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Human parainfluenza virus 3 vaccine candidates attenuated by codon-pair deoptimization are immunogenic and protective in hamsters.

Human parainfluenza virus type 3 (HPIV3) is a major pediatric respiratory pathogen lacking available vaccines or antiviral drugs. We generated live-attenuated HPIV3 vaccine candidates by codon-pair deoptimization (CPD). HPIV3 open reading frames (ORFs) encoding the nucleoprotein (N), phosphoprotein (P), matrix (M), fusion (F), hemagglutinin-neuraminidase (HN), and polymerase (L) were modified singly or in combination to generate 12 viruses designated Min-N, Min-P, Min-M, Min-FHN, Min-L, Min-NP, Min-NPM, Min-NPL, Min-PM, Min-PFHN, Min-MFHN, and Min-PMFHN. CPD of N or L severely reduced growth in vitro and was not further evaluated. CPD of P or M was associated with increased and decreased interferon (IFN) response in vitro, respectively, but had little effect on virus replication. In Vero cells, CPD of F and HN delayed virus replication, but final titers were comparable to wild-type (wt) HPIV3. In human lung epithelial A549 cells, CPD F and HN induced a stronger IFN response, viral titers were reduced 100-fold, and the expression of F and HN proteins was significantly reduced without affecting N or P or the relative packaging of proteins into virions. Following intranasal infection in hamsters, replication in the nasal turbinates and lungs tended to be the most reduced for viruses bearing CPD F and HN, with maximum reductions of approximately 10-fold. Despite decreased in vivo replication (and lower expression of CPD F and HN in vitro), all viruses induced titers of serum HPIV3-neutralizing antibodies similar to wt and provided complete protection against HPIV3 challenge. In summary, CPD of HPIV3 yielded promising vaccine candidates suitable for further development.

Animals

Clinical and Epidemiological Insights into a Parainfluenza Virus Type 3 Outbreak in Multiple Myeloma Patients.

Human parainfluenza virus type 3 (HPIV-3) can be responsible for mild to severe respiratory infections and hospital epidemics. We investigated an outbreak in a hematology unit. Respiratory viruses were screened using multiplex PCR. HPIV-3 quantification and whole-genome sequencing were performed on HPIV-3 positive respiratory samples. Clinical characteristics, infection progression, incidence rates of respiratory viruses within the hospital and detection of respiratory viruses were documented, along with the reinforcement of infection prevention and control (IPC) measures implemented. Between November 2022, and January 2023, HPIV-3 was identified in 20 of 113 hematology patients (17.7%), of whom 80% had multiple myeloma. A majority of HPIV-3-positive patients developed pneumonia (60%), and mortality was notably higher (35%) compared to patients who were negative (3%, p&#x2009;<&#x2009;0.0001). Respiratory HPIV-3 viral loads were similar between patients with and without pneumonia. In parallel, HPIV-3 incidence in the hospital overall was lower than in the hematology unit (p&#x2009;<&#x2009;0.0001). Air virus screening showed the detection of HPIV-3 in the air in different areas, and whole-genome sequencing confirmed the circulation of a single HPIV-3 strain. Strengthened IPC measures were associated with the containment of the outbreak. HPIV-3 has high epidemic potential in patients with multiple myeloma and causes severe infections. Our findings highlight the need for routine HPIV-3 testing in hematology units.

Humans

Whole-genome evolutionary dynamics of human parainfluenza virus type 3 in Shanghai, China, 2016-2024.

&#x2022; Fifty whole-genome sequencing revealed co-circulating HPIV-3 C3 sub-lineages C3f and C3a in Shanghai, China. &#x2022; Whole-genome phylogeny dated the HPIV-3 tMRCA to &#x223c;1925.6 and revealed two post-1990 demographic expansions. &#x2022; Recombination signals detected in the HN gene and other regions may lead to discordance in partial-gene phylogenies. &#x2022; The L gene showed the highest variability and harbored the largest number of putative positively selected sites.

Letter

Whole Genome Sequencing and Genetic Diversity of Respiratory Viruses Detected in Children With Acute Respiratory Infections: A One-Year Cross-Sectional Study in Senegal.

Acute respiratory infections (ARI) are a health priority, especially in countries with limited resources. They are a major cause of morbidity and mortality, especially among children and the elderly. In Senegal, the endemic circulation of respiratory viruses other than influenza has been demonstrated. However, there is a paucity of data exploring the genetic diversity of these viruses based on whole-genome sequencing. In this study, we present data on the genetic diversity of respiratory viruses in children under 15 years old in Senegal, including an overview of the different pathogens detected. Between November 2022 and November 2023, we collected nasopharyngeal swabs from children seen in curative consultations for symptoms of acute respiratory infections. Of the 156 children included, 73.7% tested positive for at least one pathogen. The most frequently detected virus was rhinovirus (50.0%), followed by influenza B (41.6%) and human parainfluenza virus type 3 (7.6%). Combinations of rhinovirus/influenza B, human parainfluenza virus type 2/human parainfluenza virus type 4, and rhinovirus/influenza B/adenovirus were the most frequently identified. A statistically significant association was detected between some of the viruses detected. A high genetic diversity of respiratory viruses circulating in children was revealed. The strains were phylogenetically close to various strains circulating worldwide, suggesting a global circulation of respiratory viruses. Our study provides the first complete genome sequences of human parainfluenza viruses type 2, 3, 4 and human bocavirus from Senegal and thus contributes to the enrichment of international databases on sequences from Senegal and underlines the importance of sequencing in the dynamics of pathogen circulation.

