0
Original Research: Asthma |

Reduced Antiviral Interferon Production in Poorly Controlled Asthma Is Associated With Neutrophilic Inflammation and High-Dose Inhaled Corticosteroids FREE TO VIEW

Jodie L. Simpson, PhD; Melanie Carroll, BS; Ian A. Yang, PhD; Paul N. Reynolds, PhD; Sandra Hodge, PhD; Alan L. James, MBBS, PhD; Peter G. Gibson, MBBS; John W. Upham, PhD
Author and Funding Information

FUNDING/SUPPORT: This study was funded by project grants from the National Health and Medical Research Council of Australia [Grants 569246 and 1046622].

CORRESPONDENCE TO: Jodie L. Simpson, PhD, Level 2, West Wing, Hunter Medical Research Institute, Lot 1 Kookaburra Circuit, New Lambton Heights, NSW, 2305, Australia


Copyright 2016, American College of Chest Physicians. All Rights Reserved.


Chest. 2016;149(3):704-713. doi:10.1016/j.chest.2015.12.018
Text Size: A A A
Published online

Background  Asthma is a heterogeneous chronic inflammatory disease in which host defense against respiratory viruses such as human rhinovirus (HRV) may be abnormal. This is a matter of some controversy, with some investigators reporting reduced type I interferon (IFN) synthesis and others suggesting that type I IFN synthesis is relatively normal in asthma.

Objective  The objective of this study was to examine the responsiveness of circulating mononuclear cells to HRV in a large cohort of participants with poorly controlled asthma and determine whether IFN-α and IFN-β synthesis varies across different inflammatory phenotypes.

Methods  Eligible adults with asthma (n = 86) underwent clinical assessment, sputum induction, and blood sampling. Asthma inflammatory subtypes were defined by sputum cell count, and supernatant assessed for IL-1β. Peripheral blood mononuclear cells (PBMCs) were exposed to HRV serotype 1b, and IFN-α and IFN-β release was measured by enzyme-linked immunosorbent assay.

Results  Participants (mean age, 59 years; atopy, 76%) had suboptimal asthma control (mean asthma control questionnaire 6, 1.7). In those with neutrophilic asthma (n = 12), HRV1b-stimulated PBMCs produced significantly less IFN-α than PBMCs from participants with eosinophilic (n = 35) and paucigranulocytic asthma (n = 35). Sputum neutrophil proportion and the dose of inhaled corticosteroids were independent predictors of reduced IFN-α production after HRV1b exposure.

Conclusions  Antiviral type I IFN production is impaired in those with neutrophilic airway inflammation and in those prescribed high doses of inhaled corticosteroids. Our study is an important step toward identifying those with poorly controlled asthma who might respond best to inhaled IFN therapy during exacerbations.

Figures in this Article

Asthma remains common and continues to cause significant morbidity and mortality even in countries with ready access to the best available therapies such as inhaled corticosteroids (ICS) and long-acting β2 agonists. Acute exacerbations are major contributors to the personal and societal burden of asthma, with most exacerbations linked to acute infections with human rhinoviruses (HRVs) and other respiratory viruses.

Evidence suggests that host defense against respiratory viruses is abnormal in asthma, or at least in a subset of people with asthma., Many investigators report abnormal airway and systemic immune responses to HRVs in asthma; this is thought to facilitate exaggerated lower airway inflammation in response to otherwise minor respiratory viral infections. Viral replication is higher in HRV-infected airway epithelial cells from patients with asthma compared with healthy control subjects, in association with deficient interferon (IFN)-β and IFN-λ synthesis, a finding independent of ICS treatment., HRV-activated alveolar macrophages also produce less IFN-λ than those from healthy individuals. Similarly, circulating leukocyte populations from patients with asthma are less responsive to RNA viruses and viral nucleic acids than cells from healthy control subjects,, producing less IFN-α and exhibiting reduced expression of multiple antiviral signaling pathways.

The concept of abnormal viral defenses in asthma remains controversial, with reports that airway epithelial cells from patients with asthma and healthy individuals produce equivalent IFN-β,, especially in well-controlled asthma. One study reported that blood mononuclear cells from patients with asthma exhibit normal IFN synthesis in vitro, although Durrani et al demonstrated significant reduction in virus-stimulated IFN production by blood mononuclear cells isolated from children with asthma. Moreover, this deficient antiviral response was amplified by cross-linking of the high-affinity IgE receptor (FcεRI).

It is possible therefore that deficient antiviral IFN production is confined to a subset of patients with asthma, perhaps those with the most severe disease, or highly sensitized individuals with recent allergen exposure. Such patients are the most prone to acute exacerbations. Asthma is a heterogeneous inflammatory disease that can be categorized into a number of inflammatory subtypes based on the presence or absence of sputum neutrophils and eosinophils, which vary in their inflammatory and immunological profiles and their responsiveness to corticosteroid treatment.,, This inflammatory response is orchestrated by a plethora of inflammatory mediators, including IL-6 and IL-1β, which have been described both in asthma in general,, and, more specifically, in neutrophilic asthma.,,,, Little progress has been made in assessing the relationship between these inflammatory phenotypes and antiviral IFN production. Inhaled IFN is currently being developed to ameliorate virus-induced asthma exacerbations, an approach that appears most effective in a subset of patients with difficult-to-treat asthma. There is a need to identify which patients exhibit deficient antiviral IFN production, as they should belong to the specific subgroup that might respond best to inhaled IFN.

The aim of this study was to examine systemic antiviral IFN production in a large group of patients with exacerbation-prone, poorly controlled asthma. Our hypothesis was that HRV-induced IFN-α and IFN-β production would vary across different inflammatory phenotypes.

Study Design and Participants

This cross-sectional study assessed eligible adults with asthma (n = 86) before commencement of study medication as part of a larger multicenter randomized controlled trial. The diagnosis of asthma was established using the American Thoracic Society guidelines as outlined in the supplementary material. Participants with asthma were stable but symptomatic, despite being prescribed maintenance ICS treatment with an Asthma Control Questionnaire 6 (ACQ6) score >0.75.,, Participants had no reported exacerbations or alterations in respiratory medications in the previous 4 weeks. Exclusion criteria and patient assessment details can be found in the supplementary material. Local institutional review boards approved the protocol (detail in supplementary material), and written informed consent was obtained from all patients.

