ABSTRACT
After the first phase of the COVID-19 pandemic in Europe, a new highly pathogenic variant of echovirus 11 (E11) was detected. The aim of this study was to analyze the genetic diversity of Polish E11 environmental and clinical strains circulating between 2017 and 2023 as well as compare them with E11 strains isolated from severe neonatal sepsis cases reported in Europe between 2022 and 2023. Additionally, the study explores the effectiveness of environmental monitoring in tracking the spread of new variants. For this purpose, the complete sequences of the VP1 capsid protein gene were determined for 266 E11 strains isolated in Poland from 2017 to 2023, and phylogenetic analysis was performed. In the years 2017-2023, a significant increase in the detection of E11 strains was observed in both environmental and clinical samples in Poland. The Polish E11 strains represented three different genotypes, C3, D5 and E, and were characterized by a high diversity. In Poland, the intensive circulation of the new variant E11, responsible for severe neonatal infections with a high mortality in Europe, was detected in the years 2022-2023. This investigation demonstrates the important role of environmental surveillance in the tracking of enteroviruses circulation, especially in settings with limited clinical surveillance.
Subject(s)
COVID-19 , Enterovirus B, Human , Phylogeny , SARS-CoV-2 , Poland/epidemiology , Humans , Enterovirus B, Human/genetics , Enterovirus B, Human/classification , Enterovirus B, Human/isolation & purification , COVID-19/epidemiology , COVID-19/virology , SARS-CoV-2/genetics , SARS-CoV-2/classification , SARS-CoV-2/isolation & purification , Genotype , Genetic Variation , Capsid Proteins/genetics , Infant, Newborn , Echovirus Infections/epidemiology , Echovirus Infections/virology , PandemicsABSTRACT
Echovirus 11 (E11) has gained attention owing to its association with severe neonatal infections. From 2018 to 2023, a surge in severe neonatal cases and fatalities linked to a novel variant of genotype D5 was documented in China, France, and Italy. However, the prevention and control of E11 variants have been hampered by limited background data on the virus circulation and genetic variance. Therefore, the present study investigated the circulating dynamics of E11 and the genetic variation and molecular evolution of genotype D5 through the collection of strains from the national acute flaccid paralysis (AFP) and hand, foot, and mouth disease (HFMD) surveillance system in China during 2000-2022 and genetic sequences published in the GenBank database. The results of this study revealed a prevalent dynamic of E11 circulation, with D5 being the predominant genotype worldwide. Further phylogenetic analysis of genotype D5 indicated that it could be subdivided into three important geographic clusters (D5-CHN1: 2014-2019, D5-CHN2: 2016-2022, and D5-EUR: 2022-2023). Additionally, variant-specific (144) amino acid mutation sites and positive-selection pressure sites (132, 262) were identified in the VP1 region. Cluster-specific recombination patterns were also identified, with CVB5, E6, and CVB4 as the major recombinant viruses. These findings provide a preliminary landscape of E11 circulation worldwide and basic scientific data for further study of the pathogenicity of E11 variants.
Subject(s)
Enterovirus B, Human , Evolution, Molecular , Genetic Variation , Genotype , Phylogeny , China/epidemiology , Humans , Enterovirus B, Human/genetics , Enterovirus B, Human/classification , Enterovirus B, Human/isolation & purification , Infant, Newborn , Echovirus Infections/virology , Echovirus Infections/epidemiology , Hand, Foot and Mouth Disease/virology , Hand, Foot and Mouth Disease/epidemiology , InfantABSTRACT
OBJECTIVES: A new variant of echovirus 11 (E11) infection is a major health concern in neonates. Here, we describe the clinical and virological characteristics of enterovirus (EV) infections in children hospitalized with acute respiratory infection in Southern Italy. METHODS: Between July 2022 and August 2023, 173 EV infections were identified. Demographic and clinical characteristics, comorbidities, and coinfections were analyzed. Genotypes were identified by sequencing of VP1. Whole-genome sequencing of five E11 strains was performed. RESULTS: Case numbers peaked in July 2022, November-December 2022, and June-July 2023. Coxsackievirus A2 was identified in 36.7%, coxsackievirus B5 in 13.8%, echovirus E11 in 9.2%, and EV-D68 in 6.4% of cases. No child had critical symptoms or a severe infection. The only neonate infected by E11 recovered fully after 5 days in hospital. Phylogenetic analysis revealed that four E11 strains were closely related to divergent lineage I E11 strains identified in France and Italy. CONCLUSIONS: The new variant of E11 was identified in children in Southern Italy. Although the cases were mild, the data suggest that transmission routes and host factors are likely to be main drivers for the development of potentially severe diseases. Systematic epidemiological/molecular surveillance will help us better understand the clinical impact of EV infections and develop preventive strategies.
