Mechanisms of allergy and clinical immunology
In vitro susceptibility to rhinovirus infection is greater for bronchial than for nasal airway epithelial cells in human subjects

https://doi.org/10.1016/j.jaci.2009.03.010Get rights and content

Background

Human rhinoviruses (HRVs) characteristically cause upper respiratory tract infection, but they also infect the lower airways, causing acute bronchitis and exacerbating asthma.

Objective

Our purpose was to study ex vivo the differences in the response to HRV infection of nasal and bronchial epithelial cultures from the same healthy and asthmatic individuals using conditions favoring development of fully differentiated, pseudostratified mucociliary epithelium.

Methods

Cells from the inferior turbinates and bronchial tree of 5 healthy and 6 asthmatic individuals were cultured at an air-liquid interface. Cultures were infected with HRV-16, and after 48 hours, the degree of infection was measured.

Results

Baseline median transepithelial resistance was lower in human bronchial epithelial (HBE) cell cultures than in human nasal epithelial (HNE) cell cultures (195 Ω.cm2 [95% CI, 164-252] vs 366 Ω.cm2 [95% CI, 234-408], respectively; P < .01). Virus replicated more easily in HBE cells than in HNE cells based on virus shedding in apical wash (log tissue culture infective dose of 50%/0.1 mL = 2.0 [95% CI, 1.0-2.5] vs 0.5 [95% CI, 0.5-1.5], P < .01) and on a 20- to 30-fold greater viral load and number of infected cells in HBE cell cultures than in HNE cell cultures. The increases in expression of RANTES and double-stranded RNA-dependent protein kinase were greater in HBE cell cultures than in HNE cell cultures, as were the concentrations of IL-8, IL-1α, RANTES, and IP-10 in basolateral medium. However, no significant differences between asthmatic and healthy subjects (including IFN-β1 expression) were found.

Conclusions

Differentiated nasal epithelial cells might have mechanisms of increased resistance to rhinovirus infection compared with bronchial epithelial cells. We could not confirm previous reports of increased susceptibility to HRV infection in epithelial cells from asthmatic subjects.

Section snippets

Study subjects

Six subjects with allergic asthma and 5 healthy nonallergic subjects were studied (Table I). Allergic status was based on the presence of a positive skin prick test response (wheal 3 mm greater than the negative control with erythema) to common allergens.20 Asthmatic subjects had a diagnosis of asthma and bronchial hyperresponsiveness (PC20 methacholine, <8 mg/mL),20 were allergic to at least 1 allergen, and had mild-to-moderate disease, as defined by the National Institutes of Health

Epithelial cell cultures

After 2 to 3 weeks of growth at an air-liquid interface, all HNE (n = 70) and HBE (n = 84) cell cultures appeared fully confluent under the inverted microscope and showed properties resembling those of native epithelium, including an apical mucus layer and ciliated pseudostratified epithelium (Fig 1), as well as a permeability barrier, as demonstrated by a “dry” apical surface and the development of Rte, which is indicative of tight junction formation.22 Cultures also showed vectorial transport

Discussion

The 2 major findings of this study are that bronchial epithelial cells are more susceptible to infection with HRV-16 than nasal epithelial cells from the same subjects and that cultures of either airway site from asthmatic subjects did not have a higher response to HRV infection than cultures from healthy subjects. The first finding rejects our hypothesis that the predominance of upper airway symptoms with most rhinovirus infections is due to a greater susceptibility of nasal versus bronchial

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    Supported by National Institutes of Health grant P01AI050496. S. Favoreto and P. C. Avila are currently supported by the Ernest S. Bazley Grant to Northwestern University.

    Disclosure of potential conflict of interest: W. E. Finkbeiner has received research support from the Cystic Fibrosis Foundation and the National Institutes of Health. G. M. Dolganov has provided legal consultation or expert witness testimony on the topic of gene expression assays. J. H. Widdicombe has received research support from Cystic Fibrosis Research, Inc. H. A. Boushey has received research support from GlaxoSmithKline and honoraria from Genentech and Novartis and has served as a member and chair of the Health Effect's Institute's review committee. P. C. Avila has received research support from Genentech. The rest of the authors have declared that they have no conflict of interest.

    H. A. Boushey and P. C. Avila contributed equally to this work.

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