Human Rhinovirus Infection Research Articles

Burden of human rhinovirus infections in influenza like illnesses in Kenya

Background: There have been several recent reports of respiratory outbreaks with associated mortality due to human rhinovirus (HRV) infection. Studies on viral etiologies of Influenza-like illness have shown that many patients are infected with more than one viral agent with frequencies of co-infection being as high as 20%. From these studies, human rhinovirus has been shown as one of the most prominent respiratory viruses that co-infect patients with influenza. In Kenya, the ongoing Influenza surveillance has provided data that now allows for exploration of other respiratory viruses that cause influenza-like illnesses. Currently, there is scanty information on the patterns of HRV circulation throughout the country. Methods & Materials: This study utilized a retrospective cross-sectional descriptive design. It involved the use of 517 archived samples from the ongoing country-wide influenza Surveillance protocol. The archived nasopharyngeal specimens were collected from a study population comprising persons from two months of age onwards who attended outpatient clinics in the year 2008, in hospitals located in 8 different regions in Kenya presenting with influenza like illnesses. Viral RNA was extracted from the samples followed by real-time RTPCR assays with HRV-specific primers for screening for HRV. Results: A total of 131 (25%) of the samples were positive for HRV. The highest prevalence (33%) was recorded from samples obtained from the Coast region of the country, followed by Western region (32.7%) while Eastern region had the lowest prevalence (11%). Children aged between 2 months -7 years were mostly infected while the age group ≥60 years was least affected. There was no significant difference in HRV infection in relation to gender. Conclusion: The study notes that HRV infection is a significant viral etiology of morbidity throughout the country. However, disparities were observed between different regions. The more humid regions in the country (Western and Coast) had the highest HRV prevalence. At the same time, children below 7 years were at a greater risk of HRV infection. Determination of the molecular characteristics of the rhinovirus strains to enable specific identification of the species and the evolutionary relationships is ongoing.

Asthma: NHLBI Workshop on the Primary Prevention of Chronic Lung Diseases.

Asthma is a common disease with enormous public health costs, and its primary prevention is an ambitious and important goal. Understanding of how host and environmental factors interact to cause asthma is incomplete, but persistent questions about mechanisms should not stop clinical research efforts aimed at reducing the prevalence of childhood asthma. Achieving the goal of primary prevention of asthma will involve integrated and parallel sets of research activities in which mechanism-oriented studies of asthma inception proceed alongside clinical intervention studies to test biologically plausible prevention ideas. For example, continued research is needed, particularly in young children, to uncover biomarkers that identify asthma risk and provide potential targets of intervention, and to improve understanding of the role of microbial factors in asthma risk and disease initiation. In terms of clinical trials that could be initiated now or in the near future, we recommend three interventions for testing: (1) preventing asthma through prophylaxis against respiratory syncytial virus and human rhinovirus infections of the airway; (2) immune modulation, using prebiotics, probiotics, and bacterial lysates; and (3) prevention of allergen sensitization and allergic inflammation, using anti-IgE. These interventions should be tested while other, more universal prevention measures that may promote lung health are also investigated. These potential universal lung health measures include prevention of preterm delivery; reduced exposure of the fetus and young infant to environmental pollutants, including tobacco smoke; prevention of maternal and child obesity; and management of psychosocial stress.

Infants 1-90 Days Old Hospitalized With Human Rhinovirus Infection

Human rhinovirus (HRV) is a common cause of respiratory illness in children. The impact of HRV infection on 1- to 90-day-old infants is unclear. We hypothesized that HRV infection would be clinically similar to respiratory syncytial virus (RSV) infection in the hospitalized infants. We conducted a retrospective study of hospitalized infants, who were 1-90 days old, with HRV or RSV within the Southern California Kaiser Permanente network over a 1-year period (August 2010 to October 2011). We identified 245 hospitalized infants who underwent respiratory virus testing. HRV was found in 52 infants (21%) compared to 79 infants (32%) with RSV (P = 0.008). Infants with HRV infection experienced longer hospital stays compared to those with RSV (median length of stay 4 days vs. 3 days, P = 0.009) and had fewer short hospital stays ≤3 days (P = 0.029). There was a trend in infants with HRV infection to be younger (P = 0.071) and have more fevers (P = 0.052). Recent advances in diagnostics allow for identification of a broad range of viral pathogens in infants. Compared to RSV, HRV was associated with longer hospital stays. Additional studies and improved, more specific testing, methods are needed to further define the effects of HRV infection in infants 1-90 days old.

