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PubMed2004

Prospective population-based study of viral lower respiratory tract infections in children under 3 years of age (the PRI.DE study).

UNLABELLED: Population-based incidence data from Europe on the disease burden of lower respiratory tract infections (LRTI) due to respiratory syncytial viruses (RSV), parainfluenza viruses (PIV) and influenzaviruses (IV) are lacking, especially with respect to the disease burden. In a 2-year prospective multicentre study of children aged <3 years in Germany, we registered population-based cases as outpatients (n=2386), inpatients (n=2924), and nosocomially-acquired (n=141). Nasopharyngeal secretions were tested for viral RNA. The annual incidence for physician visits per 100 children for all LRTI was 28.7, RSV 7.7, PIV 3.8 and IV 1.1. Annual hospitalisation rates per 10(5) children were for all LRTI 2941, RSV 1117, PIV 261 and IV 123. Annual nosocomial cases per 10(5) hospital days were for all LRTI 79, RSV 29, PIV 9 and IV 1.5. All five children (0.27%) who died had an underlying disease and four were nosocomially acquired. CONCLUSION: Hospitalisation rates due to lower respiratory tract infections in healthy children were similar to those reported elsewhere; the rates for outpatient visits were approximately ten times higher.

Child, Preschool↗
PubMed1985

Surveillance of respiratory viral infections by rapid immunofluorescence diagnosis, with emphasis on the epidemiological development of respiratory syncytial virus infections.

Surveillance of certain respiratory viral infections by applying immunofluorescence (IF) examinations to samples of nasopharyngeal secretions has been evaluated using a simplified procedure for the preparation of cell smears. Samples from 711 children living in different parts of Norway were examined during the winter 1982/83 and a positive diagnosis was made for 290 children (41%). Temporal epidemic peaks were observed for respiratory syncytial virus (RSV), parainfluenza virus type 3 and influenza virus. On the other hand, the monthly number of negative samples was almost constant throughout the period. Differences in timing of RSV outbreaks were observed between two regions in Norway. Compared to rapid IF diagnosis, RSV notifications obtained by serological examinations were delayed by several weeks. Rapid virus diagnosis by IF examinations with our simplified procedure for preparation of nasopharyngeal samples seems to be suitable for the epidemiological surveillance of respiratory viral infections, both for its simplicity of preparation of the samples and for its accuracy in defining the time of the actual virus infection. Nevertheless, the method is not without pitfalls; a close cooperation between those who take the specimens and the laboratory is essential, and the IF examinations should be performed by an experienced microscopist.

Antibodies, Viral↗
PubMed1996

Serological study of responses to selected pathogens causing respiratory tract infection in the institutionalized elderly.

In a prospective 2-year study, serological responses to selected pathogens were analyzed in 224 episodes of fever attributable to respiratory tract infection (51.8%) or of unknown source (48.2%) in 131 residents of two long-term-care facilities. A serological response was identified in 45 episodes (20.1%): Chlamydia pneumoniae (14 episodes), Haemophilus influenzae type b (1), influenza virus type A (14), respiratory syncytial virus (RSV;2), parainfluenza virus type 3 (7), C. pneumoniae and H. influenzae (3), C. pneumoniae and influenza virus type A (2), C. pneumoniae and RSV (1), and C. pneumoniae and parainfluenza virus type 3 (1). No serological responses to Chlamydia psittaci, Chlamydia trachomatis, parainfluenza virus types 1 and 2, influenza virus type B, or Mycoplasma pneumoniae were seen. Vaccination did not affect the duration of fever in those residents with serologically confirmed influenza A. Serologically confirmed C. pneumoniae infection was detected in 9.4% of all febrile episodes. Serological responses to a second agent were detected in 33% of the patients with C. pneumoniae infections, and these dual infections were associated with an underlying malignancy (P = .02). C. pneumoniae should be recognized as a potential pathogen when choosing empirical antimicrobial therapy for respiratory tract infection in residents of long-term-care facilities.

Aged↗
PubMed2001

Human parainfluenza virus-associated hospitalizations among children less than five years of age in the United States.

