Malvisi Michela, Palazzo Fiorentina, Morandi Nicola, Lazzari Barbara, Williams John L, Pagnacco Giulio, Minozzi Giulietta
Parco Tecnologico Padano, Lodi, Italy.
Department of Veterinary Medicine, University of Milan, Milan, Italy.
PLoS One. 2016 Oct 19;11(10):e0164461. doi: 10.1371/journal.pone.0164461. eCollection 2016.
Paratuberculosis in cattle is a chronic granulomatous gastroenteritis caused by Mycobacterium avium subsp. paratubercolosis (MAP) which is endemic worldwide. In dairy herds, it is responsible for huge economic losses. However, current diagnostic methods do not detect subclinical infection making control of the disease difficult. The identification of MAP infected animals during the sub-clinical phase of infection would play a key role in preventing the dissemination of the pathogen and in reducing transmission. Gene expression and circulating microRNA (miRNA) signatures have been proposed as biomarkers of disease both in the human and veterinary medicine. In this paper, gene expression and related miRNA levels were investigated in cows positive for MAP, by ELISA and culture, in order to identify potential biomarkers to improve diagnosis of MAP infection. Three groups, each of 5 animals, were used to compare the results of gene expression from positive, exposed and negative cows. Overall 258 differentially expressed genes were identified between unexposed, exposed, but ELISA negative and positive groups which were involved in biological functions related to inflammatory response, lipid metabolism and small molecule biochemistry. Differentially expressed miRNA was also found among the three groups: 7 miRNAs were at a lower level and 2 at a higher level in positive animals vs unexposed animals, while 5 and 3 miRNAs were respectively reduced and increased in the exposed group compared to the unexposed group. Among the differentially expressed miRNAs 6 have been previously described as immune-response related and two were novel miRNAs. Analysis of the miRNA levels showed correlation with expression of their target genes, known to be involved in the immune process. This study suggests that miRNA expression is affected by MAP infection and play a key role in tuning the host response to infection. The miRNA and gene expression profiles may be biomarkers of infection and potential diagnostic of MAP infection earlier than the current ELISA based diagnostic tests.
牛副结核病是由副结核分枝杆菌(MAP)引起的一种慢性肉芽肿性肠胃炎,该病在全球范围内流行。在奶牛群中,它会造成巨大的经济损失。然而,目前的诊断方法无法检测到亚临床感染,这使得疾病的控制变得困难。在感染的亚临床阶段识别出感染MAP的动物,对于防止病原体传播和减少传播将起到关键作用。基因表达和循环微小RNA(miRNA)特征已被提议作为人类和兽医学中疾病的生物标志物。在本文中,通过酶联免疫吸附测定(ELISA)和培养,对MAP呈阳性的奶牛的基因表达和相关miRNA水平进行了研究,以确定潜在的生物标志物,从而改善MAP感染的诊断。使用三组,每组5只动物,比较阳性、暴露但ELISA阴性和阴性奶牛的基因表达结果。总体而言,在未暴露、暴露但ELISA阴性和阳性组之间共鉴定出258个差异表达基因,这些基因参与了与炎症反应、脂质代谢和小分子生物化学相关的生物学功能。在三组中也发现了差异表达的miRNA:与未暴露动物相比,阳性动物中有7种miRNA水平较低,2种水平较高,而与未暴露组相比,暴露组中有5种和3种miRNA分别减少和增加。在差异表达的miRNA中,有6种先前被描述为与免疫反应相关,2种是新的miRNA。对miRNA水平的分析表明,它们与其已知参与免疫过程的靶基因的表达相关。这项研究表明,miRNA表达受MAP感染影响,并在调节宿主对感染的反应中起关键作用。miRNA和基因表达谱可能是感染的生物标志物,并且可能比目前基于ELISA的诊断测试更早地诊断MAP感染。