Rhinovirus Exacerbates House-Dust-Mite Induced Lung Disease in Adult Mice

Human rhinovirus is a key viral trigger for asthma exacerbations. To date, murine studies investigating rhinovirus-induced exacerbation of allergic airways disease have employed systemic sensitisation/intranasal challenge with ovalbumin. In this study, we combined human-rhinovirus infection with a clinically relevant mouse model of aero-allergen exposure using house-dust-mite in an attempt to more accurately understand the links between human-rhinovirus infection and exacerbations of asthma. Adult BALB/c mice were intranasally exposed to low-dose house-dust-mite (or vehicle) daily for 10 days. On day 9, mice were inoculated with human-rhinovirus-1B (or UV-inactivated human-rhinovirus-1B). Forty-eight hours after inoculation, we assessed bronchoalveolar cellular inflammation, levels of relevant cytokines/serum antibodies, lung function and responsiveness/sensitivity to methacholine. House-dust-mite exposure did not result in a classical TH2-driven response, but was more representative of noneosinophilic asthma. However, there were significant effects of house-dust-mite exposure on most of the parameters measured including increased cellular inflammation (primarily macrophages and neutrophils), increased total IgE and house-dust-mite-specific IgG1 and increased responsiveness/sensitivity to methacholine. There were limited effects of human-rhinovirus-1B infection alone, and the combination of the two insults resulted in additive increases in neutrophil levels and lung parenchymal responses to methacholine (tissue elastance). We conclude that acute rhinovirus infection exacerbates house-dust-mite-induced lung disease in adult mice. The similarity of our results using the naturally occurring allergen house-dust-mite, to previous studies using ovalbumin, suggests that the exacerbation of allergic airways disease by rhinovirus infection could act via multiple or conserved mechanisms.

Human rhinovirus infection during naturally occurring COPD exacerbations

Human rhinovirus (HRV) infection is an important trigger of exacerbations of chronic obstructive pulmonary disease (COPD) but its role in determining exacerbation frequency phenotype or the time-course of HRV infection in naturally occurring exacerbations is unknown. Sputum samples from 77 patients were analysed by real-time quantitative PCR for both HRV (388 samples), and Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis (89 samples). Patients recorded worsening of respiratory symptoms on daily diary cards, from which exacerbations were identified. HRV prevalence and load at exacerbation presentation were significantly higher than in the stable state (prevalence 53.3% versus 17.2%, respectively; p<0.001) but 0% by day 35 post-exacerbation. HRV load was higher in patients with cold symptoms (p=0.046) or sore throats (p=0.006) than those without. 73% of bacterium-negative but HRV-positive exacerbations were bacterium-positive by day 14. Patients with HRV detected at exacerbation had a higher exacerbation frequency (interquartile range) of 3.01 (2.02-5.30) per year compared with patients without HRV (2.51 (2.00-3.51)) (p=0.038). HRV prevalence and load increased at COPD exacerbation, and resolved during recovery. Frequent exacerbators were more likely to experience HRV infection. Secondary bacterial infection is common after HRV infection, and provides a possible mechanism for exacerbation recurrence and a potential target for novel therapies.

Migration of the contractile phenotype of human airway smooth muscle cells in response to supernatants from rhinovirus infected human bronchial epithelial cells

Rationale Human rhinovirus (HRV) infections during early childhood are associated with a significantly increased risk of developing asthma in subsequent years [1]. There is published evidence that airway remodeling is present in pre-school children, often before the diagnosis of asthma is established [1]. It is thought that this increased risk relates to the fact that HRV infections facilitate airway remodeling in asthma [2]. A feature of airway remodeling is the proximity of airway smooth muscle (ASM) to the subepithelial region among other pathological changes [3]. Smooth muscle is also known to exist in two distinct phenotypes: secretory and contractile [4]. We have recently shown that HRV infection of Human Bronchial Epithelial Cells (HBEC), both in vitro and in vivo, results in the up-regulation of a number of airway remodeling mediators [5]. We now sought to determine which ASM phenotype (contractile or secretory) results in migration to supernatants from HRV infected HBE cells.

