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）：本提案重点关注静脉内暴露后 HIV-1 在体内传播的机制特征。 细胞间感染已被证明是一种独特的病毒传播机制，在体外更有效、更快速。目前，病毒传播的无细胞模式与细胞间病毒传播模式的作用尚未在体内进行检验。 这项研究希望揭示胃肠外传播过程中病毒传播的主要模式，我们假设这种传播是由细胞间接触而不是无细胞病毒介导的。 深入了解 HIV-1 在体内如何传播将有助于加强治疗药物的开发，这些治疗药物可以有效阻止所有形式的病毒传播，最终防止疾病进展。 艾滋病病毒。 我们将直接可视化活组织中受感染 CD4+ T 细胞的细胞相互作用。使用人源化小鼠 HIV-1 感染模型，我们计划描述静脉吸毒者 (IDU) 静脉内 HIV 暴露模型传播后病毒建立的初始位点。 为了实现 HIV 传播的可视化，将通过流式细胞术和新型多光子实时成像技术来评估荧光标记的 HIV-1 克隆的使用。 该项目要调查的三个领域是： 1) 静脉注射细胞相关病毒或无细胞病毒后，表征 HIV-1 的器官间传播。 我们计划通过使用双光子实时成像和流式细胞术来定义人源化小鼠模型中 HIV-1 器官间转移的顺序事件。 在此过程中，我们将阐明病毒运输的机制，以显示肺部定位和细胞迁移对病毒传播的影响。 2) 确定共同传播的模式以区分两种传播模式。我们的目标是利用双重感染策略，使用不同的荧光 HIV-1 变体来表征 HIV-1 细胞间传播在共传播中的功能。 为了分析不同传播方式的双重感染的传播，我们将对人源化小鼠组织进行双光子显微镜和流式细胞术。 3) 检查体内病毒通过病毒学突触的转移。 病毒的突触转移已在体外很容易得到表征；然而，体内稳定突触形成的存在尚未观察到。 我们的目标是使用人源化小鼠淋巴器官的延时双光子活体成像来可视化病毒蛋白通过细胞间接触的转移。