Infection of duck with avian influenza H5N1 and H5N2 viruses
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ABSTRACT: Transcriptional profiling was carried out on lung and ileum samples at 1dpi and 3dpi from ducks infected with either low pathogenic (H5N2) or highly pathogenic (H5N1) avian influenza. Infected birds were compared to control birds at each time point.
Project description:Transcriptional profiling was carried out on lung and ileum samples at 1dpi and 3dpi from chickens infected with either low pathogenic (H5N2) or highly pathogenic (H5N1) avian influenza. Infected birds were compared to control birds at each time point.
Project description:Transcriptional profiling was carried out on lung and ileum samples at 1dpi and 3dpi from quail infected with either low pathogenic (H5N2) or highly pathogenic (H5N1) avian influenza. Infected birds were compared to control birds at each time point.
Project description:Infectious bronchitis is a highly contagious respiratory disease of poultry caused by the coronavius infectious bronchitis virus (IBV). Until recently is was thought that coronavirus virions were composed of the structural proteins nucleocapsid, envelope, spike and membrane proteins, but investigations of TGEV and SARS-CoV have shown the proteome of coronavirus virions also includes viral non-structural and group specific proteins as well as host cell proteins. To study the proteome of IBV virions, virus was grown in embryonated chicken eggs and purified by sucrose gradient ultracentrifugation. Purified virus was analysed using sensitive gel-free proteomic techniques to determine the proteome of IBV. Analysis of three preparations of purified IBV yielded a list of 39 proteins commonly associated with the IBV virion. Three of these proteins were the viral structural proteins spike, membrane and nucleocapsid, but none of the viral non-strucutral or groups specific proteins could be identified. The other 35 proteins commonly associated to the IBV virion were all found to be host cell proteins. These proteins were classified into 12 categories using pantherdb (pantherdb.org). These proteins were involved in a diverse range of functions such as cytoskeletal proteins, nucleic acid binding proteins and chaperone proteins. Some of these proteins were unique to this study, whilst others were found to be orthologous to proteins identified in the SARS-CoV protein, and indeed some were also identified in association with virions from a number of other RNA and DNA viruses.
Project description:The underlying molecular mechanisms of pathogenesis and outcome of disease to different pathotypes of H5N1 influenza infection in ducks remain unclear. For that, we studied genome wide host gene expression of lung tissues infected with A/duck/India/02CA10/2011(AD2011) H5N1 virus and A/duck/Tripura/103597/2008 (AD2008) H5N1 virus in ducks using custom designed microarray. AD2011 is highly pathogenic whereas AD2008 is low pathogenic to ducks. Comparative analysis of differentially expressed genes revealed that 688 genes were commonly expressed, 877 and 1556 genes are uniquely expressed to infection with AD2011 and AD2008 virus isolate, respectively. The up-regulation of cytokines genes OAS, IL1B, IL17, IFITM2, CCL4, CXCR4, STAT3, TGFB1 and TGFB2 in the lungs tissues may cause high mortality in ducks infected with AD2011 virus. The expression of important antiviral immune genes IFIT5, IFITM5, RSAD2, EIF2AK2 (PKR), Mx, β-defensins, TRIM23 and SLC16A3 to AD2008 infection, but not in AD2011 infection, cause the host may fine-tune their innate immune responses and prevent from cytokines storms and tissue damage. Several immune related Gene ontology (GO) terms and immune pathways activated were qualitatively similar but quantitatively different to both virus infections. Based on these findings, we conclude that subtle differences in host immune responses may determine the different outcome of H5N1 infection in ducks. Agilent Custom Duck Gene Expression 8X60k (AMADID: G4102A_059612) designed by Genotypic Technology Private Limited , Labeling kit: Agilent Quick-Amp labeling Kit (p/n5190-0442)
Project description:Influenza A Virus (IAV) is a recurring respiratory virus with antiviral therapies of limited use. Understanding host proteins essential for IAV infection can identify targets for alternative host-directed therapies (HDTs). Using affinity purification-mass spectrometry and global phosphoproteomic and protein abundance analyses with three IAV strains (pH1N1, H3N2, H5N1) in three human cell types (A549, NHBE, THP-1), we mapped 332 IAV-human protein-protein interactions and identified 13 IAV-modulated kinases. Whole exome sequencing of patients who experienced severe influenza revealed several genes, including the structural scaffold protein AHNAK, with predicted loss-of-function variants that were also identified in our proteomic analyses. Of our identified host factors, 54 significantly altered IAV infection upon siRNA knockdown, and two factors, COPB1 and AHNAK, were also essential for productive infection by SARS-CoV-2. Finally, 16 compounds targeting our identified host factors suppressed IAV replication, with three targeting ATP6V1A, CDK2 and FLT3 showing pan-antiviral activity across influenza and coronavirus families. This study provides a comprehensive network model of IAV infection in human cells, identifying functional host targets for pan-viral HDT. This project includes the global proteomic data (abundance and phosphorylation), the AP-MS data has been submitted separately as its own dataset and has its own dataset identifier.