Cyclic stretch augments human rhinovirus induced inflammatory responses in airway epithelial cells

Background Structural cells of the airways are subject to normal mechanical stretch during respiration [1]. Mechanical stretch acts as a mechanism of cell activation, and mechanotransduction pathways have been shown to activate pro-inflammatory genes [2]. Human rhinovirus (HRV) infections are a major cause of asthma exacerbations, and mechanical forces are likely to be more pronounced during asthma exacerbations [3]. Moreover, smoking is associated with worse clinical outcomes in asthma. Previous studies have shown that HRV infection, cigarette smoke extract (CSE), or mechanical stretch each induce CXCL8 production in bronchial epithelial cells [4-6]. In this study, we sought to determine whether mechanical stretch interacts with HRV infection and CSE to further upregulate airway inflammation.

Human rhinovirus infection of human bronchial epithelial cells results in migration of human bronchial fibroblast cells

Background Recent studies have demonstrated that structural changes in the airways characteristic of asthma, collectively referred to as airway remodeling, occur in young children even prior to the diagnosis of asthma. Young children who experience human rhinovirus (HRV)-associated wheezing illness within the first three years of life are at increased risk for the subsequent development of asthma. This association, together with evidence that HRVinfected epithelial cells release a number of growth factors and cytokines, has led to the hypothesis that HRV infection may be involved in the pathogenesis of airway remodeling. Thickening of the lamina reticularis found below the true basement membrane in the human airway is a characteristic feature of airway remodeling in asthma. The human bronchial fibroblast (HBF) is believed to contribute to this thickening by moving closer to the laminar reticularis and producing matrix proteins. We hypothesized that HRV infection of human bronchial epithelial (HBE) cells induces production of chemoattractants that can induce HBF migration.

Pre-existing human rhinovirus infection modulates host response to secondary bacterial infections

Background Secondary bacterial infections following viral infections of the airways are well documented and are associated with increased severity of respiratory disease compared to virus or bacterial infections alone. Human rhinovirus (HRV) infections are the most common causes of exacerbations in individuals with chronic airways diseases such as asthma and COPD. Moreover, bacterial colonization is commonly found in the airways of patients experiencing exacerbations of these chronic airways diseases and linked to increased severity and duration of these exacerbations. The mechanisms underlying the increased prevalence of secondary bacterial infections and the association with more severe outcomes following viral infections is not known. It has been suggested that viral infection of the airways cause dysregulation of innate host defense mechanisms, such as, impaired antimicrobial peptide expression of the airways. Antimicrobial peptides are key components of the innate immune response after infection and are important in efficient clearance of microbial colonization to prevent infection. We sought to determine whether HRV modulates the innate host defense response to secondary bacterial infections of the airways.

Human rhinovirus infection of human bronchial epithelial cells results in migration of human airway smooth muscle cells

Background Young children who experience human rhinovirus (HRV)-associated wheezing illnesses are more likely to develop subsequent asthma [1]. This has led to the hypothesis that HRV infection may be involved in the pathogenesis of airway remodeling in asthma [2]. Increased airway smooth muscle (ASM) mass, in which ASM cells are in close proximity to the subepithelial region, is a characteristic feature of airway remodeling [3]. We have shown that HRV infection of human bronchial epithelial (HBE) cells, both in vitro and in vivo, results in the upregulation of airway remodeling mediators [4]. We now sought to determine whether HRV infection of HBE cells is associated with airway smooth muscle (ASM) chemotaxis.

