Detection and Characterization of Blood-borne Prions

血源性朊病毒的检测和表征

• 批准号: 9277356
• 负责人: Candace K. Mathiason
• 金额: $ 45.47万
• 依托单位: COLORADO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-06-01 至 2019-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9277356
• 关键词: Address; Animals; B-Lymphocytes; Biological; Biological Assay; Biology; Blood; Blood Cells; Blood Platelets; Blood Transfusion; Blood specimen; Brain; Cattle; Cell Fraction; Cells; Centrifugation; Chronic Wasting Disease; Chronology; Collection; Creutzfeldt-Jakob Syndrome; Deer; Dendritic Cells; Detection; Development; Disease; Early Diagnosis; Exhibits; Generations; Goals; Goat; Government Agencies; Hamsters; Harvest; Health; Hematogenous; Hematogenous Spread; Human; In Vitro; Infection; Intervention; Laboratories; Lethal Dose 50; Leukocytes; Methodology; Methods; Outcome; Pathogenesis; Patients; Phase; Phenotype; Population; PrP; Primates; Prion Diseases; Prions; Property; Research; Rodent; Safety; Sampling; Scrapie; Sheep; Sorting - Cell Movement; Surveillance Program; System; Terminal Disease; Testing; Therapeutic; Time; Transgenic Mice; Variant; Whole Blood; Work; cell type; cervid; cost; cost effective; effective therapy; farmer; improved; in vitro Assay; in vivo; in vivo Model; magnetic beads; prevent; protein misfolding; protein misfolding cyclic amplification; prototype; public health relevance; repository; trafficking; transmission process

DESCRIPTION (provided by applicant): The commonalities between human variant Creutzfeldt-Jakob Disease (vCJD), cervid chronic wasting disease (CWD) and sheep scrapie are quite remarkable. Hematogenous prions exhibiting association with the lymphoreticular system (LRS) are recapitulated in both human and animal transmissible spongiform encephalopathies (TSEs). In particular, vCJD, CWD and scrapie are all characterized by a long asymptomatic phase of disease from which infectious blood-borne prions efficiently transmit from one host to another. Because these TSEs traffic and disseminate similarly within susceptible hosts, the use of animal prion diseases provides a unique opportunity to evaluate the biology of blood-borne prions and their spread. The long-term goal of this research in my laboratory is to develop intervention strategies to detect, mitigate and prevent hematogenous prion spread. As a step towards this goal, we propose the use of animal TSEs to better define and understand the mechanisms associated with trafficking, dissemination and transmission of blood-borne prions. The use of animal TSEs will allow what has not- and cannot- be done with humans infected with vCJD, namely collect serial longitudinal blood samples from minutes after infection to terminal disease. The unifying hypothesis for this work is that blood-borne prions circulate within the host on/in specific cell phenotypes soon after infection and are responsible for early dissemination within the host and covert transmission between hosts. To test this hypothesis, we: 1) adapted fast, sensitive and cost effective in vitro conversion assays for the detection of blood-borne prions; 2) assembled a unique repository of longitudinal blood samples from natural and experimental TSE-infected cervids, rodents and primates; and 3) established facilities and collaborators with the capacity to house and assess TSEs in cervid and rodent in vivo models. We propose to address two specific aims, to 1) determine the temporal status of prionemia in TSE- infected hosts and 2) quantitatively determine which blood compartments and cell phenotypes harbor prions. We will employ blood, from naturally and experimentally infected animals (CWD-infected cervids, experimentally vCJD, BSE and CWD-infected primates and TME-infected hamsters) to investigate the efficacy of our recently established PrP conversion assay, whole blood real-time quaking induced conversion (wbRT- QuIC). The outcome of this work will establish the temporal status of prionemia as well as discern which compartments and specific cell phenotypes harbor prions. These results will be compared, where appropriate, to protein misfolding cyclic amplification and bioassay methods in place in my laboratory. These aims will provide what has long been sought in the prion field; elucidating the mechanisms of prion trafficking and dissemination throughout the host. This will permit timely detection, and thus, more effective therapies for human TSEs and other human protein misfolding diseases.

描述（申请人提供）：人类变异型克雅氏病（vCJD）、宫颈慢性消耗病（CWD）和羊痒病之间的共同点非常显着。与淋巴网状系统（LRS）相关的血源性朊病毒在人类和动物传染性海绵状脑病（TSE）中都有体现。特别是，vCJD、CWD 和痒病的特点都是疾病的长期无症状期，传染性血源性朊病毒从一个宿主有效地传播到另一个宿主。由于这些 TSE 在易感宿主体内的传播和传播方式类似，因此动物朊病毒疾病的使用为评估血源性朊病毒的生物学及其传播提供了独特的机会。我实验室这项研究的长期目标是制定干预策略来检测、减轻和预防血源性朊病毒传播。作为实现这一目标的一步，我们建议使用动物 TSE 来更好地定义和理解与血源性朊病毒的贩运、传播和传播相关的机制。动物 TSE 的使用将实现对感染 vCJD 的人类无法做到、也不可能做到的事情，即收集从感染后几分钟到疾病末期的连续纵向血液样本。这项工作的统一假设是，血源性朊病毒在感染后不久在宿主内以特定细胞表型循环，并负责宿主内的早期传播和宿主之间的秘密传播。为了检验这一假设，我们：1）采用快速、灵敏且具有成本效益的体外转换测定法来检测血源性朊病毒； 2) 建立了一个独特的纵向血液样本库，其中包含自然和实验感染 TSE 的鹿科动物、啮齿动物和灵长类动物； 3) 建立有能力在鹿科动物和啮齿动物体内模型中容纳和评估 TSE 的设施和合作者。我们建议解决两个具体目标，1）确定 TSE 感染宿主中朊病毒血症的时间状态，2）定量确定哪些血液区室和细胞表型含有朊病毒。我们将使用来自自然和实验感染动物（CWD 感染的鹿科动物、实验性 vCJD、BSE 和 CWD 感染的灵长类动物以及 TME 感染的仓鼠）的血液来研究我们最近建立的 PrP 转换测定、全血实时颤动的功效诱导转化（wbRT-QuIC）。这项工作的结果将确定朊病毒血症的时间状态，并辨别哪些区室和特定细胞表型含有朊病毒。在适当的情况下，这些结果将与我实验室现有的蛋白质错误折叠循环扩增和生物测定方法进行比较。这些目标将提供朊病毒领域长期以来所寻求的东西；阐明朊病毒在整个宿主体内贩运和传播的机制。这将有助于及时检测，从而为人类 TSE 和其他人类蛋白质错误折叠疾病提供更有效的治疗。