Sputum Induction and Analysis

Spirometry (CPFS/D USB Spirometer, BreezeSuite v7.1, MGC Diagnostics) was performed, venous blood samples were collected, and sputum was induced by hypertonic saline (4.5%) inhalation as described by Gibson et al.

Sputum was processed as described, with further detail in the e-Appendix 1. Asthma subtypes were defined using sputum eosinophils and neutrophils as previously described with the cut point for eosinophilic asthma being a sputum eosinophil count ≥3% and the cut point for neutrophilic asthma being a sputum neutrophil count >61%.

Isolation and Culture of Peripheral Blood Mononuclear Cells

Peripheral blood mononuclear cells (PBMCs) were isolated from heparinized blood by density gradient centrifugation as described; see Supplementary Methods for further details. All cultures were performed in RPMI 1640 medium (Life Technologies) with 5% autologous plasma and penicillin/streptomycin/glutamine (Life Technologies). PBMCs were cultured with either medium alone or live HRV1b (multiplicity of infection, 0.1) for 24 hours. Supernatant was harvested and assayed by enzyme-linked immunosorbent assay (ELISA). C-X-C motif ligand (CXCL) 10 (also known as IFN-γ-induced protein 10 [IP-10]), CXCL8, and IL-6 were measured by sandwich ELISA using paired antibody sets (BD Biosciences). The limit of detection was 3.9 pg/mL. Commercially available sandwich ELISA kits were used to assay IFN-α (PBL Assay Science) and IFN-β (elisakit.com). The limits of detection were 12.5 pg/mL and 1.9 pg/mL, respectively. In samples in which the cytokine concentration was below the lower limit of detection for that assay, it was not possible to determine whether the true cytokine was zero, so the concentration was arbitrarily assigned to be half the lower limit of detection.

Rhinovirus Generation and Titration

Rhinovirus was propagated by passage in Ohio HeLa cells as outlined previously, followed by purification with iodixanol solution (Sigma-Aldrich). The 50% tissue culture infective dose was determined by titration in Ohio HeLa cells as described by Carroll et al.

Effects of IL-1β and IL-6 on Rhinovirus-Stimulated Interferon Production

PBMCs from 9 healthy volunteers (median age, 35 years; range, 25–61 years) were incubated with IL-6 or IL-1β (BD Biosciences) for 4 hours before addition of HRV1b (multiplicity of infection, 0.1). After 24 hours, supernatant was harvested and assayed by ELISA.

Statistical Analyses

The results are expressed as mean ± SD for continuous variables and median with interquartile range when data were not normally distributed, with further detail on the analysis methods described in the Supplementary Methods.

Study participants comprised 86 adult patients with asthma (mean age, 59 years) who were predominantly atopic (76%) with a mean ACQ6 score of 1.7 (Table 1). All except 3 participants were taking ICS in combination with long-acting β2 agonists.

Table Graphic Jump Location
Table 1 Participant Characteristics

Data are presented as median (q1, q3), unless indicated otherwise. ACQ6 = asthma control questionnaire 6; ICS = inhaled corticosteroid.

Based on induced sputum cell count, 35 participants (41%) each had an eosinophilic asthma phenotype, 35 (41%) had paucigranulocytic, 12 (14%) had a neutrophilic phenotype, and 4 (5%) had a mixed eosinophilic/neutrophilic phenotype. Because of the small number of participants with mixed granulocytic asthma, they were not included in this analysis. The clinical characteristics along with the peripheral blood lymphocyte and monocyte counts according to inflammatory phenotype are shown in e-Table 1. Participants with neutrophilic asthma had a significantly lower FEV1 % predicted and few peripheral blood lymphocytes compared with participants with eosinophilic asthma. As expected, participants with neutrophilic asthma had a significantly higher total number of sputum leukocytes and proportions of neutrophils (Table 2). There was no difference in smoking status or smoking pack-years between inflammatory phenotypes (P > .05, data not shown). Sputum IL-1β was assessed in 80 samples (when sufficient supernatant was available) and was significantly higher in participants with neutrophilic asthma than in those with eosinophilic and paucigranulocytic asthma. Serum IL-6 was assessed in 85 participants; results varied significantly across the four inflammatory phenotypes, with the highest concentrations observed in the neutrophilic and mixed granulocytic asthma subtypes (Table 2).

Table Graphic Jump Location
Table 2 Inflammatory Cell Counts and Mediators by Asthma Inflammatory Phenotype
a P < .008 vs eosinophilic asthma.
b P < .008 vs paucigranulocytic asthma.

Data are median (q1, q3).

Response of Peripheral Blood Mononuclear Cells to Human Rhinovirus 1b Exposure

The cytokine release from all participants is shown in e-Figure 1. PBMCs from all participants responded to HRV1b exposure, releasing IFN-α and CXCL10 in excess of media alone. HRV1b-activated PBMCs released IFN-β in 90% of participants, IL-6 in 71%, IL-1β in 60%, and CXCL8 in 46%. These percentages were similar across the different inflammatory phenotypes. The median values and first and third quartiles for PBMCs incubated with medium alone are shown in e-Table 2.

Figure 1 shows the cytokine levels released in response to HRV1b stimulation, corrected for baseline (unstimulated) cytokine values. This was calculated by subtracting the baseline cytokine result from the HRV1b-stimulated result. Therefore a zero data point indicates that no additional release of cytokine was observed above the unstimulated levels measured from the control wells. Further details of the data are shown in e- Figure 1. HRV1b-activated PBMCs from participants with neutrophilic asthma released significantly less IFN-α and IFN-β than PBMCs from those with eosinophilic and paucigranulocytic asthma (post-hoc rank-sum P = .002 and P = .022, respectively). Although HRV1b-activated PBMCs from participants with neutrophilic asthma also tended to release less CXCL10, CXCL8, and IL-6 than PBMCs from participants with other inflammatory subtypes, these differences were not statistically significant.

