New Animal and Culture Models to rapidly evaluate infectivity of the vCJD Agent

新的动物和培养模型可快速评估 vCJD 病原体的感染性

基本信息

批准号：
7642506
负责人：
laura MANUELIDIS
金额：
$ 20.69万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-06-20 至 2010-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7642506
关键词：
Address Amino Acid Sequence Animal Model Animals Biological Assay Biology Blood Blood specimen Brain Breeding Cattle Cell Line Cells Characteristics Chronic Coculture Techniques Complex Creutzfeldt-Jakob Syndrome Cultured Cells Data Development Diagnosis Diagnostic Epidemic Experimental Animal Model Experimental Models Goals Human In Vitro Individual Infection Infectious Agent Laboratories Link Meat Milk Modeling Molecular Mus Nature Nerve Degeneration Pathogenesis Peptide Hydrolases Peptide Sequence Determination Persons Pilot Projects Population Prevention Prion Diseases Prions Property Public Health Recombinants Resistance Resolution Rodent Model Sampling Sampling Studies Scrapie Scrapie Virus Sheep Specificity Staging Structure Testing Time Tissue Transplantation Tissues Titrations Transfusion Variant Virulence Virulent Virus-like particle Western Blotting Work animal tissue base bone meal brain tissue feeding in vitro Model innovation mind control mouse model nervous system disorder novel particle rapid detection superinfection tissue culture transmission process

项目摘要

DESCRIPTION (provided by applicant): The broad and long-term objective of this project is to clarify the nature of infectious vCJD agent that is linked to epidemic BSE: how infectivity spreads, how it can be accurately and rapidly titered, and how this vCJD agent can be identified with certainty. The approaches and models under development here are critical for public health and prevention. They also lay the groundwork for resolving the essential molecular components and fundamental structure of these infectious agents. There are many distinct strains of TSE agents, and vCJD in particular is problematic because of its proven virulence for many species, including humans. It has also been able to transmit from person to person by transfusion from asymptomatic donors, a further reason for concern. The vCJD agent, moreover, retains its characteristics and identity after passage in many different species, and therefore animal models of vCJD are highly relevant for addressing the spread and diagnosis of this infectious agent in the human population. This R21 seeks to establish new experimental animal and tissue culture models to define the vCJD infectious agent more thoroughly, particularly its ability to spread in different tissues where it is not detectable by late pathological markers such as abnormal host prion protein (PrP). We also seek to find faster ways to detect this agent using innovative co-culture strategies. Based on preliminary data at Yale, we have developed the quickest animal model of vCJD to date, and this mouse model can yield critical information for vCJD agent-specific features of spread. Additionally, limited pilot studies here strongly suggest the vCJD agent can infect and replicate in simplified tissue cultures of murine origin. This culture model opens new opportunities for the resolution of the infectious agent that are not possible using complex degenerating brain tissue. This application aims 1) to verify the agent-specificity of both the animal and culture vCJD models, 2) to further develop innovative superinfection tests in culture for strain-specific diagnosis of the vCJD agent, and 3) to use direct culture and superinfection approaches to rapidly reveal infectivity in samples such as blood that are verifiably infectious, but that lack pathological PrP. 4) We will also examine vCJD-infected cultures ultrastructurally to find if they contain the 25nm viruslike particles that have been documented in many TSE-infected, but not control brains. We have discovered comparable viruslike particles in cultures with high titers of scrapie infectivity, and thus we suspect such particles can also aid in the diagnosis, prevention and fundamental understanding of these inevitably lethal infections.

描述（由申请人提供）：该项目的广泛和长期目标是澄清与流行性 BSE 相关的传染性 vCJD 病原体的性质：传染性如何传播，如何准确、快速地测定其滴度，以及这种 vCJD 如何进行代理可以被确定地识别。这里正在开发的方法和模型对于公共卫生和预防至关重要。他们还为解析这些传染原的基本分子成分和基本结构奠定了基础。 TSE 病原体有许多不同的菌株，尤其是 vCJD，因为它已被证明对包括人类在内的许多物种具有毒力，因此问题尤其严重。它还能够通过无症状捐赠者的输血在人与人之间传播，这是令人担忧的另一个原因。此外，vCJD 病原体在许多不同物种中传代后仍保留其特征和特性，因此 vCJD 动物模型对于解决这种传染原在人群中的传播和诊断具有高度相关性。该 R21 旨在建立新的实验动物和组织培养模型，以更彻底地定义 vCJD 传染原，特别是其在不同组织中传播的能力，而在这些组织中，晚期病理标记物（例如异常宿主朊病毒蛋白 (PrP)）无法检测到它。我们还寻求使用创新的共培养策略找到更快的方法来检测这种物质。根据耶鲁大学的初步数据，我们开发了迄今为止最快的 vCJD 动物模型，该小鼠模型可以提供有关 vCJD 病原体特定传播特征的关键信息。此外，有限的试点研究强烈表明，vCJD 病原体可以在小鼠来源的简化组织培养物中感染和复制。这种培养模型为解决感染原提供了新的机会，而使用复杂的退化脑组织是不可能实现的。该应用的目的是 1) 验证动物和培养 vCJD 模型的病原体特异性，2) 进一步开发创新的培养物重复感染测试，用于 vCJD 病原体的菌株特异性诊断，以及 3) 使用直接培养和重复感染方法快速揭示血液等样本中具有传染性但缺乏病理性 PrP 的传染性。 4) 我们还将对 vCJD 感染的培养物进行超微结构检查，以确定它们是否含有 25 nm 病毒样颗粒，这些颗粒已在许多 TSE 感染者的大脑中记录下来，但在对照大脑中却没有。我们在具有高滴度痒病感染性的培养物中发现了类似的病毒样颗粒，因此我们怀疑这些颗粒也可以帮助诊断、预防和基本了解这些不可避免的致命感染。