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1.
PLoS Pathog ; 20(4): e1012159, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38662650

RESUMO

Human enteroviruses are the most common human pathogen with over 300 distinct genotypes. Previous work with poliovirus has suggested that it is possible to generate antibody responses in humans and animals that can recognize members of multiple enterovirus species. However, cross protective immunity across multiple enteroviruses is not observed epidemiologically in humans. Here we investigated whether immunization of mice or baboons with inactivated poliovirus or enterovirus virus-like-particles (VLPs) vaccines generates antibody responses that can recognize enterovirus D68 or A71. We found that mice only generated antibodies specific for the antigen they were immunized with, and repeated immunization failed to generate cross-reactive antibody responses as measured by both ELISA and neutralization assay. Immunization of baboons with IPV failed to generate neutralizing antibody responses against enterovirus D68 or A71. These results suggest that a multivalent approach to enterovirus vaccination is necessary to protect against enterovirus disease in vulnerable populations.


Assuntos
Anticorpos Antivirais , Reações Cruzadas , Infecções por Enterovirus , Vacina Antipólio de Vírus Inativado , Animais , Camundongos , Reações Cruzadas/imunologia , Anticorpos Antivirais/imunologia , Infecções por Enterovirus/imunologia , Infecções por Enterovirus/prevenção & controle , Infecções por Enterovirus/virologia , Vacina Antipólio de Vírus Inativado/imunologia , Vacina Antipólio de Vírus Inativado/administração & dosagem , Vacinas de Partículas Semelhantes a Vírus/imunologia , Anticorpos Neutralizantes/imunologia , Papio/imunologia , Humanos , Poliovirus/imunologia , Feminino , Formação de Anticorpos/imunologia , Enterovirus/imunologia , Camundongos Endogâmicos BALB C , Enterovirus Humano D/imunologia
2.
J Virol ; 98(7): e0039724, 2024 Jul 23.
Artigo em Inglês | MEDLINE | ID: mdl-38869283

RESUMO

Enterovirus D68 (EV-D68) is an emerging pathogen that can cause severe respiratory and neurologic disease [acute flaccid myelitis (AFM)]. Intramuscular (IM) injection of neonatal Swiss Webster (SW) mice with US/IL/14-18952 (IL52), a clinical isolate from the 2014 EV-D68 epidemic, results in many of the pathogenic features of human AFM, including viral infection of the spinal cord, death of motor neurons, and resultant progressive paralysis. In distinction, CA/14-4231 (CA4231), another clinical isolate from the 2014 EV-D68 outbreak, does not cause paralysis in mice, does not grow in the spinal cord, and does not cause motor neuron loss following IM injection. A panel of chimeric viruses containing sequences from IL52 and CA4231 was used to demonstrate that VP1 is the main determinant of EV-D68 neurovirulence following IM injection of neonatal SW mice. VP1 contains four amino acid differences between IL52 and CA4231. Mutations resulting in substituting these four amino acids (CA4231 residues into the IL52 polyprotein) completely abolished neurovirulence. Conversely, mutations resulting in substituting VP1 IL52 amino acid residues into the CA4231 polyprotein created a virus that induced paralysis to the same degree as IL52. Neurovirulence following infection of neonatal SW mice with parental and chimeric viruses was associated with viral growth in the spinal cord. IMPORTANCE: Emerging viruses allow us to investigate mutations leading to increased disease severity. Enterovirus D68 (EV-D68), once the cause of rare cases of respiratory illness, recently acquired the ability to cause severe respiratory and neurologic disease. Chimeric viruses were used to demonstrate that viral structural protein VP1 determines growth in the spinal cord, motor neuron loss, and paralysis following intramuscular (IM) injection of neonatal Swiss Webster (SW) mice with EV-D68. These results have relevance for predicting the clinical outcome of future EV-D68 epidemics as well as targeting retrograde transport as a potential strategy for treating virus-induced neurologic disease.


