Rhinitis, sinusitis, and upper airway disease
Cigarette smoke combined with Toll-like receptor 3 signaling triggers exaggerated epithelial regulated upon activation, normal T-cell expressed and secreted/CCL5 expression in chronic rhinosinusitis

https://doi.org/10.1016/j.jaci.2008.09.033Get rights and content

Background

Chronic rhinosinusitis (CRS) is characterized by persistent mucosal inflammation and frequent exacerbations.

Objective

To determine whether innate epithelial responses to cigarette smoke or bacterial or viral pathogens may be abnormal in CRS leading to an inappropriate inflammatory response.

Methods

Primary nasal epithelial cells (PNECs) were grown from middle turbinate biopsies of 9 healthy controls and 11 patients with CRS. After reaching 80% to 90% confluence, PNECs were exposed to medium or cigarette smoke extract (CSE) 5% (vol/vol) for 1 hour, washed, then stimulated with staphylococcal lipoteichoic acid, LPS, or double-stranded RNA (dsRNA). After 24 hours, gene expression was quantified by QRT-PCR.

Results

At baseline, PNECs revealed elevated TNF-α and growth-related oncogene-α (a C-X-C chemokine)/CXCL1 (GRO-α) (4-fold increase, P = .02; and 16-fold increase, P = .004, respectively) in subjects with CRS compared with controls with normal levels of IL-1β, IL-6, IL-8/CXCL8, human β-defensin-2, monocyte chemoattractant protein 2/CCL8, monocyte chemoattractant protein 3/CCL7, and regulated upon activation, normal T-cell expressed and secreted (RANTES)/CCL5. Immunostaining of nasal biopsies, however, revealed comparable epithelial staining for TNF-α, GRO-α, and RANTES. There were no differences in mRNA induction by CSE, TNF-α, lipoteichoic acid, LPS, or dsRNA alone. The combination of CSE+dsRNA induced exaggerated RANTES (12,115-fold vs 1500-fold; P = .03) and human β-defensin-2 (1120-fold vs 12.5-fold; P = .05) in subjects with CRS. No other genes were differentially induced. Furthermore, CSE+dsRNA induced normal levels of IFN-β, IFN-λ1, and IFN-λ2/3 mRNA in subjects with CRS.

Conclusion

Cigarette smoke extract plus dsRNA induces exaggerated epithelial RANTES expression in patients with CRS. We propose that an analogous response to cigarette smoke plus viral infection may contribute to acute exacerbations and eosinophilic mucosal inflammation in CRS.

Section snippets

Methods

The study was approved by the Institutional Review Board of both Massachusetts General Hospital and the Massachusetts Eye and Ear Institute. Each subject gave informed consent and completed a questionnaire, allergy skin testing, and a rhinoscopic examination. Healthy controls (N = 9) included men and women between the ages of 21 and 70 years in good general health. Exclusion criteria included a history of smoking, nasal or sinus disease, use of nasal or sinus medications, asthma, allergic

Summary of subject characteristics

The demographics of the healthy controls (HCs) and subjects with CRS are summarized in Table I. HCs were younger (age, 28 ± 10.8 years vs CRS, 44 ± 7.4 years) but had a similar sex distribution (female:male ratio, 7:2 and 8:3, respectively). Four subjects with CRS had asthma. Positive allergy skin test results were found in 5 of 9 HCs (despite a negative history of allergic rhinitis) and 7 of 11 subjects with CRS. Serum cotinine (a metabolite of nicotine) levels averaged 0.07 ng/mL in HCs and

Discussion

In this study, we investigated the potential role of the sinonasal epithelium in the pathogenesis of CRS by examining inflammatory gene expression in PNEC cultures. We identified a mild increase in baseline epithelial expression of TNF-α and GRO-α in subjects with CRS. HCs and subjects with CRS responded similarly to CSE, TNF-α, LTA (TLR-2 stimulus), and LPS (TLR-4 stimulus) as well as CSE+LTA and CSE+LPS. dsRNA was the strongest stimulus of inflammatory gene expression. Furthermore, patients

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    Supported by a grant from the Flight Attendants Medical Research Institute.

    Disclosure of potential conflict of interest: D. L. Hamilos has received honoraria from Merck, Critical Therapeutics, Genentech, and Novartis; has consulting arrangements with GlaxoSmithKline, Novartis, Accentia, Schering-Plough, and Sinexus; has received research support from the Flight Attendants Medical Research Institute and Merck; and has received gifts from the GG Monks Foundation to purchase equipment. E. H. Holbrook has received research support from the National Institutes of Health-National Institutes of Deafness and Other Communication Disorders. S. T. Gray has served as a member of the American Academy of Otolaryngology and the American Rhinologic Society. N. Busaba is on the speakers' bureau for Sanofi-Aventis and Schering-Plough. The rest of the authors have declared that they have no conflict of interest.

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