RSV-induced Cytoskeletal Inflammation

We studied RSV infection in an appropriate in vitro model of respiratory epithelium, a pseudostratified and fully differentiated mucociliary epithelium of normal human bronchial epithelial (NHBE) cells. RSV infection increased actin cytoskeleton without compromising adherent-, tight-, and tricellular-junctions as well as ciliary functions and epithelial tissue barrier integrity. This increased cytoskeleton depends on actin polymerization and the induction of proinflammatory cytokines and chemokines. Thus, we observed a novel signature “increased cytoskeleton” termed “cytoskeletal inflammation” in RSV-infected respiratory epithelium that presumably lacks classical antigen presenting cells, such as resident dendritic cells and macrophages. Our results suggest that RSV-induced cytoskeletal inflammation is a noncanonical earliest host response to the pathogen and contributes to airway inflammation. Overall design: A four-week pseudostartified highly differentiated respiratory epithelium establsihed by 3D air-liquid inteerface culturing of priamry normal human bronchial epithelial (NHBE) cells. The respiratory epithelium was infected with RSV expressing green fluorescing protein (RSV-GFP) at a multiplicity of infection of 4 (MOI=4) for six days. Total RNA was extracted with DNAse-treatment from the mock-infected and RSV-GFP infected respiratory epithelium for whole genome trasncriptome analysis by RNAseq.