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PubMed2006

Epidemiological profile and clinical associations of human bocavirus and other human parvoviruses.

BACKGROUND: Human bocavirus (HBoV) and PARV4 are newly discovered human parvoviruses. HBoV, which was first detected in respiratory samples, has a potential role in the development of human respiratory disease. The present study compared the frequencies, epidemiological profiles, and clinical backgrounds of HBoV and PARV4 infections with those of other respiratory virus infections, by evaluating diagnostic samples referred to the Specialist Virology Laboratory (SVL) at the Royal Infirmary of Edinburgh (Edinburgh, United Kingdom). METHODS: Anonymized samples and study subject information were obtained from the respiratory sample archive of the SVL. Samples were screened for HBoV, PARV4, B19, respiratory syncytial virus (RSV), adenoviruses, influenza viruses, and parainfluenza viruses by use of nested polymerase chain reaction. RESULTS: HBoV infection was detected in 47 (8.2%) of 574 study subjects, ranking third in prevalence behind RSV infection (15.7%) and adenovirus infection (10.3%). Peak incidences of HBoV were noted among infants and young children (age, 6-24 months) during the midwinter months (December and January) and were specifically associated with lower respiratory tract infections. HBoV infections were frequently accompanied by other respiratory viruses (frequency, 43%), and they were more prevalent among individuals infected with other respiratory viruses (17%), frequently adenovirus or RSV. All respiratory samples were negative for PARV4. CONCLUSIONS: In the present study, HBoV was a frequently detected, potential respiratory pathogen, with a prevalence and an epidemiological profile comparable to those of RSV. Identification of HBoV infections may be clinically important in the future.

Female↗
PubMed1999

Conventional respiratory viruses recovered from immunocompromised patients: clinical considerations.

Lower respiratory tract infection is the most common complication in the immunocompromised patient. From January 1991 to December 1995, 785 consecutive patients with suspected respiratory tract infections were studied. One hundred ninety-nine viruses were isolated from 182 (23%) of 785 bronchoalveolar lavage fluid specimens. Cytomegalovirus was isolated from 131 patients, herpes simplex virus was recovered from 31, and conventional respiratory viruses (CRVs) were recovered from 36. There were 9 influenza A viruses, 2 influenza B viruses, 7 parainfluenza viruses, 5 respiratory syncytial viruses, 5 adenoviruses, 6 enteroviruses, and 3 rhinoviruses. We identified 22 patients from whom a CRV was the only microorganism recovered; 13 patients developed pneumonia, 10 had acute respiratory failure, 5 required support with mechanical ventilation, and 5 (23%) died. In conclusion, CRVs are frequent causes of respiratory illnesses and are associated with high rates of morbidity and mortality among immunocompromised patients.

Adenoviridae↗
PubMed1998

Impact of RNase L overexpression on viral and cellular growth and death.

The biologic actions of interferons (IFNs) are complex and involve multiple biochemical mechanisms, including the 2-5A system, a regulated RNA decay pathway. The 2-5A system is implicated in the antipicornavirus activity of IFN and in the control of apoptosis. To further investigate involvement of the 2-5A system in the control of viral and cellular growth and death, human RNase L cDNA was stably expressed in murine 3T3 cells from a constitutive cytomegalovirus (CMV) promoter. A clonal cell line, 3T3/pLZ, was isolated that overexpressed RNase L by >100-fold compared with levels of the endogenous murine RNase L. Interestingly, human RNase L levels in 3T3/pLZ cells decreased 3-fold as cells entered a confluent, growth arrest state, suggesting autoregulation. Overexpression of human RNase L greatly enhanced both the cell growth inhibitory activity of IFN and the proapoptotic activity of staurosporine. Furthermore, high levels of RNase L suppressed the replication of diverse viruses: encephalomyocarditis virus, vesicular stomatitis virus, human parainfluenza virus-3, and vaccinia virus. Additional reductions in viral growth were obtained by treating 3T3/pLZ cells with IFN (a + beta) before infections. These results directly demonstrate the anticellular and antiviral potential of the 2-5A system.

3T3 Cells↗
PubMedDate not supplied

Nutritional effects on response of children in developing countries to respiratory tract pathogens: implications for vaccine development.

