Zhang Chengcheng, Wu Huimin, Li Ting, Wu Jianqiang, Zhang Xiaorong, Wu Yantao
College of Veterinary Medicine, Yangzhou University; Jiangsu Co-Innovation Center for the Prevention and Control of Important Animal Infectious Disease and Zoonoses, Yangzhou, Jiangsu 225009, China.
College of Veterinary Medicine, Yangzhou University; Jiangsu Co-Innovation Center for the Prevention and Control of Important Animal Infectious Disease and Zoonoses, Yangzhou, Jiangsu 225009, China.
Poult Sci. 2025 Jun 26;104(9):105493. doi: 10.1016/j.psj.2025.105493.
Avian reovirus (ARV) typically induces viral arthritis or tenosynovitis in chickens. The ARV structural protein σB plays a crucial role in viral replication and regulates cellular signaling pathways through interactions with host proteins. In this study, we demonstrate that both ARV infection and ARV σB protein can activate intracellular Wnt signaling pathways and induce inflammatory responses based on qPCR and Western blot analyses in HD11 cells. Interestingly, ARV infection can inhibit β-catenin ubiquitination and upregulate its protein expression levels. To further investigate the mechanism of this phenomenon, the quantity of ARV replication and the expression of inflammation cytokine IL-1β were both significantly increased when overexpression of Wnt14 protein. Conversely, shRNA-mediated knockdown of endogenous Wnt14 expression substantially suppressed ARV replication and virus-induced inflammatory responses. Furthermore, the inflammatory response was concomitantly attenuated in parallel with the suppression of ARV replication in the condition of pretreated with Wnt inhibitor. Finally, the direct interaction of ARV σB and Wnt14 protein was confirmed by using immunoprecipitation, glutathione S-transferase (GST)-pulldown assay. We further observed the colocalization of σB and Wnt14 protein by laser scanning microscopy techniques. The cellular Wnt signaling pathway regulated by ARV may mediated through this direct interaction between σB protein and Wnt14. In summary, our research provides new insights into the functional role of σB protein as well as elucidating pathogenic mechanisms associated with ARV infection-particularly its relationship with inflammatory responses.
禽呼肠孤病毒（ARV）通常会在鸡中引发病毒性关节炎或腱鞘炎。ARV结构蛋白σB在病毒复制中起关键作用，并通过与宿主蛋白相互作用调节细胞信号通路。在本研究中，基于对HD11细胞的qPCR和蛋白质印迹分析，我们证明ARV感染和ARV σB蛋白均可激活细胞内Wnt信号通路并诱导炎症反应。有趣的是，ARV感染可抑制β-连环蛋白的泛素化并上调其蛋白表达水平。为进一步探究此现象的机制，当Wnt14蛋白过表达时，ARV复制量和炎症细胞因子IL-1β的表达均显著增加。相反，shRNA介导的内源性Wnt14表达敲低显著抑制了ARV复制和病毒诱导的炎症反应。此外，在用Wnt抑制剂预处理的条件下，炎症反应随着ARV复制的抑制而同时减弱。最后，通过免疫沉淀、谷胱甘肽S-转移酶（GST）下拉试验证实了ARV σB与Wnt14蛋白的直接相互作用。我们通过激光扫描显微镜技术进一步观察到σB和Wnt14蛋白的共定位。ARV调节的细胞Wnt信号通路可能通过σB蛋白与Wnt14之间的这种直接相互作用介导。总之，我们的研究为σB蛋白的功能作用提供了新见解，并阐明了与ARV感染相关的致病机制，特别是其与炎症反应的关系。
