Pisoni G, Moroni P, Genini S, Stella A, Boettcher P J, Cremonesi P, Scaccabarozzi L, Giuffra E, Castiglioni B
Università degli Studi di Milano, Dipartimento di Patologia Animale, Igiene e Sanità Pubblica Veterinaria, via Celoria 10, 20133 Milan, Italy.
Vet Immunol Immunopathol. 2010 Jun 15;135(3-4):208-17. doi: 10.1016/j.vetimm.2009.11.016. Epub 2009 Dec 21.
To study gene expression within the mammary glands of dairy goats with mastitis, mRNA was collected from milk somatic cells (MSCs) of left udder halves challenged with Staphylococcus aureus and right udder halves infused with PBS, as control, at different time points (0, 12, 24 and 48h post-infection). Transcriptional profiles were investigated using bovine cDNA microarrays; of the total 288 differentially expressed genes identified with ANOVA analysis (False Discovery Rate=0.05, 1.5-fold change), 26, 36 and 16 genes were down-regulated at 12, 24 and 48h post-infection, respectively, while 60, 141 and 9 genes were up-regulated at the same corresponding time points. The expression profiles clearly changed at 24h post-infection with 177 genes significantly altered, corresponding to a 10-fold increase of S. aureus bacterial count in milk from infected udders. Differential expression of selected genes (CD2BP2, BCAP31, MHCII, FOSL2, MAPK13, ILT5 and JUNB) was also confirmed by real-time PCR at the different time points considered, showing high correlation with the microarray measurements and high reliability of the microarray analyses. The most readily inducible classes of genes in caprine MSCs infected with S. aureus were pro-inflammatory cytokines, chemokines and their receptors; IL-1alpha, lymphotoxin alpha, granulocyte chemotactic protein (CXCL6), and IL-2 receptor gamma were all up-regulated in infected udders versus healthy controls. This study identified a number of differentially expressed genes induced by S. aureus intramammary infection and demonstrates the intricacy of the patterns of gene expression that influence host response to a complex pathogen of significant relevance to both human and veterinary medicine.
为研究患乳腺炎的奶山羊乳腺内的基因表达情况，在不同时间点（感染后0、12、24和48小时），从用金黄色葡萄球菌攻击的左半侧乳房的乳体细胞（MSC）中收集mRNA，并从注入PBS作为对照的右半侧乳房中收集mRNA。使用牛cDNA微阵列研究转录谱；在通过ANOVA分析鉴定出的总共288个差异表达基因中（错误发现率=0.05，变化倍数为1.5倍），分别在感染后12、24和48小时有26、36和16个基因下调，而在相同的相应时间点有60、141和9个基因上调。感染后24小时表达谱明显变化，有177个基因显著改变，这与感染乳房的牛奶中金黄色葡萄球菌细菌计数增加10倍相对应。在考虑的不同时间点，通过实时PCR也证实了所选基因（CD2BP2、BCAP31、MHCII、FOSL2、MAPK13、ILT5和JUNB）的差异表达，显示出与微阵列测量高度相关以及微阵列分析的高可靠性。在感染金黄色葡萄球菌的山羊MSC中最易诱导的基因类别是促炎细胞因子、趋化因子及其受体；与健康对照相比，感染乳房中的IL-1α、淋巴毒素α、粒细胞趋化蛋白（CXCL6）和IL-2受体γ均上调。本研究鉴定了许多由金黄色葡萄球菌乳腺内感染诱导的差异表达基因，并证明了影响宿主对与人类和兽医学都有重要意义的复杂病原体反应的基因表达模式的复杂性。
