Early Response to Agents Causing Flu Like Symptoms

描述(申请人提供)：我们的申请将解决诊断区分普通流感病原体和生物恐怖分子病原体的需要。我们的方法是使用微阵列来检测暴露后几个小时内血细胞RNA的变化。将识别一组区分病原体和另一种病原体的基因，这些基因将被用于制造一个小型微阵列，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。 在接触病原体的几个小时内，白细胞会通过改变自己的基因表达来做出反应。来自白血球的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)，但有一个重要的子集是特异的，并有可能用于区分暴露于一个病原体和另一个病原体之间。初步数据显示，对几种生物恐怖分子毒剂的反应具有特定的基因表达模式。我们将使用培养的人淋巴细胞：单核细胞(4：1)来监测对4种引起流感样症状的不同病毒(甲型流感、副流感1、鼻病毒和呼吸道合胞病毒)的mRNA反应。我们将建立一个对这些病毒具有诊断性和特异性的基因表达反应数据库。这些发现将通过与人类受试者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.