Measles in children who have malignant disease.

A review study examined the clinical course of measles diagnosed in children being treated for malignant disease in Newcastle upon Tyne during 1973-86. Of the 17 cases diagnosed, five were fatal. Factors associated with a favourable outcome were a typical rash and Koplik's spots, which were accompanied by a detectable serum antibody response and the disappearance of measles giant cells from nasopharyngeal secretions. Pneumonitis severe enough to require assisted ventilation was invariably fatal. Pneumonitis and encephalitis were the main complications. Treatment included immunoglobulin, interferon, and ribavirin, but none could clearly be shown to be effective. The comparatively low mortality in this series may have been due to the extensive use of the fluorescent antibody technique in Newcastle during the study period and therefore detection of less severe cases as compared with other reports.

Virus infections in childhood malignant disease.

The viruses isolated during infective episodes over a 5-year period of 93 children with acute lymphoblastic leukemia (ALL) and 107 children with other malignancies have been studied. Viruses were identified by the fluorescent antibody technique (FAT), electron microscopy, and culture. Viruses were isolated on 204 occasions. Rhinovirus and RSV occurred significantly more often in the patients with ALL, and adenovirus more commonly in the solid tumor patients. Viruses were responsible for 4 of the 69 deaths. Multiple different virus isolation during the same infective episode was significantly more common than in a control group. Where paired sera were available, only 10 out of 27 children showed at least a fourfold rise to the isolated virus. A rise in complement-fixing antibody is confirmed as a very unreliable method of detecting virus infection in immunosuppressed children. Viruses are important pathogens in children with cancer and assume greater relevance now that over 60% of malignant disease in childhood can be cured.

Monoclonal antibodies for the rapid diagnosis of influenza A and B virus infections by immunofluorescence.

Mouse monoclonal antibodies, directed against antigenic sites on influenza A and B viruses and found to be type-specific in an immunoassay, were assessed for use as diagnostic reagents in an indirect immunofluorescence assay on nasopharyngeal secretions. The influenza A antibodies were directed against nucleoprotein or matrix protein antigens and the influenza B antibodies against nucleoprotein and haemagglutinin antigens. The influenza A anti-matrix monoclonal antibody was found to give a strong intranuclear particulate fluorescence in normal baboon kidney cells and cells from nasopharyngeal secretions negative for influenza A virus, including those from a patient infected with respiratory syncytial virus. Pools of the remaining monoclonal antibodies gave satisfactory results on 25 specimens from patients with influenza A H1N1 and H3N2 subtypes and 12 from patients with influenza B.

The development of monoclonal antibodies to respiratory syncytial virus and their use in diagnosis by indirect immunofluorescence.

Twelve clones of murine hybridoma cells secreting antibody specific for respiratory syncytial (RS) virus were classified into four groups on the basis of their pattern of staining of unfixed RS virus-infected HEp-2 cells in an indirect immunofluorescence test. Three of the groups reacted with virus antigens present on the membrane of the cells, whilst the fourth group failed to stain most live cells, suggesting specificity for an antigen expressed internally. Representative monoclonals from the membrane antigen staining groups immunoprecipitated the 86K glycoprotein (G), 50K plus 19K glycoprotein (F1,2) and a 23K non-glycosylated protein (VP23). A representative monoclonal from the fourth group that appeared to stain an internally expressed protein immunoprecipitated the virion 34K phospho-protein (P). All four monoclonals stained acetone-fixed tissue culture cells infected with either the Long strain of RS virus or with strains isolated in Newcastle during the 1965, 1972, and 1983 winter epidemics. The anti-fusion protein antibody stained acetone-fixed cells from all of 26 nasopharyngeal secretions from infants with RS virus infection. The anti-G glycoprotein antibody and the anti-VP23 antibody stained cells from secretions poorly or not at all, whilst the anti-P protein antibody stained cells in half the secretions tested but reacted with only a small proportion of cells in comparison with the anti-F or polyclonal antibodies. A pool of all four monoclonals produced more intense staining than the anti-F monoclonal alone and gave a more clearly defined staining reaction than the polyclonal antiserum used for routine diagnosis in over half the secretions. These results indicate that monoclonal antibodies will be of value in the diagnosis of RS virus by indirect immunofluorescence if care is taken in the selection of a suitable pool.

