Project description:West Nile virus (WNV) is a mosquito-borne RNA flavivirus and the cause of more than 31,000 cases in the USA from 1999-2011 including 1, 262 fatalities. WNV infections are typically asymptomatic, but some patients, especially the elderly and immunocompromised, may experience severe neurological disease and even death. Control of WNV infection by the immune system is multifactorial. We profiled antibody, cytokine responses and gene expression from a stratified cohort of WNV subjects to define immune responses that contribute to disease severity and outcome. Differential gene expression by human PBMCs from asymptomatic and severe patients with WNV infection were generated by microarray.
Project description:West Nile virus (WNV) is a mosquito-borne RNA flavivirus which has caused more than 31,000 cases in the USA from 1999-2011 including 1,262 fatalities. WNV infections are typically asymptomatic, but some patients, especially the elderly and immunocompromised, may experience severe neurological disease and even death. Control of WNV infection by the immune system is multifactorial. We profiled antibody and cytokine responses from a stratified cohort of WNV subjects to define immune responses that contribute to disease severity. While antibody levels were not significantly different between asymptomatic and severely ill subjects in our cohort, subjects with severe infections had lower levels of serum IL-4. Further, we detected 158 genes that were differentially expressed by asymptomatic and severely infected cohorts and using cluster analysis correlated WNV susceptibility with IL-4 related gene expression pathways. Our results suggest an important contribution for IL4 in more severe responses to WNV.
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:Neurotropic flavivirus Japanese encephalitis virus (JEV) and West Nile virus (WNV) are amongst the leading causes of encephalitis. Using label-free quantitative proteomics, we identified proteins differentially expressed upon JEV (gp-3, RP9) or WNV (IS98) infection of human neuroblastoma cells. Both viruses were associated with the up-regulation of immune response (IFIT1/3/5, ISG15, OAS, STAT1, IRF9) and the down-regulation of SSBP2, involved in gene expression, as well as PAM, involved in neuropeptide amidation. Proteins associated to membranes, involved in extracellular matrix organization and collagen metabolism represented major clusters down-regulated by JEV and WNV. Moreover, transcription regulation and mRNA processing clusters were also heavily regulated by both neurotropic flaviviruses. If the proteome of neuroblastoma cells infected by JEV or WNV was significantly modulated in the presence of mosquito saliva, both viruses showed distinct patterns. Mosquito saliva favored the modulation of proteins associated with gene regulation in JEV infected neuroblastoma cells while it was the modulation of proteins associated with protein maturation, signal transduction and ion transporters in the case of WNV infected neuroblastoma cells.
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.