Figure 1
Figure Jump LinkFigure 1 Inflammatory cytokine levels in supernatant from human rhinovirus 1b-activated, cultured peripheral blood mononuclear cells are shown (data corrected for baseline unstimulated levels). Bars represent median levels for IFN-α (A), IFN-β (B), CXCL10 (C), CXCL8 (D), IL-1β (E), and IL-6 (F). The number of participants for each group is below the inflammatory subtype in each panel; the dotted line shows the zero level for each cytokine. Results for all three groups were compared by Kruskal-Wallis test and indicated by the probability value in each panel. Significant differences between any two groups (P < .05 by post-hoc rank-sum test) are indicated by * compared with eosinophilic asthma and # compared with paucigranulocytic asthma. CXCL = C-X-C motif ligand; Eos = eosinophilic asthma; IFN = interferon; Neut = neutrophilic asthma; Pauci = paucigranulocytic asthma.Grahic Jump Location
Clinical and Inflammatory Associations With Human Rhinovirus 1b-Induced Interferon-α Release

IFN-α release from HRV1b-activated PBMCs was significantly inversely related to the dose of ICS (Fig 2A), asthma control (as assessed by ACQ6 score, Fig 2B), proportion of sputum neutrophils (Fig 2C), serum IL-6 (Fig 2D), and sputum IL-1β (Fig 2E) and was positively associated with atopy (as assessed by allergy skin test [Fig 2F]) and the number of peripheral blood lymphocytes (Fig 2G) before randomization. There was no significant association with age, sex, or smoking pack-years.

Figure 2
Figure Jump LinkFigure 2 Scatter plots of IFN production stimulated by human rhinovirus 1b selected for ICS total daily dose (A), mean ACQ-6 score (B), sputum neutrophil % (C), serum IL-6 (D), sputum IL-1β (E), atopy (F), blood lymphocytes (G), and blood monocytes (H). Spearman correlation coefficient (r) and probability values are as indicated. The comparison between atopic and nonatopic participants was assessed using Mann-Whitney test. ACQ-6 = asthma control questionnaire 6; ICS = inhaled corticosteroid. See Figure 1 legend for expansion of other abbreviation.Grahic Jump Location

Using stepwise multiple linear regression, the proportion of sputum neutrophils and the dose of ICS were the only significant independent predictors of IFN-α release after correction for age and sex (Table 3). The model included 79 data points and explained 27% of the variance in released IFN-α with P < .0001.

Table Graphic Jump Location
Table 3 Univariate and Stepwise Multivariate Regression Analysis
a Peripheral blood lymphocytes were assessed 2-4 weeks before randomization.

The model included 79 data points and explained 27% of the variance in IFN-β levels and was highly significant at P < .0001. ACQ6 = asthma control questionnaire 6; ICS = inhaled corticosteroid.

Effect of IL-1β on the Response of Peripheral Blood Mononuclear Cells to Human Rhinovirus 1b

Because neutrophilic asthma has been linked to overexpression of the IL-1β pathway, and because we observed that serum IL-6 (Fig 2) and sputum IL-1β were inversely associated with HRV-induced IFN-α in the current study, we next explored the potential impact of IL-1β on the ability of HRV1b-activated PBMCs to synthesize type I IFN in vitro. Preexposure of PBMCs from healthy donors to IL-1β significantly reduced IFN-α and IFN-β release after HRV exposure (Fig 3). In contrast, IL-6 had no effect on HRV-stimulated IFN-α and IFN-β release (data not shown).

Figure 3
Figure Jump LinkFigure 3 Effects of IL-1β on rhinovirus-stimulated IFN-α (A) and IFN-β (B) production from healthy control peripheral blood mononuclear cells. HRV = human rhinovirus. See Figure 1 legend for expansion of other abbreviation.Grahic Jump Location

The key finding to emerge from the present study is that HRV1b-activated PBMCs from adults with neutrophilic asthma have a significantly lower capacity to produce both IFN-α and IFN-β than PBMCs from adults with eosinophilic and paucigranulocytic asthma. Using stepwise multiple linear regressions, we identified both sputum neutrophil proportion and ICS dose as independent predictors of type I IFN production.

To the best of our knowledge this is the largest study to date of antiviral IFN production in poorly controlled asthma and the first to systematically examine the relationship between type I IFN production and asthma inflammatory phenotypes. IFN-α and IFN-β release was lowest in those participants with neutrophilic asthma, a phenotype that made up 14% of this cohort. These participants were characterized by an elevated total number of sputum leukocytes, increased sputum neutrophil proportions, and significantly elevated sputum IL-1β. Release of several other proinflammatory chemokines and cytokines by HRV-activated PBMC also tended to be lower in participants with neutrophilic asthma, although the differences did not reach statistical significance. Serum IL-6 as a marker of systemic inflammation was readily detectable across all participants, regardless of inflammatory phenotypes, but was highest in those with elevated sputum neutrophils.

IFN-α and IFN-β form part of the first line of defense against viral infections. These IFNs are produced both by airway resident cells and by circulating, bone marrow-derived cells that provide a reservoir that can be rapidly recruited to the lung during viral infection. HRV-stimulated PBMCs readily produce IFN-α and IFN-β, the vast majority of which arises from plasmacytoid dendritic cells and to a lesser extent from monocytes., In vivo, acute respiratory viral infections induce specific gene expression signatures in peripheral blood, and emerging data indicate that HRV-stimulated PBMCs exhibit gene expression signatures highly correlated with results obtained during in vivo HRV infections. Reduced IFN-α release from PBMCs has been reported in children with asthma, in adults with asthma, and in women with asthma both during pregnancy and postpartum. Wright et al recently reported an asthma-specific impairment in toll-like receptor-9–dependent IFN-α production by PBMCs that was not observed in those with COPD or healthy control subjects. This finding is particularly interesting given that the asthma and COPD groups that were studied were both taking high doses of ICS and the participants with asthma were of similar severity to those studied in the present study. Airway epithelial cells from children with severe asthma also show impaired type I IFN production, whereas some studies report that airway epithelial cells from adults with well-controlled asthma exhibit type I IFN production that is similar to that of healthy subjects. One recent study indicated that impaired type I IFN synthesis in asthma was confined to BAL cells, whereas IFN synthesis by rhinovirus-activated blood mononuclear cells was similar in patients with asthma and in healthy subjects. Most of the participants in that study had relatively well-preserved lung function and mild eosinophilia in BAL fluid; no participants appeared to have had neutrophilic airway inflammation. Our findings extend these observations, to show that patients with asthma who are most likely to have impaired type I IFN production are those with a neutrophilic inflammatory subtype. Our findings contrast with a smaller study of 25 patients with severe asthma that did not find a relationship between sputum differential leukocyte counts and IFN-α production; sample size and statistical power may explain the discrepancy between the two studies.

