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: 8264398
• 负责人: Nicholas James Haley
• 金额: $ 10.51万
• 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/8264398
• 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; 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; immunocytochemistry; 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（由申请方提供）：感染鹿慢性消耗病（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，通常超出常规检测的检测限。此外，朊病毒疾病的发病机制和传播机制尚未明确定义。发展灵敏的死前检测慢性消耗病和了解传播是至关重要的，最终控制朊病毒疾病的人和动物。我们建议研究CWD的鹿作为vCJD/朊病毒传播的模型。CWD PrPres已被证明存在于各种排泄物中，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。我们提出了三个目标，探讨朊病毒疾病的发病机制和传播：（1）在目标1中，我们将评估外周组织的早期积累的慢性消耗病朊病毒使用敏感的扩增试验，sPMCA。这些结果将与使用更传统的检测方法（如免疫组织化学）获得的结果进行比较。(2)在目标2中，我们将寻求通过使用生物测定和PMCA在多个时间点从感染的鹿科动物收集的样品来确定唾液和尿液中朊病毒脱落的动力学。将结果与外周组织中的早期检测结果（目标1）以及当前死前检测的灵敏度和特异性进行比较。(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来研究生物样品中的组织和细胞来源的感染性朊病毒。这些研究结果将有助于了解慢性消耗病、疯牛病和vCJD的发病机制和传播潜力。 公共卫生相关性（由申请人提供）：朊病毒疾病干预策略的一个关键限制是缺乏关于传播机制和动力学以及适当的死前检测的知识。此外，对排泄物（如尿液和唾液）中朊病毒的生物学性质知之甚少，识别朊病毒传播中涉及的组织和细胞类型与TSE的发病机制和诊断相关。