RESUMEN
Previously we reported on the HPIV2 genotype distribution in Croatia 2011-2014. Here we expand this period up to 2017 and confirm that G1a genotype has replaced G3 genotype from the period 2011-2014. Our hypothesis was that the G1a-to-G3 genotype replacement is an antibody-driven event. A cross-neutralisation with anti-HPIV2 sera specific for either G1a or G3 genotype revealed the presence of genotype-specific antigenic determinants. By the profound, in silico analyses three potential B cell epitopic regions were identified in the hemagglutinin neuraminidase (regions 314-361 and 474-490) and fusion protein (region 440-484). The region identified in the fusion protein does not show any unique site between the G1a and G3 isolates, five differentially glycosylated sites in the G1a and G3 genotype isolates were identified in epitopic regions of hemagglutinin neuraminidase. All positively selected codons were found to be located either in the region 314-316 or in the region 474-490 what indicates a strong positive selection in this region and reveals that these regions are susceptible to evolutionary pressure possibly caused by antibodies what gives a strong verification to our hypothesis that neutralising antibodies are a key determinant in the inherently complex adaptive evolution of HPIV2 in the region.
Asunto(s)
Anticuerpos Neutralizantes/fisiología , Virus de la Parainfluenza 2 Humana/genética , Infecciones por Rubulavirus/virología , Adolescente , Distribución por Edad , Animales , Anticuerpos Antivirales/fisiología , Niño , Preescolar , Chlorocebus aethiops , Croacia/epidemiología , Epítopos de Linfocito B/química , Epítopos de Linfocito B/inmunología , Femenino , Genotipo , Cobayas , Proteína HN/inmunología , Humanos , Lactante , Funciones de Verosimilitud , Persona de Mediana Edad , Virus de la Parainfluenza 2 Humana/clasificación , Virus de la Parainfluenza 2 Humana/inmunología , Filogenia , ARN Viral/química , ARN Viral/genética , Recurrencia , Infecciones por Rubulavirus/epidemiología , Infecciones por Rubulavirus/inmunología , Estaciones del Año , Alineación de Secuencia , Células VeroRESUMEN
The dynamics and evolution of the human parainfluenza virus type 2 (HPIV2) in Croatia, and also globally, are largely unknown. Most HPIV2 infections are treated symptomatically outside the hospital setting. Thus, the diagnosis is missing making it difficult to follow the genetic variation and evolution of the HPIV2. This study explores hospitalized HPIV2 cases in Croatia during 4-year period (2011-2014). Most cases in this period were reported in October or November (68.75%) and most of patients were under 2 years of age (81.25%). For molecular analyses, we used the F and HN gene sequences and showed that although both regions are equally suitable for phylogenetic analyses it would be advantageous to use regions longer than 2 kb for HPIV2 analyses of isolates which are spatially and temporally closely related. We show here that the dominant cluster in this area was cluster G3 while only one strain isolated in this period was positioned in the distant cluster G1a. Further monitoring of the HPIV2 will determine whether cluster G3 will remain dominant or it will be overruled by cluster G1a. This will be important for the surveillance of virus circulation in population and significance of the viral infection. J. Med. Virol. 88:1733-1741, 2016. © 2016 Wiley Periodicals, Inc.