The finding that ICS dose was inversely associated with IFN synthesis use (Table 3) warrants discussion. Bratke et al showed that toll-like receptor-9 mediated IFN-α secretion from plasmacytoid dendritic cells in asthma was lower in those taking ICS than in those not taking ICS. It is conceivable that systemic absorption of high doses of ICS might inhibit the capacity of HRV-stimulated PBMCs to synthesize IFN-α. We have shown that such inhibition can occur in vitro, but only at steroid concentrations of 10−8 M or more; such concentrations are unlikely to have been present systemically in the present study. Moreover, as discussed earlier, others have noted that ICS use in asthma, but not in COPD, is associated with deficient IFN-α production by PBMCs. We propose that the association between high-dose ICS and reduced IFN-α production is indirect, in that patients with noneosinophilic airway inflammation are often refractory to standard ICS doses, but are treated with high ICS doses nonetheless. High-dose ICS use is probably a marker of asthma severity, and direct pharmacological effects of ICS on IFN production are probably minor. A future study to examine the effects of ICS dose reduction on airway inflammation and capacity for virus-induced IFN synthesis would shed light on these uncertainties.

The mechanisms leading to reduced IFN-α and IFN-β synthesis by circulating leukocytes in neutrophilic asthma remain uncertain. One possibility is that patients with neutrophilic asthma carry an intrinsic (possibly germ line) defect in IFN-α or IFN-β synthesis, affecting all cell types, which contributes to the pathogenesis of neutrophilic airway inflammation. Although an interesting concept, currently there is little supporting evidence. Alternatively, neutrophilic airway inflammation may alter the function of circulating cells. Systemic inflammation appears to be prominent in neutrophilic asthma, relative to other asthma phenotypes, and perhaps this may alter the function of immune cell progenitors in bone marrow. In the present study we observed that IFN-α synthesis was inversely proportional to sputum IL-1β and serum IL-6 by univariate analysis (Table 3), although this association was no longer significant by stepwise multiple linear regression. Initial experiments with cells from healthy donors showed that IL-1β, but not IL-6, was able to inhibit HRV-stimulated IFN production in vitro (Fig 3). Although this provides proof of concept that such an effect may occur in vivo, further studies are required.

There are a number of limitations of the current study that should be acknowledged. The cross-sectional design means we are limited in our ability to assign cause and effect. At this stage we do not know the clinical relevance of our findings, and it will be important for future studies to determine whether the subset of asthma patients with the lowest capacity for IFN-α and IFN-β synthesis are at higher risk of asthma exacerbations. Our cohort are a group of older adults with poorly controlled asthma, and further studies are required to understand whether these differences apply to younger adults and children, although the nature of neutrophilic asthma is that it is associated with older age and therefore is less prevalent in younger adults and children with asthma.

Although a recent study found no relationship between impaired IFN-α release in severe asthma and exacerbation frequency, that study may have been underpowered to assess exacerbations as an end point. It is also possible that the mechanism by which sputum eosinophilia is linked to asthma exacerbations may be independent of type I IFN. Neutrophilic airway inflammation has been linked to poor asthma outcomes, in particular more severe airflow obstruction, and our results suggest that this may be partly related to low type I IFN production. A prospective study is needed to examine the link between type I IFN and virus-associated asthma exacerbations and secondly, to explore the underlying mechanisms. Although we have studied HRV 1b, which is an HRV type A, evidence suggests responses to other serotypes of rhinovirus may be different,, and future studies will also need to examine IFN production in response to a wider range of HRVs. Finally, we recognize the absence of a group of age-matched healthy subjects in this study, but justify this on the basis that the primary aim of the study was to examine variations in IFN-α and IFN-β production between different asthma phenotypes, rather than comparing asthma with healthy subjects. An important strength of our study is the inclusion of a large number of carefully characterized participants with poorly controlled asthma.

In conclusion, antiviral IFN-α and IFN-β production was impaired in patients with neutrophilic airway inflammation and in those prescribed high doses of ICS. Our study is therefore an important step toward identifying those with poorly controlled asthma who might respond best to inhaled IFN during exacerbations.

Author contributions: J. L. S. had full access to the data and takes responsibility for the integrity and accuracy of the analysis. J. L. S., M. C., I. A. Y., P. N. R., S. H., A. L. J., P. G. G., and J. W. U. all made substantial contributions to the conception and design, or acquisition of data, or analysis and interpretation of data. Each also has drafted the submitted article or revised it critically for important intellectual contents and provided approval of the version to be published.

Financial/nonfinancial disclosures: None declared.

Role of sponsors: The sponsor had no role in the design of the study, the collection and analysis of the data, or the preparation of the manuscript.

Other contributions: The authors would like to acknowledge the AMAZES group who collected and processed the sputum samples and the patients who participated in this study.

Additional information: The e-Appendix, e-Figure, and e-Tables can be found in the Supplemental Materials section of the online article.

Beran D. .Zar H.J. .Perrin C. .Menezes A.M. .Burney P. . Forum of International Respiratory Societies working group collaboration Burden of asthma and chronic obstructive pulmonary disease and access to essential medicines in low-income and middle-income countries. Lancet Respir Med. 2015;3:159-170 [PubMed]journal. [CrossRef] [PubMed]
 
Gehlhar K. .Bilitewski C. .Reinitz-Rademacher K. .Rohde G. .Bufe A. . Impaired virus-induced interferon-alpha2 release in adult asthmatic patients. Clin Exp Allergy. 2006;36:331-337 [PubMed]journal. [CrossRef] [PubMed]
 
Wark P.A. .Johnston S.L. .Bucchieri F. .et al Asthmatic bronchial epithelial cells have a deficient innate immune response to infection with rhinovirus. J Exp Med. 2005;201:937-947 [PubMed]journal. [CrossRef] [PubMed]
 
Contoli M. .Message S.D. .Laza-Stanca V. .et al Role of deficient type III interferon-lambda production in asthma exacerbations. Nat Med. 2006;12:1023-1026 [PubMed]journal. [CrossRef] [PubMed]
 
