Project description:Purpose of this experiment was to further understand how innate immune defenses impact host response and West Nile virus tissue tropism. This study examined host-transcriptional response to West Nile virus in permissive and nonpermissive tissues using wildtype mice and mice with genetically altered interferon signaling pathways.
Project description:High throughput sequencing was performed using Illumina HiSeq to identify differentially regulated genes in Culex mosquitoes after West Nile virus infection.
Project description:Individual variations in immune status and function determine responses to infection and contribute to disease severity and outcome. Patients exhibit considerable variation in clinical responses to infection with West Nile virus. We have undertaken a comprehensive characterization of the immune responses of a stratified cohort of patients with a history of West Nile virus infection to identify key mechanisms of resistance and susceptibility. We provide molecular profiles of cellular mechanisms of primary human immune cells defined through multifaceted interrogation including multiplexed gene expression analysis integrated with highly sensitive multidimensional flow cytometry. The availability of reliably curated patient cohorts and data-sharing and data mining techniques of high-throughout investigations should accelerate identification of critical elements of immune resistance to important pathogens
Project description:The purpose is to obtain samples for transcriptional analysis in triplicate wells using wild type West Nile virus (WNV NY99 clone 382; WNVWT) and mutant virus (WNVE218A) in mouse cortical neurons. This data set comprises two complete biological replicate experiments conducted in the same conditions and with data processed independently. Primary cortical neurons from C57Bl/6J mouse embryos (age E15) are infected with plasmid-derived wild type West Nile virus NY99 clone 382 (WNVWT) or plasmid-derived isogenic E218A mutant West Nile virus NY99 clone 382 (WNVE218A) with multiplicity of infection (MOI) 250. Three technical replicates were performed at each of 1, 8, 12 and 24 hrs post infection. Time matched mocks done in triplicate are treated with mockulum: cell media concentrated through ultracentrifugation and diluted as virus. mRNA is sampled at all time points; microRNA is sampled at 12 hours post-infection. There were two independent biological replicates of the entire procedure, distinguished by sample name prefixes ('WCN002' and 'WCN003') and the biological_replicate characteristic field.
Project description:We profile the peripheral blood of patients infected with West Nile Virus with divergent disease-trajectories (West Nile Encephalitis, West Nile Fever, and asymptomatic) during relatively acute infection and at a convalescent timepoint (~90-days later) using single-cell RNA sequencing in an effort to uncover determinants of disease progression and flesh out the landscape of infection. In the blood of the infected patients, stratified cell-states involved in acute viral infection resolve into more homogenous states at the follow-up blood draws, A dramatic shared transcriptional shift between the primary blood-draws during acute infection and the 90-day follow-ups in all observed compartments allows us to highlight multiple cell-type and cell-state-specific patterns of gene expression.
Project description:Individual variations in immune status and function determine responses to infection and contribute to disease severity and outcome. Patients exhibit considerable variation in clinical responses to infection with West Nile virus. We have undertaken a comprehensive characterization of the immune responses of a stratified cohort of patients with a history of West Nile virus infection to identify key mechanisms of resistance and susceptibility. We provide molecular profiles of cellular mechanisms of primary human immune cells defined through multifaceted interrogation including multiplexed gene expression analysis integrated with highly sensitive multidimensional flow cytometry. The availability of reliably curated patient cohorts and data-sharing and data mining techniques of high-throughout investigations should accelerate identification of critical elements of immune resistance to important pathogens Differential gene expression by human PBMCs and macrophages from asymptomatic and severe patients with WNV infection or Poly I:C were generated by microarray.
Project description:The purpose is to obtain samples for transcriptional analysis in triplicate wells using wild type West Nile virus (WNV NY99 clone 382; WNVWT) and mutant virus (WNVE218A) in mouse granule cell neurons. This data set comprises two complete biological replicate experiments conducted in the same conditions and with data processed independently. Granule cell neurons from day 6 C57Bl/6J mouse pups are infected with plasmid-derived wild type West Nile virus NY99 clone 382 (WNVWT) or plasmid-derived isogenic E218A mutant West Nile virus NY99 clone 382 (WNVE218A) with multiplicity of infection (MOI) 250. Three technical replicates were performed at each of 1, 8, 12 and 24 hrs post infection. Time matched mocks done in triplicate are treated with mockulum: cell media concentrated through ultracentrifugation and diluted as virus. mRNA is sampled at all time points; microRNA is sampled at 12 hours post-infection. There were two independent biological replicates of the entire procedure, distinguished by sample name prefixes ('WGCN002' and 'WGCN003') and the biological_replicate characteristic field.
Project description:The purpose is to obtain samples for transcriptional analysis in triplicate wells using wild type West Nile virus (WNV NY99 clone 382; WNVWT) and mutant virus (WNVE218A) in mouse granule cell neurons. This data set comprises two complete biological replicate experiments conducted in the same conditions and with data processed independently. Granule cell neurons from day 6 C57Bl/6J mouse pups are infected with plasmid-derived wild type West Nile virus NY99 clone 382 (WNVWT) or plasmid-derived isogenic E218A mutant West Nile virus NY99 clone 382 (WNVE218A) with multiplicity of infection (MOI) 250. Three technical replicates were performed at each of 1, 8, 12 and 24 hrs post infection. Time matched mocks done in triplicate are treated with mockulum: cell media concentrated through ultracentrifugation and diluted as virus. mRNA is sampled at all time points; microRNA is sampled at 12 hours post-infection. There were two independent biological replicates of the entire procedure, distinguished by sample name prefixes ('WGCN002' and 'WGCN003') and the biological_replicate characteristic field.