Early Response to Hemorrhagic Fever-Causing Arenaviruses

描述（由申请人提供）：我们的申请将满足诊断需求，以识别强毒性出血热病毒，拉沙热病毒（LASV）的感染。在进入人体的几个小时内，病原体引起循环白细胞的反应。病原体改变细胞基因表达的能力可以通过分析细胞RNA来监测。我们的方法是将白细胞的标准化培养物暴露于病原体中，并通过基因微阵列监测细胞mRNA表达的变化。筛选药剂的初步研究揭示了几种病原体特异性基因表达的变化。在这里，我们建议将我们的分析扩展到引起出血热的沙粒病毒，拉沙热病毒。我们将测试这样一个假设，即我们可以区分引起出血热的危险病毒和不引起疾病的密切相关病毒。通过监测不同时间点的基因表达反应，我们将确定病原体特异性基因表达信号出现的最早时间。这些基因表达变化可以通过分析人类和猴子外周血单个核细胞（PBMCs）的RNA得到验证，并将作为暴露于危险病原体的诊断标记。将鉴定出区分拉沙热病毒感染与相关无毒沙粒病毒感染的基因组，这些基因将用于制作更小的微阵列，以进一步简化诊断。这种诊断将消除在确定病原体之前等待症状或病原体复制的需要。这将使对生物恐怖袭击作出更适当和快速的反应成为可能。

DESCRIPTION (provided by applicant): Our application will address the need for diagnostics to identify infection with a virulent hemorrhagic fever virus, Lassa fever virus (LASV). Within hours of entering the body, pathogens elicit responses from circulating white blood cells. The pathogen's capacity to alter cellular gene expression can be monitored by analysis of cellular RNA. Our approach has been to expose a standardized culture of white blood cells to a pathogen and to monitor changes in cellular mRNA expression on gene microarrays. Preliminary studies with select agents revealed several pathogen-specific changes in gene expression. Here we propose to expand our analyses to a hemorrhagic fever-causing arenavirus, Lassa Fever virus. We will test the hypothesis that we can discriminate between a dangerous hemorrhagic fever-causing virus and closely related viruses that do not cause disease. By monitoring gene expression responses at different timepoints we will determine the earliest time at which pathogen-specific gene expression signals arise. These gene-expression changes can be validated by the analysis of RNA from human and monkey peripheral blood mononuclear cells (PBMCs), and will serve as diagnostic markers of exposure to dangerous pathogens. Sets of genes will be identified that discriminate infection with Lassa Fever virus from infection with a related avirulent arenavirus, and these genes will be used to make smaller microarrays to further streamline diagnosis. Such diagnostics would eliminate the need to wait for symptoms or for pathogen replication before identifying the pathogen. This would enable more appropriate and rapid responses to a bioterrorist attack.