BACKGROUND: Human parainfluenza viruses 1 through 3 (HPIV-1-3) are important causes of respiratory tract infections in young children. This study sought to provide current estimates of HPIV-1-3-associated hospitalizations among US children. METHODS: Hospitalizations for bronchiolitis, bronchitis, croup and pneumonia among children age <5 years were determined for the years 1979 through 1997 using the National Hospital Discharge Survey. Average annual hospitalizations during the last 4 years of the study for each of these four diseases were multiplied by the proportions of each disease associated with HPIV-1-3 infection (as previously reported in hospital-based studies) to estimate hospitalizations potentially associated with HPIV-1-3 infections. Seasonal trends in HPIV-1-3-associated hospitalizations were compared with HPIV detections in the National Respiratory and Enteric Virus Surveillance System, which prospectively monitors respiratory viral detections throughout the United States. RESULTS: The proportions of hospitalizations associated with HPIV infection for each disease varied widely in the 6 hospital-based studies we selected. Consequently our annual estimated rates of hospitalization were broad: HPIV-1, 0.32 to 1.59 per 1,000 children; HPIV-2, 0.10 to 0.86 per 1,000 children; and HPIV-3, 0.48 to 2.6 per 1,000 children. Based on these data HPIV-1 may account for 5,800 to 28,900 annual hospitalizations; HPIV-2 for 1,800 to 15,600 hospitalizations; and HPIV-3 for 8,700 to 52,000 hospitalizations. CONCLUSIONS: We provide broad, serotype-specific estimates of US childhood hospitalizations associated with HPIV infections. More precise estimates of HPIV-associated hospitalizations would require large prospective studies of HPIV-associated diseases by more sensitive viral testing methods, such as polymerase chain reaction techniques.

Bronchiolitis, Viral↗
PubMed1999

Effect of inserting paramyxovirus simian virus 5 gene junctions at the HN/L gene junction: analysis of accumulation of mRNAs transcribed from rescued viable viruses.

Simian parainfluenza virus 5 (SV5) is a prototype of the Paramyxoviridae family of nonsegmented negative-sense RNA viruses. The single-stranded RNA genomes of these viruses contain a series of tandemly linked genes separated by intergenic (IG) sequences flanked by gene-end (GE) and gene-start (GS) sequences. The viral RNA polymerase (vRNAP) complex is thought to enter the genome at its 3' end, and synthesis of mRNAs is thought to occur by a stop-start mechanism in a sequential and polar manner, with transcriptional attenuation occurring primarily at the intergenic regions. As a result, multiple nonoverlapping mRNA species are generated for each single entry of the vRNAP. To investigate the functions of GE, IG, and GS sequences in transcription, we constructed plasmids containing cDNAs of the full-length SV5 genome in which the gene junction sequences (GE, IG, and GS sequences) located between the hemagglutinin-neuraminidase (HN) and the polymerase (L) genes were replaced with the counterpart sequences from other gene junctions. By using reverse genetics, we recovered viable viruses from each cDNA construct, although their growth characteristics varied. Analysis of the HN and L mRNAs by quantitative RNase protection assay indicated that the ratios of HN to L mRNAs varied over a fourfold range. The alteration of the gene junction sequences also permitted examination of the hypothesized requirement for hexamer nucleotide position of the GS sites. The recovery of infectious viruses with transcription initiation sites that occurred at nucleotide positions 1, 2, 3, 5, and 6 of the hexamer suggest that the requirement is nonstringent.

Animals↗
PubMed1989

Complete nucleotide sequence of the hemagglutinin-neuraminidase (HN) mRNA of mumps virus and comparison of paramyxovirus HN proteins.

The complete nucleotide sequence of the hemagglutinin-neuraminidase protein (HN) mRNA of the virulent SBL-1 strain of mumps virus has been determined. The mRNA contains 1887 nucleotides excluding the poly(A). The protein encoded by the mRNA has 582 amino acids and a membrane anchorage domain near the amino terminus. The calculated molecular mass (64 kDa) of the protein is in good agreement with that of the unglycosylated HN protein (63 kDa) identified in tunicamycin treated mumps virus infected cells (Herrler and Compans, 1983). The predicted sequence has nine potential N-glycosylation sites out of which two contain a cysteine residue and one has a proline residue as the variable amino acid X in the glycosylation site (Asn-X-Ser or Asn-X-Thr) and therefore, may not be utilized. One potential glycosylation site is in the cytoplasmic region which may not also be glycosylated. Comparison of the mumps HN protein sequence with the HN protein sequences of Sendai virus, simian-virus 5 (SV5), parainfluenza virus type 3 and Newcastle disease virus (NDV) shows two major homology regions, one region near the middle of the protein and the other in the second half of the molecule. In terms of percentage amino acid homology, the HN proteins of mumps virus, SV5 and NDV are closely related to each other but distinct from Sendai virus and parainfluenza virus type 3 HN proteins.