Subject(s)
Enterovirus B, Human , Genotype , Phylogeny , Respiratory Tract Infections , Whole Genome Sequencing , Humans , Italy/epidemiology , Infant , Respiratory Tract Infections/virology , Respiratory Tract Infections/epidemiology , Male , Child, Preschool , Female , Enterovirus B, Human/genetics , Enterovirus B, Human/classification , Enterovirus B, Human/isolation & purification , Infant, Newborn , Child , Enterovirus Infections/epidemiology , Enterovirus Infections/virology , Hospitalization , Child, Hospitalized , Enterovirus/genetics , Enterovirus/classification , Enterovirus/isolation & purification , Echovirus Infections/epidemiology , Echovirus Infections/virology , Acute Disease , Coinfection/virology , Coinfection/epidemiologyABSTRACT
Echovirus 11 (E11) has gained attention owing to its association with severe neonatal infections. Due to the limited data available, the World Health Organization (WHO) considers public health risk to the general population to be low. The present study investigated the genetic variation and molecular evolution of E11 genomes collected from May to December 2023. Whole genome sequencing (WGS) was performed for 16 E11 strains. Phylogenetic analysis on WG showed how all Italian strains belonged to genogroup D5, similarly to other E11 strains recently reported in France and Germany all together aggregated into separate clusters. A cluster-specific recombination pattern was also identified using phylogenetic analysis of different genome regions. Echovirus 6 was identified as the major recombinant virus in 3Cpro and 3Dpol regions. The molecular clock analysis revealed that the recombination event probably occurred in June 2018 (95% HPD interval: Jan 2016-Jan 2020). Shannon entropy analyses, within P1 region, showed how 11 amino acids exhibited relatively high entropy. Five of them were exposed on the canyon region which is responsible for receptor binding with the neonatal Fc receptor. The present study showed the recombinant origin of a new lineage of E11 associated with severe neonatal infections.
Subject(s)
Echovirus Infections , Enterovirus B, Human , Genome, Viral , Genotype , Phylogeny , Recombination, Genetic , Humans , Infant, Newborn , Genome, Viral/genetics , Enterovirus B, Human/genetics , Enterovirus B, Human/classification , Enterovirus B, Human/isolation & purification , Echovirus Infections/virology , Echovirus Infections/epidemiology , Genetic Variation , Whole Genome Sequencing , Evolution, Molecular , Italy/epidemiologyABSTRACT
New circulating Enterovirus (EV) strains often emerge through recombination. Upsurges of recombinant non-polio enteroviruses (NPEVs) associated with neurologic manifestations such as EVA71 or Echovirus 30 (E30) are a growing public health concern in Europe. Only a few complete genomes of EVs circulating in Spain are available in public databases, making it difficult to address the emergence of recombinant EVs, understand their evolutionary relatedness and the possible implication in human disease. We have used metagenomic (untargeted) NGS to generate full-length EV genomes from CSF samples of EV-positive aseptic meningitis cases in Southern Spain between 2015 and 2018. Our analyses reveal the co-circulation of multiple Enterovirus B (EV-B) types (E6, E11, E13 and E30), including a novel E13 recombinant form. We observed a genetic turnover where emergent lineages (C1 for E6 and I [tentatively proposed in this study] for E30) replaced previous lineages circulating in Spain, some concomitant with outbreaks in other parts of Europe. Metagenomic sequencing provides an effective approach for the analysis of EV genomes directly from PCR-positive CSF samples. The detection of a novel, disease-associated, recombinant form emphasizes the importance of genomic surveillance to monitor spread and evolution of EVs.
