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. 2010 Dec;43(6):652-61.
doi: 10.1165/rcmb.2009-0244OC. Epub 2010 Jan 15.

Interleukin-13-induced mucous metaplasia increases susceptibility of human airway epithelium to rhinovirus infection

Marrah E Lachowicz-Scroggins  1 , Homer A Boushey, Walter E Finkbeiner, Jonathan H Widdicombe
Affiliations

Interleukin-13-induced mucous metaplasia increases susceptibility of human airway epithelium to rhinovirus infection

Marrah E Lachowicz-Scroggins et al. Am J Respir Cell Mol Biol. 2010 Dec.

Abstract

Infection of airway epithelium by rhinovirus is the most common cause of asthma exacerbations. Even in mild asthma, airway epithelium exhibits mucous metaplasia, which increases with increasing severity of the disease. We previously showed that squamous cultures of human airway epithelium manifest rhinoviral infection at levels many times higher than in well-differentiated cultures of a mucociliary phenotype. Here we tested the hypothesis that mucous metaplasia is also associated with increased levels of rhinoviral infection. Mucous metaplasia was induced with IL-13, which doubled the numbers of goblet cells. In both control (mucociliary) and IL-13- treated (mucous metaplastic) cultures, goblet cells were preferentially infected by rhinovirus. IL-13 doubled the numbers of infected cells by increasing the numbers of infected goblet cells. Furthermore, IL-13 increased both the maturity of goblet cells and the probability that a goblet cell would be infected. The infection of cells other than goblet cells was unaltered by IL-13. Treatment with IL-13 did not alter the levels of rhinovirus receptor ICAM-1, nor did the proliferative effects of IL-13 enhance infection, because rhinovirus did not colocalize with dividing cells. However, the induction of mucous metaplasia caused changes in the apical membrane structure, notably a marked decrease in overall ciliation, and an increase in the overall flatness of the apical surface. We conclude that mucous metaplasia in asthma increases the susceptibility of airway epithelium to infection by rhinovirus because of changes in the overall architecture of the apical surface.