Cell-mediated immunity in respiratory syncytial virus disease.

Systemic cell-mediated responses to respiratory syncytial (RS) virus were detected, using a whole blood transformation assay, in 10 of 28 infants and children with RS virus infections during the period 1-14 days postadmission. Cell-mediated responses were unrelated to the age of the patient or the severity of illness. No correlation was found between cellular responses and fourfold or greater rises in antibody titre to RS virus, as determined by a membrane immunofluorescence technique. Patients under 6 months of age had significantly lower levels of IgA and IgG antibody to RS virus compared to older patients, although cell-mediated responses were similar in both groups. The presence of cell-mediated reactivity to RS virus was also demonstrated in 5 of 95 samples of cord blood examined, and cellular responses failed to correlate with the levels of IgG antibody to RS virus.

Viral diagnoses using the rapid immunofluorescence technique and epidemiological implications of acute respiratory infections among children in different European countries.

From November 1978 to October 1981, a total of 7716 specimens of nasopharyngeal secretions were examined by the rapid immunofluorescence technique to determine the frequency of infections caused by the respiratory syncytial virus (RSV), influenza virus A, and parainfluenza viruses 1 and 3. The tests were carried out in six different virus laboratories located in Newcastle upon Tyne (England), Copenhagen, Oslo, Stockholm, Turku (Finland), and Vienna; laboratories in Lisbon and Paris participated in the study for shorter periods. The specimens were collected from infants and children less than 6 years of age who had been admitted to hospital with an acute respiratory infection. Standardized techniques and quality controlled reagents were used. At least one of the above viruses was detected in 1927 (25%) of the specimens: RSV in 1475, influenza virus A in 123, parainfluenza virus 1 in 110, and parainfluenza virus 3 in 237 specimens. Respiratory syncytial virus dominated in all centres, but in some Scandinavian centres distinct outbreaks due to this virus occurred only once or twice during the 3 years' study period. Three outbreaks of RSV were observed in Newcastle, but here an unprecedented delay of the first winter's epidemic occurred. The delay was associated with prolonged school closures in the area, and with a very early outbreak of influenza. Parainfluenza virus 3, which was predominantly a summer virus in Newcastle, was most frequently encountered during the colder months of the year in the other centres.

Fatal dissemination of cytomegalovirus after bone marrow transplantation.

The clinical course and necropsy findings are described of an 11-year-old child found to have active cytomegalovirus infection at the time of bone marrow transplantation for acute lymphoblastic leukaemia. Attention is drawn to the presence of primitive mononuclear cells of uncertain origin in the regenerating bone marrow.

Complement fixing antibody responses to virus infection in children with acute lymphoblastic leukaemia.

Thirty children with acute lymphoblastic leukaemia (ALL) were studied and had a virus isolated. Only 50% produced a significant rise in complement fixing (CF) antibody titre compared to 100% of normal children. The failure to produce antibodies was unpredictable. CF antibodies are not a reliable guide to virus infections in children with ALL.

Virus infections in children with acute lymphoblastic leukaemia.

The pattern of virus isolation and illness was studied in 64 children with acute lymphoblastic leukaemia (ALL) during periods of apparent infection and when the children were well. The virus isolation rate of 2.2 viruses per child a year is similar to that previously found in normal children. In only 32% of children with symptoms were viruses found and 14.5% had viruses isolated when asymptomatic. The children with ALL appear to be more vulnerable to multiple virus infections and to excrete the virus for longer periods. This may be due to failure of production of both local and systemic antibodies. The failure in the past to recognise the true importance of virus infections in ALL may have been due to inadequate diagnostic techniques.

Cell-free and cell-bound antibody in nasal secretions from infants with respiratory syncytial virus infection.

Twenty-two infants under 9 months of age hospitalized with bronchiolitis or pneumonia due to respiratory syncytial virus (RSV) were serially sampled to determine the pattern of secretory antibody response. Using double labeling techniques, we found several types of immunoglobulin in secretions: cell-free antibody to RSV of the immunoglobulin A (IgA), immunoglobulin G (IgG), and immunoglobulin M (IgM) classes; and immunoglobulins of all three classes bound to RSV-infected cells shed from the nasal epithelium (presumably cell-bound antibody to RSV). IgA attached to RSV-infected epithelial cells was almost always detected in the first available nasal sample (day 1 or 2 of hospitalization). In contrast, cell-free anti-RSV IgA first appeared an average of 3.5 days later at a time when virus antigen was disappearing from the secretion. IgG and IgM attached to RSV-infected cells appeared more irregularly. The titer of cell-free anti-RSV IgM was often higher than that of IgA early in the illness and declined as the infection resolved. Cell-free anti-RSV IgG was usually present earlier than IgA and rose during convalescence.

