Marcelino Isabel, Holzmuller Philippe, Coelho Ana, Mazzucchelli Gabriel, Fernandez Bernard, Vachiéry Nathalie
CIRAD, UMR ASTRE, F-97170 Petit-Bourg, Guadeloupe, France.
ASTRE, CIRAD, INRAE, Université de Montpellier (I-MUSE), 34000 Montpellier, France.
Microorganisms. 2021 May 26;9(6):1144. doi: 10.3390/microorganisms9061144.
The Rickettsiales , the causal agent of the fatal tick-borne disease Heartwater, induces severe damage to the vascular endothelium in ruminants. Nevertheless, induced pathobiology remains largely unknown. Our work paves the way for understanding this phenomenon by using quantitative proteomic analyses (2D-DIGE-MS/MS, 1DE-nanoLC-MS/MS and biotin-nanoUPLC-MS/MS) of host bovine aorta endothelial cells (BAE) during the in vitro bacterium intracellular replication cycle. We detect 265 bacterial proteins (including virulence factors), at all time-points of the replication cycle, highlighting a dynamic bacterium-host interaction. We show that infection modulates the expression of 433 host proteins: 98 being over-expressed, 161 under-expressed, 140 detected only in infected BAE cells and 34 exclusively detected in non-infected cells. Cystoscape integrated data analysis shows that these proteins lead to major changes in host cell immune responses, host cell metabolism and vesicle trafficking, with a clear involvement of inflammation-related proteins in this process. Our findings led to the first model of infection in host cells in vitro, and we highlight potential biomarkers of infection in endothelial cells (such as ROCK1, TMEM16K, Albumin and PTPN1), which may be important to further combat Heartwater, namely by developing non-antibiotic-based strategies.
立克次氏体目是致命的蜱传疾病“心水病”的病原体，会对反刍动物的血管内皮造成严重损害。然而，其引发的病理生物学机制在很大程度上仍不为人知。我们的研究通过对体外细菌细胞内复制周期中的宿主牛主动脉内皮细胞（BAE）进行定量蛋白质组分析（二维差异凝胶电泳-串联质谱、一维电泳-纳升液相色谱-串联质谱和生物素-纳升超高效液相色谱-串联质谱），为理解这一现象铺平了道路。我们在复制周期的所有时间点检测到了265种细菌蛋白（包括毒力因子），突出了细菌与宿主之间动态的相互作用。我们发现感染会调节433种宿主蛋白的表达：98种蛋白过度表达，161种蛋白表达不足，140种蛋白仅在感染的BAE细胞中检测到，34种蛋白仅在未感染的细胞中检测到。Cystoscape综合数据分析表明，这些蛋白会导致宿主细胞免疫反应、宿主细胞代谢和囊泡运输发生重大变化，炎症相关蛋白在此过程中明显发挥了作用。我们的研究结果得出了体外宿主细胞感染的首个模型，并且我们突出了内皮细胞感染的潜在生物标志物（如ROCK1、TMEM16K、白蛋白和蛋白酪氨酸磷酸酶N1），这对于进一步对抗心水病可能很重要，具体而言是通过开发非抗生素策略来实现。