Malnutrition and respiratory disease commonly occur together, and the immunologic defects caused by malnutrition may affect clinical responses and the impact of vaccines. These defects include impaired maturation and function of T lymphocytes, diminished secretory immune responses, poor humoral antibody responses to polysaccharide antigens, deficient complement activity (especially the alternative pathway), and impaired antimicrobial mechanisms by phagocytic cells. In addition, protein-energy malnutrition often leads to very high serum levels of IgE as a result of altered T cell regulation of IgE production and may contribute to IgE-mediated pathologic responses to respiratory syncytial virus or parainfluenza virus infection. Correction of these deficits may improve host response to infection and enhance vaccine-related protection. Both vitamin A and iron deficiency states have been epidemiologically associated with increased morbidity due to respiratory infection. The impact of preventive therapy with either vitamin A or iron is still uncertain, but such therapy has the potential for diminishing the incidence of respiratory infections and their consequences.

Child↗
PubMed1987

Epstein-Barr virus, cytomegalovirus, and other viral infections in children after liver transplantation.

We studied 51 consecutive pediatric patients for the frequency and morbidity of viral infections after liver transplantation. The incidence of primary (67%) and reactivation (48%) Epstein-Barr virus (EBV) infections and reactivation (88%) cytomegalovirus (CMV) infection was comparable to that seen in adult transplant recipients. However, fewer pediatric than adult transplant recipients experienced primary CMV infection (P less than .01). Five (38%) of 13 CMV infections were symptomatic and included hepatitis, pneumonitis, enteritis, and mononucleosis. Two of 14 patients with primary EBV infection subsequently developed, at two months and two years after initial infection, an EBV-associated lymphoproliferative syndrome, and one of 10 patients with reactivated EBV infection developed a possible EBV-associated febrile encephalopathy. Other viruses causing infection in these children included herpes simplex virus, varicella-zoster virus, adenovirus, parainfluenza virus, respiratory syncytial virus, and rotavirus.

Adolescent↗
PubMed1981

Pneumonia: an eleven-year study in a pediatric practice.

Data from an 11-year study of acute lower respiratory tract illness were analyzed to provide added insight into the causes and patterns of occurrence of pneumonia in an outpatient pediatric population. Over 80% of all episodes of pneumonia occurred in children less than seven years of age; the peak attack rate was observed in 2-4-year-old children. A virus or Mycoplasma pneumoniae was isolated from 24% of children with pneumonia; 57% of all episodes occurred during outbreaks for which a viral or mycoplasmal cause could usually be identified. Respiratory syncytial virus (RSV), parainfluenza viruses types 1 and 3, adenoviruses, influenza A viruses and M. pneumoniae accounted for 86% of all isolates. Attack rates of pneumonia in preschool children were highest during periods of RSV and influenza A virus prevalence; M. pneumoniae was the most frequent isolate from school aged children. The data contained in this report augment the understanding of the causes of pneumonia in children and provide a data base for scientists interested in the prevention of lower respiratory tract illness in children.

Adolescent↗
PubMed1999

Respiratory viruses in adult liver transplant recipients.

BACKGROUND: The contribution of respiratory viruses to respiratory disease in adult liver transplant (LT) recipients has not been studied. We performed a prospective audit to document the incidence of respiratory syncytial viruses ([RSVs], parainfluenza virus, influenza virus, and adenovirus) after LT, and to determine their contribution to respiratory disease in this setting. METHODS: Consecutive adult recipients were followed for 8 months after LT. Throat swabs were collected weekly for up to 12 weeks after LT, and virological surveillance was performed using conventional techniques (direct immunofluorescence and cell culture). A polymerase chain reaction assay for RSV was subsequently performed on selected specimens. Clinical data, including episodes of respiratory disease, were also recorded. RESULTS: During the study period, 51 patients received 53 LT. Five patients died, but no viruses were isolated from these patients at any stage. A total of 323 swabs were examined by conventional techniques and yielded 35 viral isolates (10.8%). Herpes simplex virus (type 1) accounted for 33 isolates, none of which were associated with respiratory disease. Two of 323 swabs (0.62%), in 2 patients, yielded respiratory viruses (both RSV); both patients had self-limiting, mild, upper respiratory tract symptoms. In these 2 patients, the polymerase chain reaction assay was more sensitive than conventional techniques and was able to detect extended RSV excretion. Of 51 recipients, 31 (61%) were always negative for viruses. Of 51 recipients, 10 developed respiratory failure, but no respiratory viruses were isolated from any of these patients. CONCLUSIONS: Respiratory viruses are rarely isolated from adult recipients after LT and are not associated with serious morbidity or with mortality. Routine surveillance for respiratory viruses in this patient population is not justified on the basis of this study.

