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Decoding Protein Networks During Porcine Epidemic Diarrhea Virus (PEDV) Infection Through Proteomics

出版	Université de Montréal, 2019
URL	http://books.google.com.hk/books?id=qhuwzQEACAAJ&hl=&source=gbs_api

註釋Porcine epidemic diarrhea virus (PEDV) is responsible for severe economic losses. The PEDV epidemics have destroyed more than 10% of the US swine population in the past 3 years. Unfortunately, the insufficient understanding of virus-host interactions impedes the development of an effective vaccine against PEDV. Virus-host interactions are highly dynamic and may involve multiprotein complexes. Growing evidence indicates that extracellular microvesicles (EMV) and composition of the viral particles play an important role in viral pathogenesis and modulation of host immune responses to infection. Additionally, it could be expected that the composition of porcine epidemic diarrhea (PED) virions is cell type dependent, due to the differential incorporation or association of host cell proteins into or with virions. Consequently, the characterization of the proteomic profiles of the EMV, produced by the PEDV-infected cells, and identification of the host proteins that are specifically encapsidated into the virions are important for our further understanding of virus-host interactions. To accomplish this objective, we produced and purified PEDV virions and EMV and analyzed their protein composition using a proteomic approach. In order to investigate the spatial-temporal regulation of viral infection and due to the low overall infectivity of the virus, a certain optimization of the PEDV infection was needed. To this end, we synchronized and increased virus entry into the cells. This allowed us to study the proteomic patterns of the PEDV-infected cells in a time-resolved mode. We found that PEDV infection affected the abundance of various host proteins associated with microvesicles produced by the infected cells. More precisely, our proteomic data revealed that proteins involved in nucleic acids binding, metabolic processes and immune response pathways were among the most affected by the PEDV infection. Interestingly, host proteins involved in cell cycle regulation and cytoskeletal system also were affected in abundance, which is not surprising since several investigators have reported that cytoskeletal proteins are actively participating in moving the viral components to the assembly site, and that many viruses manipulate DNA repair and cell cycle. The present study has demonstrated the incorporation of numerous cellular proteins into the PED virions. Additionally, we demonstrated that treatment of PEDV virions with polycations (positively charged molecules) induced a nine-fold increase in the efficiency of viral entry and infection. Thus, polycations can be used for the optimization of PEDV infection and improved vaccine production. To the best of our knowledge, this is the first study of the composition of PED virions and microvesicles produced by PEDV infection.