Experimental Creutzfeldt Jacob Disease

实验性克雅氏病

• 批准号: 7372582
• 负责人: laura MANUELIDIS
• 金额: $ 49.9万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1978
• 资助国家: 美国
• 起止时间: 1978-12-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7372582
• 关键词: Ambush; Animal Model; Animals; Biological; Biological Assay; Brain; Cell Cycle Arrest; Cell Line; Cells; Clinical; Code; Complex; Creutzfeldt-Jakob Syndrome; Cultured Cells; DNA; Daily; Detection; Development; Diagnostic; Disease; Drug Design; Encephalopathies; End Point; Epidemic; Evaluation; Generations; Hamsters; Health; Human; Hypothalamic structure; Immune response; In Situ; Infection; Infectious Agent; Label; Lead; Life; Longevity; Methods; Modeling; Molecular; Monitor; Morphology; Mus; Nature; Nerve Degeneration; Neuroblastoma; Neurodegenerative Disorders; Nucleic Acids; Numbers; Peptides; Pharmaceutical Preparations; Preventive; Prions; Progress Reports; Property; Proteins; RNA; Relative (related person); Research; Resolution; Scrapie; Screening procedure; Source; Staging; Statistically Significant; Structure; Sum; Testing; Therapeutic; Time; Titrations; Vaccination; Vaccines; Viral; Virion; Virulence; Virus; Virus-like particle; Western Blotting; cell type; day; improved; neuropathology; novel; novel diagnostics; particle; pathogen; prevent; research study; tissue culture; tissue processing; virus characteristic

DESCRIPTION (provided by applicant): TSEs are caused by a group of related infectious agents that produce neurodegenerative disease in many species. Some agents, such as BSE, have increased in virulence with broad epidemic spread worldwide. These infectious agents can provoke many innate immune responses at very early asymptomatic stages, and only later does one find secondary patholgocial changes in prion protein (PrP). This, as well the existence of many different strains of agent, point to a causal virus with non-host coding sequences. Previous research on our CJD animal models revealed 25nm virus like particles in highly infectious brain fractions. To better resolve the fundamental structure and composition of TSE infectious particles in-situ, and to elucidate novel biological features of different strains, we transmitted various TSE agents to cultured cells. In these highly infectious cells, that show no neurodegenerative changes, we identified 25nm particles in ordered arrays that are characteristic for viruses. They do not contain PrP, and hence are not prions. These particle arrays, moreover, correspond well with those found in a variety of infected brains, as well as with the isolated 25nm particles in brain fractions. The TSE-specific virus like arrays were identified in two different cell types infected with two very different TSE agents (FU-CJD and 22L-scrapie). We propose to develop rapid live cell culture assays of infectivity for these two TSE agents. This rapid assay of infectivity will be verified by parallel animal titrations. Currently, assays of TSE infectious agents involve long and expensive animal titrations, and a rapid cell assay is critical for improved purifications of infectious particles from secondary pathological products. We will compare cell and animal assays to understand the respective limitations and advantages of each, and then use them for fundamental studies. Increasingly purified particles, and their molecular components, will be applied to susceptible cells to demonstrate which specific structures or components are the causal infectious pathogen by Koch's principles. The isolation of highly infectious particles and the definition of their molecular components can lead to sensitive new diagnostic screening methods for preventing further spread of TSE diseases, and possibly to a vaccine with non-host TSE components. Rapid infectivity assays can also benefit the design of drugs to ambush the progressive replication of these agents before irreversible neurodegeneration begins.

描述（由申请人提供）：TSE是由一组与许多物种产生神经退行性疾病的相关传染药引起的。在全球范围内，一些代理人（例如BSE）的毒力增加了。这些传染性药物可以在很早的无症状阶段引起许多先天免疫反应，直到后来才发现prion蛋白（PRP）的继发性病理改变。这是许多不同菌株的存在，指向具有非宿主编码序列的因果病毒。对我们的CJD动物模型的先前研究表明，在高度感染性脑部分中，有25nm的病毒类似于颗粒。为了更好地解决TSE感染颗粒的基本结构和原位的基本结构和组成，并阐明了不同菌株的新生物学特征，我们将各种TSE剂传输到培养的细胞。在这些高度传染性的细胞中，没有显示神经退行性的变化，我们以具有病毒特征的有序阵列确定了25nm颗粒。它们不包含PRP，因此不是Prions。此外，这些粒子阵列与在各种感染的大脑中发现的阵列以及脑部分离的25nm颗粒。 TSE特异性病毒阵列被鉴定出在两种不同的TSE剂（FU-CJD和22L-SCRAPIE）感染的两种不同的细胞类型中鉴定出来。我们建议为这两种TSE药物开发快速的活细胞培养试验，以感染性。这种感染性的快速测定将通过平行动物滴定来验证。当前，TSE感染剂的测定涉及长而昂贵的动物滴定，快速细胞测定对于改善次级病理产物的传染性颗粒的纯化至关重要。我们将比较细胞和动物测定，以了解每个细胞的局限性和优势，然后将其用于基本研究。越来越纯化的颗粒及其分子成分将应用于易感细胞，以证明哪些特定结构或成分是科赫原理是因果感染病原体。高度传染性颗粒的分离及其分子成分的定义会导致敏感的新诊断筛查方法，以防止TSE疾病进一步扩散，甚至可能是具有非宿主TSE成分的疫苗。快速感染分析​​还可以使药物的设计受益，以在不可逆的神经变性开始之前伏击这些药物的进行性复制。