Adenoviridae Infections↗
PubMed2004

Outbreak of human metapneumovirus infection in norwegian children.

INTRODUCTION: Human metapneumovirus (hMPV) was recently discovered in children with acute respiratory tract infection. We have studied the occurrence of hMPV and report clinical findings of 50 hMPV-infected children who were hospitalized during an outbreak in Norway. METHODS AND POPULATION: During 5 months from November 15, 2002 to April 14, 2003 we collected nasopharyngeal aspirate specimens from 236 children admitted because of respiratory tract infection (RTI). Samples were analyzed for influenza virus A/B, parainfluenza viruses 1, 2 and 3 and respiratory syncytial virus by direct immunofluorescence assays and cell culture. Rhinovirus, adenovirus and hMPV were identified by polymerase chain reaction. RESULTS: Human metapneumovirus was identified in 50 of 236 children (21%). Most (41 of 50) hMPV-infected children were hospitalized between November 15 and January 15, and during these 2 months hMPV was the most common isolate (41 of 72 isolates; 57%). Respiratory syncytial virus was identified in 36 children (15%), among whom 34 were admitted after the hMPV outbreak. The median age of hMPV-infected children was 12 months (range, 1 to 115 months), and one-half of the children had an underlying chronic disease. The most common symptoms were fever (86%), cough (90%), dyspnea (80%), wheezing (56%), rhinorrhea (44%), anorexia (48%) and vomiting (36%). Eight (16%) had an upper respiratory tract infection (rhinopharyngitis, n = 6; laryngitis, n = 2), 24 (48%) had bronchiolitis and 17 (34%) had pneumonia. Two-thirds with a lower RTI also had signs of upper RTI. Fourteen (28%) children needed supplemental oxygen, 1 was treated with continuous positive airway pressure and 2 were ventilated mechanically. CONCLUSION: Human metapneumovirus was the most common virus isolate during the winter season 2002 to 2003 in children hospitalized for respiratory tract infection. Upper respiratory tract infections and mild to severe bronchiolitis were most common, but a relatively high proportion of hospitalized children developed severe pneumonia.

Bronchiolitis, Viral↗
PubMed1988

Molecular cloning and characterization of six genes, determination of gene order and intergenic sequences and leader sequence of mumps virus.

mRNA isolated from mumps virus-infected Vero cells was converted into cDNA and cloned into the PstI site of the plasmid pBR322. After screening with 32P-labelled cDNA synthesized from poly(A)+ RNA of uninfected or mumps virus-infected Vero cells, five different groups of virus-specific clones were obtained. The virus specificity of the clones was confirmed by Northern blot analysis, in which the cDNA inserts from the five different groups hybridized to mRNAs of about 2100, 1500, 1450, 2000 and 2200 nucleotides. By the use of oligonucleotides synthesized on the basis of sequences obtained from the five cDNA clones and mRNAs, the sequence of the intergenic and surrounding areas was determined. During genome sequencing, a separate gene was identified between the fusion protein (F) gene and the haemagglutinin-neuraminidase protein (HN) gene. Using oligonucleotides synthesized on the basis of the new gene sequence, cDNA clones with poly(A) were isolated from the cDNA library. The gene order was determined to be 3' NC-P-M-F-SH-HN-L 5' (where NC, P, M, SH, and L represent the genes for the nucleocapsid, phosphoprotein or polymerase-associated, matrix or membrane, small hydrophobic and large proteins respectively). There is one nucleotide between the P and M (A), M and F (A), and HN and L genes (G), two between the NC and P (AA) and SH and HN (3'-CG) genes, and seven between the F and SH genes (3' GAUUUUA) as intergenic sequence. The leader sequence at the 3' end of the genome has been determined by sequencing the dicistronic leader-NC mRNA using oligonucleotide primers. The sequence from the 3' terminus to the NC gene start of the mumps virus genome is similar in length (55 nucleotides) to that present in Sendai virus, Newcastle disease virus and parainfluenza virus type 3, and the first five nucleotides are conserved in all negative-stranded RNA virus genomes sequenced to date.