Subject(s)
Enterovirus B, Human/genetics , Enterovirus Infections/virology , Genome, Viral , Meningitis, Aseptic/virology , RNA, Viral/genetics , Adolescent , Adult , Enterovirus B, Human/classification , Enterovirus B, Human/isolation & purification , Enterovirus Infections/cerebrospinal fluid , Enterovirus Infections/epidemiology , Female , Genotype , Humans , Male , Meningitis, Aseptic/cerebrospinal fluid , Meningitis, Aseptic/epidemiology , Phylogeny , RNA, Viral/cerebrospinal fluid , Sequence Analysis, DNA , Spain/epidemiology , Young AdultABSTRACT
Human enteroviruses (EVs) comprise more than 100 types of coxsackievirus, echovirus, poliovirus and numbered enteroviruses, which are mainly transmitted by the faecal-oral route leading to diverse diseases such as aseptic meningitis, encephalitis, and acute flaccid paralysis, among others. Since enteroviruses are excreted in faeces, wastewater-based epidemiology approaches are useful to describe EV diversity in a community. In Uruguay, knowledge about enteroviruses is extremely limited. This study assessed the diversity of enteroviruses through Illumina next-generation sequencing of VP1-amplicons obtained by RT-PCR directly applied to viral concentrates of 84 wastewater samples collected in Uruguay during 2011-2012 and 2017-2018. Fifty out of the 84 samples were positive for enteroviruses. There were detected 27 different types belonging to Enterovirus A species (CVA2-A6, A10, A16, EV-A71, A90), Enterovirus B species (CVA9, B1-B5, E1, E6, E11, E14, E21, E30) and Enterovirus C species (CVA1, A13, A19, A22, A24, EV-C99). Enterovirus A71 (EV-A71) and echovirus 30 (E30) strains were studied more in depth through phylogenetic analysis, together with some strains previously detected by us in Argentina. Results unveiled that EV-A71 sub-genogroup C2 circulates in both countries at least since 2011-2012, and that the C1-like emerging variant recently entered in Argentina. We also confirmed the circulation of echovirus 30 genotypes E and F in Argentina, and reported the detection of genotype E in Uruguay. To the best of our knowledge this is the first report of the EV-A71 C1-like emerging variant in South-America, and the first report of EV-A71 and E30 in Uruguay.
Subject(s)
Enterovirus A, Human/genetics , Enterovirus B, Human/genetics , Genetic Linkage/genetics , Capsid Proteins/genetics , Capsid Proteins/metabolism , Enterovirus A, Human/classification , Enterovirus A, Human/isolation & purification , Enterovirus B, Human/classification , Enterovirus B, Human/isolation & purification , Enterovirus C, Human/classification , Enterovirus C, Human/genetics , Enterovirus C, Human/isolation & purification , Genotype , Humans , Phylogeny , RNA, Viral/chemistry , RNA, Viral/genetics , RNA, Viral/metabolism , Seasons , South America , Uruguay , Wastewater/virologyABSTRACT
In order to discover the causes of a coxsackievirus B4 (CV-B4)-associated hand, foot, and mouth disease (HFMD) outbreak and to study the evolutionary characteristics of the virus, we sequenced isolates obtained during an outbreak for comparative analysis with previously sequenced strains. Phylogenetic and evolutionary dynamics analysis was performed to examine the genetic characteristics of CV-B4 in China and worldwide. Phylogenetic analysis showed that CV-B4 originated from a common ancestor in Shandong. CV-B4 strains isolated worldwide could be classified into genotypes A-E based on the sequence of the VP1 region. All CV-B4 strains in China belonged to genotype E. The global population diversity of CV-B4 fluctuated substantially over time, and CV-B4 isolated in China accounted for a significant increase in the diversity of CV-B4. The average nucleotide substitution rate in VP1 of Chinese CV-B4 (5.20 × 10-3 substitutions/site/year) was slightly higher than that of global CV-B4 (4.82 × 10-3 substitutions/site/year). This study is the first to investigate the evolutionary dynamics of CV-B4 and its association with an HFMD outbreak. These findings explain both the 2011 outbreak and the global increase in CV-B4 diversity. In addition to improving our understanding of a major outbreak, these findings provide a basis for the development of surveillance strategies.
