Porcine rotavirus (PoRV), particularly group A, is one of the most important swine pathogens, causing substantial economic losses in the animal husbandry industry. To improve understanding of host responses to PoRV
infection, we applied isobaric tags for relative and absolute quantification (iTRAQ) labeling coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS) to quantitatively identify the differentially expressed
proteins in PoRV-infected IPEC-J2 cells and confirmed the differentially accumulated
proteins (
DAPs) expression differences by performing RT-qPCR and Western blot analysis. Herein, in PoRV- and mock-infected IPEC-J2 cells, relative quantitative data were identified for 4724
proteins, 223 of which were
DAPs (125 up-accumulated and 98 down-accumulated). Bioinformatics analyses further revealed that a majority of the
DAPs are involved in numerous crucial biological processes and signaling pathways, such as metabolic process, immune system process,
amino acid metabolism, energy metabolism, immune system, MHC class I
peptide loading complex, Hippo signaling pathway, Th1 and Th2 cell
differentiation, antigen processing and presentation, and tubule
bicarbonate reclamation. The cellular localization prediction analysis indicated that these
DAPs may be located in the Golgi apparatus, nucleus, peroxisomal, cytoplasm, mitochondria, extracellular, plasma membrane, and endoplasmic reticulum (ER). Expression levels of three up-accumulated (VAMP4, IKBKE, and TJP3) or two down-accumulated (SOD3 and DHX9)
DAPs upon PoRV
infection, were further validated by RT-qPCR and Western blot analysis. Collectively, this work is the first time to investigate the
protein profile of PoRV-infected IPEC-J2 cells using quantitative proteomics; these findings provide valuable information to better understand the mechanisms underlying the host responses to PoRV
infection in piglets. SIGNIFICANCE: The proteomics analysis of this study uncovered the target associated with PoRV-induced innate immune response or cellular damage, and provided relevant insights into the molecular functions, biological processes, and signaling pathway in these targets. Out of these 223
DAPs, the expression levels of three up-accumulated (VAMP4, IKBKE, and TJP3) and two down-accumulated (SOD3 and DHX9)
DAPs upon PoRV
infection, have been further validated using RT-qPCR and Western blot analysis. These outcomes could uncover how PoRV manipulated the cellular machinery, which could further our understanding of PoRV pathogenesis in piglets.