Induction of human airway epithelial to mesenchymal transition upon rhinovirus infection

Background Structural changes of the airway, collectively referred to as airway remodeling are believed to be the underlying cause of the airway hyperresponsiveness that is characteristic of asthma. Airway remodeling is characterized by thickening of the subepithelial membrane, goblet cell hyperplasia, angiogenesis, increased smooth muscle mass, and epithelial fragility. Thickening of the subepithelial membrane, due to increased deposition of matrix proteins by fibroblasts/myofibroblasts, has been observed in children even prior to the formal diagnosis of asthma. Recent data have shown that young children who experience episodes of human rhinovirus (HRV)-induced wheezing in early childhood are at increased risk of subsequently developing asthma. The primary site of infection of HRV is the airway epithelium. Recent evidence suggests that the molecular reprogramming of epithelial cells through a process called epithelial to mesenchymal transition (EMT) may contribute to increases in fibroblast/myofibroblast in the asthmatic airway. Therefore, we hypothesize that HRV infection plays a role in early airway remodeling by triggering EMT to produce fibroblasts/myofibroblasts that cause thickening of the subepithelial membrane by depositing matrix proteins. Methods The BEAS-2B human bronchial epithelial cell line was grown in 6-well plates in bronchial epithelial growth medium (BEGM). Prior to experiments, cells were grown for 24 h in BEGM from which hydrocortisone was removed. Cells were exposed to medium (control), or purified HRV-16 alone or in the presence of various growth factors for 120 hours with media and growth factor replacement at 48 and 96 hours. Cell lysates were collected and then analyzed by western blot for protein expression of epithelial and mesenchymal markers. Results BEAS-2B cells that were infected with HRV in the presence of epidermal growth factor (EGF) showed decreased expression of the epithelial marker E-cadherin, and increased expression of the mesenchymal marker vimentin.

Transcriptional and Epigenetic Modulation of Human Rhinovirus–Induced CXCL10 Production by Cigarette Smoke

Human rhinovirus (HRV) triggers exacerbations of asthma and chronic obstructive pulmonary disease. Cigarette smoking is the primary risk factor for the development of chronic obstructive pulmonary disease, and 25% of individuals with asthma smoke. Smokers experience both longer and more severe colds. We previously showed that cigarette smoke extract (CSE) inhibited HRV-induced expression of a range of epithelial antiviral molecules. Here, we use CXCL10 as a model antiviral gene to examine the mechanisms by which CSE inhibits epithelial antiviral immunity. HRV-induced CXCL10 transcription depends on activation of NF-ĸB and IFN-regulatory factor-1 (IRF-1), and we now also implicate two signal transducer and activator of transcription (STAT) consensus sequences in the CXCL10 promoter in HRV-induced CXCL10 expression. CSE inhibited HRV-induced activation and nuclear translocation/binding of both NF-ĸB, and IRF-1 to their respective recognition sequences in the CXCL10 promoter. HRV also induced formation of complexes at the STAT region in the CXCL10 promoter, and HRV-induced activation of STAT-1 was inhibited by CSE. In addition, CSE inhibited HRV-induced chromatin accessibility around the transcriptional start site of the CXCL10 promoter. Although CSE inhibited HRV-induced expression of both the viral double-stranded RNA sensors, retinoic acid-inducible gene-I and melanoma differentiation-associated gene (MDA) 5, only specific short interfering RNA (siRNA) to MDA5, but not nontargeting siRNA, or siRNA to retinoic acid-inducible gene-I, inhibited HRV-induced CXCL10 induction. We conclude that CSE reduces chromatin accessibility and inhibits viral signaling via NF-ĸB, IRF-1, STAT-1, and MDA5. Thus, we show that CSE can simultaneously modulate multiple pathways linked to innate immune responses to HRV infection.

Molecular detection of human rhinoviruses in respiratory samples: a comparison of Taqman probe-, SYBR green I- and BOXTO-based real-time PCR assays