Amino Acid Sequence↗
PubMed2003

GeneScan reverse transcription-PCR assay for detection of six common respiratory viruses in young children hospitalized with acute respiratory illness.

A reverse transcription-PCR (RT-PCR) assay based on automated fluorescent capillary electrophoresis and GeneScan software analysis was developed to detect six common respiratory viruses in clinical specimens from young children. Assays for human respiratory syncytial virus (HRSV); human parainfluenza viruses 1, 2, and 3 (HPIV1, -2, and -3, respectively); and influenza A and B viruses were incorporated into a single standard assay format. The optimized assay panel was used to test 470 respiratory specimens obtained from 462 children hospitalized with acute respiratory illness that had been previously tested by viral culture (405 specimens) or direct immunofluorescence staining (DIF) (65 specimens). Of 93 specimens positive for respiratory viruses by culture or DIF, 86 (92%) were positive by RT-PCR, including 66 HRSV, 2 HPIV2, 5 HPIV3, 3 influenza A virus, and 10 influenza B virus specimens. An additional 119 respiratory viruses were identified by RT-PCR in 116 patients for whom results were negative by viral isolation or DIF. We conclude that the GeneScan RT-PCR panel can markedly improve detection of acute respiratory virus infections in young children.

Acute Disease↗
PubMed1978

The use of polyethylene glycol in concentration and purification of several bovine viruses.

The polyethylene glycol (PEG) precipitation method was used for the concentration and purification of eight bovine viruses. Good results were obtained from four viruses, parainfluenza--3 virus, bovine enterovirus, bovine adenovirus, and bovine parvovirus. No satisfactory results of concentration were obtained from bovine reovirus, bovine viral diarrhea virus, and infectious bovine rhinotracheitis virus. The failure of concentration of the four viruses seems to be ascribed rather to the resuspending of virus from the virus--PEG precipitate than to the precipitation of virus from infective culture fluid. This method can be applied as the initial step to the concentration of parainfluenza--3 virus, bovine enterovirus, bovine adenovirus, and bovine parvovirus from a large volume of material, since it is simple, rapid, and inexpensive.

Adenoviridae↗
PubMed1979

Interaction between 6/94 virus, a parainfluenza type 1 strain, and human leukocytes.

6/94 virus, a parainfluenza type 1 virus recovered by lysolecithin fusion of multiple sclerosis brain cell cultures with CV-1 cells, replicated in monocyte macrophages and lymphocytes from normal human donors and from a patient with multiple sclerosis. In macrophage cultures, hemadsorption-positive cells and high levels of infectious virus became apparent within 24 to 48 h after infection, persisted for 6 days, and then began to decrease. Phytohemagglutinin-stimulated macrophages yielded similar titers of virus, but the levels were maintained for a longer period of time. Macrophage-produced virus appeared to be infectious for other macrophages in the same culture. Both unstimulated and phytohemagglutinin-stimulated lymphocytes also supported virus replication. Significantly higher titers were produced in the stimulated cultures, T cell-enriched populations producing more virus than unseparated populations whether stimulated or unstimulated. The presence or absence of antibodies to the virus in the donors did not appear to influence the levels of virus obtained in any of the leukocyte cultures. However, an increase in blastic forms after 6/94 virus infection was noted in lymphocytes from donors with antibodies as revealed morphologically and by increased incorporations of tritiated thymidine. Furthermore, 6/94 virus-infected lymphocytes, unlike Sendai virus-infected lymphocytes, were able to respond well to mitogenic stimulation by phytohemagglutinin.

Antibodies, Viral↗
PubMed1985

In vitro protective effect of bacteria-derived bovine alpha interferon I1 against selected bovine viruses.

We used bacteria-derived bovine alpha-interferon I1 (Bo IFN-alpha I1) to study its antiviral effect in a bovine turbinate cell line on bovine diarrhea virus, infectious bovine rhinotracheitis virus, parainfluenza 3 virus, and pseudorabies virus. We based our study upon replicate tests for each strain by using a block titration system with various concentrations of Bo IFN-alpha I1 against various concentrations of virus. The data were compiled in two-axis tables (replicate X concentration) and were statistically analyzed by the Spearman-Kärber method. An increase in the concentration of Bo IFN-alpha I1 enhanced its protective effect against every test virus strain. Bo IFN-alpha I1 had a marked in vitro effect on the bovine diarrhea viral strains. It demonstrated less protection against the pseudorabies and parainfluenza 3 viruses. Its effectiveness against the two infectious bovine rhinotracheitis viral strains was lesser and of a low order.

