College of Veterinary Medicine, Gansu Agricultural University, Lanzhou, 730070, PR China.
Shaanxi Meili-OH Animal Health Co., Ltd, Xi'an, 712034, PR China.
BMC Vet Res. 2024 Jan 5;20(1):10. doi: 10.1186/s12917-023-03852-5.
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.
马立克氏病病毒（MDV）GX0101 株是首例报道的重组鸡传染性喉气管炎病毒 2 型（GaHV-2）野毒株。然而，MDV 感染鸡的脾脏蛋白质组仍不清楚。在本研究中，将 28 只 1 日龄 SPF 鸡腹腔注射含有 2000PFU GX0101 的鸡胚成纤维细胞（CEF）。另外，4 只 1 日龄 SPF 鸡作为对照组，腹腔内给予等量的 CEF。在不同时间点（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、实时 PCR 和无标记定量分析。结果表明，血清中 MDV 相关基因 meq 和 gB 的表达在 45dpi 时达到峰值。在 30 和 45dpi 时剖检心脏、肝脏和脾脏，苏木精-伊红染色显示 45dpi 时病毒感染破坏了正常的组织结构。特别是脾脏结构严重受损，白髓中的淋巴细胞明显减少。此外，采用液相色谱-质谱（LC-MS）和无标记技术分析 30 和 45dpi 实验组和对照组脾脏蛋白质组图谱的差异。蛋白质组学分析发现，与未感染的脾脏组织相比，分别有 1660 和 1244 个差异表达蛋白（DEPs）在 30 和 40dpi 时表达上调。GO 分析表明，这些 DEPs 参与细胞器组织、细胞成分发生、细胞成分组装、阴离子结合、小分子结合、金属离子结合、阳离子结合、细胞质、核部分等过程。此外，KEGG 分析表明，以下途径与 MDV 诱导的炎症、细胞凋亡和肿瘤有关：Wnt、Hippo、AMPK、cAMP、Notch、TGF-β、PI3K-Akt、Rap1、Ras、钙、NF-κB、PPAR、cGMP-PKG、细胞凋亡、VEGF、mTOR、FoxO、TNF、JAK-STAT、MAPK、朊病毒病、T 细胞受体和 B 细胞受体。我们最终筛选出 674 个与脾脏组织中 MDV 感染相关的 DEPs。本研究提高了我们对 MDV 在脾脏中反应机制的理解。