BackgroundHuman Rhinoviruses (HRV) are major causative agents of acute respiratory tract infections in all age group and important contributing factors of childhood morbidity and mortality. Clinical presentation is poorly specific and the great antigenic and genetic variability of HRVs renders the biological diagnosis complex. Here, we have evaluated several molecular diagnostic protocols, including Taqman probe-based and intercalating agent-based RT-PCR assays.Methods5,627 respiratory samples sent to the laboratory of Virology of the University Hospitals of Marseille, France, from March 2011 to February 2012, were tested using a real-time RT-PCR assay in the 5’NCR of the rhinoviral genome that associated a Taqman probe and the detection of DNA-BOXTO-dye complexes. A sample of 500 BOXTO-positive samples were further tested using the same probe assay (without BOXTO), and a SYBR Green assay (using the same amplification primers). The specific amplification of HRV sequences was assessed by NGS amplicon sequencing.ResultsThe Taqman probe RT-PCR assay identified 696/5,627 samples (12,4%) as HRV-positive. BOXTO-positive samples included all probe-positive samples and 1,913 additional samples, of which only 24.3% were confirmed by sequencing. The SYBR Green assay was more specific (16/550 samples were probe-negative/SYBR Green-positive, all confirmed by 5′NCR sequencing), but 3/500 samples were probe-positive/SYBR Green-negative.ConclusionsOur results highlight the difficulty in detecting HRVs in clinical samples using a single molecular detection system. Amongst the 3 systems tested, the best compromise was obtained with the SYBR Green assay, which, by comparison with our probe-based assay provided an improved sensitivity without altering the detection specificity. Interestingly, a majority of probe-negative/BOXTO- or SYBR Green-positive samples were not associated with mutations in the sequence targeted by the probe. Sequence-based modifications of the secondary structure of the HRV 5′NCR may be associated with a limited access to the probe hybridisation region. Further investigations may identify a test combining a probe based- and an intercalating agent-based detection, which will significantly improve the diagnosis of HRV infections.

Human Rhinovirus and Disease Severity in Children

To evaluate retrospectively human rhinovirus (HRV) infections in children up to 5 years old and factors involved in disease severity. Nasopharyngeal aspirates from 434 children presenting a broad range of respiratory infection symptoms and severity degrees were tested for presence of HRV and 8 other respiratory viruses. Presence of host risk factors was also assessed. HRV was detected in 181 (41.7%) samples, in 107 of them as the only agent and in 74 as coinfections, mostly with respiratory syncytial virus (RSV; 43.2%). Moderate to severe symptoms were observed in 28.9% (31/107) single infections and in 51.3% (38/74) coinfections (P = .004). Multivariate analyses showed association of coinfections with lower respiratory tract symptoms and some parameters of disease severity, such as hospitalization. In coinfections, RSV was the most important virus associated with severe disease. Prematurity, cardiomyopathies, and noninfectious respiratory diseases were comorbidities that also were associated with disease severity (P = .007). Our study showed that HRV was a common pathogen of respiratory disease in children and was also involved in severe cases, causing symptoms of the lower respiratory tract. Severe disease in HRV infections were caused mainly by presence of RSV in coinfections, prematurity, congenital heart disease, and noninfectious respiratory disease.

Analysis Of Circulating Rhinovirus-Specific CD4+ T Cells Using Novel MHC Class II Tetramers Reveals Marked Expansion Of Effector Memory Cells In Infected Subjects

Little is known about T cell mechanisms of human rhinovirus (HRV) infection, despite the link to asthma. Our objective was to provide proof-of-concept for elucidating properties of circulating virus-specific CD4+ T cells in HRV-infected individuals using novel MHC II/peptide tetramers. HRV-39 seronegative, non-allergic subjects who were HLA-DR*0401-positive were infected intranasally with HRV-39. PBMCs isolated 28 days before inoculation, at day 0, and days 5 and 21 post-infection, were stained with PE-labeled HLA-DR*0401 tetramers containing HRV-39 epitopes. Cells were enriched using an anti-PE column, then stained for surface markers and analyzed by flow cytometry. A discrete population of tetramer-positive CD4+ T cells was detected at each time point using tetramers containing two novel epitopes of HRV-39 capsid proteins. Before infection, circulating HRV-specific T cells constituted 0.2-1.2% of CD4+ T cells following enrichment, corresponding to a precursor frequency of 4/10,000 to 3/1,000 CD4+ T cells. Five days post-infection, numbers increased to 1.2-10% of enriched cells (precursor frequency = 3/10,000 to 6/1,000 CD4+ T cells). Additionally, virus-specific T cells with a central memory phenotype (CCR7+ CD45RO+) decreased, whereas effector memory (CCR7- CD45RO+) and lung-homing (CCR5+) cells increased. At day 21, HRV-specific T cell numbers contracted, or else were further expanded among a subset of subjects. In subjects infected with HRV-39, circulating virus-specific CD4+ T cells expand and convert to an effector memory phenotype. Our findings demonstrate the feasibility of using tetramers containing epitopes of viral capsid proteins to accurately enumerate and phenotype HRV-specific CD4+ T cells in infected subjects.

