Department of Veterinary Medicine, University of Perugia, 06126 Perugia, Italy.
Department of BioScience and Technology for Food, Agriculture, and Environment, University of Teramo, 64100 Teramo, Italy.
Viruses. 2024 Mar 6;16(3):408. doi: 10.3390/v16030408.
Hepatitis E represents an emerging zoonotic disease caused by the Hepatitis E virus (HEV), for which the main route of transmission is foodborne. In particular, infection in humans has been associated with the consumption of contaminated undercooked meat of pig origin. The aim of this study was to apply comparative proteomics to determine if porcine liver protein profiles could be used to distinguish between pigs seropositive and seronegative for HEV. Preliminarily, an ELISA was used to evaluate the presence of anti-HEV antibodies in the blood serum of 136 animals sent to slaughter. Among the analyzed samples, a seroprevalence of 72.8% was estimated, and it was also possible to identify 10 animals, 5 positive and 5 negative, coming from the same farm. This condition created the basis for the quantitative proteomics comparison between homogeneous animals, in which only the contact with HEV should represent the discriminating factor. The analysis of the proteome in all samples of liver exudate led to the identification of 554 proteins differentially expressed between the two experimental groups, with 293 proteins having greater abundance in positive samples and 261 more represented in negative exudates. The pathway enrichment analysis allowed us to highlight the effect of the interaction between HEV and the host biological system in inducing the potential enrichment of 69 pathways. Among these, carbon metabolism stands out with the involvement of 41 proteins, which were subjected to interactomic analysis. This approach allowed us to focus our attention on three enzymes involved in glycolysis: glucose-6-phosphate isomerase (GPI), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and fructose-bisphosphate aldolase A (ALDOA). It therefore appears that infection with HEV induced a strengthening of the process, which involves the breakdown of glucose to obtain energy and carbon residues useful for the virus's survival. In conclusion, the label-free LC-MS/MS approach showed effectiveness in highlighting the main differences induced on the porcine liver proteome by the interaction with HEV, providing crucial information in identifying a viral signature on the host metabolism.
戊型肝炎是一种由戊型肝炎病毒（HEV）引起的新兴人畜共患病，其主要传播途径为食源性传播。特别是，人类感染与食用受污染的未煮熟的猪源性肉有关。本研究旨在应用比较蛋白质组学来确定猪肝蛋白谱是否可用于区分 HEV 血清阳性和血清阴性的猪。初步采用 ELISA 法评估了 136 头送往屠宰的动物血清中抗-HEV 抗体的存在情况。在分析的样本中，估计血清阳性率为 72.8%，还可以识别来自同一农场的 10 头动物，其中 5 头为阳性，5 头为阴性。这种情况为同种动物之间的定量蛋白质组比较奠定了基础，只有与 HEV 的接触才应是区分因素。对所有肝渗出液样本的蛋白质组分析导致在两组实验样本之间鉴定出 554 种差异表达的蛋白质，其中 293 种蛋白质在阳性样本中丰度更高，而 261 种蛋白质在阴性渗出液中丰度更高。途径富集分析使我们能够突出 HEV 与宿主生物系统相互作用诱导潜在富集 69 种途径的影响。其中，碳代谢引人注目，涉及 41 种蛋白质，这些蛋白质进行了互作分析。这种方法使我们能够将注意力集中在参与糖酵解的三种酶上：葡萄糖-6-磷酸异构酶（GPI）、甘油醛-3-磷酸脱氢酶（GAPDH）和果糖-1,6-二磷酸醛缩酶 A（ALDOA）。因此，感染 HEV 似乎诱导了该过程的加强，该过程涉及葡萄糖的分解以获得能量和对病毒生存有用的碳残基。总之，无标记 LC-MS/MS 方法在突出 HEV 与猪肝蛋白质组相互作用诱导的主要差异方面显示出有效性，为宿主代谢中鉴定病毒特征提供了关键信息。
