title: Adenoviral vectors stimulate innate immune responses in macrophages through cross-talk with epithelial cells.
abstract: Although adenovirus vectors (Ads) have been widely utilized for gene delivery, their clinical application has been hampered by host immune responses. It has been shown that macrophages can induce inflammatory response against Ads in vivo, but they are not easily activated by Ads in vitro, suggesting their activation requires interaction with other cells. In this study, we investigated the interaction between macrophages and epithelial cells during Ad infection. Ad infection of the macrophage-epithelial cell co-culture resulted in rapid and drastic changes in the cell culture such as decrease in pH within 24h, indicating macrophage activation. Ad infected co-culture showed several characteristics of inflammation including cytotoxicity, induction of pro-inflammatory cytokines, and generation of nitric oxide and reactive oxygen species. These signs of macrophage activation and inflammation were observed exclusively in the co-culture and were absent or significantly weaker in the macrophage mono-culture suggesting that there was a synergistic response by the interaction between macrophages and epithelial cells. We found that inhibition of NF-κB activation significantly reduced the inflammatory responses in the co-culture. Furthermore, we show that only the macrophages adjacent to epithelial cells were activated during Ad infection demonstrating that the interaction between macrophages and epithelial cells are crucial for Ad-induced inflammatory response.
authors.affiliation: Physiology and Experimental Medicine Program, Toronto, Ontario, Canada.
authors.forename: Benjamin H
authors.initials: BH
authors.lastname: Lee
authors.forename: Rahul
authors.initials: R
authors.lastname: Kushwah
authors.forename: Jing
authors.initials: J
authors.lastname: Wu
authors.forename: Philip
authors.initials: P
authors.lastname: Ng
authors.forename: Nades
authors.initials: N
authors.lastname: Palaniyar
authors.forename: Sergio
authors.initials: S
authors.lastname: Grinstein
authors.forename: Dana J
authors.initials: DJ
authors.lastname: Philpott
authors.forename: Jim
authors.initials: J
authors.lastname: Hu
mesh_terms: D000256:Adenoviridae / Q000235:genetics* / Q000502:physiology; D000818:Animals; D015153:Blotting, Western; D002450:Cell Communication / Q000276:immunology; D002460:Cell Line; D002470:Cell Survival / Q000276:immunology; D018925:Chemokines / Q000235:genetics / Q000378:metabolism; D018920:Coculture Techniques; D016207:Cytokines / Q000235:genetics / Q000378:metabolism; D003602:Cytotoxicity, Immunologic / Q000276:immunology; D004847:Epithelial Cells / Q000276:immunology* / Q000378:metabolism / Q000821:virology; D005434:Flow Cytometry; D005822:Genetic Vectors / Q000235:genetics*; D054884:Host-Pathogen Interactions; D018836:Inflammation Mediators / Q000378:metabolism; D008262:Macrophage Activation / Q000276:immunology; D008264:Macrophages / Q000276:immunology* / Q000378:metabolism / Q000821:virology; D016328:NF-kappa B / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism; D052247:Nitric Oxide Synthase Type II / Q000235:genetics / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D018925:Chemokines; D016207:Cytokines; D018836:Inflammation Mediators; D016328:NF-kappa B; D017382:Reactive Oxygen Species; D009569:Nitric Oxide; D052247:Nitric Oxide Synthase Type II