Animals↗
PubMed1975

Hydrocephalus produced by the 6/94 virus; A parainfluenza type 1 isolate from multiple sclerosis brain tissue.

The 6/94 virus, parainfluenza type 1 isolate from multiple sclerosis brain tissue, produced hydrocephalus in newborn Syrian hamsters. All animals developed clinical disease and died within a week. Ependymal cells lining the aqueduct of Sylvius became necrotic and fused, resulting in obstructive hydrocephalus. The 6/94 virus antigen was seen in ependyma and meninges. Paramyxovirus nucleocapsids were seen within cytoplasm of ependymal cells. Virus was recovered from hamster brains for only two days. Infectious virus could be recovered from brains grown in vitro as explants for 21 days. No evidence of rising hemagglutination-inhibiting antibody was noted for up to one month after infection. Intraperitoneal or subcutaneous injection of 6/94 virus did not produce hydrocephalus. HA2 virus and the temperature sensitive mutant of HA2 virus failed to produce hydrocephalus, while Sendai virus caused lesions similar to those of 6/94 virus.

Animals↗
PubMed1975

Immunoglobulins, antibodies and inhibitors of parainfluenza 3 virus in respiratory secretions of sheep.

Virus neutralising and haemagglutination inhibiting (HI) activities were monitored in the serum, nasal secretions and tracheo-bronchial secretions of lambs infected with Parainfluenza virus type 3 (PI3). HI activity was found in the secretions of both control and infected lambs, whereas neutralising antibody was found only in nasal secretions from infected lambs. Fractionation revealed that most of the HI activity in the secretions was due to a large molecular weight protein which also inhibited the haemagglutination (HA) of Parainfluenza virus type 1 (PI1) and type 2 (PI2) and Newcastle disease virus (NDV). This inhibitory activity was partially sensitive to receptor destroying enzyme (RDE). IgA antibodies specific for PI3 were also found in the respiratory secretions. However no increase in IgA levels was detected in the nasal secretions of the infected lambs. It is suggested that in certain reports non-specific inhibitors present in the nasal secretions of calves, may have been confused with PI3 specific IgA antibody.

Animals↗
PubMed2004

Conserved glycine residues in the fusion peptide of the paramyxovirus fusion protein regulate activation of the native state.

Hydrophobic fusion peptides (FPs) are the most highly conserved regions of class I viral fusion-mediating glycoproteins (vFGPs). FPs often contain conserved glycine residues thought to be critical for forming structures that destabilize target membranes. Unexpectedly, a mutation of glycine residues in the FP of the fusion (F) protein from the paramyxovirus simian parainfluenza virus 5 (SV5) resulted in mutant F proteins with hyperactive fusion phenotypes (C. M. Horvath and R. A. Lamb, J. Virol. 66:2443-2455, 1992). Here, we constructed G3A and G7A mutations into the F proteins of SV5 (W3A and WR isolates), Newcastle disease virus (NDV), and human parainfluenza virus type 3 (HPIV3). All of the mutant F proteins, except NDV G7A, caused increased cell-cell fusion despite having slight to moderate reductions in cell surface expression compared to those of wild-type F proteins. The G3A and G7A mutations cause SV5 WR F, but not NDV F or HPIV3 F, to be triggered to cause fusion in the absence of coexpression of its homotypic receptor-binding protein hemagglutinin-neuraminidase (HN), suggesting that NDV and HPIV3 F have stricter requirements for homotypic HN for fusion activation. Dye transfer assays show that the G3A and G7A mutations decrease the energy required to activate F at a step in the fusion cascade preceding prehairpin intermediate formation and hemifusion. Conserved glycine residues in the FP of paramyxovirus F appear to have a primary role in regulating the activation of the metastable native form of F. Glycine residues in the FPs of other class I vFGPs may also regulate fusion activation.

Amino Acid Sequence↗
PubMed2003

Evaluation of three Copan viral transport systems for the recovery of cultivatable, clinical virus isolates.

