Identification of the initial cells infected by West Nile virus ex vivo and in vivo

离体和体内西尼罗河病毒感染的初始细胞的鉴定

基本信息

批准号：
10595385
负责人：
Jean Kyou Lim
金额：
$ 55.75万
依托单位：
ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-21 至 2027-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10595385
关键词：
Arbovirus Encephalitis Architecture Bite Blood CD4 Positive T Lymphocytes Cell Lineage Cell surface Cells Collecting Cell Culicidae Data Dendritic Cells Deposition Development Engineering Enterobacteria phage P1 Cre recombinase Event Flow Cytometry Gastrointestinal tract structure Genome Germ Cells Hematopoietic Human Immune response Immunocompetent Immunophenotyping In Situ Hybridization In Vitro Individual Infection Injections Label Lymphoid Lymphoid Cell Lymphoid Tissue MicroRNAs Modeling Mus Myelogenous Myeloid Cells Neuraxis Neurons Organ Pathogenesis Peripheral Phenotype Physiological Population Primary Cell Cultures Production RNA Site Skin Spleen Stains System T-Lymphocyte Subsets Target Populations Testing Time Tissue Model United States Vaccines Viral Pathogenesis Viremia Virulent Virus Virus Replication West Nile viral infection West Nile virus Wild Type Mouse base cell type cellular targeting chemokine cytokine draining lymph node human pathogen in vivo in vivo Model keratinocyte lymph nodes macrophage monocyte mouse model mutant neutrophil single-cell RNA sequencing

Arbovirus Encephalitis Architecture Bite Blood CD4 Positive T Lymphocytes Cell Lineage Cell surface Cells Collecting Cell Culicidae Data Dendritic Cells Deposition Development Engineering Enterobacteria phage P1 Cre recombinase Event Flow Cytometry Gastrointestinal tract structure Genome Germ Cells Hematopoietic Human Immune response Immunocompetent Immunophenotyping In Situ Hybridization In Vitro Individual Infection Injections Label Lymphoid Lymphoid Cell Lymphoid Tissue MicroRNAs Modeling Mus Myelogenous Myeloid Cells Neuraxis Neurons Organ Pathogenesis Peripheral Phenotype Physiological Population Primary Cell Cultures Production RNA Site Skin Spleen Stains System T-Lymphocyte Subsets Target Populations Testing Time Tissue Model United States Vaccines Viral Pathogenesis Viremia Virulent Virus Virus Replication West Nile viral infection West Nile virus Wild Type Mouse base cell type cellular targeting chemokine cytokine draining lymph node human pathogen in vivo in vivo Model keratinocyte lymph nodes macrophage monocyte mouse model mutant neutrophil single-cell RNA sequencing

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项目摘要

PROJECT SUMMARY: West Nile virus (WNV) is a highly virulent human pathogen of the central nervous system (CNS) and the most common cause of epidemic encephalitis in the United States. There are no vaccines or specific antiviral treatments available for WNV-infected individuals, and development of such an arsenal requires a more complete understanding of its pathogenesis. WNV is primarily transmitted through the bit of an infected mosquito. Once deposited into the skin, the virus replicates in a local draining lymph node prior to viremia, which allows the virus to spread to various organs, including the CNS, where neurons are the major targets of infection. Currently, it is unclear which cells are infected outside of the CNS, but numerous in vitro studies suggest that myeloid cells, such as monocytes/macrophages, are likely the primary targets. To begin understanding this in a more physiologically relevant model, we utilized an ex vivo human lymphoid tissue model, where WNV replicates robustly. We immunophenotyped the WNV-infected cells and found that a significant proportion were CD4+ T cells. To further validate these findings, we generated WNV mutant strains that contain cell-specific microRNA (miR) targets (miR-Ts) to restrict WNV replication in a cell-specific manner. We engineered lymphoid- or myeloid-specific miR-Ts into the genome of WNV and found that WNV strains unable to replicate in myeloid cells were able to replicate nearly identically to wild-type WNV strains, while WNV strains incapable of replicating in lymphoid cells were unable to replicate in this same model. Therefore, we hypothesize that lymphoid cells are essential initial cellular targets for WNV replication in the periphery. In this application, we seek to identify the cells that are essential for WNV replication in the human lymphoid tissue ex vivo (Aim 1) as well as in mice (Aim 2).

项目摘要：西尼罗河病毒（WNV）是中枢神经系统（CNS）和美国最常见的流行脑炎原因。没有疫苗或特定的疫苗可用于WNV感染的个体可用的抗病毒治疗，这种武器库的发展需要对其发病机理的更完整理解。 WNV主要通过感染的蚊子。一旦沉积到皮肤中，病毒就会在局部排水淋巴结中复制病毒血症，允许病毒扩散到包括CNS在内的各种器官，神经元为CNS 感染的主要靶标。目前，尚不清楚哪些细胞在中枢神经系统之外被感染，但许多细胞都在体外研究表明，髓样细胞（例如单核细胞/巨噬细胞）可能是主要靶标。为了开始在一个更加生理相关的模型中理解这一点，我们利用了一个体内人类淋巴组织模型，其中WNV稳健地复制。我们对WNV感染的细胞进行了免疫表型并发现很大的比例是CD4+ T细胞。为了进一步验证这些发现，我们生成了 WNV突变菌株包含细胞特异性microRNA（miR）靶标（miR-ts）以限制WNV复制以特异性的方式。我们在WNV的基因组中设计了淋巴样或髓样特异性miR-TS 并发现无法在髓样细胞中复制的WNV菌株能够复制几乎相同野生型WNV菌株，而WNV菌株无法在淋巴样细胞中复制在同一模型中复制。因此，我们假设淋巴样细胞是必不可少的初始细胞细胞靶标在外围复制。在此应用程序中，我们试图识别细胞对于人类淋巴组织中的WNV复制至关重要（AIM 1）以及小鼠（目标2）。