Analysis of cellular factors limiting productive JC virus infections

项目摘要 JCV和BKV是多瘤病毒家族的成员，感染了很大一部分人类人口。 在正常情况下，这两种病毒都会在肾脏中建立终身的、无症状的、持续的感染， 并经常在尿液和循环血液中检测到，没有明显的后果。而在大多数情况下 JCV和BKV是无害的，它们可以在免疫抑制的患者中重新激活，从而导致严重的 疾病。然而，这两种感染的结果都非常不同：BKV感染会导致炎症 导致肾病和出血性膀胱炎的肾脏反应和组织损伤，而JCV 重新激活会导致大脑中致命的神经退化，但只有少数几个肾病病例。影响因素 控制病毒生产性感染和持续感染之间的平衡是未知的，就像 控制不同细胞对相似多瘤病毒反应的机制。 这个应用侧重于了解相关原始人对JCV的细胞反应的基础 细胞类型，肾近端小管上皮细胞(RPTE)。与BKV相比，BKV的感染可引起广泛的 细胞死亡在5dpi左右，释放约40个感染粒子/细胞，JCV建立了低水平 感染RPTE，引起最小的细胞病变，持续至少3周，不会导致完全 生殖性感染。用JCV接种RPTE会诱导干扰素反应，这可能 防止病毒扩散。 这项研究将使用人类原代细胞来确定影响感染结果的因素：多产 或者是执着。在RPTE中JCV感染对干扰素反应的激活将使用两种方法进行评估 分子和功能方法，以确定途径的关键成分，以响应 控制病毒和限制病毒感染。然后，将单细胞转录物应用于RPTE以评估 研究细胞亚群对感染的反应，并确定在限制JCV复制方面发挥作用的基因。 最后，将BKV和JCV感染RPTE后观察到的反应进行比较 用来识别细胞基因，区分流产和繁殖感染的细胞。 这项应用试图了解临床相关的人类原代细胞对JCV的独特反应 来自肾脏的，驱动这些反应的机制，以及相同的细胞如何在不同的 流行到感染BKV。这项研究将增加我们对细胞管理原理的理解 对病毒感染的反应，以及这些反应如何影响感染的结果。

Project Summary JCV and BKV are members of the Polyomavirus family that infect a large portion of the human population. Under normal conditions both viruses establish a lifelong, asymptomatic, persistent infection in the kidneys, and are frequently detected in urine and circulating blood without apparent consequences. While in most cases JCV and BKV are harmless, they can reactivate in immunosuppressed patients, and then induce serious diseases. However, the outcome from either infection is very different: BKV infection induces an inflammatory response and tissue damage in the kidney that results in nephropathy and hemorrhagic cystitis, while JCV reactivation causes fatal neurodegeneration in the brain, but only a few nephropathy cases. The factors governing the equilibrium between viral productive infection and persistent infection are unknown, as are the mechanisms controlling the different cellular responses to similar polyomaviruses. This application focuses on understanding the basis for cellular responses to JCV in a relevant primary human cell type, renal proximal tubule epithelial cells (RPTE). In contrast with BKV, whose infection induces extensive cell death around 5 dpi and the release of about 40 infectious particles/cell, JCV establishes a low-level infection in RPTE, causing minimal cytopathic effects and persisting for at least 3 weeks without inducing a full productive infection. Inoculation of RPTE with JCV results in induction of the interferon response, which could prevent viral expansion. This research will use human primary cells to identify the factors that influence infection outcomes: productive or persistent. Activation of the interferon response by JCV infection in RPTE will be assessed using both molecular and functional approaches to identify components of the pathway that are critical for responding to the virus and for limiting viral infection. Then, single cell transcriptomics will be applied to RPTE to assess the response of cell subpopulations to infection and to identify genes that play a role in restricting JCV replication. Finally, a comparison between the responses observed upon infection of RPTE with either BKV or JCV will be used to identify cellular genes that distinguish abortively versus productively infected cells. This application seeks to understand the unique responses to JCV in a clinically relevant human primary cell from the kidneys, the mechanisms that drive those responses and how the same cells respond in a different fashion to infection with BKV. This research will increase our understanding of principles governing cellular responses to viral infection, and how these responses influence the outcome of infection.