Humans

Lymphocyte response to virus antigens in systemic lupus erythematosus.

The cell-mediated immune response of lymphocytes to rubella, measles, parainfluenza types 1, 2, and 3, varicella-zoster and herpes virus type 1 virus antigens was evaluated in 15 SLE patients and 15 matched controls by incorporating 3H-thymidine in whole blood cultures as a measure of blastic transformation. SLE patients were less responsive than normal individuals to six of eight virus antigens tested. Culture of washed SLE cells in AB plasma did not reverse the hyporesponsiveness. The results indicated that a functional impairment of the circulating lymphocytes appeared to be responsible for the in vitro hyporesponsiveness of SLE patients to virus antigens.

ABO Blood-Group System

Maintenance of viability and comparison of identification methods for influenza and other respiratory viruses of humans.

A comparison of Hanks balanced salt solution, veal infusion broth (VIB), and charcoal viral transport medium for maintaining viability of type A influenza virus indicated approximately equal survival of virus on all three media at -70 and 4 degrees C, whereas at 25 degrees C virus survived best in VIB. VIB supplemented with bovine serum albumin was used as transport medium in a community-wide surveillance of febrile respiratory disease for influenza viruses. Unfrozen throat swab specimens were placed in VIB and stored at 4 degrees C for up to 5 days without effect on isolation frequencies of either type A or type B influenza virus or type 1 or type 3 parainfluenza virus. Comparison of indirect immunofluorescence with hemadsorption for detection of type A influenza virus in rhesus monkey kidney cultures revealed a requirement for at least five fluorescing cells to eliminate false positive indirect immunofluorescence tests and at least 3 days of incubation to eliminate false negative tests when compared with hemadsorption at later times. Detection frequencies for the two methods after 2 and 3 days of incubation were not significantly different.

Culture Media

[Interfering and interferonogenic activity of attenuated and original para-influenza viral strains].

A higher interfering activity of attenuated (vaccine) strains of parainfluenza virus types 1, 2 and 3 which had undergone a long-term adaptation to cell cultures as compared to the original viruses was established. The interferon-inducing activity of vaccine parainfluenza virus types 2 and 3 was also higher than that of the original viruses. The high interfering and interferon-inducing activity of attenuated parainfluenza virus strains of type 2 and 3 correlated with reduced virulence of these variants for man. These tests may be used for the assessment of virulence of parainfluenza virus strains type 2 and 3 in the laboratory.

Cells, Cultured

Inhibition of multiplication of parainfluenza 3 virus in prostaglandin-treated WISH cells.

Influence of prostaglandins E2 (PGE2) and F2alpha (PGF 2alpha) on multiplication of myxovirus parainfluenza 3 was investigated. At concentrations of 0.01--1 mug/ml prostaglandins had no direct cytotoxic effects. PGE2 and PGF2alpha inhibited multiplication of parainfluenza 3 virus at concentrations 0.1--10 mug/ml. The inhibitory effect was most pronounced when the prostaglandins were added to medium for the whole period of virus multiplication (48 hours) while little or no effect was found when prostaglandins were added before virus inoculation or for 2 hours after infection. It is suggested that prostaglandins exert an inhibitory effect on the replication phase of the virus by influencing the growth of the WISH cells.

Amnion

Elevated parainfluenza virus type 1 antibody in patients with subacute sclerosing panencephalitis.

Parainfluenza virus hemagglutination inhibition (HI) antibodies were determined 3 times in the sera of 9 patients with subacute sclerosiing panencephalitis (SSPE) and 20 healthy controls matched for age and place of residence. Serum antibody against parainfluenza virus type 1 was significantly elevated in SSPE patients as compared with controls, whereas antibodies against type 2 and 3 were found to be in normal ranges. Higher titres of parainfluenza virus type 1 antibody might depend on: (1) dual viral infection, (2) cross-reaction between antigens of SSPE virus and parainfluenza virus type 1, and (3) non-specific activation of latent virus type 1 genome. The latter explanation seems to be particularly interesting since the parainfluenza type 1 antibody titres remained constant despite the clinical progression. This finding is comparable to the elevated titres against Epstein-Barr virus of adenovirus which have been found occasionally in this disease.

Adolescent

[Respiratory tract infections of viral origin in infants. Examination of 103 serum pairs in autumn and following spring].

