Expression data of influenza A infected human macrophages

Human disease caused by highly pathogenic avian influenza (HPAI) H5N1 can lead to a rapidly progressive viral pneumonia leading to acute respiratory distress syndrome. There is increasing evidence suggests a role for virus-induced cytokine dysregulation in contributing to the pathogenesis of human H5N1 disease. The key target cells for the virus in the lung are the alveolar epithelium and alveolar macrophages, and previous data has shown that compared to seasonal human influenza viruses, equivalent infecting doses of H5N1 viruses markedly up-regulate pro-inflammatory cytokines in both primary cell types in vitro. The dysregulation of H5N1-induced host responses is therefore important for understanding the viral pathogenesis. We used microarrays to analyze and compare the gene expression profiles in primary human macrophages after influenza A virus infection. Peripheral-blood leucocytes were separated from buffy coats of three healthy blood donors and cells were differentiated for 14 days before use. Differentiated macrophages were infected with H1N1 and H5N1 at a multiplicity of infection (MOI) of two. Total RNA was extracted from cells after 1, 3, and 6h post-infection, and gene expression profiling was performed using an Affymetrix Human Gene 1.0 ST microarray platform.

高致病性禽流感（highly pathogenic avian influenza, HPAI）H5N1感染人类后引发的疾病，可导致快速进展性病毒性肺炎，进而诱发急性呼吸窘迫综合征。越来越多的研究证据表明，病毒诱导的细胞因子失调在人类H5N1感染疾病的发病机制中扮演重要角色。该病毒在肺部的主要靶细胞为肺泡上皮细胞与肺泡巨噬细胞；既往研究数据显示，与季节性人类流感病毒相比，同等感染剂量的H5N1病毒在体外原代细胞中可显著上调促炎细胞因子的表达水平。因此，解析H5N1诱导的宿主应答失调机制，对于阐明该病毒的致病机理具有重要意义。本研究采用基因芯片技术，分析并比较了甲型流感病毒（influenza A virus）感染后人原代巨噬细胞的基因表达谱。研究人员从3名健康献血者的血沉棕黄层（buffy coat）中分离外周血白细胞，并在使用前将其诱导分化14天。以感染复数（multiplicity of infection, MOI）为2的感染比例，分别用H1N1与H5N1病毒感染分化成熟的巨噬细胞。分别于感染后1、3、6小时提取细胞总RNA，并通过Affymetrix Human Gene 1.0 ST基因芯片平台完成基因表达谱分析。