Subject(s)
Capsid Proteins/genetics , Enterovirus B, Human/classification , Hand, Foot and Mouth Disease/virology , Polymorphism, Single Nucleotide , China , Enterovirus B, Human/genetics , Enterovirus B, Human/isolation & purification , Evolution, Molecular , Humans , Molecular Typing , Mutation Rate , Phylogeny , Sequence Analysis, RNAABSTRACT
Human enteroviruses are the most prevalent causes of aseptic meningitis worldwide. However, despite such predominancy, defining the enteroviral etiology of aseptic meningitis remains a diagnostic dilemma for the clinician in Iran. Therefore, this study was conducted to characterize the prevalence and clinical significance of enteroviral aseptic meningitis as well as the predominant enterovirus serotypes among patients with aseptic meningitis in the South of Iran.Cerebrospinal fluid (CSF) specimens were obtained from 73 patients with aseptic meningitis (52.1% males and 47.9% females), ages ranging from 1 month to 88 years. Following the extraction of nucleic acid, the detection of enteroviruses was performed by RT-PCR, targeting the 5' untranslated region of the genome, and sequencing. Enteroviruses were found in 46.6% of samples (34/73). The most predominant serotype was echovirus 30, followed by coxsackievirus B5 and poliovirus type 1 Sabin strain. The enterovirus infections were more prevalent among female patients (58.8%) and those below 5 years of age (52.9%). Although enterovirus infections were observed throughout the year, the infections were more prevalent during autumn with fever as the predominant clinical symptom. The outcomes revealed that enteroviruses are significant causes of aseptic meningitis in the South of Iran, while suspected cases of aseptic meningitis are usually monitored by bacterial culture and biochemical testing of CSF samples. Therefore, the etiology remains unknown in most cases. Molecular detection of viral pathogens should be included as a common approach in the screening of patients with aseptic meningitis to prevent unnecessary treatment and to improve clinical management.
Subject(s)
Enterovirus B, Human/genetics , Enterovirus Infections/epidemiology , Meningitis, Aseptic/epidemiology , Meningitis, Viral/epidemiology , Poliomyelitis/epidemiology , Poliovirus/genetics , Adolescent , Adult , Age Factors , Aged , Aged, 80 and over , Child , Child, Preschool , Enterovirus B, Human/classification , Enterovirus B, Human/isolation & purification , Enterovirus Infections/cerebrospinal fluid , Enterovirus Infections/diagnosis , Enterovirus Infections/virology , Female , Genome, Viral , Humans , Infant , Infant, Newborn , Iran/epidemiology , Male , Meningitis, Aseptic/cerebrospinal fluid , Meningitis, Aseptic/diagnosis , Meningitis, Aseptic/virology , Meningitis, Viral/cerebrospinal fluid , Meningitis, Viral/diagnosis , Meningitis, Viral/virology , Middle Aged , Molecular Epidemiology , Phylogeny , Poliomyelitis/cerebrospinal fluid , Poliomyelitis/diagnosis , Poliomyelitis/virology , Poliovirus/classification , Poliovirus/isolation & purification , Prevalence , RNA, Viral/geneticsABSTRACT
Type B coxsackieviruses (CV-B) frequently infect the central nervous system (CNS) causing neurological diseases notably meningitis and encephalitis. These infections occur principally among newborns and children. Epidemiological studies of patients with nervous system disorders demonstrate the presence of infectious virus, its components, or anti-CV-B antibodies. Some experimental studies conducted in vitro and in vivo support the potential association between CV-B and idiopathic neurodegenerative diseases such as amyotrophic lateral sclerosis and psychiatric illness such as schizophrenia. However, mechanisms explaining how CV-B infections may contribute to the genesis of CNS disorders remain unclear. The proposed mechanisms focus on the immune response following the viral infection as a contributor to pathogenesis. This review describes these epidemiological and experimental studies, the modes of transmission of CV-B with an emphasis on congenital transmission, the routes used by CV-B to reach the brain parenchyma, and plausible mechanisms by which CV-B may induce CNS diseases, with a focus on potential immunopathogenesis.
