Plum Island Animal Disease Center, Agricultural Research Service, US Department of Agriculture, Greenport, NY 11944, United States.
Virus Res. 2010 Jul;151(1):10-8. doi: 10.1016/j.virusres.2010.03.007. Epub 2010 Mar 17.
Infection of domestic swine with highly virulent, classical swine fever virus (CSFV) strain Brescia, causes lethal disease in all infected animals. However, the molecular mechanisms involved in modulating the host cellular processes and evasion of the immune response have not been clearly established. To gain insight into, the early host response to CSFV, we analyzed the pattern of gene expression in infected swine macrophages, using custom designed swine microarrays. Macrophages, the target cell for CSFV infection, were isolated from primary cultures of peripheral blood mononuclear cells, allowing us to utilize identical uninfected macrophages at the same time points as CSFV-infected macrophages, allowing only genes induced by CSFV to be identified. First, microarray probes were optimized by screening 244,000 probes for hybridization with RNA from infected and uninfected macrophages. Probes that hybridized and passed quality control standards were used to design a 44,000 probe microarray for this study. Changes in expression levels of 79 genes (48 up- and 31 down-regulated) during the first 48h post-infection were observed. As expected many of the genes with an altered pattern of expression are involved in the development of an innate immune response. Several of these genes had differential expression in an attenuated strain NS4B.VGIv, suggesting that some of these differences are responsible for virulence. The observed gene expression profile might help to explain the immunological and pathological changes associated with infection of pigs with CSFV Brescia.
感染具有高致病性、经典猪瘟病毒（CSFV）毒株 Brescia 的家猪，会导致所有感染动物致命性疾病。然而，调节宿主细胞过程和逃避免疫反应的分子机制尚未明确建立。为了深入了解 CSFV 对宿主的早期反应，我们使用定制的猪微阵列分析了感染猪巨噬细胞中的基因表达模式。巨噬细胞是 CSFV 感染的靶细胞，从外周血单核细胞的原代培养物中分离出来，这使我们能够在相同的时间点使用与 CSFV 感染的巨噬细胞相同的未感染巨噬细胞，从而仅鉴定出由 CSFV 诱导的基因。首先，通过筛选 244000 个探针与感染和未感染的巨噬细胞 RNA 的杂交，优化了微阵列探针。杂交并通过质量控制标准的探针用于设计本研究的 44000 个探针微阵列。在感染后最初的 48 小时内，观察到 79 个基因（48 个上调和 31 个下调）的表达水平发生变化。正如预期的那样，许多表达模式发生改变的基因参与先天免疫反应的发展。其中一些基因在减毒菌株 NS4B.VGIv 中存在差异表达，表明其中一些差异与毒力有关。观察到的基因表达谱可能有助于解释与 CSFV Brescia 感染猪相关的免疫学和病理学变化。
