Respiratory syncytial virus and sudden infant death.
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Specimens of 387 nasopharyngeal suction smears obtained from 354 children hospitalized with acute respiratory infections during an eight-month period were examined for the presence of respiratory syncytial (RS) virus by the indirect immunofluorescence antibody technique (IFAT) and by conventional tissue culture infectivity techniques. Respiratory syncytial virus was identified in nasopharyngeal suction smear specimens from 123 of these specimens (32%) with the use of both techniques. Of the specimens positive on tissue culture 92% were also positive for RS virus by IFAT. However, eight specimens positive for RS virus by tissue culture were negative by IFAT, although three of the specimens were technically unsuitable. Six percent of the specimens negative for RS virus by tissue culture were positive for RS virus antigen when tested by IFAT. Using IFAT, identification of RS virus could be accomplished within four to six hours, whereas isolation by tissue culture took an average period of ten days. These data suggest that IFAT is a reliable means for the rapid diagnosis of RS virus infection in infants and children.
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Five temperature-sensitive (ts) mutants of respiratory syncytial (RS) virus (ts-1, ts-1 NG-1, ts-1 NG-16, ts-2, and ts-7), previously evaluated forinfectivity and virulence in chimpanzees and owl monkeys, were also assayed for in vivo genetic stability. None of the five mutants tested was completely stable genetically. Thus, virus which had lost some or all of the ts property was recovered from each infected chimpanzee. Significantly, each ts-1 NG-1 isolate retained some degree of temperature sensitivity and hence was not true wild-type virus. Clonal analysis of viruses shed by ts-1, ts-1 NG-1, ts-1 NG-16, or ts-7 infected chimpanzees indicated that in most instances only a minority of the virus shed was altered genetically. Of five chimpanzees infected with the ts-2 mutant, three shed only ts virus, and the remaining two chimpanzees shed only ts+ virus. Such ts+ virus proved to be avirulent when evaluated in chimpanzees or owl monkeys, indicating that loss of the ts property did not restore virulence. Based upon these findings, the ts-2 mutant appears to be a suitable candidate for clinical trials in man.
Shedding of respiratory syncytial virus (RSV) in nasopharyngeal aspirates (NPA) of hospitalized children with acute respiratory infection was studied using direct antigen detection by time-resolved fluoroimmunoassay, rapid identification of infectious virus in centrifugally inoculated cell cultures by immunoperoxidase staining and conventional virus culture. Sequential NPAs, in which also local RSV-specific IgA response was measured, were collected from children with proven RSV infection. The shedding pattern was similar for both infectious virus and viral antigen. The overall agreement of the three methods was good (81%) in diagnostic specimens collected on admission, but markedly reduced (46%) in follow-up specimens. Secretory IgA was abundant in specimens giving discrepant or negative results only. The proportion of patients who shed RSV was high (> or = 87%) in the first week after onset of symptoms, and decreased sharply in the second week. An opposite temporal pattern was found in the proportion of patients with detectable RSV-IgA in their secretions. Sequentially isolated strains were antigenically stable as determined by their reactivity with a large panel of monoclonal antibodies. The findings suggest that RSV shedding should be monitored by using more than one method for virus detection.
Eleven hundred and thirty-three clinical specimens submitted to the laboratory for diagnosis of respiratory virus infections were tested by direct immunofluorescence (DIF) for respiratory syncytial virus (RSV), by shell vial culture, and by conventional cell culture. The shell vial cultures were stained with 8 different monoclonal antibodies both 1 day and 3-7 days after inoculation. In order to limit the cost and the workload, mixtures of monoclonal antibodies were used. Coverslips with HEp-2 cells were incubated with a mixture of FITC-labeled monoclonal antibody to RSV and nonlabeled monoclonal antibody to adenovirus. When no RSV positive IF staining was observed after the first incubation step, the same coverslip was incubated once more with FITC-labeled anti-mouse antibody. A positive reaction at this stage indicated the presence of adenovirus. Similarly, cultures of tertiary monkey kidney cells were investigated with a mixture of two FITC-labeled monoclonals to the influenza viruses A and B and three nonlabeled monoclonals to the parainfluenza viruses 1, 2 and 3. If influenza virus or parainfluenza virus was detected, the exact type was determined by staining different parts of a duplicate coverslip. Shell vial cultures for cytomegalovirus (CMV) were always performed separately on human embryonic lung fibroblasts. Using this approach, we detected RSV (n = 248), CMV (n = 42), parainfluenza virus (n = 31), influenza virus (n = 28), and adenovirus (n = 6), in most cases after only one day of culture. For RSV, the sensitivity of the shell vial method was too low (74%) to allow omission of DIF (sensitivity 95%).(ABSTRACT TRUNCATED AT 250 WORDS)
For the last five years, the Brussels Institute of Hygiene and Epidemiology has been involved in the surveillance of acute respiratory infections (ARI). The four indicators used (number of encounters of ARI by GP's/100 encounters, virus isolations, absenteeism and mortality) are discussed. A regression procedure is applied to the data collected by a sentinel network of general practitioners (GP's). This procedure permits the baseline to be visualized and an epidemic threshold to be determined in order to recognize early an influenza outbreak. The traditional use of flu-like illnesses as an indicator might be improved by the addition of non-specific ARI which are more precocious, especially in children. The criteria for an accurate definition of an influenza epidemic are discussed. The same mathematical model can be used for the analysis of mortality linked with an outbreak. It shows that the last epidemic in the winter 1989-1990 was responsible for about 4900 deaths directly or indirectly related to influenza.