Asunto(s)
Variación Genética , Virus de la Parainfluenza 2 Humana/genética , Infecciones por Rubulavirus/virología , Animales , Niño , Preescolar , Chlorocebus aethiops , Croacia/epidemiología , Femenino , Proteína HN/genética , Hospitalización , Humanos , Lactante , Masculino , Virus de la Parainfluenza 2 Humana/clasificación , Virus de la Parainfluenza 2 Humana/aislamiento & purificación , Filogenia , Infecciones por Rubulavirus/epidemiología , Células Vero , Proteínas Virales de Fusión/genéticaAsunto(s)
Herpes Zóster/epidemiología , Gripe Humana/epidemiología , Visita a Consultorio Médico/estadística & datos numéricos , Infecciones por Picornaviridae/epidemiología , Infecciones por Respirovirus/epidemiología , Australia/epidemiología , Herpes Zóster/diagnóstico , Herpes Zóster/virología , Herpesvirus Humano 3/clasificación , Herpesvirus Humano 3/genética , Herpesvirus Humano 3/aislamiento & purificación , Humanos , Subtipo H1N1 del Virus de la Influenza A/clasificación , Subtipo H1N1 del Virus de la Influenza A/genética , Subtipo H1N1 del Virus de la Influenza A/aislamiento & purificación , Virus de la Influenza B/clasificación , Virus de la Influenza B/genética , Virus de la Influenza B/aislamiento & purificación , Gripe Humana/diagnóstico , Gripe Humana/virología , Mucosa Nasal/virología , Virus de la Parainfluenza 1 Humana/clasificación , Virus de la Parainfluenza 1 Humana/genética , Virus de la Parainfluenza 1 Humana/aislamiento & purificación , Virus de la Parainfluenza 2 Humana/clasificación , Virus de la Parainfluenza 2 Humana/genética , Virus de la Parainfluenza 2 Humana/aislamiento & purificación , Virus de la Parainfluenza 3 Humana/clasificación , Virus de la Parainfluenza 3 Humana/genética , Virus de la Parainfluenza 3 Humana/aislamiento & purificación , Infecciones por Picornaviridae/diagnóstico , Infecciones por Picornaviridae/virología , Infecciones por Respirovirus/diagnóstico , Infecciones por Respirovirus/virología , Rhinovirus/clasificación , Rhinovirus/genética , Rhinovirus/aislamiento & purificación , Vigilancia de GuardiaRESUMEN
BACKGROUND: Although human parainfluenza type 2 (HPIV-2) virus is an important respiratory pathogen, a little is known about strains circulating in Saudi Arabia. FINDINGS: Among 180 nasopharyngeal aspirates collected from suspected cases in Riyadh, only one sample (0.56%) was confirmed HPIV-2 positive by nested RT-PCR. The sample that was designated Riyadh 105/2009 was used for sequencing and phylogenetic analysis of the most variable virus gene; the haemagglutinin-neuramindase (HN). Comparison of HN gene of Riyadh 105/2009 strain and the relevant sequences available in GenBank revealed a strong relationship with Oklahoma-94-2009 strain. Phylogenetic analysis indicated four different clusters of HPIV-2 strains (G1-4). Twenty-three amino acid substitutions were recorded for Riyadh 105/2009, from which four are unique. The majority of substitutions (n=18) had changed their amino acids characteristics. By analyzing the effect of the recorded substitutions on the protein function using SIFT program, only two located at positions 360 and 571 were predicted to be deleterious. CONCLUSIONS: The presented changes of Riyadh 105/2009 strain may possess potential effect on the protein structure and/or function level. This is the first report that describes partial characterization of Saudi HPIV-2 strain.
Asunto(s)
Genes Virales , Proteína HN/genética , Virus de la Parainfluenza 2 Humana/genética , Filogenia , Secuencia de Aminoácidos , Sustitución de Aminoácidos , Preescolar , Bases de Datos de Ácidos Nucleicos , Femenino , Humanos , Virus de la Parainfluenza 2 Humana/clasificación , Virus de la Parainfluenza 2 Humana/enzimología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Arabia Saudita , Alineación de Secuencia , Homología de Secuencia de Aminoácido , Programas Informáticos , Especificidad de la EspecieRESUMEN
Human parainfluenza viruses (PIVs) cause acute respiratory illness in children, the elderly, and immunocompromised patients. PIV3 is a common cause of bronchiolitis and pneumonia, whereas PIV1 and 2 are frequent causes of upper respiratory tract illness and croup. To assess how PIV1, 2, and 3 differ with regard to replication and induction of type I interferons, interleukin-6, and relevant chemokines, we infected primary human airway epithelium (HAE) cultures from the same tissue donors and examined replication kinetics and cytokine secretion. PIV1 replicated to high titer yet did not induce cytokine secretion until late in infection, while PIV2 replicated less efficiently but induced an early cytokine peak. PIV3 replicated to high titer but induced a slower rise in cytokine secretion. The T cell chemoattractants CXCL10 and CXCL11 were the most abundant chemokines induced. Differences in replication and cytokine secretion might explain some of the differences in PIV serotype-specific pathogenesis and epidemiology.