In autumn 1973 und about four months later in spring 1974 blood samples were taken from the same 103 children aged from a few months to five years and coming from different parts of Bavaria. We examined the samples for complement-fixing antibodies against the influenza virus types A, B and C, the parainfluenza virus types 1, 2 and 3, the adenoviruses and the respiratory-syncytial virus, and for neutralising antibodies against the ECHO virus type 25. At the commencement of the investigation the most frequently detected antibodies were those against adenoviruses (66.0%), ECHO virus type 25 (22.3%), influenza virus type A (15.5%) AND THE RESPIRATORY-SYNCYTIAL VIRUS (10.7%). Over autumn and winter in the four month interval between the drawing of blood samples the most frequently observed infections were those caused by the respiratory-syncytial virus (35.9%), the adenoviruses (13.6%) and the influenza viruses A and B (6.8%). By comparing the results of two blood tests from the same individual infections could be recognised more often than would have been the case if comparing the results of two different groups of children examined in autumn and spring.

Adenoviridae

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.

Adolescent

In vitro and in vivo effect of 1-beta-D-ribofuranosyl-1,2,4-triazole-3-carboxamide (ribavirin) on types 1 and 3 parainfulenza virus infections.

1-beta-D-ribofuranosyl-1,2,4-triazole-3-carboxamide (ribavirin) had significant in vitro activity against type 1 parainfluenza (Sendai) and type 3 parainogenic (HA-1) viruses. Activity was manifested as inhibition of both viral cytopathogenic effect and of recoverable virus or viral hemagglutinin titer. The minimum Sendai virus inhibitory concentration was determined to be approximately 3.2 mug/ml. Previous studies had determined the minimum concentration inhibiting HA-1 virus was approximately 1-10 mug/ml. The effect of time of addition of ribavirin to virus-infected cells was determined; maximal activity was seen when the drug added just prior to either virus or within 4-8 h after each virus, although anti-Senadi viral effects were still apparent when ribavirin was added as late as 24 h after the virus. Ribavirin had no effect on adsorption of HA-1 or Sendai virus to cells. Lethal Sendai virus infections of mice were significantly inhibited by multiple intraperitoneal ribavirin treatment, starting either 4 h before or up to 24 h after virus inoculation. Therapy starting 48, 72 or 96 h after virus exposure had a moderate degree of efficacy. Treatment using an aerosol chamber also was of moderate effectiveness, although the procedure was considered traumatic to the animals. A nonlethal, principally upper respiratory tract infection of hamsters induced by the HA-1 virus was inhibited by ribavirin therapy. Treatment administered intraperitoneally, per os or by aerosol chamber resulted in reduced 23-day antibody titers to the virus, presumably because of reduction of virus in the animal. In a separate experiment, intraperitoneal ribavirin therapy resulted in a 1 log10 or less reduction in virus titer in nasal washings from HA-1 virus-infected hamsters, whereas, when the drug was administered intranasally in a dry powder aerosol spray, nasal virus titers were reduced up to 2 log10 and a moderate virus-induced lung consolidation was completely inhibited.

Adsorption

Parainfluenzavirus upper respiratory tract illnesses in partially immune adult human subjects: a study at an Antarctic station.

Outbreaks of respiratory tract illnesses (RTI) in adult humans during October and November 1975 at McMurdo Station, Antarctica, were investigated by viral isolation and serologic procedures. The recovery of viral agents was enhanced by use of cell cultures in the field. Recoveries of parainfluenzaviruses types 1 and 3 and rhinoviruses were made from 10 of 39 nasal washings. Parainfluenzaviruses types 1 and 3 accounted for 50 and 30 per cent, respectively, of the total viruses recovered during October and November 1975. Acute and convalescent sera collected from 32 adult humans were examined for antiviral antibody by hemagglutination inhibition (HI) and radioimmunoassay (RIA) techniques. Serologic responses (HI and RIA) confirmed that parainfluenzaviruses were the predominent cause of RTI at McMurdo Station during that time. The temporal relationship between parainfluenzaviral diseases occurring in US communities and at McMurdo suggests that these viruses are transported to the Antarctic by personnel originating within the US. Standardization of the RIA allowed sequential assay of large numbers of sera using multiple preparations of radiolabeled indicator antibody, while ensuring the reproducibility of antiviral antibody titers to within one twofold dilution between subsequent labeled antibody preparations. The RIA detected both lower levels of virus specific antibody and more serologic responses than could be detected by HI.

Adult

Epidemiology of herpesvirus and respiratory virus infections. Part 1. Serologic findings.

The majority of the population group studied had complement-fixation antibodies toward the following viruses: influenza type A, respiratory syncytial, cytomegalovirus, and Epstein-Barr. The herpesvirus infections (cytomegalovirus and Epstein-Barr virus) seemed to be prevalent. Only low incidences of antibodies were found toward adenovirus, influenza type B, influenza type C, and parainfluenza type II. A total of 70 aucte virus infections (increases of antibody titer) were diagnosed in 49 patients. Besides cytomegalovirus, no particular virus infection occurred in a large number. Only 11 of the 70 acute virus infections diagnosed serologically were accompanied by clinical signs of disease

Adenoviridae