Moser Ralf J, Reverter Antonio, Lehnert Sigrid A
CSIRO Livestock Industries, St Lucia 4067, Australia.
Vet Immunol Immunopathol. 2008 Feb 15;121(3-4):260-74. doi: 10.1016/j.vetimm.2007.10.007. Epub 2007 Oct 16.
The gene expression profile of peripheral blood leukocytes (PBL) from extreme performing pigs after infection with Actinobacillus pleuropneumoniae was analysed using a custom complementary DNA (cDNA) microarray and quantitative reverse transcription-PCR (qRT-PCR). Four high performing animals with low disease-score (HP), three low performing animals with high disease-score (LP) and one medium performing animal with medium disease-score (MP) were selected for microarray profiling. PBL RNA from these eight pigs collected before and at 24h after APP infection, was examined. The study identified 92 genes that were up-regulated and four genes that were down-regulated in PBL RNA from HP pigs compared to LP pigs. The majority of differentially expressed (DE) genes were identified by virtue of their elevated expression in the HP animals at 24h post-infection. A large number of annotated DE genes are involved in innate immune response pathways. The gene expression profile of 10 DE candidate genes was further explored across the entire pig population in the same infection trial using qRT-PCR. Considerable animal-to-animal variation in PBL gene expression was observed, especially in the LP group. The qRT-PCR analysis suggested that only one true LP pig might be present in this study, which contributes significantly to the differential expression profile of the selected genes in HP animals following APP infection. This study has therefore identified a set of genes which could serve as molecular indicators for an effective immune response to APP in pigs and which could also serve as source for gene marker development in molecular genetics studies of heritable immune traits.
利用定制的互补DNA（cDNA）微阵列和定量逆转录聚合酶链反应（qRT-PCR）分析了感染胸膜肺炎放线杆菌后极端高性能猪外周血白细胞（PBL）的基因表达谱。选择了4只疾病评分低的高性能动物（HP）、3只疾病评分高的低性能动物（LP）和1只疾病评分为中等的中等性能动物（MP）进行微阵列分析。检测了这8头猪在感染APP之前和感染后24小时采集的PBL RNA。该研究确定，与LP猪相比，HP猪的PBL RNA中有92个基因上调，4个基因下调。大多数差异表达（DE）基因是通过它们在感染后24小时在HP动物中表达升高而鉴定出来的。大量注释的DE基因参与先天免疫反应途径。在同一感染试验中，使用qRT-PCR在整个猪群中进一步探索了10个DE候选基因的基因表达谱。观察到PBL基因表达在动物之间存在相当大的差异，尤其是在LP组。qRT-PCR分析表明，本研究中可能只存在一头真正的LP猪，这对APP感染后HP动物中所选基因的差异表达谱有显著贡献。因此，本研究确定了一组基因，这些基因可作为猪对APP有效免疫反应的分子指标，也可作为遗传性免疫性状分子遗传学研究中基因标记开发的来源。