Retrospective and prospective serological surveys to determine the prevalence of respiratory syncytial virus (RSV) and adenovirus (ADV) infections in children with respiratory diseases were carried out from 1985 to 1988 at the University College Hospital, Ibadan, Nigeria. 306 sera from subjects between 3 months and 12 years old were assayed by complement fixation test for antibodies to both viruses. Second samples were available from 42 of the subjects for antibody conversion testing. The rate varied with age, period of the year and virus. Overall prevalences of 23.5% and 18.3% were obtained for RSV and ADV respectively. 2.6% of the subjects had antibodies to both viruses. Among 42 paired samples tested, 61.9% and 33.3% showed positive antibody conversion to RSV and ADV respectively. Although there was evidence of active circulation of both viruses throughout the year, infection with RSV was higher from September to January, while ADV infection was significantly higher during February and March. Antibody prevalence to both viruses increased with age up to a peak of 39.1% for RSV in children 5-6 years old and 32% for ADV in children 3-4 years old. None of the subjects aged 10 years and above was positive for both viruses. Both RSV and ADV are important agents of acute respiratory infection in children in Nigeria.
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Cystic fibrosis is the most common lethal or semi-lethal genetic disease in Caucasians of Central European origin, among whom it is inherited as an autosomal recessive trait at a frequency approximately 10 times that expected from recurrent mutation alone. A decreased sialic acid content has been observed in cell surface glycoproteins on cystic fibrosis fibroblasts and in numerous soluble glycoprotein preparations from cystic fibrosis homozygotes. Sialic acid residues on cell surface glycoconjugates play an essential role in the binding and infectivity of myxoviruses and paramyxoviruses, including those causing pandemic influenza. It is suggested that increased resistance to these viruses conferred by similar but quantitatively smaller alterations in sialoglycoconjugate structure in cystic fibrosis heterozygotes may have provided a selective advantage to maintain the high frequency of the cystic fibrosis gene in Caucasian populations.
To better understand the recovery process of infants with lower respiratory tract disease due to respiratory syncytial virus, the production of interferon by 129 children (ages 10 days to 24 months) with RSV infection was compared to that of 20 children with influenza (ages 1 to 36 months), and 37 children with parainfluenza virus infection (ages 4 to 66 months). Interferon assays of 285 nasal washes from children with RSV revealed that interferon production occurred in only 5 (4%) of the children. Significantly more children infected with infleunza virus, 55% (P less than 0.001), and parainfluenza virus, 30% (P less than 0.001), produced interferon. In addition, the quantity of interferon produced by children with RSV (geometric mean titer = 2) was significantly less than that of children with influenza (GMT = 26.8, P less than 0.001) and parainfluenza virus (GMT = 23.5, P less than 0.001). In the children infected with RSV, in constrast to those with influenza, interferon detection was not associated with diminished shedding of virus.
Interferon levels in nasal secretions of infants under one year of age, and hospitalized with lower repiratory tract disease, were measured during two respiratory infection seasons. In the first year serial secretions from 50 infants with respiratory syncytial virus infection were examined. Undetectable or low levels of interferon were found in all samples, and mean levels did not fluctuate significantly in relation to disease and recovery. This was in contrast to anti-RSV IgA, which appeared and increased in concentration as virus shedding decreased and stopped. In the second year secretions were obtained from nine infants with influenza A virus infection as well as from 13 with RSV. All those with influenza developed measurable interferon in secretions (geometric mean titer 138 units/ml), which was acid and heat stable, and trypsin sensitive (type I interferon). RSV infection again stimulated very low levels (geometric mean 5 units/ml). The lack of correlation of interferon concentration with cessation of RSV shedding suggests either that it is not involved in recovery or that low levels are adequate. On the other hand, it appears that the young infant is fully capable of a brisk local interferon response, at least to infection by influenza A.
The possibility that cell-mediated immunity might play a role in the pathogenesis of infection with respiratory syncytial virus was evaluated in a study of 39 infants. Infection with RSV was confirmed by identification of virus in nasopharyngeal secretions using immunofluorescence, and by tissue culture infectivity. CMI, as determined by a whole blood lymphocyte transformation technique, was evaluated in samples taken 0 to 10 and 20 to 60 days after the onset of illness. Patients diagnosed as having RSV-induced bronchiolitis or recurrence of asthma had evidence of significantly (P less than 0.01) higher degree of CMI in the 0 to 10-day period than patients with RSV pneumonia or upper respiratory illness. Higher CMI activity in the 20 to 60-day period was also seen in patients with more severe illness, with moderate-to-severe degree of hypoxia. A positive correlation was observed between the degree of LTF activity in samples taken 20 to 60 days after the onset of illness ard subsequent episodes of wheezing. Eleven patients had one or more episodes of wheezing in the first six months after RSV infection. LTF activity in samples taken during the 20 to 60-day period from these patients was significantly higher (P less than 0.02) than LTF activity in corresponding samples from six patients who were free of wheezing in the six months after RSV infection. The results suggest that alterations of RSV-specific cell-mediated immune mechanisms may result in an increased tendency toward airway reactivity on primary and subsequent exposure to RSV and possibly to other agents.