Epidemiology of respiratory viral infection among paediatric inpatients over a six-year period in north-east England.

During a 6-year, hospital-based study at Newcastle upon Tyne five consecutive winter epidemics of respiratory syncytial (R.S.) virus infection occurred; the virus was identified in 1428 cases, showing that 1 in 50 live-births were admitted to hospital with R.S. virus infection. Epidemics were inversely related to temperature and to number of hours of sunshine. Parainfluenzaviruses, the second largest group of pathogens, were identified in 543 cases; most infections by this group were due to parainfluenzavirus type 3, which accounted for admission to hospital of 1 in 300 live-births. Epidemics of parainfluenza type 3 showed a summer peak (there was often a second peak in autumn) and a positive correlation with temperature and number of hours of sunshine. Epidemics of parainfluenza 1 and 2 occurred together, but only every 2 years. Influenza A epidemics occurred every winter; they coincided with, and had the same climatic correlations as, epidemics of R.S. virus infections. Influenza A was the second most frequently identified virus, and was associated with the admission to hospital of 1 in 100 to 1 in 500 live-births. Knowledge of the epidemiology of respiratory viral infections may help in the planning of preventative measures.

Respiratory viruses and cot death.

Respiratory viruses and histological appearances of the lung were studied prospectively in an unselected series of 104 children who died between 1 week and 2 years of age. Thirty-one of the cases were cot deaths. Seven of these showed evidence of active virus infection in the lower respiratory tract. Similar evidence was found in two children who died from known causes and did not have a severe respiratory illness terminally. Although in some cases of cot death respiratory viruses may be responsible for a severe and rapidly overwhelming illness, the present results are compatible with an alternative hypothesis-namely, that minor respiratory illness may trigger sudden apnoea.

The coating of respiratory syncytial (RS) virus-infected cells in the respiratory tract by immunoglobulins.

During the course of 26 respiratory syncytial (RS) virus infections, infected cells in the respiratory tract become coated with immunoglobulins (IgA, IgG, and IgM), IgA being predominant. Methods are described for detecting both intracellular virus and coating immunoglobulin using a double staining technique and immunofluorescence. IgA coating antibody appears in small amounts very early in the illness. The relationship of coating antibody to pathogenesis, prevention, and recovery from RS virus infection is discussed.

Diagnosis of human coronavirus infection by immunofluorescence: method and application to respiratory disease in hospitalized children.

Rabbit antisera were prepared against coronavirus strains 229E and OC43 and used successfully to detect viral antigen in epithelial cells shed from the nasopharynx of symptomatic volunteers who had received coronavirus inocula three to four days before. The same serologic reagents were applied to nasopharyngeal secretion cells obtained from 106 infants and children hospitalized with respiratory tract disease and apparently not infected with conventional respiratory viruses. No coronavirus infections were detected by this method. It appears that coronavirus OC43 or 229E infections were not common in children in Tyneside hospitals during the period of study. However, fluorescence is a useful method for detection of coronavirus infections in symptomatic human subjects.

Comparison of immunofluorescence and immunoperoxidase methods for viral diagnosis at a distance: a WHO collaborative study.

Nasopharyngeal secretions collected in Newcastle were examined in both Newcastle and Stockholm for the presence of influenza virus type A and respiratory syncytial (RS) virus by immunofluorescence and immunoperoxidase techniques. A total of 139 specimens were examined in this way and the agreement between the two centres for immunofluorescence was 94% for influenza virus A, 95% for respiratory syncytial virus, and 95% for negative specimens. This technique can therefore be used for examining specimens taken at great distances from the collecting laboratory. The results of the immunoperoxidase technique were unsatisfactory mainly because of the presence of endogenous peroxidase. Measures taken to remove this also destroyed RS virus antigen. It is premature to introduce the immunoperoxidase technique for rapid virus diagnosis, but in due course, when the problems outlined in this article are solved, it could become a useful technique.