Abstract
BackgroundBovine 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.MethodsWe 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.ResultsThere 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.ConclusionsThese 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.
Highlights
Bovine viral diarrhea (BVD) which is caused by Bovine viral diarrhea virus (BVDV), is an acute, contagious disease
Virus inoculation When the bottom surface was covered by the above cultured MBDK cells, we changed the Dulbecco’s modified Eagle’s medium (DMEM) with 10% fetal bovine serum (FBS) to the medium without serum and the cells were infected with BVDV NADL (100 Tissue culture infective dose (TCID50)/ 0.1 ml)
Identification of differentially expressed proteins by isobaric tags for relative and absolute quantitation (iTRAQ) LC–MS/MS analysis Considering the dynamic changes in host response to viral infection, we collected the whole cell lysates of Madin-Darby bovine kidney (MDBK) cells at 12 hpi, 24 hpi, and 48 hpi In this study, the differentially expressed proteins were detected and their relative rates of change were analyzed
Summary
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. With the development of proteomics technology, it has outstanding advantages in studying virus-host cell interactions. From the study of intracellular protein composition, overall horizontal activity patterns and protein interactions, high-throughput analysis of multiple species samples by proteomics technology allows for the constitutive expression, quality assessment levels and modification status of proteins in the samples. In this way, the function of proteins and the potential relationship between proteins can be revealed and new proteins can be discovered. Combined with bio-informatics to reveal the physiological and pathological functions of cells, it can qualitatively and quantitatively analyze certain key proteins
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