Assuntos
Proteínas do Capsídeo , Viroses do Sistema Nervoso Central , Modelos Animais de Doenças , Enterovirus Humano D , Infecções por Enterovirus , Mielite , Doenças Neuromusculares , Animais , Enterovirus Humano D/patogenicidade , Enterovirus Humano D/genética , Enterovirus Humano D/fisiologia , Mielite/virologia , Camundongos , Infecções por Enterovirus/virologia , Infecções por Enterovirus/patologia , Doenças Neuromusculares/virologia , Doenças Neuromusculares/patologia , Proteínas do Capsídeo/genética , Proteínas do Capsídeo/metabolismo , Viroses do Sistema Nervoso Central/virologia , Viroses do Sistema Nervoso Central/patologia , Humanos , Medula Espinal/virologia , Medula Espinal/patologia , Neurônios Motores/virologia , Neurônios Motores/patologia , Animais Recém-Nascidos , Virulência , Paralisia/virologia
3.
J Virol ; 98(7): e0055624, 2024 Jul 23.
Artigo em Inglês | MEDLINE | ID: mdl-38888347

RESUMO

Enterovirus D68 (EV-D68) is a picornavirus associated with severe respiratory illness and a paralytic disease called acute flaccid myelitis in infants. Currently, no protective vaccines or antivirals are available to combat this virus. Like other enteroviruses, EV-D68 uses components of the cellular autophagy pathway to rewire membranes for its replication. Here, we show that transcription factor EB (TFEB), the master transcriptional regulator of autophagy and lysosomal biogenesis, is crucial for EV-D68 infection. Knockdown of TFEB attenuated EV-D68 genomic RNA replication but did not impact viral binding or entry into host cells. The 3C protease of EV-D68 cleaves TFEB at the N-terminus at glutamine 60 (Q60) immediately post-peak viral RNA replication, disrupting TFEB-RagC interaction and restricting TFEB transport to the surface of the lysosome. Despite this, TFEB remained mostly cytosolic during EV-D68 infection. Overexpression of a TFEB mutant construct lacking the RagC-binding domain, but not the wild-type construct, blocks autophagy and increases EV-D68 nonlytic release in H1HeLa cells but not in autophagy-defective ATG7 KO H1HeLa cells. Our results identify TFEB as a vital host factor regulating multiple stages of the EV-D68 lifecycle and suggest that TFEB could be a promising target for antiviral development against EV-D68. IMPORTANCE: Enteroviruses are among the most significant causes of human disease. Some enteroviruses are responsible for severe paralytic diseases such as poliomyelitis or acute flaccid myelitis. The latter disease is associated with multiple non-polio enterovirus species, including enterovirus D68 (EV-D68), enterovirus 71, and coxsackievirus B3 (CVB3). Here, we demonstrate that EV-D68 interacts with a host transcription factor, transcription factor EB (TFEB), to promote viral RNA(vRNA) replication and regulate the egress of virions from cells. TFEB was previously implicated in the viral egress of CVB3, and the viral protease 3C cleaves TFEB during infection. Here, we show that EV-D68 3C protease also cleaves TFEB after the peak of vRNA replication. This cleavage disrupts TFEB interaction with the host protein RagC, which changes the localization and regulation of TFEB. TFEB lacking a RagC-binding domain inhibits autophagic flux and promotes virus egress. These mechanistic insights highlight how common host factors affect closely related, medically important viruses differently.


Assuntos
Autofagia , Fatores de Transcrição de Zíper de Leucina e Hélice-Alça-Hélix Básicos , Enterovirus Humano D , Infecções por Enterovirus , Replicação Viral , Fatores de Transcrição de Zíper de Leucina e Hélice-Alça-Hélix Básicos/metabolismo , Fatores de Transcrição de Zíper de Leucina e Hélice-Alça-Hélix Básicos/genética , Humanos , Enterovirus Humano D/fisiologia , Enterovirus Humano D/metabolismo , Enterovirus Humano D/genética , Infecções por Enterovirus/metabolismo , Infecções por Enterovirus/virologia , Proteases Virais 3C/metabolismo , Lisossomos/metabolismo , RNA Viral/metabolismo , RNA Viral/genética , Proteínas Virais/metabolismo , Proteínas Virais/genética , Mielite/metabolismo , Mielite/virologia , Ligação Proteica , Células HEK293 , Doenças Neuromusculares , Viroses do Sistema Nervoso Central
4.
J Virol ; 98(2): e0199423, 2024 Feb 20.
Artigo em Inglês | MEDLINE | ID: mdl-38240591