Subject(s)
Brain/virology , Central Nervous System Diseases/virology , Central Nervous System Infections/virology , Coxsackievirus Infections/diagnosis , Enterovirus B, Human/isolation & purification , Antibodies, Viral , Central Nervous System Diseases/epidemiology , Central Nervous System Diseases/etiology , Child , Coxsackievirus Infections/pathology , Enterovirus B, Human/immunology , Humans , Infant, NewbornABSTRACT
EV-B93 is a novel serotype within the Enterovirus B species and is uncommon worldwide. Currently, only one full-length genomic sequence (the prototype strain) has been deposited in the GenBank database. In this study, three EV-B93 were identified, including one from an acute flaccid paralysis (AFP) patient (named 99052/XZ/CHN/1999, hereafter XZ99052) and two from healthy children (named 99096/XZ/CHN/1999 and 99167/XZ/CHN/1999, hereafter XZ99096 and XZ99167, respectively) from Tibet in 1999 during the polio eradication program. The identity between the nucleotide and amino acid sequences of the Tibet EV-B93 strain and the EV-B93 prototype strain is 83.2%-83.4% and 96.8%-96.9%, respectively. The Tibet EV-B93 strain was found to have greater nucleotide sequence identity in the P3 region to another enterovirus EV-B107 as per a phylogenetic tree analysis, which revealed that recombination occurred. Seroepidemiology data showed that EV-B93 has not produced an epidemic in Tibet and there may be susceptible individuals. The three Tibet EV-B93 strains are temperature-resistant with prognosticative virulence, suggesting the possibility of a potential large-scale outbreak of EV-B93. The analyzed EV-B93 strains enrich our knowledge about this serotype and provide valuable information on global EV-B93 molecular epidemiology. What is more, they permit the appraisal of the serotype's potential public health impact and aid in understanding the role of recombination events in the evolution of enteroviruses.
Subject(s)
Enterovirus B, Human/genetics , Enterovirus Infections/virology , Genome, Viral/genetics , Paralysis/virology , Child, Preschool , Disease Outbreaks/prevention & control , Enterovirus B, Human/isolation & purification , Enterovirus B, Human/pathogenicity , Enterovirus Infections/epidemiology , Feces/virology , Female , Humans , Infant , Male , Molecular Typing , Paralysis/epidemiology , Phylogeny , RNA, Viral/genetics , Recombination, Genetic , Sequence Analysis, RNA , Seroepidemiologic Studies , Tibet/epidemiologyABSTRACT
Due to the risk of poliovirus importation from Ukraine in 2015, a combined surveillance program monitoring the circulation of enteroviruses (EVs) in healthy children from at-risk areas and in the environment was conducted in Romania. Virological testing of stool samples collected from 155 healthy children aged from two months to six years and of 186 sewage water samples collected from different areas was performed. A total of 58 (37.42%) stool samples and 50 (26.88%) sewage water samples were positive for non-polio EVs, but no poliovirus was detected. A high level of circulation of echovirus (E) types 6 and 7 and coxsackievirus (CV) type B5 was observed.
Subject(s)
Enterovirus B, Human/isolation & purification , Enterovirus/isolation & purification , Feces/virology , Sewage/virology , Child , Child, Preschool , Enterovirus/classification , Enterovirus/genetics , Enterovirus B, Human/genetics , Enterovirus Infections/virology , Environment , Environmental Monitoring/methods , Healthy Volunteers , Humans , Infant , Limit of Detection , Logistic Models , Molecular Typing/methods , Phylogeny , Poliovirus/genetics , Poliovirus/isolation & purification , Romania , Wastewater/virologyABSTRACT
BACKGROUND: Little is known about the association between genetic susceptibility and the severity of hand, foot, and mouth disease (HFMD) infected with coxsackievirus A6 (CV-A6). METHODS: Three hundred and sixty-four CV-A6 HFMD patients were enrolled, including 115 severe and 249 mild patients. A genome-wide association study (GWAS) was performed involving eight DNA pools of 115 severe and 115 mild CV-A6 HFMD patients pair-matched by age and gender. Differences in relative allele signal scores of SNPs in Illumina Human OmniZhongHua-8 BeadChips were compared between the two groups. The tag SNPS for potentially functional SNPs or their high linked SNPs were selected for individual genotyping in all 364 patients and assessed for their associations with severe CV-A6 HFMD using multivariable logistic regression analyses. RESULTS: The top 30 significant SNPs obtained from pooled DNA GWAS analysis were checked for biological functions and their high linkage disequilibrium (LD) SNPs. Four tag SNPs (rs1558206, rs6927647, rs9375728 and rs10879355) were selected for further individual genotyping in 364 CV-A6 patients. Only SNP rs10879355 was associated with severe CV-A6 HFMD, with CC genotype having a greater risk of severe illness than TT+TC genotypes (OR=2.48, 95%CI: 1.34, 4.56). SNP rs4290270 is in complete LD with rs10879355 in Chinese Han children. CONCLUSIONS: This is the first report that one potentially functional SNP rs4290270 in the TPH2 gene may be associated with the risk of severe CV-A6 HFMD.
