Airway inflammation and illness severity in response to experimental rhinovirus infection in asthma

Jie Zhu, Simon D Message, Yusheng Qiu, Patrick Mallia, Tatiana Kebadze, Marco Contoli, Christine K Ward, Elliot S Barnathan, Mary Ann Mascelli, Onn M Kon, Alberto Papi, Luminita A Stanciu, Peter K Jeffery, Sebastian L Johnston, Jie Zhu, Simon D Message, Yusheng Qiu, Patrick Mallia, Tatiana Kebadze, Marco Contoli, Christine K Ward, Elliot S Barnathan, Mary Ann Mascelli, Onn M Kon, Alberto Papi, Luminita A Stanciu, Peter K Jeffery, Sebastian L Johnston

Abstract

Background: The nature of bronchial mucosal inflammation and its physiologic and clinical significance in rhinovirus-induced asthma exacerbations is unclear. We investigated bronchial mucosal inflammatory response and its association with physiologic and clinical outcomes in an experimental model of rhinovirus-induced asthma exacerbations.

Methods: We used immunohistochemistry methods to detect phenotypes of inflammatory cells infiltrating the bronchial mucosa before and after experimental rhinovirus infection in 10 subjects with asthma and 15 normal subjects.

Results: Compared with baseline, rhinovirus infection significantly increased the number of epithelial (P = .005) and subepithelial (P = .017) neutrophils in subjects with asthma only and subepithelial CD68+ macrophages in both subjects with asthma (P = .009) and normal subjects (P = .018) but more so in those with asthma (P = .021). Numbers of CD45+, CD68+, and CD20+ cells; neutrophils; and eosinophils at day 4 postinfection were positively associated with virus load (r = 0.50-0.72, P = .016-0.03). At acute infection in subjects with asthma, CD4+ cells correlated with chest symptom scores (r = 0.69, P = .029), the fall in the 10% fall in FEV1 (PC10) correlated with neutrophils (r = -0.89, P = .029), the PC10 correlated inversely with CD4+ (r = -0.67, P = .023) and CD8+ cells (r = -0.65, P = .03), the 20% fall in FEV1 was inversely associated with CD20+ cells (r = -0.65, P = .03), and higher epithelial CD8+ cell counts were significantly associated with a greater maximum fall in FEV1 (r = -0.72, P = .03), whereas higher subepithelial mast cell counts were significantly associated with a lower maximum percent fall in peak expiratory flow (r = 0.8, P = .024).

Conclusions: In subjects with asthma, rhinovirus infection induces bronchial mucosal neutrophilia and more severe monocyte/macrophage infiltration than in normal subjects. Airway neutrophils, eosinophils, and T and B lymphocytes during infection are related to virus load and physiologic and clinical severity, whereas mast cells are related to greater lung function.

Figures

Figure 1.
Figure 1.
Immunohistochemistry-stained cells are seen as red fuchsin positivity. A and B, Subject with asthma at day 4 rhinovirus 16 infection demonstrating more elastase-positive neutrophils (A) and CD68+ monocytes/macrophages (B) in bronchial epithelium and subepithelium. C and D, Subject with asthma at baseline showing fewer neutrophils (C) and CD68+ cells (D). E, Negative control sample shows an absence of signal. Scale bar = 20 μm for all images.
Figure 2.
Figure 2.
Cell counts in bronchial biopsy specimens of subjects with and without asthma at baseline and days 4 and 6 of rhinovirus 16 infection. A, CD45+ cells. B, Epithelial neutrophils. C, Subepithelial neutrophils. D, CD68+ cells. E, CD4+ cells. Data are presented as the number of positive cells per square millimeter of subepithelium or per 0.1 mm2 of epithelium. △ and ▲ show individual counts, and horizontal bars show median values (Wilcoxon matched pairs and Mann-Whitney U tests).
Figure 3.
Figure 3.
A and B, Changes in counts of epithelial (A) and subepithelial (B) neutrophils and CD68+ cells from baseline to d 4 postinfection in bronchial biopsy specimens from subjects with and without asthma. Data are presented as change in the number of positive cells per 0.1 mm2 of epithelium or per square millimeter of subepithelium. ○ and ● show individual changes in counts, and horizontal bars show median values (Mann-Whitney U test).
Figure 4.
Figure 4.
A-C, Correlations between the numbers of subepithelial CD45+ cells (A) at d 4 and peak sputum virus load at d 3 after infection and neutrophils (B) and CD68+ cells (C) at d 4 and BAL virus load at d 4 in all subjects with asthma (●) and normal subjects (○) (Spearman rank correlation).
Figure 5.
Figure 5.
A-F, In subjects with asthma alone, correlations between total chest symptom score d 0 to 14 after the rhinovirus 16 infection period and counts of epithelial CD4+ cells (A), between fall in PC10 (log2) and counts of subepithelial neutrophils at d 4 (B), between PC10 at d 6 and counts of subepithelial CD3+ (C), CD4+ (D), and CD8+ (E) cells at d 4; and between PC20 at d 6 and counts of subepithelial CD20+ cells at d 4 (F) (Spearman rank correlation, n = 10 for all). PC10 = 10% fall in FEV1; PC20 = 20% fall in FEV1.
Figure 6.
Figure 6.
A, B, In subjects with asthma at d 4 after rhinovirus infection, correlations between maximum fall in FEV1 (%) and counts of epithelial CD8+ cells (A) and between maximum fall in PEF (%) and counts of subepithelial tryptase-positive mast cells (B) (Spearman rank correlation). PEF = peak expiratory flow.
Figure 7.
Figure 7.
A, B, Correlations between serum IgE levels and counts of subepithelial EG2+ eosinophils in asthma at baseline (A) and d 4 after rhinovirus infection (B) (Spearman rank correlation).

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Source: PubMed

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