Asunto(s)
Bronquios/inmunología , Bronquios/virología , Virus de la Parainfluenza 1 Humana/fisiología , Virus de la Parainfluenza 2 Humana/fisiología , Virus de la Parainfluenza 3 Humana/fisiología , Tráquea/inmunología , Tráquea/virología , Células Cultivadas , Quimiocinas/biosíntesis , Citocinas/biosíntesis , Citocinas/genética , Interacciones Huésped-Patógeno/genética , Interacciones Huésped-Patógeno/inmunología , Humanos , Interleucina-6/biosíntesis , Interleucina-6/genética , Cinética , Virus de la Parainfluenza 1 Humana/clasificación , Virus de la Parainfluenza 1 Humana/inmunología , Virus de la Parainfluenza 1 Humana/patogenicidad , Virus de la Parainfluenza 2 Humana/clasificación , Virus de la Parainfluenza 2 Humana/inmunología , Virus de la Parainfluenza 2 Humana/patogenicidad , Virus de la Parainfluenza 3 Humana/clasificación , Virus de la Parainfluenza 3 Humana/inmunología , Virus de la Parainfluenza 3 Humana/patogenicidad , Polimorfismo de Nucleótido Simple , Mucosa Respiratoria/inmunología , Mucosa Respiratoria/virología , Serotipificación , Especificidad de la Especie , Replicación ViralRESUMEN
BACKGROUND: Human parainfluenza viruses (hPIV) are respiratory pathogens responsible for upper and lower respiratory tract infections. In most labs, the clinical diagnosis of hPIV is routinely done using techniques based on the detection of viral antigens such as immunofluorescence assay or/and viral isolation. STUDY DESIGN: Five hPIV-2 isolated from respiratory samples exhibited unusual phenotypic and antigenic characteristics. These isolates showed important syncytial cytopathic effect and failed to react with one specific monoclonal antibody. These variant strains were subsequently compared with hPIV-2 prototype strain by cellular and molecular techniques. RESULTS: Both variant and prototype strains showed similar growth kinetics. Observation of plaque formation and syncytia assay indicated a more important fusogenic activity for the variant strains. Sequencing of fusion (F) and hemagglutinin-neuraminidase (HN) genes showed differences between the "atypical" hPIV-2 isolates and the Greer hPIV-2 prototype strain. These differences were analyzed with molecular modelling and structure prediction soft wares. A potential new glycosylation site in HN, in addition to minor changes observed in the predicted structure for the variant strains could explain their antigenic variation. Genetic changes in the fusion peptide and the cleavage site of F could also explain the difference observed in the fusion activity. CONCLUSIONS: Continuous global viral surveillance is essential to monitor antigenic changes that may occur in nature particularly with regards to the implementation of diagnostic assays. The differences observed in F and HN between the prototype strain and clinical hPIV-2 variants could also provide new data for the analysis of Paramyxovirus fusion mechanisms and their pathogenesis.
