Analysis of factors controlling coxsackievirus population dynamics in the intestine

项目摘要 胃肠道(GI)是一个动态的生态系统，调节营养吸收、感染、 共生，以及在充满活力的动态平衡中的豁免权。胃肠道包括物理、化学、分子和 抵抗感染的免疫屏障。尽管存在这些障碍，包括柯萨奇病毒B3(CVB3)在内的肠道病毒， 成功地通过GI屏障启动感染，在宿主中传播，导致各种疾病 纸巾，最后在粪便中脱落。限制胃肠道的基本分子和解剖学特征 或者使宿主容易受到肠道病毒感染，这一点才刚刚开始被理解。此外，主机 屏障极大地影响病原菌的种群动态。这从根本上说很重要，因为主机内 肠道病毒种群动态直接影响发病机制。然而，胃肠道对CVB3的影响 种群动态完全没有被研究过。该项目的长期目标是定义 影响CVB3进化和发病机制的肠道分子和细胞压力。眼前的目标 本项目的主要目的是：1)精确定位胃肠道中参与CVB3复制的动力学、组织和细胞 肠道；2)评估CVB3种群动态，包括建立肠道中的种群大小；3)测试其作用 宿主因素，包括干扰素和性别，对肠道CVB3复制和种群动态的影响。这些遗嘱 具体目标：具体目标1：对柯萨奇病毒B3感染的细胞类型进行分类 通过A)通过细胞分选识别特定的感染细胞类型，B)确定 使用报告病毒的感染部位和动力学，C)评估免疫细胞在肠道感染中的作用 通过耗尽特定的细胞。具体目标2：审查柯萨奇病毒3型的种群动态和传播途径 雄性和雌性小鼠通过A)开发和验证等基因条形码CVB3毒株文库，B)创建 用组织和特定细胞病毒测序研究CVB3种群动态的时空图谱 C)消耗免疫细胞，评估CVB3的传播和传播。《知识》 在这项工作中获得的将是理解胃肠道在调节CVB3遗传学中的作用的关键一步， 进化，以及发病机制。

Project Summary The gastrointestinal (GI) tract is a dynamic ecosystem, regulating nutrient absorption, infection, commensalism, and immunity in vibrant homeostasis. The GI tract includes physical, chemical, molecular, and immune barrier to infection. Despite these barriers, enteroviruses, including coxsackievirus B3 (CVB3), successfully transit the GI barrier to initiate infection, spread throughout the host causing disease in diverse tissues, and finally be shed in stool. The essential molecular and anatomic features of the GI tract that restrict or render hosts susceptible to enterovirus infection are only beginning to be understood. Additionally, host barriers dramatically affect pathogen population dynamics. This is fundamentally important because intrahost enterovirus population dynamics directly affect pathogenesis. However, the effect of the GI tract on CVB3 population dynamics is completely unexplored. The long-term objectives for this project is to define the intestinal molecular and cellular pressures that effect CVB3 evolution and pathogenesis. The immediate goals of this project are to: 1) precisely map the kinetics, tissues, and cells involved in CVB3 replication in the GI tract; 2) assess CVB3 population dynamics, including founding population size, in the intestine; 3) test the role of host factors, including interferon and sex, on intestinal CVB3 replication and population dynamics. These will be accomplished in the following specific aims: Specific Aim 1: Categorize the cell types CVB3 infects in the intestine of male and female mice by A) identifying specific infected cell types by cell sorting, B) determining infection site and kinetics using reporter viruses, C) assessing the role of immune cells on intestinal infection by depleting specific cells. Specific Aim 2: Examine CVB3 population dynamics and routes of dissemination in male and female mice by A) developing and validating a library of isogenic barcoded CVB3 strains, B) creating a spatio-temporal atlas of CVB3 population dynamics by sequencing viruses from tissues and specific cell types, and C) depleting immune cells and assessing spread and transmission of CVB3. The knowledge gained in this work will be a critical step to understanding the role of the GI tract in modulating CVB3 genetics, evolution, and pathogenesis.