Early Response to Agents Causing Flu Like Symptoms

描述（由申请人提供）：我们的申请将解决区分常见流感样病原体与生物恐怖分子制剂的诊断需求。我们的方法是使用微阵列检测暴露后数小时内血细胞RNA的变化。一组基因将被识别出来，这些基因将被用来制作一个小的微阵列，以进一步简化诊断。这种方法将消除等待症状或病原体复制的需要。 在暴露于病原体的数小时内，白色血细胞通过改变其基因表达做出反应。来自白色血细胞的RNA将被转录成cDNA，并与监测细胞mRNA表达变化的标准化基因微阵列杂交。虽然大多数反应是非特异性的（如干扰素对病原体核酸的反应），但一个重要的子集是特异性的，并可能用于区分暴露于一种病原体和另一种病原体。初步数据表明，特定的基因表达模式，在几个生物恐怖分子代理。我们将使用培养的人淋巴细胞：单核细胞（4：1）监测对4种不同病毒（甲型流感病毒、副流感病毒1型、鼻病毒和呼吸道合胞病毒）的mRNA反应。我们将建立一个基因表达反应的数据库，这些数据库对这些病毒具有诊断性和特异性。这些发现将通过与来自人类受试者的PBMC中的基因表达进行比较来验证。

DESCRIPTION (provided by applicant): Our application will address the need for diagnostics to discriminate common flu-like pathogens from bioterrorist agents. Our approach is to use microarrays to detect changes in blood cell RNA within hours of exposure. Sets of genes will be identified that discriminate one pathogen from another, and these genes will be used to make a small microarray to further streamline diagnosis. Such an approach will eliminate the need to wait for symptoms or for pathogen replication. Within hours of exposure to a pathogen, white blood cells respond by altering their gene expression. RNA from white blood cells will be transcribed into cDNA and hybridized to standardized gene microarrays that monitor changes in cellular mRNA expression. Although the majority of the responses are non-specific (such as interferon responses to pathogen nucleic acid), an important subset is specific and potentially diagnostic for discriminating between exposure to one pathogen and another. Preliminary data demonstrate specific gene expression patterns in response to several bioterrorist agents. We will use cultured human lymphocytes: monocytes (4:1) to monitor mRNA responses to 4 different viruses causing flu-like symptoms (Influenza A, Parainfluenza 1, Rhinovirus, and Respiratory Syncytial virus). We will establish a database of gene expression responses that are diagnostic and specific for those viruses. These findings will be validated by comparison with gene expression in PBMC from human subjects.