Richard G. Hegele

Epidemiology of respiratory viruses in patients hospitalized with near-fatal asthma, acute exacerbations of asthma, or chronic obstructive pulmonary disease

PURPOSE We compared the prevalence and spectrum of common respiratory viruses among patients with near-fatal asthma, acute exacerbations of asthma, or chronic obstructive pulmonary disease (COPD), and the relation of these findings to acute respiratory symptoms. METHODS We obtained adequate samples of respiratory secretions from 17 patients hospitalized with near-fatal asthma, 29 with acute asthma, and 14 with COPD. We used a polymerase chain reaction-based method to test for six common respiratory viruses in samples from endotracheal tube aspirates from patients with near-fatal asthma, and from induced sputum specimens from patients with acute asthma or COPD. Respiratory symptoms (runny nose, sore throat, fever, chills, malaise, and cough) were recorded. Quiescent-phase induced sputum specimens were examined from patients who were initially virus positive. RESULTS Viral nucleic acids were detected in 52% (31/60) of acute-phase specimens and 7% (2/29) of quiescent-phase specimens examined (P <0.001), with similar proportions of virus-positive patients during the acute phase in the three groups: 59% (10/17) of those with near-fatal asthma, 41% (12/29) with acute asthma, and 64% (9/14) with COPD. Picornavirus (47% [n = 8]) and adenovirus (24% [n = 4]) were most commonly identified in near-fatal asthma, whereas influenza virus (36% [n = 5]) predominated in COPD. Virus-positive patients had a significantly increased frequency of runny nose, sore throat, fever, chills, and malaise (odds ratio = 4.1 to 18; P = 0.02 to 0.001). CONCLUSION Respiratory viruses are associated with hospitalizations for near-fatal asthma, acute asthma, and COPD, with some differences in the spectrum of viruses involved in the different groups of patients. Respiratory viruses are a target for the prevention and perhaps the treatment of these conditions.

Persistent Airway Inflammation after Resolution of Respiratory Syncytial Virus Infection in Rats

Neurogenic inflammation is markedly potentiated in airways that are infected with respiratory syncytial virus (RSV). Aims of this study were to determine whether this potentiation persists after the virus is cleared, investigate the mechanism of postviral potentiation, and define whether prophylaxis with a MAb against the RSV fusion protein (palivizumab) prevents this effect. Thirty days after inoculation, no evidence of active RSV infection was found in the airway epithelium by plaque assay or immunostaining and no viral nucleic sequences were detected by PCR, yet capsaicin-induced plasma extravasation in the airways that were infected 30 d earlier with RSV was still significantly larger compared with pathogen-free controls. Substance P content in lung tissues and capsaicin-induced release of this peptide from sensory nerves were significantly increased at 30 d. The administration of palivizumab 24 h before virus inoculation prevented the development of abnormal neurogenic inflammatory responses. Our data suggest that the airways remain abnormally susceptible to the proinflammatory effects of sensory nerves after RSV infection is cleared, as a result of changes in sensory innervation, and that this abnormality can be prevented by passive prophylaxis against RSV.

ERK MAP kinase-activated Arf6 trafficking directs coxsackievirus type B3 into an unproductive compartment during virus host-cell entry.

Clathrin- and caveolae-mediated endocytosis have been implicated in the productive entry of many viruses into host cells. ADP-ribosylation factor 6 (Arf6)-dependent endocytosis is another endocytosis pathway that traffics from the cell surface and it is the only Arf that traffics at the plasma membrane. However, little is known about Arf6-dependent trafficking during virus entry. This study showed that coxsackievirus type B3 (CVB3) associated with decay-accelerating factor in non-polarized HeLa cells can be redirected into non-productive compartments by Arf6-dependent internalization, thus restricting infection. Overexpression of wild-type (WT) and constitutively active (CA) Arf6 in HeLa cells resulted in a 2.3- and 3.6-fold decrease in infection, respectively. A dominant-negative inhibitor of Arf6 recovered restriction of infection by WT-Arf6 and CA-Arf6. RNA interference of endogenous Arf6 resulted in a 3.3-fold increase in CVB3 titre in HeLa cells. It was shown that coxsackie-adenovirus receptor (CAR) ligation by virus or CAR-specific antibody could activate extracellular signal-regulated kinase (ERK) of the mitogen-activated protein kinase family and lead to Arf6-mediated viral restriction. In the absence of ERK activation, CVB3 internalization into early endosomes was inhibited and subsequent infection was reduced, but Arf6-mediated restriction was also abolished. In conclusion, receptor-mediated signalling enhances CVB3 entry whilst also activating non-productive pathways of virus entry; thus, virus infection is an equilibrium of productive and non-productive pathways of entry.