Roponen M. .Yerkovich S.T. .Hollams E. .Sly P.D. .Holt P.G. .Upham J.W. . Toll-like receptor 7 function is reduced in adolescents with asthma. Eur Respir J. 2010;35:64-71 [PubMed]journal. [CrossRef] [PubMed]
 
Pritchard A.L. .White O.J. .Burel J.G. .Carroll M.L. .Phipps S. .Upham J.W. . Asthma is associated with multiple alterations in anti-viral innate signalling pathways. PLoS One. 2014;9:e106501- [PubMed]journal. [CrossRef] [PubMed]
 
Lopez-Souza N. .Favoreto S. .Wong H. .et al In vitro susceptibility to rhinovirus infection is greater for bronchial than for nasal airway epithelial cells in human subjects. J Allergy Clin Immunol. 2009;123:1384-1390.e2 [PubMed]journal. [CrossRef] [PubMed]
 
Bochkov Y.A. .Hanson K.M. .Keles S. .Brockman-Schneider R.A. .Jarjour N.N. .Gern J.E. . Rhinovirus-induced modulation of gene expression in bronchial epithelial cells from subjects with asthma. Mucosal Immunol. 2010;3:69-80 [PubMed]journal. [CrossRef] [PubMed]
 
Sykes A. .Macintyre J. .Edwards M.R. .et al Rhinovirus-induced interferon production is not deficient in well controlled asthma. Thorax. 2014;69:240-246 [PubMed]journal. [CrossRef] [PubMed]
 
Durrani S.R. .Montville D.J. .Pratt A.S. .et al Innate immune responses to rhinovirus are reduced by the high-affinity IgE receptor in allergic asthmatic children. J Allergy Clin Immunol. 2012;130:489-495 [PubMed]journal. [CrossRef] [PubMed]
 
Edwards M.R. .Regamey N. .Vareille M. .et al Impaired innate interferon induction in severe therapy resistant atopic asthmatic children. Mucosal Immunol. 2013;6:797-806 [PubMed]journal. [CrossRef] [PubMed]
 
Pavord I.D. .Brightling C.E. .Woltmann G. .Wardlaw A.J. . Non-eosinophilic corticosteroid unresponsive asthma. Lancet. 1999;353:2213-2214 [PubMed]journal. [CrossRef] [PubMed]
 
Simpson J.L. .Scott R. .Boyle M.J. .Gibson P.G. . Inflammatory subtypes in asthma: assessment and identification using induced sputum. Respirology. 2006;11:54-61 [PubMed]journal. [CrossRef] [PubMed]
 
Wenzel S.E. .Schwartz L.B. .Langmack E.L. .et al Evidence that severe asthma can be divided pathologically into two inflammatory subtypes with distinct physiologic and clinical characteristics. Am J Respir Crit Care Med. 1999;160:1001-1008 [PubMed]journal. [CrossRef] [PubMed]
 
Neveu W.A. .Allard J.B. .Dienz O. .et al IL-6 is required for airway mucus production induced by inhaled fungal allergens. J Immunol. 2009;183:1732-1738 [PubMed]journal. [CrossRef] [PubMed]
 
Kay A.B. . Immunomodulation in asthma: mechanisms and possible pitfalls. Curr Opin Pharmacol. 2003;3:220-226 [PubMed]journal. [CrossRef] [PubMed]
 
Shahid S.K. .Kharitonov S.A. .Wilson N.M. .Bush A. .Barnes P.J. . Increased interleukin-4 and decreased interferon-gamma in exhaled breath condensate of children with asthma. Am J Respir Crit Care Med. 2002;165:1290-1293 [PubMed]journal. [CrossRef] [PubMed]
 
Hastie A.T. .Moore W.C. .Meyers D.A. .Vestal P.L. .Li H. .Peters S.P. .Bleecker E.R. . National Heart, Lung, and Blood Institute Severe Asthma Research Program Analyses of asthma severity phenotypes and inflammatory proteins in subjects stratified by sputum granulocytes. J Allergy Clin Immunol. 2010;125:1028-1036.e13 [PubMed]journal. [CrossRef] [PubMed]
 
Wood L.G. .Baines K.J. .Fu J. .Scott H.A. .Gibson P.G. . The neutrophilic inflammatory phenotype is associated with systemic inflammation in asthma. Chest. 2012;142:86-93 [PubMed]journal. [CrossRef] [PubMed]
 
Chu D.K. .Al-Garawi A. .Llop-Guevara A. .et al Therapeutic potential of anti-IL-6 therapies for granulocytic airway inflammation in asthma. Allergy Asthma Clin Immunol. 2015;11:14- [PubMed]journal. [CrossRef] [PubMed]
 
Baines K.J. .Simpson J.L. .Bowden N.A. .Scott R.J. .Gibson P.G. . Differential gene expression and cytokine production from neutrophils in asthma phenotypes. Eur Respir J. 2010;35:522-531 [PubMed]journal. [CrossRef] [PubMed]
 
Simpson J.L. .Phipps S. .Baines K.J. .Oreo K.M. .Gunawardhana L. .Gibson P.G. . Elevated expression of the NLRP3 inflammasome in neutrophilic asthma. Eur Respir J. 2014;43:1067-1076 [PubMed]journal. [CrossRef] [PubMed]
 
Djukanović R. .Harrison T. .Johnston S.L. . INTERCIA Study Groupet al The effect of inhaled IFN-β on worsening of asthma symptoms caused by viral infections. A randomized trial. Am J Respir Crit Care Med. 2014;190:145-154 [PubMed]journal. [CrossRef] [PubMed]
 
Juniper E.F. .O’Byrne P.M. .Roberts J.N. . Measuring asthma control in group studies: Do we need airway calibre and rescue β2-agonist use? Respir Med. 2001;95:319-323 [PubMed]journal. [CrossRef] [PubMed]
 
Juniper E.F. .Svensson K. .Mörk A.-C. .Ståhl E. . Measurement properties and interpretation of three shortened versions of the asthma control questionnaire. Respir Med. 2005;99:553-558 [PubMed]journal. [CrossRef] [PubMed]
 
Leite M. .Ponte E.V. .Petroni J. .D’Oliveira Júnior A. .Pizzichini E. .Cruz Á.A. . Evaluation of the asthma control questionnaire validated for use in Brazil. J Bras Pneumol. 2008;34:756-763 [PubMed]journal. [CrossRef] [PubMed]
 