RESUMO

Following the successful control of poliovirus, the re-emergence of respiratory enterovirus D68 (EV-D68), a prominent non-polio enterovirus, has become a serious public health concern worldwide. Host innate immune responses are the primary defense against EV-D68 invasion; however, the mechanism underlying viral evasion of the antiviral activity of interferons (IFN) remains unclear. In this study, we found that EV-D68 inhibited type I IFN signaling by cleaving signal transducer and activator of transcription 1 (STAT1), a crucial factor in cellular responses to interferons and other cytokines. We observed that the prototype and circulating EV-D68 strains conserved their ability to induce STAT1 cleavage and attenuate IFN signal transduction. Further investigation revealed that EV-D68 3C protease cleaves STAT1 at the 131Q residue. Interestingly, not all enterovirus-encoded 3C proteases exhibited this ability. EV-D68 and poliovirus 3C proteases efficiently induced STAT1 cleavage; whereas, 3C proteases from EV-A71, coxsackievirus A16, and echoviruses did not. STAT1 cleavage also abolished the nuclear translocation capacity of STAT1 in response to IFN stimulation to activate downstream signaling elements. Overall, these results suggest that STAT1, targeted by viral protease 3C, is utilized by EV-D68 to subvert the host's innate immune response.IMPORTANCEEnterovirus D68 (EV-D68) has significantly transformed over the past decade, evolving from a rare pathogen to a potential pandemic pathogen. The interferon (IFN) signaling pathway is an important defense mechanism and therapeutic target for the host to resist viral invasion. Previous studies have reported that the EV-D68 virus blocks or weakens immune recognition and IFN production in host cells through diverse strategies; however, the mechanisms of EV-D68 resistance to IFN signaling have not been fully elucidated. Our study revealed that EV-D68 relies on its own encoded protease, 3C, to directly cleave signal transducer and activator of transcription 1 (STAT1), a pivotal transduction component in the IFN signaling pathway, disrupting the IFN-mediated antiviral response. Previous studies on human enteroviruses have not documented direct cleavage of the STAT1 protein to evade cellular immune defenses. However, not all enteroviral 3C proteins can cleave STAT1. These findings highlight the diverse evolutionary strategies different human enteroviruses employ to evade host immunity.


Assuntos
Proteases Virais 3C , Enterovirus Humano D , Interferon Tipo I , Transdução de Sinais , Humanos , Proteases Virais 3C/metabolismo , Antígenos Virais/metabolismo , Antivirais/farmacologia , Cisteína Endopeptidases/metabolismo , Enterovirus Humano D/fisiologia , Interações Hospedeiro-Patógeno , Evasão da Resposta Imune , Imunidade Inata , Interferon Tipo I/metabolismo , Peptídeo Hidrolases/metabolismo , Proteólise , Fator de Transcrição STAT1/metabolismo , Proteínas Virais/metabolismo
5.
J Virol ; 98(2): e0190923, 2024 Feb 20.
Artigo em Inglês | MEDLINE | ID: mdl-38289118

RESUMO

Pyroptosis, a pro-inflammatory programmed cell death, has been implicated in the pathogenesis of coronavirus disease 2019 and other viral diseases. Gasdermin family proteins (GSDMs), including GSDMD and GSDME, are key regulators of pyroptotic cell death. However, the mechanisms by which virus infection modulates pyroptosis remain unclear. Here, we employed a mCherry-GSDMD fluorescent reporter assay to screen for viral proteins that impede the localization and function of GSDMD in living cells. Our data indicated that the main protease NSP5 of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) blocked GSDMD-mediated pyroptosis via cleaving residues Q29 and Q193 of GSDMD. While another SARS-CoV-2 protease, NSP3, cleaved GSDME at residue G370 but activated GSDME-mediated pyroptosis. Interestingly, respiratory enterovirus EV-D68-encoded proteases 3C and 2A also exhibit similar differential regulation on the functions of GSDMs by inactivating GSDMD but initiating GSDME-mediated pyroptosis. EV-D68 infection exerted oncolytic effects on human cancer cells by inducing pyroptotic cell death. Our findings provide insights into how respiratory viruses manipulate host cell pyroptosis and suggest potential targets for antiviral therapy as well as cancer treatment.IMPORTANCEPyroptosis plays a crucial role in the pathogenesis of coronavirus disease 2019, and comprehending its function may facilitate the development of novel therapeutic strategies. This study aims to explore how viral-encoded proteases modulate pyroptosis. We investigated the impact of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and respiratory enterovirus D68 (EV-D68) proteases on host cell pyroptosis. We found that SARS-CoV-2-encoded proteases NSP5 and NSP3 inactivate gasdermin D (GSDMD) but initiate gasdermin E (GSDME)-mediated pyroptosis, respectively. We also discovered that another respiratory virus EV-D68 encodes two distinct proteases 2A and 3C that selectively trigger GSDME-mediated pyroptosis while suppressing the function of GSDMD. Based on these findings, we further noted that EV-D68 infection triggers pyroptosis and produces oncolytic effects in human carcinoma cells. Our study provides new insights into the molecular mechanisms underlying virus-modulated pyroptosis and identifies potential targets for the development of antiviral and cancer therapeutics.