Subject(s)
Hand, Foot and Mouth Disease/genetics , Tryptophan Hydroxylase/genetics , Alleles , Child , Child, Preschool , China/epidemiology , China/ethnology , Enterovirus B, Human/genetics , Enterovirus B, Human/isolation & purification , Enterovirus B, Human/physiology , Female , Genetic Predisposition to Disease , Genome-Wide Association Study , Genotype , Hand, Foot and Mouth Disease/epidemiology , Hand, Foot and Mouth Disease/ethnology , Hand, Foot and Mouth Disease/virology , Humans , Infant , Male , Polymorphism, Single NucleotideABSTRACT
Enterovirus B83 (EV-B83) is a new member of the enterovirus B group. Currently, there are only two full-length genomic sequences of EV-B83 in the GenBank database and few VP1 region sequences. The aetiology and epidemiology of EV-B83 is unclear. 24 stool specimens were collected from twelve AFP patients and 298 stool specimens were collected from 298 healthy children in support of polio eradication activities in Tibet in 1999. Two polioviruses (isolated by L20B cell) and one non-polio enterovirus (isolated by RD cell) were isolated from AFP patients and nine polioviruses (isolated by L20B cell) and 90 non-polio enteroviruses (isolated by RD cell) were isolated from health children. Through molecular typing, we confirmed that the six of non-polio enteroviruses belong to EV-B83. The sequence similarity between the VP1 region of the Tibet isolates and that of the EV-B83 prototype strain was 80%. The maximum-likelihood phylogenetic tree of the partial VP1 region in EV-B83 demonstrated that EV-B83 formed four genotypes globally during the evolution process. The six Tibet EV-B83 strains formed the D genotype alone. Recombination analysis of Tibet EV-B83 showed that CV-B4, CV-A9, EV-B80, and EV-B106 may act as recombinant donors in multiple regions. The serum neutralization test showed that the antibody-positive rate was 58.8% and GMT was 1:19.70, which was higher than the previously reported results of EV-B106 and EV-B80. Temperature sensitivity test results showed that the six Tibet EV-B83 strains were temperature-insensitive with stronger virulence and potential infectivity, which was consistent with the results of the serum neutralization test. This study enriched the genome-wide sequence, epidemiological characteristics, and provided basic data for the follow-up study of EV-B83.
Subject(s)
Antibodies, Viral/blood , Capsid Proteins/genetics , Enterovirus B, Human/genetics , Enterovirus Infections/epidemiology , Genome, Viral , Phylogeny , Case-Control Studies , Child , Enterovirus B, Human/classification , Enterovirus B, Human/immunology , Enterovirus B, Human/isolation & purification , Enterovirus Infections/immunology , Enterovirus Infections/virology , Feces/virology , Female , Genotype , Humans , Infant , Male , Molecular Epidemiology , Molecular Typing , Neutralization Tests , Phenotype , Poliomyelitis/epidemiology , Poliomyelitis/immunology , Poliomyelitis/virology , Poliovirus/classification , Poliovirus/genetics , Poliovirus/immunology , Poliovirus/isolation & purification , Tibet/epidemiologyABSTRACT
BACKGROUND: Studies in prospective cohorts have suggested that enterovirus infections are associated with the appearance of islet autoantibodies that precede later appearance of type 1 diabetes (T1D). It was shown that in addition to an antibody-mediated anti-coxsackievirus (CV)-B neutralizing activity of serum from patients with T1D, there was also enhancing anti-CV-B activity in vitro. In this study, the patterns of enhancing and neutralizing anti-CV activities were analysed from consecutive serum samples collected from children who were followed from birth until they developed T1D in the Finnish Type 1 Diabetes Prediction and Prevention (DIPP) study and compared to those in non-diabetic control children. METHODS: The titres of serum neutralizing activity were analysed against those CVs which were detected in the stools in these children (CV-B3, CV-B5 or CV-A4) using plaque assay. The enhancing activity of these serum samples was analysed by measuring interferon-alpha (INF-α) production in cultures of peripheral blood mononuclear cells (PBMC) inoculated with a mixture of these viruses and diluted serum. RESULTS: A sustained anti-CV enhancing activity was observed in consecutive serum samples in patients with T1D. The pattern of responses differed between children who developed T1D and control children. In patients, the anti-CV enhancing activity was predominant or even exclusive over the neutralizing activity, whereas in controls the enhancing and neutralising activities were more balanced or the neutralizing activity was largely predominant. CONCLUSIONS: Evaluating the anti-enterovirus neutralizing and enhancing activity of serum samples can be useful to investigate further the relationship between enteroviruses and the development of T1D.
