PMCA Detection of CWD Infection in Cervid and Non-Cervid Species

描述(申请人提供)：鹿和麋鹿的慢性消耗性疾病(CWD)是一种新出现的高度传染性的普恩病毒疾病，现已在18个州、2个加拿大省份和韩国得到确认。我们已经证明，受感染的鹿在唾液、血液、尿液和粪便以及产生这些体液和排泄物的组织中藏匿和释放高水平的传染性病毒，从而导致通过直接接触和环境污染容易传播。我们还表明CWD可以感染一些非鹿类物种，因此CWD对家畜和人类构成的潜在风险尚不清楚。血液、唾液、鼻液、牛奶或排泄物中携带的Prion是在邻近的外周组织部位产生或修饰的，可能与脑中产生的有微妙的不同，或者可能被适应于粘膜感染，但仍是悬而未决的问题。Pron疾病与人类神经退行性疾病(如阿尔茨海默病和帕金森病)之间的发病机制越来越相似，这增加了CWD作为一种可传递的蛋白质错误折叠疾病的相关性。这项工作的总体目标是通过解决以下问题来阐明CWD Prion从粘膜分泌和排泄组织部位传播的过程：(A)暴露后CWD Prion脱落的动力学和大小是什么？(B)在生物化学方面，排泄出来的普恩病毒与中枢神经系统中的普恩病毒是否不同？(C)外周上皮组织或中枢神经系统组织，或两者兼而有之，是排泄普恩的来源？以及(D)排出的普恩病毒是否适合通过自然/跨粘膜途径水平传播？这项建议的具体目的是：(1)确定CWD Prion在鹿和宫颈切割小鼠体内的开始和脱落的一致性；(2)比较排泄物和CNS Prion的生化和生物物理性质；(3)确定外周组织支持CWD Prion复制的能力；(4)确定排泄物和CNS Prion对蛋白酶敏感的感染部分；以及(5)比较排泄物和CNS Prion的粘膜感染性。理解能够有效地传播和脱落普里子的机制将有助于阐明水平可传递的普恩是如何进化和成功的，这也是这一提议的基础。了解传染性错误折叠蛋白(普恩)是如何产生、贩运、脱落和传播的，将有助于预防、治疗和管理与这些病原体及其引起的疾病相关的风险。

DESCRIPTION (provided by applicant): Chronic wasting disease (CWD) of deer and elk is an emerging highly transmissible prion disease now recognized in 18 States, 2 Canadian provinces, and Korea. We have shown that Infected deer harbor and shed high levels of infectious prions in saliva, blood, urine, and feces, and in the tissues generating those body fluids and excreta, thereby leading to facile transmission by direct contact and environmental contamination. We have also shown that CWD can infect some non-cervid species, thus the potential risk CWD represents to domestic animal species and to humans remains unknown. Whether prions borne in blood, saliva, nasal fluids, milk, or excreta are generated or modified in the proximate peripheral tissue sites, may differ in subtle ways from those generated in brain, or may be adapted for mucosal infection remain open questions. The increasing parallels in the pathogenesis between prion diseases and human neurodegenerative conditions, such as Alzheimer's and Parkinson's diseases, add relevance to CWD as a transmissible protein misfolding disease. The overall goal of this work is to elucidate the process of CWD prion transmission from mucosal secretory and excretory tissue sites by addressing these questions: (a) What are the kinetics and magnitude of CWD prion shedding post-exposure? (b) Are excreted prions biochemically distinct, or not, from those in the CNS? (c) Are peripheral epithelial or CNS tissues, or both, the source of excreted prions? and (d) Are excreted prions adapted for horizontal transmission via natural/trans-mucosal routes? The specific aims of this proposal are: (1) To determine the onset and consistency of CWD prion shedding in deer and cervidized mice; (2); To compare the biochemical and biophysical properties of excretory vs. CNS prions; (3) To determine the capacity of peripheral tissues to support replication of CWD prions; (4) To determine the protease- sensitive infectious fraction of excreted vs. CNS prions; and (5) To compare the mucosal infectivity of excretory vs. CNS prions. Understanding the mechanisms that enable efficient prion dissemination and shedding will help elucidate how horizontally transmissible prions evolve and succeed, and is the basis of this proposal. Understanding how infectious misfolded proteins (prions) are generated, trafficked, shed, and transmitted will aid in preventing, treating, and managing the risks associated with these agents and the diseases they cause.