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Protein Profile of Extracellular Vesicles Released by Chicken Tracheal Cells Upon Avian Influenza Virus Infection and Toll-Like Receptor Ligand Stimulation

2021 CPRF
During viral respiratory infections in chickens, host cell communication and interaction at the early stages of infection are required for mounting effective immune responses. The release of extracellular vesicles (EVs) from host cells is a system of mediated intercellular communication. These EVs contain an array of biomolecules, such as proteins and RNAs, which can be regulated by viral infections and may play a role in antiviral responses. Furthermore, macrophages are essential players in the host innate responses. Their functions, including phagocytosis and cytokine production, also require specific regulation. In this study, we aimed to identify the immunoregulatory roles of EVs released from tracheal cells. We hypothesized that EVs released from chicken tracheal cells regulate the functions of other immune cells, specifically macrophages. To this end, chicken tracheal cells were infected with avian influenza virus (AIV) H4N6 or stimulated with Toll-like receptor (TLR) 3 or 4 ligands. EVs were isolated 24 h post-infection/stimulation and subjected to mass spectrometry, followed by bioinformatic and functional analysis. We then treated chicken macrophages with 5ug/ml or 25ug/ml EVs and evaluated the direct effects of the treatment. In this study, a total of 140 differentially expressed proteins (i.e., > 2-fold change) were identified. Analysis of gene functions and protein-protein interactions revealed that EVs secreted by chicken tracheal cells in response to various stimuli were enriched in protein markers involved in immune responses and cell signaling pathways. Furthermore, we investigated the impact of EVs on macrophage phagocytosis, nitric oxide production and the induction of antiviral responses in chicken macrophages. The results demonstrated that AIV infection impacts the protein contents of EVs. Furthermore, the results showed that EVs regulate chicken macrophage function in the context of antiviral responses by modulating the expression of several antiviral genes and the activity of macrophages. Overall, this study highlighted the active role of EVs in the cell-to-cell communication during AIV infection in chickens. This study provided a platform to understand the initiation of antiviral responses by tracheal cells and their impact on induced immune responses.
Tags :
avian influenza,protein profile
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PhD Student

University of Montreal

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