Base Sequence↗
PubMed1992

Effects of protease inhibitors on replication of various myxoviruses.

We studied the effects of eight protease inhibitors on the multicycle replications of various orthomyxoviruses and paramyxoviruses. Among the compounds, nafamostat mesilate, camostat mesilate, gabexate mesilate, and aprotinin, which are widely used in the treatment of pancreatitis, inhibited influenza virus A and B replication at concentrations that were significantly lower than their cytotoxic thresholds in vitro. None of the protease inhibitors had activity against respiratory syncytial virus, measles virus, or parainfluenza virus type 3 at the highest concentrations tested. Camostat mesilate was found to be the most selective inhibitor. Its 50% effective concentration for influenza virus A replication was 2.2 micrograms/ml, and the selectivity index, which was based on the ratio of the 50% inhibitory concentration for host cell proliferation to the 50% effective concentration for influenza virus A replication, was 680. When the in ovo antiviral activity of the compounds was tested by using chicken embryos, camostat mesilate at a dose of 10 micrograms/g markedly reduced the hemagglutinin titers of influenza viruses A and B.

Amino Acid Sequence↗
PubMed1976

Viral infections in renal transplant recipients.

Cytomegalovirus (CMV) infections are common in renal transplant recipients. We studied 23 recipients prospectively to determine whether infections by other herpes-group and non-herpes-group viruses were also present. Sera, obtained at the time of surgery and periodically thereafter, were tested for antibody to CMV, herpes simplex virus (HSV), Epstein-Barr virus (EBV), parainfluenza viruses types 1, 2, and 3, and the viruses of measles and rubella. We found no evidence of an unusual incidence of primary or secondary infection by the non-herpesviruses tested. Rises to CMV, HSV, and EBV antibody titers occurred in 43, 38, and 32% of patients, respectively. All serological rises to herpes-group viruses occurred in patients seropositive at the time of transplantation, with the exception of three patients who experienced primary CMV infections. We conclude that reactivation of all herpes-group viruses tested may occur in transplant recipients. Morbidity was associated only with primarly CMV infection.

Adolescent↗
PubMed1983

Viral antibody screening system that uses a standardized single dilution immunoglobulin G enzyme immunoassay with multiple antigens.

We present an enzyme-linked immunosorbent assay (ELISA) system for the simultaneous determination of immunoglobulin G antibodies directed against several viruses. Antibodies to up to eight different viruses could be determined for three different sera on one microtitration plate. After subtraction of the absorbance values obtained with the control antigens, the viral antigen absorbancies were expressed as percentages of the absorbance obtained with a pooled immunoglobulin standard. This value, the relative antibody activity, was rapidly calculated by means of a computer directly connected to the ELISA photometer and was stored on magnetic disks, thereby facilitating seroepidemiological studies. The reproducibility of the relative antibody activity was calculated to at best +/- 3.6% (standard deviation) in an intraassay test and to at worst +/- 20.4% (standard deviation) in an interassay test. Each serum was analyzed only at a dilution of 1/75. The sensitivity of this single-dilution ELISA (SD-ELISA) method for the detection of titer rises was compared with those of conventional methods, mostly complement fixation but also hemagglutination inhibition. A total of 142 of 155 (92%) paired sera showing fourfold complement fixation or hemagglutination inhibition rises also showed significant results in SD-ELISA. A total of 22 of 57 (39%) significant relative antibody activity rises were significant in complement fixation or hemagglutination inhibition. Overall, up to twice as many significant titer rises could be detected with SD-ELISA. Most of these seemed to have a sound correlation with clinical data. The specificity of SD-ELISA was found to be similar to that of complement fixation, with some cross-reactions occurring between herpes simplex and varicella-zoster virus antigens and between parainfluenza viruses. We have found SD-ELISA to be a valuable clinical virological tool that supplements conventional serology.

Antibodies, Viral↗
PubMed1986

Simultaneous infections with different enteric and respiratory tract viruses.

