Identification of the Initial Targets of Transmission

识别初始传播目标

• 批准号: 10610848
• 负责人: Thomas Hope
• 金额: $ 89.45万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10610848
• 关键词: AIDS prevention; Adjuvant; Anatomy; Animals; Antigens; Area; Autopsy; Cells; Complex; Environment; Event; Goals; HIV Infections; HIV vaccine; Immune response; Infection; Inflammatory Response; Injectable; Instruction; Intervention; Label; Luciferases; Methodology; Methods; Mucous Membrane; Natural History; PET/CT scan; Pathway interactions; Phase; Physiology; Predisposition; Prevention approach; Principal Investigator; Process; Rectum; Reporter; SIV; Science; Signal Transduction; Site; Systemic infection; Techniques; Tissues; Tropism; Vagina; Viral; Viremia; Virus; Virus Replication; cell motility; env Gene Products; innovation; neutralizing antibody; next generation; novel; novel strategies; prevent; preventive intervention; programs; rectal; response; sexual HIV transmission; success; transmission process; vector

Program Director/Principal Investigator (Last, First, Middle): Hope, Thomas J. Recent advances in HIV Prevention science include the demonstration of PreP efficacy of the long acting injectable Cabotegravir and broadly neutralizing antibodies against sensitive strains. Likewise, there have been advances in HIV vaccine science with novel immunogens, adjuvants, and delivery strategies that are increasing the immune responses to the virus and their ability to prevent systemic infection. However, the fine tuning of these preventative interventions and our ability to increase their potency require a better understanding of the mechanisms of HIV sexual transmission. The primary focus of this project has been on 1) developing and optimizing methods allowing the identification of the first cells infected after a mucosal challenge; 2) the characterization of the expanding foci of infection, and 3) the definition of the cascade of events that takes place during the eclipse phase as the virus disseminates before detectable viremia. This project has uniquely impacted and advanced our understanding of the detailed natural history of the virus in the first 4 days after mucosal challenge. This success is a consequence of the innovative approach of beacon-guided necropsy where a signal is generated by the presence of infected cells. The first version of this technique utilized luciferase expressed by a replication defective dual-reporter vector. However, we have developed the next generation of this approach, which uses 64Cu labeled FAB2 probes specific for the SIV envelope protein and PET/CT as next generation beacon-guided necropsy. This approach is highly sensitive and efficient allowing the unbiased identification of multiple foci within the same animal at the whole-body level including the characterization of the interactions of the virus with host innate and inflammatory responses. The study of small foci of infection during the eclipse phase after transmission reveals a complex crosstalk between different infected cells and local tissue environment, which can vary in different areas and tissues within the same animal. This and other observations reveal target cell susceptibility, rather than the “tropism” of the viral envelope, is the key driver of early infection. It is clear that the local anatomy and physiology of virus exposed mucosal tissue has a major impact on the natural history of the virus during the eclipse phase. Through the interrogation of small tissue blocks containing replication foci, we will define the who, where, and when of early mucosal infection, including which cells are generating virus specific alarms and which cells are responding to these alarms. This will be accomplished by incorporating new approaches for the identification of cells migrating into the infected tissue site, restriction of cell mobilization, the disruption of pathways involved in innate and inflammatory responses, and short and long term responses within the infected tissues. Advancing the goals and focus of this project will result in a substantial increase in our understanding of the earliest cascade events in vaginal and rectal transmission. In turn, clarifying this process, and the impact of local anatomy and physiology on virus expansion and dissemination, will clearly advance HIV prevention science. RELEVANCE (See instructions): Developing interventions to prevent HIV infections requires a more complete understanding of how transmission is initiated and progresses to cause systemic infection. The studies proposed here will leverage state-of-the-art methodologies that can identify foci of viral replication that can be defined and characterized to guide optimization of HIV prevention approaches.

项目总监/首席研究员（最后、第一、中间）：Hope, Thomas J. HIV 预防科学的最新进展包括证明长效药物的 PreP 功效 同样，还有可注射的 Cabotegravir 和针对敏感菌株的广泛中和抗体。 凭借新型免疫原、佐剂和递送策略，HIV 疫苗科学不断取得进展 增强对病毒的免疫反应及其预防全身感染的能力。 对这些预防性干预措施的微调以及我们提高其效力的能力需要更好的 该项目的主要重点是了解艾滋病毒性传播的机制。 1）开发和优化方法，以识别感染后第一个被感染的细胞 粘膜挑战；2) 感染灶扩大的特征，以及 3) 感染的定义 当病毒在可检测到之前传播时，在日食阶段发生的一系列事件 该项目独特地影响并增进了我们对详细自然历史的理解。 这一成功是创新的结果。 信标引导尸检方法，通过受感染细胞的存在产生信号。 该技术的第一个版本利用了由复制缺陷双报告载体表达的荧光素酶。 然而，我们已经开发了这种方法的下一代，它使用 64Cu 标记的 FAB2 探针 专门针对 SIV 包膜蛋白和 PET/CT 作为下一代信标引导尸检。 该方法高度灵敏且高效，可以公正地识别区域内的多个病灶 同一动物的全身水平，包括病毒与宿主相互作用的特征 先天性和炎症反应的研究。 传播揭示了不同感染细胞和局部组织环境之间复杂的串扰， 这一观察结果和其他观察结果表明，同一动物的不同区域和组织可能会有所不同。 靶细胞的敏感性，而不是病毒包膜的“向性”，是早期感染的关键驱动因素。 很明显，病毒暴露的粘膜组织的局部解剖学和生理学对 通过对小组织块的询问来了解病毒在日食阶段的自然历史。 包含复制灶，我们将定义早期粘膜感染的人员、地点和时间，包括 哪些细胞正在产生病毒特定的警报以及哪些细胞正在对这些警报做出反应。 通过采用新方法来识别迁移到感染细胞中的细胞来完成 组织部位、细胞动员的限制、先天性和炎症相关途径的破坏 反应，以及受感染组织内的短期和长期反应，推进目标和重点。 这个项目的开展将大大增加我们对最早的级联事件的了解 进而阐明这一过程以及局部解剖学和直肠传播的影响。 生理学对病毒扩张和传播的研究，将明显推进艾滋病毒预防科学。 相关性（参见说明）： 制定预防艾滋病毒感染的干预措施需要更全面地了解如何 传播开始并进展导致全身感染。 利用最先进的方法来识别病毒复制的焦点，这些复制焦点可以被定义和 旨在指导艾滋病毒预防方法的优化。

项目成果

Quantitative Immunofluorescent Imaging of Immune Cells in Mucosal Tissues.

粘膜组织中免疫细胞的定量免疫荧光成像。

• 发表时间: 2022
• 作者: Buchanan, Lane B;Shao, Zhongtian;Jiang, Yuan Chung;Lai, Abbie;Hope, Thomas J;Carias, Ann M;Prodger, Jessica L
• 通讯作者: Prodger, Jessica L