Asunto(s)
Proteína HN/genética , Virus de la Parainfluenza 2 Humana/fisiología , ARN Viral , Infecciones por Rubulavirus/virología , Proteínas Virales de Fusión/genética , Adulto , Secuencia de Aminoácidos , Animales , Variación Antigénica , Línea Celular , Niño , Glicosilación , Proteína HN/química , Proteína HN/inmunología , Haplorrinos , Humanos , Modelos Moleculares , Datos de Secuencia Molecular , Virus de la Parainfluenza 2 Humana/clasificación , Virus de la Parainfluenza 2 Humana/genética , Virus de la Parainfluenza 2 Humana/aislamiento & purificación , Filogenia , Conformación Proteica , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Proteínas Virales de Fusión/química , Proteínas Virales de Fusión/inmunología , Ensayo de Placa ViralRESUMEN
There is a need for rapid, sensitive, and accurate diagnosis of lower respiratory tract infections in children, elderly, and immunocompromised patients, who are susceptible to serious complications. The multiplex RT-nested PCR assay has been used widely for simultaneous detection of non-related viruses involved in infectious diseases because of its high specificity and sensitivity. A new multiplex RT-PCR assay is described in this report. This approach includes nested primer sets targeted to conserve regions of human parainfluenza virus haemagglutinin, human coronavirus spike protein, and human enterovirus and rhinovirus polyprotein genes. It permits rapid, sensitive, and simultaneous detection and typing of the four types of parainfluenza viruses (1, 2, 3, 4AB), human coronavirus 229E and OC43, and the generic detection of enteroviruses and rhinoviruses. The testing of 201 clinical specimens with this multiplex assay along with other one formerly described by our group to simultaneously detect and type the influenza viruses, respiratory syncytial viruses, and a generic detection of all serotypes of adenovirus, covers the detection of most viruses causing respiratory infectious disease in humans. The results obtained were compared with conventional viral culture, immunofluorescence assay, and a third multiplex RT-PCR assay for all human parainfluenza viruses types described previously. In conclusion, both multiplex RT-PCR assays provide a system capable of detecting and identifying simultaneously 14 different respiratory viruses in clinical specimens with high sensitivity and specificity, being useful for routine diagnosis and survey of these viruses within the population.
Asunto(s)
Coronavirus/aislamiento & purificación , Paramyxovirinae/aislamiento & purificación , Picornaviridae/aislamiento & purificación , Sistema Respiratorio/virología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Coronavirus/clasificación , Coronavirus/genética , Coronavirus Humano 229E/clasificación , Coronavirus Humano 229E/genética , Coronavirus Humano 229E/aislamiento & purificación , Coronavirus Humano OC43/clasificación , Coronavirus Humano OC43/genética , Coronavirus Humano OC43/aislamiento & purificación , Enterovirus/clasificación , Enterovirus/genética , Enterovirus/aislamiento & purificación , Genes Virales , Hemaglutininas Virales/genética , Humanos , Glicoproteínas de Membrana/genética , Virus de la Parainfluenza 1 Humana/clasificación , Virus de la Parainfluenza 1 Humana/genética , Virus de la Parainfluenza 1 Humana/aislamiento & purificación , Virus de la Parainfluenza 2 Humana/clasificación , Virus de la Parainfluenza 2 Humana/genética , Virus de la Parainfluenza 2 Humana/aislamiento & purificación , Virus de la Parainfluenza 3 Humana/clasificación , Virus de la Parainfluenza 3 Humana/genética , Virus de la Parainfluenza 3 Humana/aislamiento & purificación , Virus de la Parainfluenza 4 Humana/clasificación , Virus de la Parainfluenza 4 Humana/genética , Virus de la Parainfluenza 4 Humana/aislamiento & purificación , Paramyxovirinae/clasificación , Paramyxovirinae/genética , Picornaviridae/clasificación , Picornaviridae/genética , Poliproteínas/genética , Infecciones del Sistema Respiratorio/diagnóstico , Infecciones del Sistema Respiratorio/virología , Rhinovirus/clasificación , Rhinovirus/genética , Rhinovirus/aislamiento & purificación , Sensibilidad y Especificidad , Glicoproteína de la Espiga del Coronavirus , Proteínas del Envoltorio Viral/genética , Proteínas Virales/genéticaRESUMEN
