Comparative Secretome Analyses of Mycoplasma bovis Virulent and Attenuated Strains Revealed MbovP0145 as a Promising Diagnostic Biomarker.
Comparative Secretome Analyses of Mycoplasma bovis Virulent and Attenuated Strains Revealed MbovP0145 as a Promising Diagnostic Biomarker.
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DOI：
10.3389/fvets.2021.666769
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发表时间：
2021
影响因子：
3.2
通讯作者：
Guo A
中科院分区：
文献类型：
--
作者：
Zhang H;Hu G;Lu D;Zhao G;Zhang Y;Zubair M;Chen Y;Hu C;Chen X;Chen J;Chen H;Yang L;Guo A
支原体是人类和动物中成功的病原体。牛支原体（Mycoplasma bovis，M.牛）已知是导致严重的健康并发症的原因，包括肺炎、乳腺炎和关节炎。然而，M。牛的发病机制尚不清楚。M.牛进入宿主微环境的细胞外空间后，可以影响感染并改变宿主防御信号通路。因此，本研究的目的是比较M。牛HB 0801强毒（P1）和减毒（P150）菌株，并鉴定潜在的发病相关分泌蛋白和生物标志物。将P1和P150菌株的细胞在胸膜肺炎样生物培养基中生长至对数期，然后转移至磷酸盐缓冲盐水中2小时。利用无标记定量蛋白质组学技术对培养上清进行分析，共鉴定出477个潜在的分泌蛋白。结合生物信息学预测，发现P1和P150菌株共分泌178个蛋白，其中49个由支原体核心基因编码。此外，发现79种蛋白质在P1和P150菌株之间具有不同的丰度。在这些蛋白中，有34个蛋白在P1中表达量更高，并且是唯一表达的，表明这些蛋白可能与M的毒力有关。牛分别克隆、表达了三种差异分泌蛋白MbovP0145、MbovP0725和MbovP0174，以及作为阳性对照的一种同等分泌蛋白MbovP0481和作为阴性对照的一种内膜蛋白MbovP0310，并对其抗原性、亚细胞定位、用小鼠抗血清进行免疫印迹和集落免疫印迹分析。其中MbovP0145被证实比P150菌株更高地由P1分泌，与来自自然感染和P1实验感染的牛的抗血清高度反应，而与P150免疫的小牛不反应，表明其作为诊断抗原的潜力。总之，这些发现可能代表了支原体种属中潜在分泌蛋白的最广泛的汇编，以及毒力和减毒支原体之间最大数量的差异分泌蛋白。牛菌株，并提供了新的见解M.牛的发病机制和诊断。
Mycoplasmas are successful pathogens both in humans as well as in animals. In cattle, Mycoplasma bovis (M. bovis) is known to be responsible for serious health complications, including pneumonia, mastitis, and arthritis. However, M. bovis pathogenesis remains unclear. Secreted proteins of M. bovis could influence infection and modify host defense signaling pathways after they enter their extracellular space in the host micro-environment. Therefore, this study was aimed to compare the secretomes of M. bovis HB0801 virulent (P1) and attenuated (P150) strains and identify potential pathogenesis-related secreted proteins and biomarkers. The cells of P1 and P150 strains were grown in pleuropneumonia-like organism medium to log phase and then transferred to phosphate-buffered saline for 2 h. Then, the supernatant was analyzed by using label-free quantitative proteomics, and 477 potential secreted proteins were identified. Combined with the bioinformatics prediction, we found that 178 proteins were commonly secreted by the P1 and P150 strains, and 49 of them were encoded by mycoplasmal core genes. Additionally, 79 proteins were found to have a different abundance between the P1 and P150 strains. Among these proteins, 34 were more abundant and uniquely expressed in P1, indicating a possible association with the virulence of M. bovis. Three differentially secreted proteins, MbovP0145, MbovP0725, and MbovP0174, as well as one equally secreted protein, MbovP0481, as positive control and one protein of inner membrane, MbovP0310, as negative control were, respectively, cloned, expressed, and evaluated for antigenicity, subcellular location, and the secretion nature with their mouse antisera by western blotting and colony immunoblotting assay. Among them, MbovP0145 was confirmed to be more secreted by P1 than P150 strain, highly reactive with the antisera from naturally infected and P1 experimentally infected cattle but not with the P150 vaccinated calves, indicating its potential as a diagnostic antigen. In conclusion, these findings may represent the most extensive compilation of potentially secreted proteins in mycoplasma species and the largest number of differentially secreted proteins between the virulent and attenuated M. bovis strains to date and provide new insights into M. bovis pathogenesis and diagnosis.
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影响因子：
--
作者：
Chernov VM;Chernova OA;Mouzykantov AA;Efimova IR;Shaymardanova GF;Medvedeva ES;Trushin MV
通讯作者：
Trushin MV
影响因子：
5.7
作者：
Rasheed MA;Qi J;Zhu X;Chenfei H;Menghwar H;Khan FA;Zhao G;Zubair M;Hu C;Chen Y;Chen H;Guo A
通讯作者：
Guo A
影响因子：
3.7
作者：
Maunsell F;Brown MB;Powe J;Ivey J;Woolard M;Love W;Simecka JW
通讯作者：
Simecka JW
建立抗体亲和力测试以区分接种疫苗的牛与自然感染牛支原体的牛
影响因子：
2.2
作者：
Han, Xiaoxiao;Khan, Farhan Anwar;Guo, Aizhen
通讯作者：
Guo, Aizhen
影响因子：
2.1
作者：
Sambir, Mariya;Ivanova, Larisa B.;Cabello, Felipe C.
通讯作者：
Cabello, Felipe C.