Three Copan viral transport media (VTM) were compared to the BBL Viral Culturette and Micro Test M4 systems for their ability to maintain the viability of clinical strains of enterovirus, herpes simplex virus, adenovirus and parainfluenza virus. VTM were inoculated, incubated up to 72 h at both 4 degrees C and 22 degrees C, and processed for cell culture. The concentration of infectious virus in VTM was determined by standard endpoint dilution assay. All VTM maintained viable enterovirus and adenovirus for 72 h but adenovirus from M4 and Copan CVM media were detected in cell culture at least one day sooner than from other media. Only M4 and Copan CVM media supported the recognition of low-titer herpes simplex virus in cell culture within 24 h. Parainfluenza virus was consistently detected in culture within six days when maintained in M4 and two of the Copan VTM media. Overall, the Copan CVM compared favorably to the M4 in maintaining concentrations of viable virus that could be recovered and identified rapidly in routine cell culture.

Adenoviridae↗
PubMed1986

Molecular cloning and sequence analysis of the human parainfluenza 3 virus mRNA encoding the P and C proteins.

The sequence of the mRNA encoding the phosphoprotein (P protein) of the human parainfluenza virus 3 (PF3) was determined by molecular cloning. In other Parmyxoviridae the P protein mRNA is functionally bicistronic and encodes an additional smaller nonstructural protein termed C. In this report three open reading frames (ORF) are described. These consist of a single long ORF encoding the P protein, and two shorter ORFs encoding the structural Vp18 protein (analogous to the Sendai C protein) and a putative polypeptide termed D protein. The encoded phosphoprotein consists of 603 amino acids and has a predicted molecular weight of 67,683. The C protein consists of 199 amino acids and has a predicted molecular weight of 23,288. The D protein consists of 140 amino acids and has a predicted molecular weight of 16,270. Although the D protein has not yet been demonstrated in vivo its synthesis could be demonstrated in vitro using a rabbit reticulocyte lysate system. Thus it appears that unlike the other paramyxoviruses, the PF3 P protein mRNA may be functionally tricistronic.

Amino Acid Sequence↗
PubMed1987

Structural proteins of bovine parainfluenza-3 virus.

Six structural proteins of bovine parainfluenza-3 virus (PI-3V) labeled with [35S]-methionine could be resolved by polyacrylamide gel electrophoresis (PAGE). Five structural proteins of this virus had been previously reported. The 6 proteins found in this study were: L, a 180,000 (180 kD) molecular weight (MW) large protein; P, 83 kD phosphoprotein; HN, 69 kD hemagglutinin-neuraminidase glycoprotein; NP, 66 kD nucleocapsid protein; F, 55 kD fusion glycoprotein; and M, 38 kD matrix protein. Selective labeling with [2-3H]-mannose revealed only HN and F glycoprotein bands. A cellular actin protein (43 kD), associated with many enveloped viruses, was also found as a seventh protein in bovine PI-3V.

Animals↗
PubMed1975

Vaccination against parainfluenza 1 virus (typus muris) infection in order to eradicate this virus in colonies of laboratory animals.

Parainfluenza 1 virus (typus muris), commonly known as Sendai virus, is still contaminating mouse colonies in Japan. This presents a serious problem to keep mouse colonies pathogen-free. It seems that the infection is spread by dust and the virus remains active a considerable time in the mouse breeding rooms after all mice are removed. In order to eradicate the infection of this virus, vaccination was attempted to inoculate newly born young mice successively for about six months and during and after these periods the movements of the infection were under observation. The vaccine used for this experiment was inactivated virus vaccine with mineral oil-Arlacel adjuvant. The induction of antibody level by the inoculation of this vaccine was evidenced to be very efficient and the duration of the immunity was also very satisfactory. However, after the cessation of the vaccination, the infection of the virus reappeared gradually after some period of temporary ease.

Animals↗
PubMed1985

Production of monoclonal antibodies against parainfluenza 3 virus and their use in diagnosis by immunofluorescence.

Monoclonal antibodies were produced against parainfluenza virus type 3 (PI-3) and used to identify PI-3 clinical isolates in cell culture and PI-3 antigen in cells obtained from nasopharyngeal (NP) washes of patients. Two (2E9 and 4G5) of the three monoclonal antibodies characterized reacted by immunoblotting with a 67,000-dalton PI-3 protein, and one antibody (4E5) reacted with two viral proteins in the range of 29,000 to 31,000 daltons. The three monoclonal antibodies did not cross-react by indirect immunofluorescence (IFA) with PI-1 or PI-2 and identified by IFA 18 isolates of PI-3 in cell culture. The 2E9 antibody reacted with PI-3 antigen in cells of 8 NP wash specimens that also yielded PI-3 in cell culture. Cells from 12 specimens reactive by IFA for respiratory syncytial virus, 1 specimen yielding adenovirus in cell culture, and 5 specimens yielding influenza virus were not reactive.

Animals↗