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阐明与流行病BSE相关的感染性VCJD药物的性质：感染力传播，如何准确和快速地介绍它，以及如何确定该VCJD代理。这里开发的方法和模型对于公共卫生和预防至关重要。他们还为解决这些传染性剂的基本分子成分和基本结构奠定了基础。 TSE代理有许多不同的菌株，尤其是VCJD是有问题的，因为它证明了包括人类在内的许多物种的毒力。它还能够通过从无症状捐助者的输血中从人到一个人传播，这是引起关注的另一个原因。此外，VCJD代理在通过许多不同的物种中保留其特征和身份，因此VCJD的动物模型与解决人口中这种传染性药物的传播和诊断高度相关。该R21试图建立新的实验性动物和组织培养模型，以更彻底地定义VCJD感染剂，尤其是其在不同组织中传播的能力，在不同的组织中无法通过晚期病理标记（例如异常的宿主prion蛋白（PRP））检测到它。我们还寻求使用创新的共同文化策略来找到更快的方法来检测该代理。根据耶鲁大学的初步数据，我们已经开发了迄今为止VCJD的最快动物模型，并且该鼠标模型可以为VCJD代理特定特定特征的传播特征提供关键信息。此外，这里有限的初步研究强烈表明VCJD药物可以在简化的鼠起源组织培养物中感染并复制。这种培养模型为解决复杂的退化脑组织无法解析传染剂开辟了新的机会。该应用的目的1）旨在验证动物和培养VCJD模型的代理特异性，2）进一步在培养中开发创新的超级感染测试，用于对VCJD剂的特定菌株诊断，以及3）使用直接培养和超级感染方法来快速揭示出诸如血液中缺乏感染的样本的快速感染，但缺乏病态的病态，但这些样本缺乏病理学。 4）我们还将在超大型结构上检查VCJD感染的培养物是否包含25nm病毒样颗粒，这些颗粒已在许多受TSE感染但无法控制的大脑中进行了记录。我们在具有较高的冰草感染性滴度的培养物中发现了可比较的病毒样颗粒，因此我们怀疑这种颗粒还可以帮助对这些不可避免的致命感染进行诊断，预防和基本了解。