Imaging of HTLV-1 by cryo-CLEM

通过冷冻 CLEM 进行 HTLV-1 成像

• 批准号: 10868844
• 负责人: Louis M Mansky
• 金额: $ 8.42万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-16 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10868844
• 关键词: 2019-nCoV; Address; Adult T-Cell Leukemia/Lymphoma; Behavior; Biogenesis; Biomedical Engineering; Capsid; Cell Communication; Cell Culture Techniques; Cells; Cellular biology; Cryoelectron Microscopy; Development; Disease; Electron Microscope; Electron Microscopy; Engineering; Etiology; Event; Eye; Fluorescence Microscopy; Goals; Human; Human Characteristics; Human T-Cell Leukemia Viruses; Human T-lymphotropic virus 1; Image; Imaging technology; Individual; Infection; Inflammatory; Knowledge; Lasers; Malignant Neoplasms; Morphology; Nature; Oral; Organ; Pattern; Preparation; Proteins; Pulmonary Pathology; Reproducibility; Research; Research Project Grants; Resolution; Retroviridae; Role; Sampling; Skin; Spinal Cord Diseases; Syndrome; Technology; Testing; Thyroid Gland; Tissues; Tropical Spastic Paraparesis; Viral; Virion; Virus; Virus Assembly; Virus Diseases; Virus Replication; experimental study; fluorescence imaging; high resolution imaging; human imaging; imaging study; improved; insight; instrument; light microscopy; live cell imaging; microscopic imaging; mutant; new technology; novel; particle; permissiveness; quantitative imaging; success; superresolution microscopy; transmission process; viral transmission; virological synapse; virology

RESEARCH PROJECT Abstract Human T-cell leukemia virus (HTLV-1) has been estimated to infect 15-20 million individuals worldwide and is known to be the etiological agent of an adult T-cell leukemia/lymphoma (ATLL), an inflammatory disease syndrome known as HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP), and pathologies of the lung, skin, eyes, and thyroid gland. HTLV-1 is notorious for being extremely difficult to propagate in cell culture, which has prohibited rigorous analyses of virus replication, including the steps involved in retrovirus assembly. HTLV-1 spread is known to be heavily reliant on virus infection involving cell-to-cell contacts that form what is termed the virological synapse (VS), which represents the primary means for virus spread, including events associated with oral transmission. While HTLV-1 has been previously studied in regard to virus spread via cell-cell contacts, a significant knowledge gap exists regarding the nature of virus particle assembly and transmission via the VS. In general, virus particle spread through cell-cell contacts increases the likelihood of an infection event of virus particles that may possess low particle infectivity in cells without formation of VS. Previous studies have indicated that a low proportion of mature HTLV-1 particles possess an intact capsid core, suggesting that aberrant particle morphology could help to explain the poorly infectious nature of cell-free HTLV- 1. In order to address the current knowledge gap in the field, we propose in this exploratory application to develop workflows for state-of-the-art bioengineering and quantitative imaging technologies that hold high promise in being applied to the efficient study of HTLV-1 particle assembly and spread at the VS. First, we will develop a workflow for the use of cell micropatterning technology in order to reproducibly and efficiently create cell-cell contacts and investigate the role virus budding in virus spread. Second, we propose to establish an efficient workflow in which we can view cell-cell contacts utilizing high-resolution cryo-correlative light and electron microscopy in order to investigate the role of host cell proteins in virus assembly at cell-cell contacts. Development of these workflows will allow for quantitative analysis of virus particle biogenesis at cell-cell contacts. These technologies have broad applicability in virology and the success of this research will be applicable to a variety of questions regarding virus replication and virus-host cell interactions.

研究项目 抽象的 据估计，人类T细胞白血病病毒（HTLV-1）已感染全球15-2000万人 已知是成人T细胞白血病/淋巴瘤（ATLL）的病因学药物，一种炎症性疾病 综合征称为HTLV-1相关的脊髓病/热带痉挛性瘫痪（HAM/TSP）和病理 肺，皮肤，眼睛和甲状腺。 HTLV-1因在细胞中极难传播而臭名昭著 培养，禁止对病毒复制进行严格分析，包括逆转录病毒涉及的步骤 集会。已知HTLV-1传播广泛地依赖于涉及形成细胞对细胞接触的病毒感染 所谓的病毒学突触（VS），它代表了病毒传播的主要手段，包括 与口腔传输相关的事件。虽然先前已经研究了有关病毒扩散的HTLV-1 通过细胞 - 细胞触点，存在有关病毒颗粒组装和 通过VS传输通常，病毒颗粒通过细胞细胞接触传播，增加了 病毒颗粒的感染事件可能在细胞中可能具有低颗粒感染性而没有形成V。以前的 研究表明，成熟的HTLV-1颗粒的比例较低，具有完整的衣壳芯， 提示异常的粒子形态可以帮助解释无细胞HTLV-的感染性不佳的性质 1。为了解决该领域的当前知识差距，我们建议在此探索性应用中开发 最先进的生物工程和定量成像技术的工作流，这些技术在 被应用于HTLV-1颗粒组件的有效研究并扩散在Vs。首先，我们将发展一个 使用细胞微图案技术的工作流程，以便于重复地创建细胞电池 接触并研究病毒扩散中的病毒萌芽。其次，我们建议建立一个有效的 我们可以使用高分辨率低分辨率光和电子查看细胞电池触点的工作流程 显微镜为了研究宿主细胞蛋白在细胞 - 细胞接触处病毒组装中的作用。 这些工作流程的开发将允许对细胞细胞处的病毒颗粒生物发生进行定量分析 联系人。这些技术在病毒学方面具有广泛的适用性，这项研究的成功将是 适用于有关病毒复制和病毒宿主相互作用的各种问题。