Project description:Long term cell culture adaptation of hepatitis C virus resulted in increased replication fitness in various human liver cell lines but in a moderate decrease in virus particle production upon infection of primary human hepatocytes (PHH). In order to identify molecular mechanisms conferring phenotypic differences in replicative fitness of the cell culture adapted virus strain p100pop, we infected PHH and Huh-7 cells with HCV, using the cell culture adapted strain p100pop or a Jc1 strain with similar genome organisation (Jc1-SP). Total RNA was extracted at 28 hours post inoculation and used for RNA sequencing. Transcriptome analyses, gene ontology enrichment analyses and KEGG pathway analyses revealed strong differences in the transcriptional signature of infected hepatoma cells and primary hepatocytes and the two virus strains used in this study. Wheras an innate immune response was induced in primary cells regardless of the infecting virus strain, this was not detectable in Huh-7 cells. Even though both viruses induce a similar host response in primary cells, the data indicate that the presence of cell culture adaptive mutations results in an increased expression of genes involved in the defense response to viral infection. In Huh-7 cells, differentially expressed genes associated with ER stress and apoptosis were solely enhanced upon p100pop infection but not upon Jc1-SP infection.
Project description:Huh7/5-2 cells (Binder et al., Hepatology 2007) were mock infected (DMEM) (time points 4 and 48 h) or infected with the chimeric HCV virus Jc1 (Pietschmann et al., PNAS 2006) (all time points). Multiplicity of infection was 15 (TCID50). Cells were lysed after 4, 12, 24, 48 and 72 hours post infection and total cellular RNA was prepared.
Project description:Cytosolic lipid droplets (LDs) are vital to Hepatitis C Virus (HCV) infection as the putative sites of virion assembly. To identify novel regulators of HCV particle production, we performed quantitative LD proteome analysis. Huh7.5 cells were labeled by stable isotope labeling with heavy amino acids in cell culture (SILAC) and subsequently infected with an HCV Jc1 reporter virus. After selection for HCV-infected cells, equal amounts of HCV-infected and uninfected control cells were mixed, LDs were isolated and analyzed by LC-ESI-MS/MS.
Project description:Expression profiling of prognostic liver signature in clinical fibrotic liver tissues cultured with various anti-fibrotic and chemopreventive agents
Project description:Huh7/5-2 cells (Binder et al., Hepatology 2007) were mock infected (DMEM) (time points 4 and 48 h) or infected with the chimeric HCV virus Jc1 (Pietschmann et al., PNAS 2006) (all time points). Multiplicity of infection was 15 (TCID50). Cells were lysed after 4, 12, 24, 48 and 72 hours post infection and total cellular RNA was prepared. Mock infected cells serve as controls for the infected samples. The whole experiment was repeated on independent days (2 biological replicates). The dataset was also submitted to GEO (GSE38720).
