A型产气荚膜梭菌代谢产物棕榈酸增强猪肠道冠状病毒PEDV感染

doi:10.1016/j.jia.2024.05.014

摘要/Abstract

摘要： 宿主肠道微生物群已成为宿主与肠道病毒相互作用的第三要素，并可能直接或间接地影响肠道病毒的感染和致病过程。目前，PEDV与肠道菌群相互作用及其对病毒感染作用相关机制尚不清楚，限制了病毒感染和致病机制的理解以及有效预防策略的设计。在本研究中，我们利用基于16S rRNA的Illumina MiSeq高通量测序技术对猪肠道冠状病毒猪流行性腹泻病毒（porcine epidemic diarrhea virus，PEDV）感染介导仔猪肠道菌群变化进行鉴定，进一步深入研究了变化的肠道细菌对PEDV感染作用及其分子机制。结果显示，感染PEDV的仔猪肠道微生物群落多样性显著低于健康组，而且细菌群落特征也不同。在变化的肠道细菌中，PEDV感染组A型产气荚膜梭菌的相对丰度显著增加。在健康仔猪肠道中成功分离出一株A型产气荚膜梭菌，将其命名为DQ21。猪A型产气荚膜梭菌DQ21菌株的代谢产物在猪肠上皮细胞克隆J2（IPEC-J2）细胞中显著增强PEDV的复制，同时能显著增强PEDV对哺乳仔猪的感染和致病性。利用代谢组学技术在猪A型产气荚膜梭菌DQ21代谢产物中鉴定的棕榈酸（palmitic acid，PA）在IPEC-J2细胞能显著增强PEDV复制，同时能显著增强PEDV对哺乳仔猪的感染和致病性。PA还能提高小鼠免疫血清中的中和抗体滴度。进一步发现，PA可以介导PEDV S蛋白的棕榈酰化修饰，通过增强病毒的稳定性和膜融合从而促进病毒感染。我们研究揭示，PEDV在与宿主相互作用过程中进化出一种利用肠道菌群代谢产物增强自身感染的致病模式：PEDV感染引起仔猪肠道中猪A型产气荚膜梭菌丰度上调，其代谢产物PA通过介导病毒粒子S蛋白棕榈酰化修饰增强病毒粒子稳定性和病毒膜融合，进而增强病毒感染。该研究成果为PEDV致病机制和防控提供了新的思路，为肠道冠状病毒感染提供了新的理解。

Abstract: The host intestinal microbiota has emerged as the third element in the interactions between hosts and enteric viruses, and potentially affects the infection processes of enteric viruses. However, the interaction of porcine enteric coronavirus with intestinal microorganisms during infection remains unclear. In this study, we used 16S-rRNA-based Illumina NovaSeq high-throughput sequencing to identify the changes in the intestinal microbiota of piglets mediated by porcine epidemic diarrhea virus (PEDV) infection and the effects of the alterations in intestinal bacteria on PEDV infection and its molecular mechanisms. The intestinal microbiota of PEDV-infected piglets had significantly less diversity than the healthy group and different bacterial community characteristics. Among the altered intestinal bacteria, the relative abundance of Clostridium perfringens was significantly increased in the PEDV-infected group. A strain of C. perfringens type A, named DQ21, was successfully isolated from the intestines of healthy piglets. The metabolites of swine C. perfringens type A strain DQ21 significantly enhanced PEDV replication in porcine intestinal epithelial cell clone J2 (IPEC-J2) cells, and PEDV infection and pathogenicity in suckling piglets. Palmitic acid (PA) was identified as one of those metabolites with metabolomic technology, and significantly enhanced PEDV replication in IPEC-J2 cells and PEDV infection and pathogenicity in suckling piglets. PA also increased the neutralizing antibody titer in the immune sera of mice. Furthermore, PA mediated the palmitoylation of the PEDV S protein, which improved virion stability and membrane fusion, thereby enhancing viral infection. Overall, our study demonstrates a novel mechanism of PEDV infection, with implications for PEDV pathogenicity.

Key words: Porcine epidemic diarrhea virus , Clostridium perfringens , palmitic acid , palmitoylation , infection

. A型产气荚膜梭菌代谢产物棕榈酸增强猪肠道冠状病毒PEDV感染[J]. Journal of Integrative Agriculture, DOI: 10.1016/j.jia.2024.05.014.

Shanshan Qi, Haoyang Wu, Donghua Guo, Dan Yang, Yongchen Zhang, Ming Liu, Jingxuan Zhou, Jun Wang, Feiyu Zhao, Wenfei Bai, Shiping Yu, Xu Yang, Hansong, Li, Fanbo Shen, Xingyang Guo, Xinglin Wang, Wei Zhou, Qinghe Zhu, Xiaoxu Xing, Chunqiu Li, Dongbo Sun. Metabolite of Clostridium perfringens type A, palmitic acid, enhances porcine enteric coronavirus porcine epidemic diarrhea virus infection[J]. Journal of Integrative Agriculture, DOI: 10.1016/j.jia.2024.05.014.

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