Several viruses may cause more or less severe acute respiratory infections in man, some of which are followed by systemic infection. Only for influenza and measles are licensed vaccines available at present. The protection induced by influenza vaccines, which are based on inactivated whole virus or viral subunits, depends largely on the matching of vaccine strain and circulating virus. Measles vaccines, which are based on attenuated live virus, have been quite effective in controlling the disease in vaccinated populations in the industrialized world. In developing countries, severe measles infections occur in infants from six to nine months of age, which necessitates the vaccination of children of less than six months. At that time maternal antibodies, that may interfere with the induction of protection, may still be present. Therefore, instead of using the parenteral route, the possibility to use the mucosal route of primary immunization is also investigated for vaccination with attenuated live measles vaccines. The use of inactivated measles vaccines has resulted in a state of immunity which upon exposure to the virus may induce an atypical measles syndrome including a severe pneumonia. Measles virus proteins presented in an iscom matrix have recently been shown to induce functional B and T cell responses to both the surface glycoproteins of the virus. These responses could also be induced in the presence of virus neutralizing antibodies and they proved to be protective in several animal model systems. Many of the problems that have been encountered in the development of measles vaccines, proved to be similar in the development of vaccines against other paramyxoviruses causing acute respiratory infections in man, including respiratory syncytial virus. Parenteral application of inactivated and attenuated live vaccines against these paramyxoviruses has generally had little success. Topical application of attenuated live vaccines has been more successful, and also the use of vaccinia recombinant viruses expressing foreign paramyxoviral glycoproteins has shown promising results in laboratory animals. Live vaccines based on adenovirus types 4 and 7 in oral enteric-coated vaccines, which lead to virus replication in the intestines but not in the respiratory tract have been included in military vaccination programs. The possibility to replace e.g. the E3 region with foreign DNA makes adenoviruses also suitable as cloning vectors for proteins of other respiratory viruses. Although live attenuated vaccines against some of the serotypes of rhinoviruses have shown promising results, the generation of a multivalent vaccine against this epidemiologically most significant cause of acute respiratory infections will be almost impossible, due to the multiplicity of serotypes involved.(ABSTRACT TRUNCATED AT 400 WORDS)
Respiratory syncytial virus infections are thought to be uncommon in the first month of life. During a community outbreak, we prospectively studied such infection in our neonatal units. Of 82 neonates studied, 66 were hospitalized for six days or longer, and 23 (35 per cent) acquired this virus. Four infants died, two unexpectedly. Infected infants had a significantly shorter gestation and birth weight. Illness was often atypical, with nonspecific signs, especially in infants under three weeks of age, who had significantly less lower-respiratory-tract involvement and lower quantities of virus in their nasal washes. The titer of virus shed correlated with the infants' postnatal, but not gestational, age. Infection was also acquired by 34 per cent of the staff, who appeared to be important in the spread of the virus. These findings suggest that respiratory syncytial virus may readily infect neonates, but the disease may be atypical and may be overlooked.
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To better understand acquired immunity to respiratory-syncytial-virus infections, we analyzed data from a 10-year study of respiratory illness in normal children who were followed longitudinally from early infancy. Immunity was measured in terms of failure to become infected or reduction in severity of clinical illness upon reinfection. Outbreaks of infections occurred seven times over the 10-year-period. During epidemics the attack rate for first infection was 98 per cent. The rate for second infections (75 per cent) was modestly reduced (P less than 0.001); that for third infections was 65 per cent. Age and history of infection both influenced illness. Immunity induced by a single infection had no demonstrable effect on illness associated with reinfection one year later; however, a considerable reduction in severity occurred with the third infection. These observations suggest that amelioration of illness--rather than prevention of infection--may be a realistic goal for immunoprophylaxis.
An outbreak of respiratory syncytial virus (RSV) infection occurred among 31 patients in a marrow transplant center over a 13-week period beginning in January 1990. RSV infection was also documented in 35 family members and employees. Of 18 patients with pneumonia, 14 (78%) died. None of 13 with upper respiratory infection died. Preengraftment patients tended to develop pneumonia more frequently than did engrafted patients. Early administration of ribavirin may have had a beneficial effect in patients with pneumonia. Antigenic and genomic analysis of 14 available isolates suggested that at least four different viral strains were responsible for the outbreak. One group of patients and 1 employee in spatial proximity were infected with the same strain and likely acquired their infections nosocomially. RSV infection in marrow transplant patients is a serious and life-threatening infection with a high mortality rate once pneumonia develops.