Inducing a Mucosal Barrier-Sparing Inflammatory Response in Laboratory-Grown Primary Human Nasal Epithelial Cells

Mahnaz Ramezanpour; Harrison Bolt; Alkis Psaltis; Peter-John Wormald; Sarah Vreugde

doi:10.1002/cptx.69

Inducing a Mucosal Barrier-Sparing Inflammatory Response in Laboratory-Grown Primary Human Nasal Epithelial Cells

Curr Protoc Toxicol. 2019 Jun;80(1):e69. doi: 10.1002/cptx.69. Epub 2019 Feb 4.

Authors

Mahnaz Ramezanpour¹, Harrison Bolt¹, Alkis Psaltis¹, Peter-John Wormald¹, Sarah Vreugde¹

Affiliation

¹ Department of Surgery, Otorhinolaryngology Head and Neck Surgery, Queen Elizabeth Hospital, and the University of Adelaide, Adelaide, South Australia, Australia.

PMID: 30715797
DOI: 10.1002/cptx.69

Abstract

Here we use the toll-like receptor (TLR) 3 agonist poly I:C (LMW) to induce an inflammatory response in cells of submerged and/or air-liquid interface (ALI) cultures of human nasal epithelial cells (HNECs). The inflammatory response is determined by measuring interleukin-6 (IL-6) protein levels by enzyme-linked immunosorbent assay (ELISA). The mucosal barrier integrity is determined by measuring transepithelial electrical resistance (TEER) and passage of fluorescently labeled dextrans. Stimulation with poly (I:C) LMW induces a 15- to 17-fold increase in IL-6 production by HNEC-ALI cells. © 2019 by John Wiley & Sons, Inc.

Keywords: IL-6; TLR 3; air-liquid interface cultures; chronic rhinosinusitis; immunofluorescence; poly (I:C) LMW; primary human nasal epithelial cells.

MeSH terms

Cells, Cultured
Epithelial Cells / drug effects
Epithelial Cells / immunology*
Epithelial Cells / metabolism
Guidelines as Topic
Humans
Interleukin-6 / biosynthesis*
Nasal Mucosa / cytology
Nasal Mucosa / immunology*
Nasal Mucosa / metabolism
Poly I-C / pharmacology
Primary Cell Culture / methods*
Toll-Like Receptor 3 / agonists*

Substances

IL6 protein, human
Interleukin-6
TLR3 protein, human
Toll-Like Receptor 3
Poly I-C