Project description:The M-bM-^@M-^\Spanish influenzaM-bM-^@M-^] of 1918 claimed an unprecedented number of lives, yet the determinants of virulence for this virus are still not fully understood. Here, we used functional genomics and an in vitro human lung epithelial cell infection model to define the global host transcriptional response to the eight-gene 1918 virus. To better understand the role of the 1918 virus NS1 gene, we evaluated the host response to A/Texas/36/91 (a seasonal isolate of human influenza virus) and a reassortant of A/Texas/36/91 containing the 1918 NS1 gene. A549 cells were infected at a MOI=2 with either A/Texas/36/91 (Tx/91) or A/Texas/36/91 (Tx/91) containing the NS1 gene from r1918 influenza. Virus was allowed to bind to cells for 1 h at 4M-BM-0C in serum-free infection medium supplemented with trypsin (1 g/ml). Mock-infected cells were treated with allantoic fluid instead of virus. Three replicate wells were used for each infection condition at each time point. Cells were harvested for array analysis at 2, 6, and 24hr post-infection. Cells from three individual cultures were pooled for each condition for microarray.
Project description:This study is planned to characterize the proteome profile of HPAI H5N1 virus infected chicken lung tissues to identify the molecular pathogenesis and proteomic determinant associate with disease progression in susceptible host.
Project description:A global genomics approach was used to identify patterns of immune dysregulation during H5N1 influenza virus infection as the host response, in particular hyperchemokinemia, is thought to contribute to the extreme pathology associated with this disease. Keywords: time course Ferrets were inoculated intranasally with 10(6) EID50 of either A/Vietnam/1203/04 (H5N1) or A/Panama/2007/99 (H3N2). At 2, 4 and 6 days post-infection (DPI), ferrets were euthanized and lung tissue was excised for RNA purification and subsequent gene expression analysis.
Project description:A/Vietnam/1203-CIP048_RG3/2004 (H5N1) is a PB1-F2 deletion in wild type A/Vietnam/1203/2004 (H5N1). The goal of this study was to determine the host response (C57BL/6 mouse model) to the PB1-F2 mutation at a 10^4 PFU dose. Groups of 20-week-old female C57BL/6 mice were infected with A/Vietnam/1203-CIP048_RG3/2004 (H5N1). This is a PB1-F2 deletion in wild type A/Vietnam/1203/2004 (H5N1). Infections were with 10^4 PFU or time-matched mock infections. Time points were 1, 2, 4 and 7 d.p.i. There were 1-5 animals/dose/time point. Lung samples were collected for virus load, transcriptional analysis and proteomic analysis. Weight loss and animal survival were also monitored.
Project description:Small RNAs were profiled during Sendai virus infection of human A549 cells to identify changes in microRNA abundance during the cellular antiviral response. Examination of microRNA abundance during Sendai virus infection.