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Figures

Figure 1.
Figure 1.
Induction of mucous metaplasia by IL-13. (A) Detection of mucus with fluorescent periodic acid Schiff (orange). Focal plane, ∼5 μm below apical membrane; nuclei are green. (B) Representative staining of goblet cells with monoclonal antibody B6E8 (blue). Nuclei are stained green with Yo-Pro-1. (C) Light microscopy with mucicarmine staining. Distended cell bodies of several goblet cells are marked by asterisks. Several more lie predominantly outside the plane of the section, but all can be identified by the mucus they release. Each pair of images was from cell sheets derived from the same trachea. Scale bars = 25 μm (A), 50 μm (B), and 20 μm (C).
Figure 2.
Figure 2.
Colocalization of antibodies MUC5AC and B6E8. Panels, left to right: B6E8, MUC5AC and colocalization of both. Scale bar = 10 μm.
Figure 3.
Figure 3.
Decreased ciliogenesis induced by IL-13 treatment. (A) Scanning electron micrographs of apical surface of mature cell sheets. Large apical surface area and a decrease in ciliated cells are evident after treatment with IL-13. (B) Immunofluorescence of centrin-1. Cilia are evident at the apical surface of cell sheets (green). (C) Quantification of cilia in airway epithelial cells by centrin-1 Western blot. IL-13–treated cell sheets have lower concentration of centrin-1 protein, when normalized to β-actin levels. (D) Pooled data from three experiments. Centrin-1 is detected at ∼2-fold higher level in cell lysates from control cell sheets. AU, arbitrary units as a measure of pixel density. Centrin-1 levels were normalized to β-actin. Means ± SE for 2–3 cells sheets for three cultures.
Figure 4.
Figure 4.
Detection of rhinovirus infection of airway epithelial cells by rhinovirus (RV)-16 immunohistochemistry (nuclei, green, using Yo-Pro-1; RV-16, red). (A) Squamous-cell cultures not receiving virus. (B) Squamous cultures exposed to RV-16. (C) Mucociliary cultures exposed to virus demonstrate occasional perinuclear staining of infectious particles (arrows). (D) IL-13–treated cells exposed to virus. Cell sheets are from the same tissue. Scale bar = 10 μm.
Figure 5.
Figure 5.
Rhinovirus preferentially infects goblet cells, an effect enhanced by goblet-cell metaplasia induced by IL-13. Nuclei, green; mucus, blue; virus, red. (A) Apical plane of control cell sheets shows colocalization of virus to goblet cells. (B) Center plane of control cell sheets shows no mucus staining. (C) Z-stack of control cell sheets. (D) Apical plane of IL-13–treated cell sheet shows colocalization of virus to goblet cells. (E) Center plane of IL-13–treated cell sheet shows no mucus staining. (F) Z-stack of IL-13–treated cell sheets; scale bar = 25 μm. Arrows indicate colocalization of mucus and virus. Where this occurs in IL-13–treated cells, levels of both mucus and virus are higher than in control cells. Asterisks indicate virus unassociated with mucus. Cells sheets were grown from the same trachea. Scale bar in E = 25 μm. Same scale for A, B, D, and E. Red vertical bars in z-stacks represent 25 μm.
Figure 6.
Figure 6.
Quantification of rhinovirus infection by Western blot (A) IL-13–treated cell sheets have higher levels of rhinovirus capsid proteins, when normalized to β-actin levels. (B) Pooled data from three experiments. Rhinovirus proteins VP2 and VP0 are detected at ∼3-fold higher levels in cell lysates from IL-13–treated cell sheets. AU, arbitrary units as a measure of pixel density.
Figure 7.
Figure 7.
Quantification of rhinovirus receptor ICAM-1 in airway epithelial cells. (A) Immunofluorescence of ICAM-1 in control cell sheets shows the receptor is primarily localized to the apical surface. (B) Immunofluorescence of ICAM-1 in IL-13–treated cell sheets shows similar apical localization of ICAM-1, with no apparent association with goblet-cell staining. Top: Z-projection of an x–y stack. Bottom: x–y plane at level of the apical membrane. B6E8, blue; ICAM-1, green, at apical membrane; nuclei, green, using Yo-Pro-1. Scale bar = 15 μm. (C) Squamous-cell sheets have higher levels of ICAM-1 protein levels, compared with control cell sheets when normalized to β-actin levels. Mucus (IL-13–treated) and mucociliary cell sheets have similar levels of ICAM-1 protein when normalized to β-actin levels. (D) Pooled data from three experiments. AU, arbitrary units as a measure of pixel density.
Figure 8.
Figure 8.
Immunofluorescence of Ki-67 (blue) and rhinovirus (red) shows no correlation between dividing cells and infection. Nuclei are green. Results are from a 10-day-old culture in which division was greater than at 21 days. However, virus only infected nondividing cells at all ages. Scale bar = 10 μm.
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References

    1. van Elden LJR, Sachs APE, van Loon AM, Haarman M, van de Vijver DA, Kimman TG, Zuithoff P, Schipper PJ, Verheij TJM, Nijhuis M. Enhanced severity of virus associated lower respiratory tract disease in asthma patients may not be associated with delayed viral clearance and increased viral load in the upper respiratory tract. J Clin Virol 41(2):116–121. - PMC - PubMed
    1. Papadopoulos NG, Papi A, Psarras S, Johnston SL. Mechanisms of rhinovirus-induced asthma. Paediatr Respir Rev 2004;5:255–260. - PMC - PubMed
    1. Jackson DJ, Gangnon RE, Evans MD, Roberg KA, Anderson EL, Pappas TE, Printz MC, Lee W-M, Shult PA, Reisdorf E, et al. Wheezing rhinovirus illnesses in early life predict asthma development in high risk children. Am J Respir Crit Care Med 2008;178:667–672. - PMC - PubMed
    1. Hansbro NG, Horvat JC, Wark PA, Hansbro PM. Understanding the mechanisms of viral induced asthma: new therapeutic directions. Pharmacol Ther 2008;117:313–353. - PMC - PubMed
    1. Tan WC. Viruses in asthma exacerbations. Curr Opin Pulm Med 2005;11:21–26. - PubMed

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