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， 以及免疫系统的动态平衡。胃肠道包括物理的、化学的、分子的和 抵抗感染的免疫屏障。尽管有这些障碍，肠道病毒，包括柯萨奇病毒B3（CVB 3）， 成功地通过胃肠道屏障开始感染，在宿主中传播，引起多种疾病， 组织，并最终在粪便中脱落。胃肠道的基本分子和解剖特征限制了 或使宿主易受肠病毒感染的原因才刚刚开始被了解。此外，host 屏障显著影响病原体种群动态。这一点非常重要，因为主机内部 肠道病毒种群动态直接影响致病机制。然而，胃肠道对CVB 3的影响 人口动态是完全未知的。该项目的长期目标是确定 肠道分子和细胞压力影响CVB 3演变和发病机制。近期目标 该项目的主要目的是：1）精确绘制CVB 3在GI中复制的动力学、组织和细胞 2）评估肠道中CVB 3群体动力学，包括发现群体大小; 3）测试CVB 3在肠道中的作用。 包括干扰素和性别在内的宿主因素对肠道CVB 3复制和种群动态的影响。这些将 具体目标1：将CVB 3感染的细胞类型分类， 通过A）通过细胞分选鉴定特定的感染细胞类型，B）确定 C）评估免疫细胞对肠道感染的作用 通过消耗特定的细胞。具体目标2：研究CVB 3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化的CVB 3菌株的文库，B）产生 通过对组织和特定细胞中的病毒进行测序， C）消耗免疫细胞并评估CVB 3的扩散和传播。知识 在这项工作中获得的将是理解胃肠道在调节CVB 3遗传学中的作用的关键一步， 进化和发病机制。

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.