title: Plasticity of airway epithelial cell transcriptome in response to flagellin.
abstract: Airway epithelial cells (AEC) are critical components of the inflammatory and immune response during exposure to pathogens. AECs in monolayer culture and differentiated epithelial cells in air-liquid interface (ALI) represent two distinct and commonly used in vitro models, yet differences in their response to pathogens have not been investigated. In this study, we compared the transcriptional effects of flagellin on AECs in monolayer culture versus ALI culture using whole-genome microarrays and RNA sequencing. We exposed monolayer and ALI AEC cultures to flagellin in vitro and analyzed the transcriptional response by microarray and RNA-sequencing. ELISA and RT-PCR were used to validate changes in select candidates. We found that AECs cultured in monolayer and ALI have strikingly different transcriptional states at baseline. When challenged with flagellin, monolayer AEC cultures greatly increased transcription of numerous genes mapping to wounding response, immunity and inflammatory response. In contrast, AECs in ALI culture had an unexpectedly muted response to flagellin, both in number of genes expressed and relative enrichment of inflammatory and immune pathways. We conclude that in vitro culturing methods have a dramatic effect on the transcriptional profile of AECs at baseline and after stimulation with flagellin. These differences suggest that epithelial responses to pathogen challenges are distinctly different in culture models of intact and injured epithelium.
authors: Division of Pulmonary and Critical Care Medicine, Department of Medicine, University of Washington, Seattle, Washington, United States of America; Fred Hutchinson Cancer Research Center, Seattle, Washington, United States of America.
authors: Joan G
authors: JG
authors: Clark
authors: Fred Hutchinson Cancer Research Center, Seattle, Washington, United States of America.
authors: Kyoung-Hee
authors: KH
authors: Kim
authors: Fred Hutchinson Cancer Research Center, Seattle, Washington, United States of America.
authors: Ryan S
authors: RS
authors: Basom
authors: Division of Pulmonary and Critical Care Medicine, Department of Medicine, University of Washington, Seattle, Washington, United States of America; Computational Medicine Core, Center for Lung Biology, University of Washington, Seattle, Washington, United States of America.
authors: Sina A
authors: SA
authors: Gharib
mesh_terms: D001483:Base Sequence; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000378:metabolism*; D005408:Flagellin / Q000494:pharmacology*; D005786:Gene Expression Regulation / Q000187:drug effects*; D006801:Humans; D046228:Microarray Analysis; D008969:Molecular Sequence Data; D020545:Respiratory Mucosa / Q000166:cytology*; D017423:Sequence Analysis, RNA; D059467:Transcriptome / Q000187:drug effects* / Q000235:genetics
chemical_list: D005408:Flagellin