ITRAQ-based quantitative proteomics reveals the proteome profiles of MDBK cells infected with bovine viral diarrhea virus

Abstract

Background Bovine viral diarrhea (BVD) which is caused by Bovine viral diarrhea virus (BVDV), is an acute, contagious disease. In spite of the use of vaccines and elimination projects, BVDV still causes severe economic losses to the cattle industry for the past few years. The current study presents a preliminary analysis of the pathogenic mechanisms from the perspective of protein expression levels in infected host cells at different points in time to elucidate the infection process associated with BVDV. Methods We used the isobaric tags for relative and absolute quantitation (iTRAQ) technology coupled with liquid chromatography-tandem mass spectrometric (LC–MS/MS) approach for a quantitative proteomics comparison of BVDV NADL-infected MDBK cells and non-infected cells. The functions of the proteins were deduced by functional annotation and their involvement in metabolic processes explored by KEGG pathway analysis to identify their interactions. Results There were 357 (47.6% downregulated, 52.4% upregulated infected vs. control), 101 (52.5% downregulated, 47.5% upregulated infected vs. control), and 66 (21.2% downregulated, 78.8% upregulated infected vs. control) proteins were differentially expressed (fold change > 1.5 or < 0.67) in the BVDV NADL-infected MDBK cells at 12, 24, and 48 h after infection. GO analysis showed that the differentially expressed proteins (DEPs) are mainly involved in metabolic processes, biological regulation and localization. KEGG enrichment analysis showed that some signaling pathways that involved in the regulation of BVDV NADL-infection and host resistance are significantly (P < 0.05) enriched at different stages of the BVDV NADL-infection, such as Endocytosis signaling pathway, FoxO signaling pathway, Homologous recombination signaling pathway and Lysosome pathway. Conclusions These results revealed that the DEPs in BVDV NADL-infected MDBK cells have a wide range of regulatory effects; in addition, they provide a lot of resources for the study of host cell proteomics after BVDV infection.

Transcriptomic Analysis of MDBK Cells Infected with Cytopathic and Non-Cytopathic Strains of Bovine Viral Diarrhea Virus (BVDV)

“…All of those changes enriched the lysosome pathway according to the KEGG database (Tables S10, S11 and S13). Other studies also indicated enrichment of the lysosome pathway after infection with the NADL (cp) strain [39]. The results of HCV studies indicated that there was no fusion of autophagosomes and lysosomes at an early stage of infection [40].…”

Integrative Transcriptomics and Proteomics Analysis Provide a Deep Insight Into Bovine Viral Diarrhea Virus-Host Interactions During BVDV Infection

“…Therefore, we collected the cell samples at 48 h post BVD infection for the integrated analysis of RNA-seq-based transcriptomics profiles and iTRAQ-based proteomics profiles of the BVDV-infected host cells. Although there has transcriptomic sequencing analysis for BVDV-infected host cells at 2 h, 6 h, 12 h, and 24 h post BVDV infection ( 21 ) and proteomic analysis for BVDV-infected host cells at 12 h, 24 h, and 48 h post BVDV infection ( 22 ), and a lot of DEGs and DEPs were obtained, just only one omics data type analysis is limited to correlation between mRNA and protein abundances in BVDV-infected host cells.…”

“…Using RNA-seq-based transcriptome analysis for PEDV-infected Vero cells, researchers observed that significant changes in the mTOR signaling pathway occurred at different time points after viral infection, and this was further experimentally confirmed to promote virus infection ( 20 ). Moreover, there have been several studies that report transcriptomic changes ( 21 ) and proteomic changes ( 22 ) in BVDV-infected host cells. However, only one omics data type analysis is limited to correlation between mRNA and protein abundances, such as transcriptomic data only represents the mRNA expression level, but not represents the presence of post-translational modifications, which was insufficient to predict protein expression levels from quantitative mRNA data.…”

Proteins in pregnant swine serum promote the African swine fever virus replication: an iTRAQ-based quantitative proteomic analysis

“…iTRAQ is a high-throughput screening technology commonly used in quantitative proteomics [ 44 , 45 ]. iTRAQ technology has widely been used in the study of viral infection and disease development and for screening of biological markers [ 34 , 46 , 47 ]. To understand the role of PSS in ASFV replication, BMDMs were cultured with PSS and NPSS for 24 h, and DEPs in PSS and NPSS were identified using iTRAQ technology.…”