Project description:Amphotericin B (AmB) is a polyene macrolide antifungal agent that binds to ergosterol, disrupting fungal membranes through lipid interactions. However, its effects on host cell protein composition remain unclear. In this study, A549 cells were pretreated with AmB prior to infection with the influenza A virus, and early and late endosomal fractions were isolated. DIA-based quantitative proteomics revealed that AmB alters the protein composition of endosomes during infection, suggesting that it modulates endosomal protein dynamics in virus-infected cells.
Project description:When challenged with an invading pathogen, host defense mechanisms are engaged to eliminate the pathogen (resistance) and limit tissue damage that results from host-pathogen interactions (disease tolerance). We identified Arhgdia as a driver of the molecular disease-tolerance response in epithelial cells. The effect of Arhgdia on disease tolerance and resistance was evaluated at early stages of influenza A virus (IAV) infection. Our observations indicates that Arhgdia has an effect on disease tolerance during IAV infection.
Project description:We used the microarray data to analyze host cells response on A549 cells infected with Influenza A virus (A/Singapore/478/2009 (pH1N1)) The Influenza A virus (A/Singapore/478/2009 (pH1N1)) infected A549 cells were harvested at 2, 4, 6, 8 and 10 hpi and RNA extraction was performed using standard protocol as described by Affymetrix. The aim of this experiment is to analyze host response to Influenza A virus (A/Singapore/478/2009 (pH1N1)) infection.
Project description:Virus and host factors contribute to cell-to-cell variation in viral infection and determine the outcome of the overall infection. However, the extent of the variability at the single cell level and how it impacts virus-host interactions at a systems level are not well understood. To characterize the dynamics of viral transcription and host responses, we used single-cell RNA sequencing to quantify at multiple time points the host and viral transcriptomes of human A549 cells and primary bronchial epithelial cells infected with influenza A virus. We observed substantial variability of viral transcription between cells, including the accumulation of defective viral genomes (DVGs) that impact viral replication. We show a correlation between DVGs and viral-induced variation of the host transcriptional program and an association between differential induction of innate immune response genes and attenuated viral transcription in subpopulations of cells. These observations at the single cell level improve our understanding of the complex virus-host interplay during influenza infection.
Project description:Amphotericin B (AmB), a polyene macrolide antifungal, is well known for its ability to bind ergosterol and cholesterol; however, its interactions with host proteins remain poorly characterized. In this study, we aimed to identify AmB-binding targets in A549 cells infected with the influenza A virus (H1N1) strain A/PR/8/34 (PR8). We isolated late endosomal fractions and performed a biotin–streptavidin pulldown using a biotinylated AmB probe, with specificity confirmed by competition with excess free AmB. The proteins enriched from these fractions were then analyzed by shotgun LC–MS/MS, revealing a set of late endosomal proteins that specifically interact with AmB during influenza A virus infection.
Project description:Small RNAs were profiled during influenza A virus infection of human A549 cells to identify changes in microRNA abundance during the cellular antiviral response. Examination of microRNA abundance during influenza A virus infection.
Project description:Virus-host interactions are complicated processes, and multiple cellular proteins have been reported to promote or inhibit viral replication through different mechanisms. Recent progress has implicated circular RNAs (circRNA) in cancer biology and progression; however, the role of circRNAs in viral infection remains largely unclear. Here, we detected 11,620 circRNAs in A549 cells and found that 411 of them were differentially expressed in influenza virus-infected A549 cells. We characterized a novel intronic circRNA, AIVR, that was upregulated in influenza virus-infected A549 cells, and found that silencing of AIVR significantly promoted influenza virus replication in A549 cells. We further found that AIVR predominantly localizes in the cytoplasm and works as a microRNA (miRNA) sponge. One of the miRNAs absorbed by AIVR binds the mRNA of CREBBP, which is an important component of the large nucleoprotein complex IFN-β enhanceosome that accelerates IFN-β production. AIVR-overexpression significantly increased the mRNA and protein levels of INF-β in the influenza virus-infected A549 cells. Therefore, the upregulation of AIVR is a cellular antiviral strategy, with AIVR exerting its antiviral effect by absorbing miRNA and promoting the expression of CREBBP to facilitate IFN-β production. Our study provides new insights into the roles of circRNAs in the cellular innate antiviral response.
Project description:The goal of this experiment was to determine gene expression changes during influenza A virus infection as the result of expression influenza virus inducible miRNAs in A549 cells.
