Splenic Tissue; Structure of parenchyma of lung; Synapses; T-Cell Depletion; Therapeutic Agents; Tissues; Variant; Viral; Viral Proteins; Virus; cell motility; chemokine; in vivo; insight; intravenous drug user; intravenous injection; intravital imaging; mouse model; multi-photon; novel; prevent; public health relevance; synaptogenesis; time use; trafficking; transmission process; two-photon; virological synapse

DESCRIPTION (provided by applicant): This proposal focuses on characterizing mechanisms of HIV-1 spread in vivo after intravenous exposure. Cell-to-cell infection has been shown to be a distinct mechanism of viral spread that is more efficient and rapid in vitro. Currently, the roleof cell-free versus cell-to-cell modes of viral spread has not been examined in vivo. This study hopes to reveal the dominant mode of viral spread that occurs during parenteral transmission, which we hypothesize to be mediated by cell-to-cell contacts rather than cell-free virus. Insight into how HIV-1 propagates in vivo will help to enhance the development of therapeutic agents that can efficiently block all forms of viral spread, ultimately, preventing disease progression of HIV. We will directly visualize cellular interactions of infected CD4+ T cells in live tissue. Usig a humanized mouse HIV-1 infection model, we plan to characterize the initial sites where virus establishes after intravenous HIV exposure to model transmission in intravenous drug users (IDU). For visualization of HIV transmission, the use of fluorescently-tagged HIV-1 clones will be assessed by flow cytometry and novel multi-photon live imaging techniques. The three areas to be investigated in this project are: 1) Characterize organ-to-organ spread of HIV-1 after intravenous injection of cell-associated or cell-free virus. We plan to define the sequential events of organ-to-organ transfer of HIV-1 in humanized mouse models through the use of two-photon live imaging and flow cytometry. In doing so we will elucidate the mechanism of viral trafficking to show the impact lung localization and cellular migration has on virus dissemination. 2) Determine the patterns of co-transmissions to discern between two modes of spread. We aim to utilize a dual infection strategy using distinct fluorescent HIV-1 variants to characterize the function of HIV-1 cell-to-cell spread in cotransmission. To analyze the spread of double infection with different modes of transmission, we will perform two-photon microscopy and flow cytometry on humanized murine tissues. 3) Examine virus transfer through virological synapse in vivo. Synaptic transfer of virus has been readily characterized in vitro; however, the presence of stable synapse formation in vivo has yet to be observed. We aim to visualize the transfer of viral proteins through cell-to-cell contact using time-lapse two-photon intravital imaging of lymphoid organs in humanized mice.

描述（由申请人提供）：该提案的重点是表征静脉注射后体内HIV-1扩散的机制。 细胞对细胞感染已被证明是病毒扩散的独特机制，在体外更有效，快速。当前，尚未在体内检查无细胞与细胞对病毒传播的细胞对细胞模式的作用。 这项研究希望揭示肠胃外传播期间发生的病毒传播的主要模式，我们假设这是由细胞对细胞接触而不是无细胞病毒介导的。 深入了解HIV-1如何在体内传播将有助于增强治疗剂的发展，这些治疗剂可以有效地阻止所有形式的病毒传播，最终，防止疾病的进展 艾滋病病毒。 我们将直接可视化活组织中感染的CD4+ T细胞的细胞相互作用。 USIG是一种人源化的小鼠HIV-1感染模型，我们计划表征静脉内HIV暴露于静脉吸毒者（IDU）中的静脉内HIV后病毒建立的初始部位。 为了可视化HIV传播，将通过流式细胞仪和新型的多光子实时成像技术评估荧光标记的HIV-1克隆的使用。 该项目要研究的三个领域是： 1）表征静脉注射细胞相关或无细胞病毒后HIV-1的器官到器官扩散。 我们计划通过使用两光子实时成像和流式细胞术来定义人源性小鼠模型中HIV-1的器官到器官转移的顺序事件。 通过这样做，我们将阐明病毒贩运的机制，以显示肺部定位和细胞迁移对病毒传播的影响。 2）确定偶然转移的模式，以辨别两种传播模式。我们旨在利用双重感染策略使用不同的荧光HIV-1变体来表征HIV-1细胞到细胞在共晶中传播的功能。 为了通过不同的传播模式分析双重感染的扩散，我们将对人源化鼠组织进行两光子显微镜和流式细胞仪。 3）通过体内病毒学突触检查病毒转移。 病毒的突触转移很容易在体外表征。但是，尚未观察到体内稳定突触形成的存在。 我们的目的是使用人源化小鼠中淋巴机器人的延时两光室内成像，通过细胞对细胞接触可视化病毒蛋白的转移。