马立克氏病病毒(MDV)株GX0101是首次报道的重组gallid疱疹病毒2型(GaHV-2)的田间毒株。然而,MDV感染鸡的脾蛋白质组仍不清楚.在这项研究中,对总共28只1日龄SPF鸡进行腹腔注射含有2000PFUGX0101的鸡胚成纤维细胞(CEF).此外,一个对照组,由四只一天大的SPF鸡组成,接受腹膜内等剂量的CEF。以不同的时间间隔(感染后7、14、21、30、45、60和90天;dpi)收集血液和各种组织样本进行组织病理学检查,实时PCR,和无标签的定量分析。结果表明,MDV相关基因的血清表达,meq和gb,峰值为45dpi。心脏,肝脏,在30和45dpi时解剖脾脏,他们的苏木精-伊红染色表明,病毒感染在45dpi时损害了正常的组织结构。特别是,脾脏结构严重受损,白色髓质中的淋巴细胞明显减少。此外,使用液相色谱-质谱(LC-MS)和无标记技术分析了实验组和对照组在30和45dpi时的脾蛋白质组分布差异。蛋白质组学分析在30和40dpi鉴定了1660和1244差异表达的蛋白质(DEP),分别,与未感染的脾组织相比。根据GO分析,这些DEP参与了细胞器组织等过程,细胞成分生物发生,蜂窝组件组件,阴离子结合,小分子结合,金属离子结合,阳离子结合,胞质溶胶,核部分,等。此外,KEGG分析表明,以下途径与MDV诱导的炎症有关,凋亡,肿瘤:Wnt,河马,AMPK,cAMP,缺口,TGF-β,PI3K-Akt,Rap1,拉斯,钙,NF-κB,PPAR,cGMP-PKG,细胞凋亡,VEGF,mTOR,福克斯,TNF,JAK-STAT,MAPK,朊病毒病,T细胞受体,B细胞受体.我们最终筛选了与脾组织中MDV感染相关的674个DEP。这项研究提高了我们对脾脏中MDV反应机制的理解。
Marek\'s disease virus (MDV) strain GX0101 was the first reported field strain of recombinant gallid herpesvirus type 2 (GaHV-2). However, the splenic proteome of MDV-infected chickens remains unclear. In this study, a total of 28 1-day-old SPF chickens were intraperitoneally injected with chicken embryo fibroblast (CEF) containing 2000 PFU GX0101. Additionally, a control group, consisting of four one-day-old SPF chickens, received intraperitoneal equal doses of CEF. Blood and various tissue samples were collected at different intervals (7, 14, 21, 30, 45, 60, and 90 days post-infection; dpi) for histopathological, real-time PCR, and label-free quantitative analyses. The results showed that the serum expressions of MDV-related genes, meq and gB, peaked at 45 dpi. The heart, liver, and spleen were dissected at 30 and 45 dpi, and their hematoxylin-eosin staining indicated that virus infection compromised the normal organizational structure at 45 dpi. Particularly, the spleen structure was severely damaged, and the lymphocytes in the white medulla were significantly reduced. Furthermore, liquid chromatography-mass spectrometry (LC-MS) and label-free techniques were used to analyze the difference in splenic proteome profiles of the experimental and control groups at 30 and 45 dpi. Proteomic analysis identified 1660 and 1244 differentially expressed proteins (DEPs) at 30 and 40 dpi, respectively, compared with the uninfected spleen tissues. According to GO analysis, these DEPs were involved in processes such as organelle organization, cellular component biogenesis, cellular component assembly, anion binding, small molecule binding, metal ion binding, cation binding, cytosol, nuclear part, etc. Additionally, KEGG analysis indicated that the following pathways were linked to MDV-induced inflammation, apoptosis, and tumor: Wnt, Hippo, AMPK, cAMP, Notch, TGF-β, PI3K-Akt, Rap1, Ras, Calcium, NF-κB, PPAR, cGMP-PKG, Apoptosis, VEGF, mTOR, FoxO, TNF, JAK-STAT, MAPK, Prion disease, T cell receptor, and B cell receptor. We finally screened 674 DEPs that were linked to MDV infection in spleen tissue. This study improves our understanding of the MDV response mechanism in the spleen.