The aim of this study was to compare the clinical course of severe lower respiratory tract infections associated with human parainfluenza virus types 1-3 (HPIV 1-3) in hospitalised children infected with the human immunodeficiency virus type 1 (HIV-1) versus that in hospitalised children not infected with HIV-1. Children were enrolled prospectively as part of a broader study that evaluated the aetiology of lower respiratory tract infections in HIV-1-infected and -noninfected children from March 1997 through March 1999. HPIV types 1-3 were isolated from nasopharyngeal aspirate samples that were analysed using immunofluorescein monoclonal antibody assays. Thirty percent (24 of 80) of the children from whom HPIV was isolated were infected with HIV-1. Sixty-six percent (47 of 62) and 22% (14 of 62) of the HPIV isolates that were typed were subtypes 3 and 1, respectively. The clinical presentation of severe lower respiratory tract infection was similar in both HIV-1-infected and -noninfected children, except that the former were less likely to have wheezing (4.2% vs. 28.6%, P=0.01). Furthermore, the duration of hospitalisation was longer in HIV-1-infected children than in HIV-1-noninfected children (median 11.5 days [range 1-15 days] vs. median 7.5 days [range 1-22 days]; P=0.02), and mortality was higher (5 of 24 [20.8%] infected children vs. 0 of 56 noninfected children; P=0.001). Importantly, four of five (80%) of the HIV-1-infected children who died had other concurrent illnesses or predisposing factors for severe HPIV-associated disease. HPIV-associated lower respiratory tract infection causes greater morbidity and mortality in HIV-1-infected children than in HIV-1-noninfected children; however, this may be due to other concurrent illnesses in HIV-1-infected children.
Asunto(s)
Infecciones por VIH/complicaciones , Virus de la Parainfluenza 1 Humana/fisiología , Virus de la Parainfluenza 2 Humana/fisiología , Virus de la Parainfluenza 3 Humana/fisiología , Infecciones por Paramyxoviridae/complicaciones , Infecciones por Paramyxoviridae/fisiopatología , Infecciones del Sistema Respiratorio/complicaciones , Infecciones del Sistema Respiratorio/fisiopatología , Preescolar , Femenino , Infecciones por VIH/mortalidad , Infecciones por VIH/virología , VIH-1/fisiología , Humanos , Lactante , Masculino , Virus de la Parainfluenza 1 Humana/clasificación , Virus de la Parainfluenza 2 Humana/clasificación , Virus de la Parainfluenza 3 Humana/clasificación , Infecciones por Paramyxoviridae/mortalidad , Infecciones por Paramyxoviridae/virología , Neumonía Viral/complicaciones , Neumonía Viral/mortalidad , Neumonía Viral/fisiopatología , Neumonía Viral/virología , Estudios Prospectivos , Infecciones del Sistema Respiratorio/mortalidad , Infecciones del Sistema Respiratorio/virología , Estaciones del Año , Sudáfrica/epidemiologíaRESUMEN
La laringotraqueobronquitis o crup viral es una de las entidades clínicas más frecuentes y clásicas de la patología respiratoria pediátrica. La presente revisión evalúa, a la luz de las más recientes publicaciones sobre el tema, el papel terapéutico de medidas como la aerosolterapia, las nebulizaciones de epinefrina racémica, adrenalina y fenilefrina, la administración parenteral de glucocorticoides y el empleo de una escala de puntaje para evaluar la severidad del compromiso del paciente y la respuesta al tratamiento.
Asunto(s)
Humanos , Preescolar , Niño , Crup , Virus de la Parainfluenza 2 Humana/clasificación , Virus de la Parainfluenza 2 Humana/patogenicidad , Virus de la Parainfluenza 2 Humana/fisiología , Virus de la Parainfluenza 2 Humana/clasificación , Virus de la Parainfluenza 2 Humana/patogenicidadRESUMEN
We describe the case of an 11-year-old Bolivian boy with parotitis and aseptic meningitis to demonstrate that parainfluenza virus type 2 can cause disseminated infection in a normal child. Parainfluenza virus type 2 was isolated from nasopharyngeal and CSF specimens from the patient and was confirmed to be parainfluenza virus type 2 by hemadsorption inhibition and by complement fixation. Parainfluenza virus type 2 may cause aseptic meningitis and parotitis.
