CWD: A Model of Prion Transmission via Saliva and Urine

描述(由申请人提供)：感染宫颈慢性消耗性疾病(CWD)的动物，就像人类的变异克雅氏病(VCJD)一样，其感染普恩病毒的水平通常超出了传统检测方法的检测范围。此外，PrP疾病的发病机制和传播机制还没有明确的定义。开发灵敏的CWD生前检测方法并了解传播情况，对于最终控制人和动物的普恩病毒疾病至关重要。我们建议研究CWD作为vCJD/Prion传递的模型。CWD PrPreres已被证明存在于各种排泄物中，并可在感染的鹿的各种排泄物中传播，包括唾液、血液、尿液和粪便，尽管其他生物样本也可能作为宫颈之间的传播途径。此外，这些排泄物中PrPres的可检测水平和确切来源尚未得到证实。(1)在目标1中，我们将使用一种灵敏的扩增实验sPMCA来评估CWD患者外周组织中早期聚集的Prion。这些发现将与使用免疫组织化学等更传统的分析方法得到的结果进行比较。(2)在目标2中，我们将利用生物测定和PMCA方法，通过多个时间点从感染宫颈采集样本，来确定唾液和尿液中Pron的脱落动力学。结果将与外周组织的早期检测结果(目标1)以及目前的生前检测的敏感性和特异性进行比较。(3)在目标3中，我们将使用免疫细胞和组织化学、生物测定和PMCA来研究生物样品中传染性病毒的组织和细胞来源。这些研究结果将有助于了解CWD、BSE和vCJD的发病机制和传播潜力。

DESCRIPTION (provided by applicant): The levels of infectious prions in animals infected with chronic wasting disease (CWD) in cervids, as with variant Creutzfeldt-Jakob-disease (vCJD) in human, are often beyond the limits of detection using conventional assays. In addition, the mechanisms of pathogenesis and transmission of prion diseases are not clearly defined. Development of sensitive antemortem assays for CWD and an understanding of transmission are critical for the eventual control of prion diseases of both man and animals. We propose to study CWD of cervids as a model for vCJD/prion transmission. CWD PrPres has been shown to be present and transmissible in various excreta, including saliva, blood, urine and feces of infected deer, though other biological samples may also serve as routes of transmission between cervids. In addition, detectable levels and the exact source of PrPres in these excreta have yet to be demonstrated. We propose three aims which explore the pathogenesis and transmission of prion diseases: (1) in Aim 1, we will evaluate peripheral tissues for early accumulation of CWD prions using a sensitive amplification assay, sPMCA. These findings will be compared with those achieved using more traditional assays such as immunohistochemistry. (2) In Aim 2, we will seek to determine the kinetics of prion shedding in saliva and urine through samples collected at multiple time points from infected cervids using bioassay and PMCA. Findings will be compared to results of early detection in peripheral tissues (Aim 1) as well as the sensitivity and specificity of current antemortem assays. (3) In Aim 3, we will investigate the tissue and cellular origins infectious prions in biological samples using immunocyto- and histochemistry, bioassay, and PMCA. The results of these studies will contribute greatly to the understanding of pathogenesis and transmission potential of CWD, BSE, and vCJD.