Data_Sheet_1_Autophagy Is a Potential Therapeutic Target Against Duck Tembusu Virus Infection in vivo.docx

Duck tembusu virus (DTMUV) is newly emerged in poultry and causes great losses to the breeding industry in China and neighboring countries. Effective antiviral strategies are still being studied. Autophagy is a cellular degradative pathway, and our lab's previous data show that autophagy promotes DTMUV replication in vitro. To study the role of autophagy further in vivo, we utilized ducks as the animal model to investigate the autophagy responses in DTMUV-targeted tissues. And also, we utilized autophagy regulators, including Rapamycin (Rapa) as the autophagy enhancer, 3-Methyladenine (3-MA) and Chloroquine (CQ) as the autophagy inhibitors, to adjust the host autophagic levels and then study the effects of autophagy on tissue damages and virus replication. As a result, we first found DTMUV infection trigged autophagy and autophagy regulator treatments regulated autophagy levels successfully in duck spleens and brains. Next, we found that autophagy inhibitors inhibited DTMUV replication and alleviated DTMUV-induced pathological symptoms, whereas the autophagy inducer treatment led to the opposite effects. And we also found that autophagic regulation was correlated with the expression of innate immune genes, including pattern recognition receptors, type I interferons, and cytokines, and caused different effects in different tissues. In summary, we demonstrated that autophagy facilitated DTMUV replication, aggravated the developments of pathological symptoms and possibly counteracts the host's innate immunity response in vivo.

鸭坦布苏病毒（Duck tembusu virus, DTMUV）是一种新近出现的家禽传染病病原，可给我国及周边国家的养殖业造成严重经济损失，目前高效的抗病毒防治策略仍处于研究阶段。细胞自噬（autophagy）是一种细胞内降解通路，本实验室前期研究数据显示，细胞自噬可在体外促进DTMUV的复制。为进一步探究细胞自噬在体内的功能，本研究以鸭作为动物模型，对DTMUV靶向侵染的组织中的细胞自噬应答情况进行了分析；此外，本研究使用了多种细胞自噬调控剂：其中雷帕霉素（Rapamycin, Rapa）作为细胞自噬激活剂，3-甲基腺嘌呤（3-Methyladenine, 3-MA）与氯喹（Chloroquine, CQ）作为细胞自噬抑制剂，通过调控宿主的细胞自噬水平，进而探究细胞自噬对组织损伤及病毒复制的影响。研究结果显示，DTMUV感染可触发细胞自噬，且通过细胞自噬调控剂处理可成功调节鸭脾脏与脑组织中的细胞自噬水平；进一步研究发现，细胞自噬抑制剂可抑制DTMUV复制并缓解DTMUV诱导的病理症状，而细胞自噬激活剂则会产生相反的调控效果。此外，本研究还发现，细胞自噬调控与模式识别受体、I型干扰素及细胞因子等先天免疫基因的表达密切相关，且在不同组织中呈现出不同的调控效应。综上，本研究证实细胞自噬可促进DTMUV复制，加重病理症状的发展进程，并可能在体内拮抗宿主的先天免疫应答。