Subject(s)
Antibodies, Neutralizing/immunology , Autoantibodies/immunology , Coxsackievirus Infections/immunology , Diabetes Mellitus, Type 1/epidemiology , Enterovirus B, Human/immunology , Immunoglobulin G/immunology , Leukocytes, Mononuclear/immunology , Adolescent , Antibodies, Neutralizing/blood , Autoantibodies/blood , Biomarkers/blood , Child , Child, Preschool , Coxsackievirus Infections/virology , Diabetes Mellitus, Type 1/blood , Diabetes Mellitus, Type 1/immunology , Diabetes Mellitus, Type 1/virology , Enterovirus B, Human/isolation & purification , Female , Finland/epidemiology , Follow-Up Studies , Humans , Immunoglobulin G/blood , Infant , Infant, Newborn , Leukocytes, Mononuclear/virology , Male , PrognosisABSTRACT
Human enteroviruses (EV) are the most common cause of viral meningitis in children. Human parechoviruses (HPeV) are increasingly being recognized as a cause of central nervous system (CNS) infections and sepsis-like disease in children. Both viruses belong to Picornaviridae family. The clinical picture in EV and HPeV infections is usually nonspecific. Therefore, molecular detection of both viruses is needed for etiological diagnosis. In this case report, we describe and discuss clinical and laboratory findings of two consecutive episodes of viral meningitis caused by EV and HPeV, respectively, occurring in the first month of a newborn's life.
Subject(s)
Enterovirus B, Human/genetics , Meningitis, Viral/diagnosis , Parechovirus/genetics , Picornaviridae Infections/diagnosis , RNA, Viral/genetics , Sepsis/diagnosis , Enterovirus B, Human/classification , Enterovirus B, Human/isolation & purification , Enterovirus B, Human/pathogenicity , Female , Humans , Infant, Newborn , Meningitis, Viral/pathology , Meningitis, Viral/virology , Parechovirus/classification , Parechovirus/isolation & purification , Parechovirus/pathogenicity , Picornaviridae Infections/pathology , Picornaviridae Infections/virology , Recurrence , Reverse Transcriptase Polymerase Chain Reaction , Sepsis/pathology , Sepsis/virology , Sequence Analysis, DNASubject(s)
Antiviral Agents/pharmacology , Coxsackievirus Infections/drug therapy , Coxsackievirus Infections/virology , Enterovirus B, Human/drug effects , Enterovirus B, Human/genetics , Animals , Animals, Newborn , Antiviral Agents/administration & dosage , Apomorphine/administration & dosage , Apomorphine/analogs & derivatives , Apomorphine/pharmacology , Drug Therapy, Combination , Enterovirus B, Human/isolation & purification , Mice , Mice, Inbred ICR , Microbial Sensitivity Tests , Nitriles/administration & dosage , Nitriles/pharmacology , Oxadiazoles/administration & dosage , Oxadiazoles/pharmacology , Oxazoles , RNA, Viral/drug effects , RNA, Viral/geneticsABSTRACT
The aim of the present study was to investigate the expression levels and roles of microRNA (miR)217 and miR543 in viral myocarditis, and to examine their underlying mechanisms. Coxsackievirus B3 (CVB3) was used to establish in vivo and in vitro models of viral myocarditis. The levels of miR217 and miR543 were detected using reverse transcriptionquantitative PCR. The association between miR217 and miR543 and sirtuin1 (SIRT1) was predicted and confirmed by TargetScan and dualluciferase reporter assay. Cell viability was detected using Cell Counting Kit8 assay, and cell apoptosis was measured by analyzing the expression levels of Bcl2 and Bax, and by flow cytometry. In addition, the synthesis of various proinflammatory factors was determined by ELISA. In addition, superoxide dismutase (SOD) activity and malondialdehyde (MDA) levels were measured in cardiomyocytes following transfection and CVB infection. miR217 and miR543 were found to be highly expressed in the peripheral blood of pediatric patients with viral myocarditis, in the peripheral blood and myocardial tissues of viral myocarditis mice and in CVB3infected cardiomyocytes. SIRT1 was found to be a target of both miR217 and miR543, and SIRT1 expression level was downregulated in viral myocarditis. Further analysis indicated that the reduced cell viability, increased cell apoptosis, enhanced synthesis of inflammatory factors, increased MDA content and decreased SOD activity associated with myocarditis were significantly reversed after inhibition of miR217 or miR543. Importantly, the present results showed that all the effects of miR217 and miR543 inhibition on cardiomyocytes were significantly suppressed following SIRT1 knockdown. Collectively, the present data indicated that miR217 and miR543 were significantly upregulated in viral myocarditis, and downregulation of miR217 and miR543 attenuated CVB3 infectioninduced cardiomyocyte injury by targeting SIRT1. miR217 and miR543 may be potential therapeutic targets for developing novel viral myocarditis treatments in the future.
