Functional effects of Toll-like receptor (TLR)3, 7, 9, RIG-I and MDA-5 stimulation in nasal epithelial cells

PLoS One. 2014 Jun 2;9(6):e98239. doi: 10.1371/journal.pone.0098239. eCollection 2014.

Abstract

Background: The human nasal epithelium is an important physical barrier, and a part of the innate immune defense that protect against pathogens. The epithelial cells recognize microbial components by pattern-recognition receptors (PRRs), and thereby trigger an immune response. Even though TLR3, TLR7, TLR9, RIG-I and MDA-5 are all known to respond to viral stimulation, their potential role in chronic airway inflammation triggered by local cytokine release remains to be established.

Methods: mRNA and corresponding protein expression of TLR3, TLR7, TLR9, RIG-I and MDA-5 were analyzed in nasal biopsies and various upper airway epithelial cell lines using real-time reverse transcription PCR, immunohistochemistry and flow cytometry. Ligand induced, cytokine release, was evaluated with ELISA.

Results: Nasal biopsies were found to express TLR3, TLR7, TLR9, RIG-I and MDA-5, with the most abundant expression in the surface epithelium. These receptors were verified in primary human nasal epithelial cell (HNEC) as well as in the airway epithelial cell lines Detroit-562 and FaDu. Poly(I:C) (TLR3) and R-837 (TLR7) stimulation increased secretion of IL-6 and GM-CSF from the nasal mucosa and the epithelial cell lines. CpG (TLR9) stimulation caused release of IL-8 in the nasal mucosa and in FaDu. Poly(I:C)/LyoVec (RIG-I/MDA-5) stimulation activated the secretion of IFN-β in the nasal mucosa. A corresponding release was also detected from HNEC and Detroit-562.

Conclusion: The nasal epithelium has the ability to recognize viral intrusion through TLR and RLR receptors, and the subsequent response might have a role in exacerbation of inflammatory diseases like allergic rhinitis and chronic rhinosinusitis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adolescent
  • Adult
  • Cell Line
  • DEAD Box Protein 58
  • DEAD-box RNA Helicases / genetics
  • DEAD-box RNA Helicases / metabolism*
  • Epithelial Cells / metabolism*
  • Epithelium / metabolism
  • Female
  • Humans
  • Interferon-Induced Helicase, IFIH1
  • Male
  • Nasal Mucosa / metabolism
  • Nose / cytology*
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Receptors, Immunologic
  • Toll-Like Receptors / genetics
  • Toll-Like Receptors / metabolism*
  • Young Adult

Substances

  • RNA, Messenger
  • Receptors, Immunologic
  • Toll-Like Receptors
  • RIGI protein, human
  • IFIH1 protein, human
  • DEAD Box Protein 58
  • DEAD-box RNA Helicases
  • Interferon-Induced Helicase, IFIH1

Grants and funding

The authors would like to acknowledge the support from the Swedish Medical Research Council, Grant number:2011-3357, http://www.vr.se. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.