Infants and young children with rotavirus (RV) or visualized adenovirus in their stools were tested for the simultaneous presence of a respiratory viral pathogen in their upper respiratory tract. Overall, at least 10.7% of 484 study subjects had such dual infections, including 8.3% of 385 RV-positive gastroenteritis patients and 24.3% of 37 RV-positive respiratory disease patients. Respiratory syncytial virus was present in 34.1% of 41 dual infections with RV and at least 40% of the 12 to 15 dual infections with visualized fecal adenovirus. Other pathogens found in the respiratory tract of patients with RV or visualized fecal adenovirus infections included influenza viruses, adenoviruses, parainfluenza viruses, rhinoviruses, and a cytomegalovirus.

Adenoviridae Infections↗
PubMed1989

Evaluation of a monoclonal antibody pool for rapid diagnosis of respiratory viral infections.

A pool of monoclonal antibodies (MAbP) was evaluated both as a method of cell culture confirmation and as a rapid diagnostic screen for viral infection in respiratory secretions. The MAbP was used in a two-step fluorescent staining procedure on cells harvested from cultures (phase 1) and on exfoliated nasopharyngeal or tracheal cells (phase 2). Antibodies in the MAbP were directed against respiratory syncytial virus, adenoviruses, parainfluenza virus types 1, 2, and 3, and influenza viruses A and B. Individual antiviral antibody stains were used to identify specific viruses from MAbP-positive specimens. In phase 1 (cell culture confirmation only), 241 respiratory specimens were tested. MAbP sensitivity and specificity were 91 and 94%, respectively. In phase 2, 376 respiratory specimens were evaluated by direct staining of exfoliated cells, and these results were compared with results of cell culture isolation. The sensitivity and specificity of the MAbP used in direct specimen testing were 69 and 97%, respectively. These results indicate that the MAbP is highly specific and fairly sensitive for detection of seven different respiratory viruses in one testing procedure. The MAbP is a rapid screening technique for respiratory secretions and is potentially a cost-effective approach to viral detection.

Adenoviruses, Human↗
PubMed2000

Clinical and financial benefits of rapid detection of respiratory viruses: an outcomes study.

To assess the expected benefits of rapid reporting of respiratory viruses, we compared patients whose samples were processed using standard techniques such as enzyme immunoassays, shell vial assays, and culture tube assays (year 1) to patients whose samples were processed with the same standard techniques in addition to immunofluorescent testing (FA) directly on cytocentrifuged samples (year 2). The cytospin FA screened for influenza A and B viruses, respiratory syncytial virus (RSV), parainfluenza viruses 1 to 3, and adenovirus (DAKO Diagnostics Ltd.). The specificity of the cytospin FA for all viruses was 100%. The sensitivities for influenza A virus and RSV were 90 and 98%, respectively, but the sensitivities for influenza B virus and adenovirus were unacceptable (14.3 and 0%, respectively). However, since the former viruses account for >85% of our isolates from clinical specimens, the cytospin FA is an excellent screening test since the positive result was available within hours. The mean turnaround time for all positive viruses was 4.5 days in year 1 and 0.9 day in year 2 (P = 0.001). This rapid reporting resulted in physicians having access to information sooner, enabling more appropriate treatment. The mean length of stay in the hospital for inpatients with respiratory viral isolates was 10.6 days for year 1 versus 5.3 days for year 2. Mean variable costs for these patients was $7,893 in year 1 and $2,177 in year 2. After subtracting reagent costs and technological time, the savings in variable costs was $144,332/year. Summarizing, the cytospin FA markedly decreased turnaround time and was associated with decreased mortality, length of stay, and costs and with better antibiotic stewardship.

Centrifugation↗
PubMed1989

A neutrophil-derived antiviral protein: induction requirements and biological properties.