Gibson P.G. .Wlodarczyk J.W. .Hensley M.J. .et al Epidemiological association of airway inflammation with asthma symptoms and airway hyperresponsiveness in childhood. Am J Respir Crit Care Med. 1998;158:36-41 [PubMed]journal. [CrossRef] [PubMed]
 
Subauste M.C. .Jacoby D.B. .Richards S.M. .Proud D. . Infection of a human respiratory epithelial cell line with rhinovirus. Induction of cytokine release and modulation of susceptibility to infection by cytokine exposure. J Clin Invest. 1995;96:549-557 [PubMed]journal. [CrossRef] [PubMed]
 
Carroll M.L. .Yerkovich S.T. .Pritchard A.L. .Davies J.M. .Upham J.W. . Adaptive immunity to rhinoviruses: Sex and age matter. Respir Res. 2010;11:184- [PubMed]journal. [CrossRef] [PubMed]
 
Pritchard A.L. .Carroll M.L. .Burel J.G. .White O.J. .Phipps S. .Upham J.W. . Innate IFNs and plasmacytoid dendritic cells constrain Th2 cytokine responses to rhinovirus: a regulatory mechanism with relevance to asthma. J Immunol. 2012;188:5898-5905 [PubMed]journal. [CrossRef] [PubMed]
 
Zaas A.K. .Chen M. .Varkey J. .et al Gene expression signatures diagnose influenza and other symptomatic respiratory viral infections in humans. Cell Host Microbe. 2009;6:207-217 [PubMed]journal. [CrossRef] [PubMed]
 
Gardeux V. .Bosco A. .Li J. .et al Towards a PBMC “virogram assay” for precision medicine: concordance between ex vivo and in vivo viral infection transcriptomes. J Biomed Inform. 2015;55:94-103 [PubMed]journal. [CrossRef] [PubMed]
 
Iikura K. .Katsunuma T. .Saika S. .et al Peripheral blood mononuclear cells from patients with bronchial asthma show impaired innate immune responses to rhinovirus in vitro. Int Arch Allergy Immunol. 2011;155:27-33 [PubMed]journal. [CrossRef] [PubMed]
 
Forbes R.L. .Gibson P.G. .Murphy V.E. .Wark P.A. . Impaired type I and III interferon response to rhinovirus infection during pregnancy and asthma. Thorax. 2012;67:209-214 [PubMed]journal. [CrossRef] [PubMed]
 
Wright A.K. .Mistry V. .Richardson M. .et al Toll-like receptor 9 dependent interferon-α release is impaired in severe asthma but is not associated with exacerbation frequency. Immunobiology. 2015;220:859-864 [PubMed]journal. [CrossRef] [PubMed]
 
Sykes A. .Edwards M.R. .Macintyre J. .et al Rhinovirus 16-induced IFN-alpha and IFN-beta are deficient in bronchoalveolar lavage cells in asthmatic patients. J Allergy Clin Immunol. 2012;129:1506-1514.e6 [PubMed]journal. [CrossRef] [PubMed]
 
Bratke K. .Prieschenk C. .Garbe K. .Kuepper M. .Lommatzsch M. .Virchow J.C. . Plasmacytoid dendritic cells in allergic asthma and the role of inhaled corticosteroid treatment. Clin Exp Allergy. 2013;43:312-321 [PubMed]journal. [CrossRef] [PubMed]
 
Davies J.M. .Carroll M.L. .Li H. .et al Budesonide and formoterol reduce early innate anti-viral immune responses in vitro. PLoS One. 2011;6:e27898- [PubMed]journal. [CrossRef] [PubMed]
 
Nakagome K. .Bochkov Y.A. .Ashraf S. .et al Effects of rhinovirus species on viral replication and cytokine production. J Allergy Clin Immunol. 2014;134:332-341 [PubMed]journal. [CrossRef] [PubMed]
 
Bizzintino J. .Lee W.M. .Laing I.A. .et al Association between human rhinovirus C and severity of acute asthma in children. Eur Respir J. 2011;37:1037-1042 [PubMed]journal. [CrossRef] [PubMed]
 

Figures

Figure Jump LinkFigure 1 Inflammatory cytokine levels in supernatant from human rhinovirus 1b-activated, cultured peripheral blood mononuclear cells are shown (data corrected for baseline unstimulated levels). Bars represent median levels for IFN-α (A), IFN-β (B), CXCL10 (C), CXCL8 (D), IL-1β (E), and IL-6 (F). The number of participants for each group is below the inflammatory subtype in each panel; the dotted line shows the zero level for each cytokine. Results for all three groups were compared by Kruskal-Wallis test and indicated by the probability value in each panel. Significant differences between any two groups (P < .05 by post-hoc rank-sum test) are indicated by * compared with eosinophilic asthma and # compared with paucigranulocytic asthma. CXCL = C-X-C motif ligand; Eos = eosinophilic asthma; IFN = interferon; Neut = neutrophilic asthma; Pauci = paucigranulocytic asthma.Grahic Jump Location
Figure Jump LinkFigure 2 Scatter plots of IFN production stimulated by human rhinovirus 1b selected for ICS total daily dose (A), mean ACQ-6 score (B), sputum neutrophil % (C), serum IL-6 (D), sputum IL-1β (E), atopy (F), blood lymphocytes (G), and blood monocytes (H). Spearman correlation coefficient (r) and probability values are as indicated. The comparison between atopic and nonatopic participants was assessed using Mann-Whitney test. ACQ-6 = asthma control questionnaire 6; ICS = inhaled corticosteroid. See Figure 1 legend for expansion of other abbreviation.Grahic Jump Location
Figure Jump LinkFigure 3 Effects of IL-1β on rhinovirus-stimulated IFN-α (A) and IFN-β (B) production from healthy control peripheral blood mononuclear cells. HRV = human rhinovirus. See Figure 1 legend for expansion of other abbreviation.Grahic Jump Location

Tables

Table Graphic Jump Location
Table 1 Participant Characteristics

Data are presented as median (q1, q3), unless indicated otherwise. ACQ6 = asthma control questionnaire 6; ICS = inhaled corticosteroid.

Table Graphic Jump Location
Table 2 Inflammatory Cell Counts and Mediators by Asthma Inflammatory Phenotype
a P < .008 vs eosinophilic asthma.
b P < .008 vs paucigranulocytic asthma.