Human rhinovirus-induced ISG15 selectively modulates epithelial antiviral immunity.

Human rhinovirus (HRV) infections trigger exacerbations of lower airway diseases. HRV infects human airway epithelial cells and induces proinflammatory and antiviral molecules that regulate the response to HRV infection. Interferon (IFN)-stimulated gene of 15 kDa (ISG15) has been shown to regulate other viruses. We now show that HRV-16 infection induces both intracellular epithelial ISG15 expression and ISG15 secretion in vitro. Moreover, ISG15 protein levels increased in nasal secretions of subjects with symptomatic HRV infections. HRV-16-induced ISG15 expression is transcriptionally regulated via an IFN regulatory factor pathway. ISG15 does not directly alter HRV replication but does modulate immune signaling via the viral sensor protein RIG-I to impact production of CXCL10, which has been linked to innate immunity to viruses. Extracellular ISG15 also alters CXCL10 production. We conclude that ISG15 has a complex role in host defense against HRV infection, and that additional studies are needed to clarify the role of this molecule.

Human Rhinovirus Infection in Children with Cystic Fibrosis

Nasopharyngeal swabs obtained from 78 pediatric patients with cystic fibrosis (CF), including 47 with acute pulmonary exacerbation and 31 in a stable clinical condition, were evaluated for 17 respiratory viruses. Human rhinovirus (HRV) was the most frequently detected virus in patients with pulmonary exacerbation and in those who were clinically stable (21.3% vs. 12.9%; P = 0.52). HRV-A was the main RV detected in patients with pulmonary exacerbations. However, no prevalence of particular HRV-A subtypes was found. This study highlights that RV is frequently found in the respiratory secretions of patients with CF and the impact of HRV-A appears higher than that of the other HRV types in patients with pulmonary exacerbations.

Rhinoviral infection and asthma: the detection and management of rhinoviruses by airway epithelial cells

Human rhinoviruses (HRV) have been linked to the development of childhood asthma and recurrent acute asthma exacerbations throughout life, and contribute considerably to the healthcare and economic burden of this disease. However, the ability of HRV infections to trigger exacerbations, and the link between allergic status and HRV responsiveness, remains incompletely understood. Whilst the receptors on human airway cells that detect and are utilized by most HRV group A and B, but not C serotypes are known, how endosomal pattern recognition receptors (PRRs) detect HRV replication products that are generated within the cytoplasm remains somewhat of an enigma. In this article, we explore a role for autophagy, a cellular homeostatic process that allows the cell to encapsulate its own cytosolic constituents, as the crucial mechanism controlling this process and regulating the innate immune response of airway epithelial cells to viral infection. We will also briefly describe some of the recent insights into the immune responses of the airway to HRV, focusing on neutrophilic inflammation that is a potentially unwanted feature of the acute response to viral infection, and the roles of IL-1 and Pellinos in the regulation of responses to HRV.

Synthesis and anti-rhinovirus activity of novel 3-[2-(pyridinyl)vinyl]substituted -2H-chromenes and -4H-chromen-4-ones

Human rhinoviruses (HRVs) are the most common cause of viral respiratory infections and their complications. So far, no anti-viral agent has been approved for prevention or treatment of HRV infections. Pursuing our researches on small molecules with anti-rhinovirus activity, in this paper we describe the synthesis and in vitro anti-HRV 1B and 14 properties of new [2-(2H-chromen-3-yl)vinyl]pyridines and 3-[2-(pyridinyl)vinyl]-4H-chromen-4-ones. Generally, the synthesized compounds interfered with the replication of both serotypes at the micromolar or submicromolar concentrations. Preliminary results on their mechanism of action, performed on selected (E)-2-[2-(2H-chromen-3-yl)vinyl]pyridine, indicate an interference with the early step(s) of HRV 1B and 14 replication, probably at the uncoating level.

Human Rhinovirus, Wheezing Illness, and the Primary Prevention of Childhood Asthma

Human Rhinovirus, Wheezing Illness, and the Primary Prevention of Childhood Asthma