CWD: A Model of Prion Transmission via Saliva and Urine

CWD：朊病毒通过唾液和尿液传播的模型

• 批准号: 7988242
• 负责人: Nicholas James Haley
• 金额: $ 10.27万
• 依托单位: COLORADO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-06-01 至 2015-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7988242
• 关键词: Address; Animal Model; Animals; Biological; Biological Assay; Biological Models; Biopsy; Biopsy Specimen; Blinded; Blood; Centrifugation; Chronic Wasting Disease; Clinical; Creutzfeldt-Jakob Syndrome; Deer; Detection; Development; Diagnosis; Diagnostic; Disease; Early Diagnosis; Early identification; Engineering; Enzyme-Linked Immunosorbent Assay; Epidemic; Epithelial Cells; Equine mule; Evaluation; Excretory function; Experimental Models; Feces; Future; Genitourinary system; Goals; Gold; Hamsters; Histocytochemistry; Human; Immunohistochemistry; In Vitro; Individual; Infection; Infectious Agent; Intervention; Kentucky; Kinetics; Knock-out; Knowledge; Laboratories; Liquid substance; Locales; Location; Longevity; Lymphoid Tissue; Modeling; Mus; Nature; Oral; Pathogenesis; Pathologic; Pathology; Peptide Hydrolases; Peripheral; Phase; PrP; PrPCWD; Prevalence; Prion Diseases; Prions; Resistance; Role; Route; Saliva; Salivary Glands; Sampling; Scrapie; Screening procedure; Sensitivity and Specificity; Site; Source; System; Tail; Techniques; Terminal Disease; Testing; Time; Tissues; Tonsil; Transgenic Mice; Universities; Urine; Variant; Western Blotting; Work; aqueous; base; cell type; cervid; cytochemistry; density; design; disease transmission; expression vector; in vitro Bioassay; insight; man; minimally invasive; novel; protein misfolding cyclic amplification; public health relevance; rectal; transmission process

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. PUBLIC HEALTH RELEVANCE (provided by applicant): A critical limitation of prion disease intervention strategies is the lack of knowledge regarding mechanisms and dynamics of transmission and a suitable antemortem assay. Additionally, little is known about the biological nature of prions in excreta (e.g. urine and saliva), and identifying the tissues and cell types involved in prion transmission is relevant to both the pathogenesis and diagnosis of TSE's.

描述（由申请人提供）：感染鹿科动物慢性消耗性疾病（CWD）的动物中的传染性朊病毒水平，与人类变异型克雅氏病（vCJD）一样，通常超出了常规测定的检测极限。此外，朊病毒疾病的发病机制和传播机制尚不明确。开发灵敏的 CWD 生前检测方法和了解传播情况对于最终控制人类和动物的朊病毒疾病至关重要。我们建议研究鹿的 CWD 作为 vCJD/朊病毒传播的模型。 CWD PrPres 已被证明存在于各种排泄物中并可传播，包括受感染鹿的唾液、血液、尿液和粪便，尽管其他生物样本也可能作为鹿科动物之间的传播途径。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。我们提出了三个探索朊病毒疾病发病机制和传播的目标：（1）在目标 1 中，我们将使用灵敏的扩增测定 sPMCA 评估外周组织中 CWD 朊病毒的早期积累。这些发现将与使用更传统的检测（例如免疫组织化学）所获得的结果进行比较。 (2) 在目标 2 中，我们将利用生物测定和 PMCA 在多个时间点从受感染的鹿科动物中收集样本，寻求确定唾液和尿液中朊病毒脱落的动力学。研究结果将与外周组织早期检测结果（目标 1）以及当前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化学和组织化学、生物测定和 PMCA 研究生物样品中感染性朊病毒的组织和细胞起源。这些研究的结果将极大地有助于了解 CWD、BSE 和 vCJD 的发病机制和传播潜力。 公共卫生相关性（由申请人提供）：朊病毒病干预策略的一个关键限制是缺乏有关传播机制和动力学以及合适的生前测定的知识。此外，人们对排泄物（例如尿液和唾液）中朊病毒的生物学性质知之甚少，识别参与朊病毒传播的组织和细胞类型与 TSE 的发病机制和诊断相关。