Comparison of three methods for respiratory virus detection between induced sputum and nasopharyngeal aspirate specimens in acute asthma.

Viral respiratory tract infections are associated frequently with acute exacerbations of asthma. Nasopharyngeal aspirates and bronchoalveolar lavage specimens are used extensively for detecting viral respiratory tract infections, but not sputum. The aim of the study was to determine the efficiency of viral detection in induced sputum versus nasopharyngeal aspirate obtained during acute exacerbations of asthma, comparing three laboratory methods of viral diagnosis. Paired samples of induced sputum and nasopharyngeal aspirate obtained from 32 adults admitted to hospital with acute asthma were subjected to reverse transcription-polymerase chain reaction (RT-PCR), viral culture, and immunofluorescence assay. The results show that RT-PCR was associated with significantly higher rates of viral detection than culture (P=0.005) or immunofluorescence (P=0.001), without significant differences in the rates of viral detection between induced sputum and nasopharyngeal aspirate. It is concluded that induced sputum specimens are feasible for detection of viral respiratory tract infections by RT-PCR during acute exacerbations of asthma.

Increased airway reactivity in human RSV bronchiolitis in the guinea pig is not due to increased wall thickness

Bronchiolitis due to the respiratory syncytial virus (RSV) is the most common cause of lower respiratory tract infection in the first year of life. It has been suggested that RSV infection may cause subsequent asthma, but a mechanism for this relationship has not been demonstrated. Studies examining the presence of airway reactivity in infants with RSV bronchiolitis are limited by our inability to administer provocative agents such as histamine to such ill infants. This makes a small animal model of this condition a useful tool in which to investigate the pathophysiology of RSV bronchiolitis. We, therefore, evaluated airway responsiveness in vivo and airway morphometric changes in 20 guinea pigs infected by instilling 4,000 plaque‐forming units of human RSV virus onto the nasal mucosa under halothane anaesthesia, while 20 control animals received an equivalent volume of sterile cell culture medium.

Community Study Using a Polymerase Chain Reaction Panel to Determine the Prevalence of Common Respiratory Viruses in Asthmatic and Nonasthmatic Children

We developed a sensitive polymerase chain reaction (PCR) panel, suitable for the detection of seven common respiratory viruses, to study the prevalence of viruses in nasal swabs obtained from clinically stable asthmatic children (n = 21), non-physician diagnosed asthmatic children with exercise-induced bronchoconstriction (EIB) (n = 16), and nonasthmatic, non-EIB controls (n = 33). The PCR panel detected viruses in 43/70 (61.4%) specimens but there were no significant differences in prevalence of these viruses between the three groups of children. These results indicate that clinically stable asthmatic and nonasthmatic children frequently harbor viruses in the upper respiratory tract.

Usefulness of bronchoalveolar lavage for diagnosis of acute and persistent respiratory syncytial virus lung infections in guinea pigs.

To investigate whether bronchoalveolar lavage (BAL) fluid specimens can be used to diagnose acute and persistent respiratory syncytial virus (RSV) lung infections in guinea pigs, we tested BAL fluid and lung tissue specimens for evidence of viral infection, and compared BAL cytology between infected and uninfected animals. RSV‐inoculated guinea pigs were studied during acute bronchiolitis (days 3 and 7 postinoculation), convalescence (Day 14 postinoculation), and persistent infection (Days 28 and 60 postinoculation), and were compared to the sham‐infected control animals. BAL and lung tissue specimens were cultured for virus and tested by immunocytochemistry for viral protein. A reverse transcription‐polymerase chain reaction (RT‐PCR) method was used to test for viral nucleic acid. Total and differential BAL cell counts were compared between RSV‐inoculated and control animals on each study day.