Polymorphonuclear neutrophilic granulocytes (PMN) have been implicated as playing a role in antiviral defense. In addition to having phagocytic and cytotoxic activities, PMN may produce an antiviral substance with interferon (IFN)-like activity. The product, for which the name polyferon (PF) has been coined, is produced upon direct encounter of PMN with bovine herpesvirus 1 (BHV-1)-infected bovine cells or membranes thereof. Exposure to purified virus only does not induce PF. The intimate interaction between PMN and the membranes was also revealed by electron microscopy studies. Bovine cells infected with herpes simplex virus type 1 could also induce PF production by bovine PMN, whereas cells infected with BHV-2, herpes simplex virus type 2, equine herpesvirus 1, bovine respiratory syncytial virus, bovine viral diarrhea virus, or parainfluenza virus 3 were unable to do so. Results obtained in experiments using transfected cells expressing BHV-1 glycoproteins as well as blocking experiments using BHV-1 glycoprotein-monospecific antibodies suggested that a combination of both viral product(s) and host cell factor(s) unique to bovine cells is required for induction of PF production by PMN. PF, which appeared in detectable amounts 12 to 18 h after exposure of PMN to the appropriate inducer, could not be neutralized by antibodies to bovine IFN-alpha, -beta, and -gamma. PF may nevertheless belong to the IFN family of proteins, as indicated by its ability to induce 2',5'-oligoadenyl synthetase in various cell types that are responsive to bovine IFNs and by its antiviral spectrum. It does, however, differ from the other cytokines in most immunological characteristics tested so far, including major histocompatibility complex class II antigen induction, cell migration, and cytotoxicity.

Animals↗
PubMed1995

Intracellular neutralization of influenza virus by immunoglobulin A anti-hemagglutinin monoclonal antibodies.

Traditionally, immunoglobulin A (IgA) was thought to neutralize virus by forming complexes with viral attachment proteins, blocking attachment of virions to host epithelial cells. Recently we have proposed an intracellular action for dimeric IgA, which is actively transported through epithelial cells by the polymeric immunoglobulin receptor (pIgR), in that it may be able to bind to newly synthesized viral proteins within the cell, preventing viral assembly. To this effect, we have previously demonstrated that IgA monoclonal antibodies against Sendai virus, a parainfluenza virus, colocalize with the viral hemagglutinin-neuraminidase protein within infected epithelial cells and reduce intracellular viral titers. Here we determine whether IgA can interact with influenza virus hemagglutinin (HA) protein within epithelial cells. Polarized monolayers of Madin-Darby canine kidney epithelial cells expressing the pIgR were infected on their apical surfaces with influenza virus A/Puerto Rico/8-Mount Sinai. Polymeric IgA anti-HA, but not IgG anti-HA, delivered to the basolateral surface colocalized with HA protein within the cell by immunofluorescence. Compared with those of controls, viral titers were reduced in the supernatants and cell lysates from monolayers treated with anti-HA IgA but not with anti-HA IgG. Furthermore, the addition of anti-IgA antibodies to supernatants did not interfere with the neutralizing activity of IgA placed in the basal chamber, indicating that IgA was acting within the cell and not in the extracellular medium to interrupt viral replication. Thus, these studies provide additional support for the concept that IgA can inhibit replication of microbial pathogens intracellularly.

Animals↗
PubMed2006

Human metapneumovirus infections in hospitalised infants in Spain.

BACKGROUND: Human metapneumovirus (hMPV) causes lower respiratory tract infections, particularly in young children and the elderly. METHODS: A prospective study was conducted on the clinical characteristics of infants <2 years of age admitted to hospital for respiratory infection and the characteristics of hMPV infections were compared with those of infections caused by respiratory syncytial virus (RSV). Influenza A, B and C viruses, RSV, parainfluenza viruses, and adenoviruses were simultaneously detected in clinical samples by multiple reverse transcription nested-PCR assay. The presence of hMPV was tested in all samples using two separate RT-PCR tests. RESULTS: A respiratory virus was detected in 65.9% of the 749 children included in the study. hMPV, found in 69 of the positive nasopharyngeal aspirates (14%), was the most common virus after RSV. Peak incidence was in March and over 80% of children were <12 months of age. The most common diagnoses were recurrent wheezing (49.3%) and bronchiolitis (46.4%). Oxygen therapy was required by 58% of patients, and assisted ventilation by one. Clinical characteristics in the 18 co-infections were indistinguishable from those of single infections. Fifty one hMPV single infections were compared with 88 hRSV single infections. Recurrent wheezing was diagnosed more frequently in hMPV patients. All other variables tested were similar in both groups. CONCLUSIONS: hMPV was the second most frequent virus after RSV in infants <2 years of age hospitalised for respiratory infection and was associated with lower respiratory tract infections. hMPV occurred predominantly in springtime. Co-infections were frequent and clinically similar to single infections and RSV infections.

Acute Disease↗