Subject(s)
Coxsackievirus Infections/complications , MicroRNAs/genetics , Myocarditis/genetics , Myocarditis/virology , AMP-Activated Protein Kinases/genetics , AMP-Activated Protein Kinases/metabolism , Animals , Cell Line , Child , Child, Preschool , Coxsackievirus Infections/genetics , Coxsackievirus Infections/metabolism , Coxsackievirus Infections/virology , Down-Regulation , Enterovirus B, Human/isolation & purification , Female , Humans , Infant , Male , Mice, Inbred BALB C , Myocarditis/metabolism , NF-kappa B/genetics , NF-kappa B/metabolism , Signal Transduction , Sirtuin 1/genetics , Sirtuin 1/metabolism , Up-RegulationABSTRACT
Coxsackievirus A9 (CVA9) is an enterically transmitted enterovirus and one of the most pathogenic type among human enteroviruses. CVA9 isolates use a distinctive RGD (Arg-Gly-Asp) motif within VP1 capsid protein that defines its ability to bind to integrin receptor(s) for cellular entry. To investigate CVA9 evolution and pathogenicity, genetic relationships and recombination events were analyzed between 54 novel clinical isolates of CVA9, as well as 21 previously published full length CVA9 sequences from GenBank. Samples were investigated by partial sequencing of the novel VP1 and 3Dpol genes, as well as including the corresponding areas from GenBank sequences. Phylogenetic analyses were combined with clinical data in a further attempt to analyze whether sequence evolution reflects CVA9 pathogenicity in the phylogenies. Furthermore, VP1 gene was also analyzed for receptor binding sites including the RGD motif and the putative heparan sulfate (HS) site. Analysis of the 559-nucleotide-long VP1 sequences identified six clades. Although most of the strains within each clade showed geographical clustering, the grouping pattern of the isolates in the analysis of the VP1 gene was strikingly different from grouping of 3Dpol, which suggests that recombination events may have occurred in the region encoding the nonstructural proteins. Inclusion of clinical data did not provide any evidence of symptom based phylogenetic clustering of CVA9 isolates. Amino acid sequence analysis of the VP1 polypeptide demonstrated that the RGD motif was fully conserved among the isolates while the putative HS binding site was only found in one isolate. These data suggest that integrin binding is essential for virus tropism, but do not explain the symptom repertoire.
Subject(s)
Capsid Proteins/genetics , Coxsackievirus Infections/virology , Enterovirus B, Human/genetics , Enterovirus B, Human/isolation & purification , Amino Acid Motifs/genetics , Binding Sites , Capsid Proteins/chemistry , Capsid Proteins/metabolism , Conserved Sequence , DNA-Directed RNA Polymerases/genetics , Enterovirus B, Human/classification , Evolution, Molecular , Heparitin Sulfate , Humans , Oligopeptides , Phylogeny , Protein Binding , Receptors, Virus/metabolism , Recombination, GeneticABSTRACT
Echovirus 21 (E21) belongs to the species Enterovirus B, whose members are frequently associated with acute flaccid paralysis. E21 strain 553/YN/CHN/2013 was isolated from a healthy child in Yunnan, China, in 2013. This is the first report of the complete genome sequence of E21 in China. This strain shared 81.7% nucleotide sequence identity and 96.8% amino acid sequence identity with the E21 prototype strain Farina. Although strain 553/YN/CHN/2013 belongs to the E21 serotype, the only similarity to the E21 strain was in the VP1 region, as other genomic regions, including VP2-VP4, were more similar to other EV-B members. Recombination analysis showed evidence of recombination events between E21 and other EV-B viruses. E21 strain 553/YN/CHN/2013 failed to infect suckling mice via intracerebral injection. Surveillance of E21 is very important to help forecast the potential of emerging E21 outbreaks and related diseases.