Exosomal microRNA (miRNA) expression in Blonchoalveolar lavage fluid (BALF) collected from mice infected with influenza virus and inoculated with poly(I:C).
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ABSTRACT: To clarify the profile of in BALF exosome collected from mice infected with influenza virus, we infected 100000 pfu of A/Puerto Rico/8/1934 (PR8) strain. BALF was collected at 24, 48, and 72 hour post infection (hpi). For comparison of the profile of the miRNA in BALF exosome induced by innate immune response, we also intranasally inoculated mice with 50 μg of poly(I:C) and collected BALF at 72 hour post inoculation. We found that some miRNAs were common to both influenza virus infectiona and poly(I:C) inoculation, suggesting that exosomal miRNAs in BALF may function in the innate immune response to virus infection.
Project description:To study the effects of secondary bacterial infection during 1918 pandemic H1N1 influenza virus infection, BALB/c mice were inoculated with the fully reconstructed 1918 influenza virus followed by inoculation with pneumococcus 72h later. To study the effects of secondary bacterial infection during 1918 pandemic H1N1 influenza virus infection, BALB/c mice were inoculated with the fully reconstructed 1918 influenza virus followed by inoculation with pneumococcus 72h later.
Project description:To study the effects of secondary bacterial infection during 1918 pandemic H1N1 influenza virus infection, BALB/c mice were inoculated with the fully reconstructed 1918 influenza virus followed by inoculation with pneumococcus 72h later.
Project description:The nuclear RNA exosome is an essential multi-subunit complex that controls RNA homeostasis. Congenital mutations in exosome genes are associated with neurodegenerative diseases. Here, we show that transient depletion of nuclear RNA exosome subunits in epithelial cells inhibits influenza virus replication. Similarly, viral biogenesis was suppressed in cells derived from mice with conditional ablation of the RNA exosome subunit Exosc3. Furthermore, patient-derived cells with a congenital EXOSC3 mutation were less susceptible to influenza virus infection. Using proteomics and next generation sequencing during infection, we show that the viral polymerase complex (PA, PB2, PB1) co-opts the nuclear RNA exosome complex and cellular RNAs en route to 3’ end degradation. Mechanistically, the nuclear RNA exosome coordinates the initial steps of viral transcription with RNAPII at host promoters. Exosome deficiency uncouples chromatin targeting of the viral polymerase complex and the formation of cellular:viral RNA hybrids, which are essential RNA intermediates that license transcription of antisense genomic viral RNAs. Overall, we discovered a critical nexus between an essential component of the influenza virus (polymerase) and an essential component of the cell (exosome), alteration of which leads to breakage of host-pathogen symmetry and a lose-lose scenario (viral impairment and neurodegeneration).
Project description:Exosomal microRNA (miRNA) expression in Blonchoalveolar lavage fluid (BALF) collected from mice infected with influenza virus and inoculated with poly(I:C).
Project description:We describe here transcripts induced after intraperitoneal injection of rainbow trout with 2 different viruses, both belonging to strain 23.75 of viral hemorrhagic septicemia virus (VHSV): a deleted Nv gene (dNV) virus and a wild type (wt) virus. Two days after infection, differentially expressed transcript levels from selected immune-related trout genes were studied in internal organs (spleen and head kidney). Fishes were divided in two groups (3 fishes per group). The first group was intraperitoneally injected with 100000 pfu per trout of dNV VHSV, while the second group was injected with 100000 pfu/trout of wt VHSV. All fishes were sacrificed two days post infection.
Project description:In this study broncho-alveolar lavage fluid (BALF) from mice infected with influenza A virus, PR8 (280 PFU) was analyzed for lipidomic host response in comparison with the mock infected animals. Time Points: 6, 9, 11, 13 and 19 days post infection, done in 8-10 replicates.)
Project description:In this study broncho-alveolar lavage fluid (BALF) from mice infected with influenza A virus, PR8 (200 PFU) or X31 (200,000 PFU) was analyzed for lipidomic host response in comparison with the mock infected animals. Time Points: 3, 5, 6, 7, 9, 10 and 13 days post infection.)
Project description:In this study broncho-alveolar lavage fluid (BALF) from mice infected with influenza A virus, PR8 (200 PFU and 200,000 PFU), X31 (2000,000 PFU) was analyzed for lipidomic host response in comparison with the mock infected animals. Time Points: 3, 5, 7, 9, 11, 13, 19 days post infection, done in 8-10 replicates.)
Project description:Diagnosis of influenza A infection is currently based on clinical symptoms and pathogen detection. Use of host peripheral blood gene expression data to classify individuals with influenza A virus infection represents a novel approach to infection diagnosis We used microarrays to assay peripheral blood gene expression at baseline and every 8 hours for 7 days following intranasal influenza A H1N1 or H3N2 inoculation in healthy volunteers. We determined groups of coexpressed genes that classified symptomatic influenza infection. We then tested this gene expression classifier in patients with naturally acquired respiratory illness. We experimentally inoculated healthy volunteers with intranasal influenza A H1N1 and H3N2. Symptoms were documented and peripheral blood samples drawn into PAXgene RNA tubes for RNA isolation. We further enrolled patients presenting to the Emergency Department with naturally acquired respiratory illness, and documented symptoms and collected PAXgene RNA samples for RNA isolation.
Project description:N. benthamiana plants were grown under 16 hour light/8 hour dark cycle in a plant growth room at 24°C for approximately six weeks before subjected to virus-induced gene silencing. Agrobacterium stain GV2260 (OD600=1.0) carrying silencing constructs were infiltrated into 2 fully expended leaves for inducing gene silencing. Samples were collected at 8, 10, 12, 14, and 16 days post Agro-inoculation (DPI) for RPN9-silenced plants and N-silenced control. For RPN8-silencing, samples were collected at 8, 10, and 12 DPI, and the empty vector treated plants were used as a control. Each sample was a pool of 6 silenced leaves collected from 3 individual plants. All the samples were in biological triplicates from 3 sets of independently silenced plants. Total RNA was extracted using Trizol and DNA was removed with DNase I treatment before cDNA synthesis. Keywords: Reference design