Data are median (q1, q3).

Table Graphic Jump Location
Table 3 Univariate and Stepwise Multivariate Regression Analysis
a Peripheral blood lymphocytes were assessed 2-4 weeks before randomization.

The model included 79 data points and explained 27% of the variance in IFN-β levels and was highly significant at P < .0001. ACQ6 = asthma control questionnaire 6; ICS = inhaled corticosteroid.

References

Beran D. .Zar H.J. .Perrin C. .Menezes A.M. .Burney P. . Forum of International Respiratory Societies working group collaboration Burden of asthma and chronic obstructive pulmonary disease and access to essential medicines in low-income and middle-income countries. Lancet Respir Med. 2015;3:159-170 [PubMed]journal. [CrossRef] [PubMed]
 
Gehlhar K. .Bilitewski C. .Reinitz-Rademacher K. .Rohde G. .Bufe A. . Impaired virus-induced interferon-alpha2 release in adult asthmatic patients. Clin Exp Allergy. 2006;36:331-337 [PubMed]journal. [CrossRef] [PubMed]
 
Wark P.A. .Johnston S.L. .Bucchieri F. .et al Asthmatic bronchial epithelial cells have a deficient innate immune response to infection with rhinovirus. J Exp Med. 2005;201:937-947 [PubMed]journal. [CrossRef] [PubMed]
 
Contoli M. .Message S.D. .Laza-Stanca V. .et al Role of deficient type III interferon-lambda production in asthma exacerbations. Nat Med. 2006;12:1023-1026 [PubMed]journal. [CrossRef] [PubMed]
 
Roponen M. .Yerkovich S.T. .Hollams E. .Sly P.D. .Holt P.G. .Upham J.W. . Toll-like receptor 7 function is reduced in adolescents with asthma. Eur Respir J. 2010;35:64-71 [PubMed]journal. [CrossRef] [PubMed]
 
Pritchard A.L. .White O.J. .Burel J.G. .Carroll M.L. .Phipps S. .Upham J.W. . Asthma is associated with multiple alterations in anti-viral innate signalling pathways. PLoS One. 2014;9:e106501- [PubMed]journal. [CrossRef] [PubMed]
 
Lopez-Souza N. .Favoreto S. .Wong H. .et al In vitro susceptibility to rhinovirus infection is greater for bronchial than for nasal airway epithelial cells in human subjects. J Allergy Clin Immunol. 2009;123:1384-1390.e2 [PubMed]journal. [CrossRef] [PubMed]
 
Bochkov Y.A. .Hanson K.M. .Keles S. .Brockman-Schneider R.A. .Jarjour N.N. .Gern J.E. . Rhinovirus-induced modulation of gene expression in bronchial epithelial cells from subjects with asthma. Mucosal Immunol. 2010;3:69-80 [PubMed]journal. [CrossRef] [PubMed]
 
Sykes A. .Macintyre J. .Edwards M.R. .et al Rhinovirus-induced interferon production is not deficient in well controlled asthma. Thorax. 2014;69:240-246 [PubMed]journal. [CrossRef] [PubMed]
 
Durrani S.R. .Montville D.J. .Pratt A.S. .et al Innate immune responses to rhinovirus are reduced by the high-affinity IgE receptor in allergic asthmatic children. J Allergy Clin Immunol. 2012;130:489-495 [PubMed]journal. [CrossRef] [PubMed]
 
Edwards M.R. .Regamey N. .Vareille M. .et al Impaired innate interferon induction in severe therapy resistant atopic asthmatic children. Mucosal Immunol. 2013;6:797-806 [PubMed]journal. [CrossRef] [PubMed]
 
Pavord I.D. .Brightling C.E. .Woltmann G. .Wardlaw A.J. . Non-eosinophilic corticosteroid unresponsive asthma. Lancet. 1999;353:2213-2214 [PubMed]journal. [CrossRef] [PubMed]
 
Simpson J.L. .Scott R. .Boyle M.J. .Gibson P.G. . Inflammatory subtypes in asthma: assessment and identification using induced sputum. Respirology. 2006;11:54-61 [PubMed]journal. [CrossRef] [PubMed]
 
Wenzel S.E. .Schwartz L.B. .Langmack E.L. .et al Evidence that severe asthma can be divided pathologically into two inflammatory subtypes with distinct physiologic and clinical characteristics. Am J Respir Crit Care Med. 1999;160:1001-1008 [PubMed]journal. [CrossRef] [PubMed]
 
Neveu W.A. .Allard J.B. .Dienz O. .et al IL-6 is required for airway mucus production induced by inhaled fungal allergens. J Immunol. 2009;183:1732-1738 [PubMed]journal. [CrossRef] [PubMed]
 
Kay A.B. . Immunomodulation in asthma: mechanisms and possible pitfalls. Curr Opin Pharmacol. 2003;3:220-226 [PubMed]journal. [CrossRef] [PubMed]
 
Shahid S.K. .Kharitonov S.A. .Wilson N.M. .Bush A. .Barnes P.J. . Increased interleukin-4 and decreased interferon-gamma in exhaled breath condensate of children with asthma. Am J Respir Crit Care Med. 2002;165:1290-1293 [PubMed]journal. [CrossRef] [PubMed]
 
Hastie A.T. .Moore W.C. .Meyers D.A. .Vestal P.L. .Li H. .Peters S.P. .Bleecker E.R. . National Heart, Lung, and Blood Institute Severe Asthma Research Program Analyses of asthma severity phenotypes and inflammatory proteins in subjects stratified by sputum granulocytes. J Allergy Clin Immunol. 2010;125:1028-1036.e13 [PubMed]journal. [CrossRef] [PubMed]
 
Wood L.G. .Baines K.J. .Fu J. .Scott H.A. .Gibson P.G. . The neutrophilic inflammatory phenotype is associated with systemic inflammation in asthma. Chest. 2012;142:86-93 [PubMed]journal. [CrossRef] [PubMed]
 
Chu D.K. .Al-Garawi A. .Llop-Guevara A. .et al Therapeutic potential of anti-IL-6 therapies for granulocytic airway inflammation in asthma. Allergy Asthma Clin Immunol. 2015;11:14- [PubMed]journal. [CrossRef] [PubMed]
 
