Weldearegay Yenehiwot B, Pich Andreas, Schieck Elise, Liljander Anne, Gicheru Nimmo, Wesonga Hezron, Thiaucourt Francois, Kiirika Leonard M, Valentin-Weigand Peter, Jores Joerg, Meens Jochen
Institute for Microbiology, Department of Infectious Diseases, University of Veterinary Medicine Hannover, Hannover, Germany.
Core Unit Proteomics, Hannover Medical School, Hannover, Germany.
J Proteomics. 2016 Jan 10;131:93-103. doi: 10.1016/j.jprot.2015.10.016. Epub 2015 Oct 23.
Mycoplasma mycoides subsp. mycoides (Mmm) is the causative agent of contagious bovine pleuropneumonia (CBPP), a severe pleuropneumonia in cattle. The abnormal accumulation of pleural fluid, called pleural effusion (PE), is one of the characteristics of this disease. We performed a proteomic analysis of seven PE samples from experimentally infected cattle and characterized their composition with respect to bovine and Mmm proteins. We detected a total of 963 different bovine proteins. Further analysis indicated a strong enrichment of proteins involved in antigen processing, platelet activation and degranulation and apoptosis and an increased abundance of acute phase proteins.With regard to the pathogen, up to 108 viable mycoplasma cells per ml were detected in the PE supernatant. The proteomic analysis revealed 350 mycoplasma proteins, including proteins involved in virulence-associated processes like hydrogen peroxide (H2O2) production and capsule synthesis. The bovine proteins detected will aid to characterize the inflammasome during an acute pleuropneumonia in cattle and the identified mycoplasma proteins will serve as baseline data to be compared with in vitro studies to improve our understanding of pathogenicity mechanisms. Based on our results, we named the pleural effusion an “in vivo niche” of Mmm during the acute phase of CBPP. Biological significance: This is the first study on bovine pleural effusions derived from an infectious disease and the first approach to characterize the proteome of Mycoplasma mycoides in vivo. This study revealed a high number of viable Mmm cells in the pleural effusion. The bovine pleural effusion proteome during Mmm infection is qualitatively similar to plasma, but differs with respect to high abundance of acute phase proteins. On the other hand,Mmm in its natural host produces proteins involved in capsule synthesis, H2O2 production and induction of inflammatory response, supporting previous knowledge on mechanisms underlying the survival and virulence of this pathogen while inside the natural host. This knowledge forms a profound basis for testing the identified protein candidates for diagnostics or vaccines.
丝状支原体丝状亚种(Mmm)是牛传染性胸膜肺炎(CBPP)的病原体,CBPP是一种严重的牛胸膜肺炎。胸腔积液(PE)异常积聚是该疾病的特征之一。我们对来自实验感染牛的7份PE样本进行了蛋白质组分析,并对其牛源蛋白和Mmm蛋白的组成进行了表征。我们共检测到963种不同的牛源蛋白。进一步分析表明,参与抗原加工、血小板活化与脱颗粒以及细胞凋亡的蛋白大量富集,急性期蛋白丰度增加。关于病原体,在PE上清液中每毫升检测到多达108个活支原体细胞。蛋白质组分析揭示了350种支原体蛋白,包括参与毒力相关过程如过氧化氢(H2O2)产生和荚膜合成的蛋白。检测到的牛源蛋白将有助于表征牛急性胸膜肺炎期间的炎性小体,而鉴定出的支原体蛋白将作为基线数据,与体外研究进行比较,以增进我们对致病机制的理解。基于我们的结果,我们将胸腔积液命名为CBPP急性期Mmm的“体内生态位”。生物学意义:这是第一项关于源自传染病的牛胸腔积液的研究,也是第一项在体内表征丝状支原体蛋白质组的方法。这项研究揭示了胸腔积液中有大量活的Mmm细胞。Mmm感染期间的牛胸腔积液蛋白质组在性质上与血浆相似,但在急性期蛋白高丰度方面有所不同。另一方面,Mmm在其天然宿主中产生参与荚膜合成、H2O2产生和炎症反应诱导的蛋白,支持了此前关于该病原体在天然宿主体内生存和毒力机制的认识。这些知识为测试鉴定出的蛋白质候选物用于诊断或疫苗提供了深厚的基础。