Asunto(s)
Meningitis Aséptica/complicaciones , Meningitis Aséptica/virología , Meningitis Viral/complicaciones , Virus de la Parainfluenza 2 Humana , Infecciones por Paramyxoviridae/complicaciones , Parotiditis/complicaciones , Parotiditis/virología , Niño , Humanos , Masculino , Meningitis Aséptica/líquido cefalorraquídeo , Meningitis Aséptica/diagnóstico , Meningitis Viral/líquido cefalorraquídeo , Meningitis Viral/diagnóstico , Nasofaringe/virología , Virus de la Parainfluenza 2 Humana/clasificación , Virus de la Parainfluenza 2 Humana/aislamiento & purificación , Infecciones por Paramyxoviridae/líquido cefalorraquídeo , Infecciones por Paramyxoviridae/diagnóstico , Parotiditis/líquido cefalorraquídeo , Parotiditis/diagnósticoRESUMEN
The nucleotide sequences of cDNAs of the simian virus 5 (SV5) nucleoprotein (NP) gene, and the 3' end of the genome and NP gene of SV41 were determined. The open reading frames of the SV5 and SV41 NP genes encode polypeptides with Mrs of 56,582 and 60,575, respectively, values which are consistent with those estimated by SDS-PAGE. The NP of human parainfluenza virus type 2 (hPIV-2) was more closely related to that of SV41 (amino acid sequence identity 70.5%) than that of SV5 (57.0%); the amino acid sequence identity between the NPs of SV41 and SV5 was 63.3%. The sequence of the 3' end of the genome of SV41 showed a high level of similarity to that of hPIV-2, the terminal 18 nucleotides being identical. It is concluded from these findings that SV41 is related most closely to hPIV-2, even though SV5 had been thought to be an animal type of hPIV-2.
Asunto(s)
Cápside/genética , ADN Viral/química , Virus de la Parainfluenza 2 Humana/genética , Respirovirus/genética , Proteínas del Núcleo Viral/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Cápside/química , Humanos , Macaca fascicularis , Datos de Secuencia Molecular , Sistemas de Lectura Abierta , Virus de la Parainfluenza 2 Humana/clasificación , Respirovirus/clasificación , Programas Informáticos , Células Vero , Proteínas del Núcleo Viral/químicaRESUMEN
Marked differences in the apparent Mrs of the HN, NP and F proteins of simian virus 5 (SV5) and parainfluenza virus type 2 (PF-2) were revealed by SDS-PAGE. To examine the antigenic relationships between SV5, PF-2 and other paramyxoviruses, monoclonal antibodies (MAbs) specific to PF-2 were isolated. These antibodies had specificities for the HN, NP and P proteins and together with 54 MAbs to SV5 were tested for their ability to react with SV5, PF-2, PF-3, mumps and measles virus proteins. Most of these MAbs (55 out of 60) reacted with homologous virus only. However, five reacted with both SV5 and PF-2. These antibodies had reactivities to the NP, M and P proteins. Furthermore, one of the antibodies with reactivity to the P protein also reacted with mumps virus. Although none of the 21 MAbs with specificities for the HN protein of either SV5 or PF-2 cross-reacted with heterologous virus, some antigenic similarities between the HN protein of SV5 and PF-2 could be detected. This was demonstrated by raising a series of polyclonal antisera to purified preparations of SV5 or PF-2 HN proteins in BALB/c mice, and testing for their ability to neutralize both SV5 and PF-2 and also to immunoprecipitate the HN proteins of these viruses. Surprisingly, while low levels of cross-neutralizing antibody could be detected in some sera (e.g. neutralization of SV5 1:1600 and of PF-2 1:80), other sera with similar neutralization titres against homologous virus failed to neutralize heterologous virus. Furthermore, only a minority of the anti-HN antisera showed any immune-precipitating activity against the heterologous HN protein.