Baines K.J. .Simpson J.L. .Bowden N.A. .Scott R.J. .Gibson P.G. . Differential gene expression and cytokine production from neutrophils in asthma phenotypes. Eur Respir J. 2010;35:522-531 [PubMed]journal. [CrossRef] [PubMed]
 
Simpson J.L. .Phipps S. .Baines K.J. .Oreo K.M. .Gunawardhana L. .Gibson P.G. . Elevated expression of the NLRP3 inflammasome in neutrophilic asthma. Eur Respir J. 2014;43:1067-1076 [PubMed]journal. [CrossRef] [PubMed]
 
Djukanović R. .Harrison T. .Johnston S.L. . INTERCIA Study Groupet al The effect of inhaled IFN-β on worsening of asthma symptoms caused by viral infections. A randomized trial. Am J Respir Crit Care Med. 2014;190:145-154 [PubMed]journal. [CrossRef] [PubMed]
 
Juniper E.F. .O’Byrne P.M. .Roberts J.N. . Measuring asthma control in group studies: Do we need airway calibre and rescue β2-agonist use? Respir Med. 2001;95:319-323 [PubMed]journal. [CrossRef] [PubMed]
 
Juniper E.F. .Svensson K. .Mörk A.-C. .Ståhl E. . Measurement properties and interpretation of three shortened versions of the asthma control questionnaire. Respir Med. 2005;99:553-558 [PubMed]journal. [CrossRef] [PubMed]
 
Leite M. .Ponte E.V. .Petroni J. .D’Oliveira Júnior A. .Pizzichini E. .Cruz Á.A. . Evaluation of the asthma control questionnaire validated for use in Brazil. J Bras Pneumol. 2008;34:756-763 [PubMed]journal. [CrossRef] [PubMed]
 
Gibson P.G. .Wlodarczyk J.W. .Hensley M.J. .et al Epidemiological association of airway inflammation with asthma symptoms and airway hyperresponsiveness in childhood. Am J Respir Crit Care Med. 1998;158:36-41 [PubMed]journal. [CrossRef] [PubMed]
 
Subauste M.C. .Jacoby D.B. .Richards S.M. .Proud D. . Infection of a human respiratory epithelial cell line with rhinovirus. Induction of cytokine release and modulation of susceptibility to infection by cytokine exposure. J Clin Invest. 1995;96:549-557 [PubMed]journal. [CrossRef] [PubMed]
 
Carroll M.L. .Yerkovich S.T. .Pritchard A.L. .Davies J.M. .Upham J.W. . Adaptive immunity to rhinoviruses: Sex and age matter. Respir Res. 2010;11:184- [PubMed]journal. [CrossRef] [PubMed]
 
Pritchard A.L. .Carroll M.L. .Burel J.G. .White O.J. .Phipps S. .Upham J.W. . Innate IFNs and plasmacytoid dendritic cells constrain Th2 cytokine responses to rhinovirus: a regulatory mechanism with relevance to asthma. J Immunol. 2012;188:5898-5905 [PubMed]journal. [CrossRef] [PubMed]
 
Zaas A.K. .Chen M. .Varkey J. .et al Gene expression signatures diagnose influenza and other symptomatic respiratory viral infections in humans. Cell Host Microbe. 2009;6:207-217 [PubMed]journal. [CrossRef] [PubMed]
 
Gardeux V. .Bosco A. .Li J. .et al Towards a PBMC “virogram assay” for precision medicine: concordance between ex vivo and in vivo viral infection transcriptomes. J Biomed Inform. 2015;55:94-103 [PubMed]journal. [CrossRef] [PubMed]
 
Iikura K. .Katsunuma T. .Saika S. .et al Peripheral blood mononuclear cells from patients with bronchial asthma show impaired innate immune responses to rhinovirus in vitro. Int Arch Allergy Immunol. 2011;155:27-33 [PubMed]journal. [CrossRef] [PubMed]
 
Forbes R.L. .Gibson P.G. .Murphy V.E. .Wark P.A. . Impaired type I and III interferon response to rhinovirus infection during pregnancy and asthma. Thorax. 2012;67:209-214 [PubMed]journal. [CrossRef] [PubMed]
 
Wright A.K. .Mistry V. .Richardson M. .et al Toll-like receptor 9 dependent interferon-α release is impaired in severe asthma but is not associated with exacerbation frequency. Immunobiology. 2015;220:859-864 [PubMed]journal. [CrossRef] [PubMed]
 
Sykes A. .Edwards M.R. .Macintyre J. .et al Rhinovirus 16-induced IFN-alpha and IFN-beta are deficient in bronchoalveolar lavage cells in asthmatic patients. J Allergy Clin Immunol. 2012;129:1506-1514.e6 [PubMed]journal. [CrossRef] [PubMed]
 
Bratke K. .Prieschenk C. .Garbe K. .Kuepper M. .Lommatzsch M. .Virchow J.C. . Plasmacytoid dendritic cells in allergic asthma and the role of inhaled corticosteroid treatment. Clin Exp Allergy. 2013;43:312-321 [PubMed]journal. [CrossRef] [PubMed]
 
Davies J.M. .Carroll M.L. .Li H. .et al Budesonide and formoterol reduce early innate anti-viral immune responses in vitro. PLoS One. 2011;6:e27898- [PubMed]journal. [CrossRef] [PubMed]
 
Nakagome K. .Bochkov Y.A. .Ashraf S. .et al Effects of rhinovirus species on viral replication and cytokine production. J Allergy Clin Immunol. 2014;134:332-341 [PubMed]journal. [CrossRef] [PubMed]
 
Bizzintino J. .Lee W.M. .Laing I.A. .et al Association between human rhinovirus C and severity of acute asthma in children. Eur Respir J. 2011;37:1037-1042 [PubMed]journal. [CrossRef] [PubMed]
 
NOTE:
Citing articles are presented as examples only. In non-demo SCM6 implementation, integration with CrossRef’s "Cited By" API will populate this tab (http://www.crossref.org/citedby.html).
Supporting Data

e-Online Data

Some tools below are only available to our subscribers or users with an online account.

Related Content

Customize your page view by dragging & repositioning the boxes below.

Find Similar Articles
CHEST Journal Articles
PubMed Articles
  • CHEST Journal
    Print ISSN: 0012-3692
    Online ISSN: 1931-3543