Small RNA profiling of extracellular vesicles secreted by A549 cells infected with West Nile virus and treated with interferon alpha

The goal of this study is to determine if infection with flavivirus (West Nile virus) and action of proinflamatory cytokine interferon alter incorporation of host RNAs into extracellular vesicles secreted by human cells. Overall design: Human lung carcinoma cells A545 were infected with Kunjin strain of West Nile virus at multiplicity of infection (MOI)=1 or incubated in the media containing 300u/ml IFN alpha 2a. Cells were then maintained in exosome free media for the next 24h. At 24 h post infection extracellular vesicles were isolated from culture fluids of infected or uninfected (Mock) cells using ExoQuick TC Exosomes isolation reagent. Enriched fractions of small RNAs were isolated from extacellular vesicles using mirVana miRNA isolation Kit (Ambion, USA) and used for library construction and RNA-Seq on Illumina HiSeq platform.