Assuntos
Endopeptidases , Enterovirus Humano D , Interações entre Hospedeiro e Microrganismos , Vírus Oncolíticos , Piroptose , SARS-CoV-2 , Humanos , Linhagem Celular Tumoral , COVID-19/metabolismo , COVID-19/terapia , COVID-19/virologia , Endopeptidases/genética , Endopeptidases/metabolismo , Enterovirus Humano D/enzimologia , Enterovirus Humano D/genética , Infecções por Enterovirus/metabolismo , Infecções por Enterovirus/virologia , Gasderminas/antagonistas & inibidores , Gasderminas/genética , Gasderminas/metabolismo , Terapia Viral Oncolítica , Vírus Oncolíticos/enzimologia , Vírus Oncolíticos/genética , SARS-CoV-2/enzimologia , SARS-CoV-2/genética , Proteínas Virais/genética , Proteínas Virais/metabolismo
6.
J Virol ; 98(6): e0043424, 2024 Jun 13.
Artigo em Inglês | MEDLINE | ID: mdl-38690875

RESUMO

The globally reemerging respiratory pathogen enterovirus D68 (EV-D68) is implicated in outbreaks of severe respiratory illness and associated with acute flaccid myelitis. However, there remains a lack of effective treatments for EV-D68 infection. In this work, we found that the host Toll-like receptor 7 (TLR7) proteins, which function as powerful innate immune sensors, were selectively elevated in expression in response to EV-D68 infection. Subsequently, we investigated the impact of Vesatolimod (GS-9620), a Toll-like receptor 7 agonist, on EV-D68 replication. Our findings revealed that EV-D68 infection resulted in increased mRNA levels of TLR7. Treatment with Vesatolimod significantly inhibited EV-D68 replication [half maximal effective concentration (EC50) = 0.1427 µM] without inducing significant cytotoxicity at virucidal concentrations. Although Vesatolimod exhibited limited impact on EV-D68 attachment, it suppressed RNA replication and viral protein synthesis after virus entry. Vesatolimod broadly inhibited the replication of circulating isolated strains of EV-D68. Furthermore, our findings demonstrated that treatment with Vesatolimod conferred resistance to both respiratory and neural cells against EV-D68 infection. Overall, these results present a promising strategy for drug development by pharmacologically activating TLR7 to initiate an antiviral state in EV-D68-infected cells selectively.IMPORTANCEConventional strategies for antiviral drug development primarily focus on directly targeting viral proteases or key components, as well as host proteins involved in viral replication. In this study, based on our intriguing discovery that enterovirus D68 (EV-D68) infection specifically upregulates the expression of immune sensor Toll-like receptor 7 (TLR7) protein, which is either absent or expressed at low levels in respiratory cells, we propose a potential antiviral approach utilizing TLR7 agonists to activate EV-D68-infected cells into an anti-viral defense state. Notably, our findings demonstrate that pharmacological activation of TLR7 effectively suppresses EV-D68 replication in respiratory tract cells through a TLR7/MyD88-dependent mechanism. This study not only presents a promising drug candidate and target against EV-D68 dissemination but also highlights the potential to exploit unique alterations in cellular innate immune responses induced by viral infections, selectively inducing a defensive state in infected cells while safeguarding uninfected normal cells from potential adverse effects associated with therapeutic interventions.


Assuntos
Antivirais , Enterovirus Humano D , Receptor 7 Toll-Like , Replicação Viral , Receptor 7 Toll-Like/agonistas , Receptor 7 Toll-Like/metabolismo , Humanos , Replicação Viral/efeitos dos fármacos , Enterovirus Humano D/efeitos dos fármacos , Antivirais/farmacologia , Indóis/farmacologia , Infecções por Enterovirus/virologia , Imunidade Inata/efeitos dos fármacos , Linhagem Celular , Internalização do Vírus/efeitos dos fármacos , Pteridinas
7.
J Infect Dis ; 230(4): e917-e928, 2024 Oct 16.
Artigo em Inglês | MEDLINE | ID: mdl-38547499

RESUMO

Enterovirus D68 (EV-D68) infections are associated with severe respiratory disease and acute flaccid myelitis (AFM). The European Non-Polio Enterovirus Network (ENPEN) aimed to investigate the epidemiological and genetic characteristics of EV-D68 infections and its clinical impact during the fall-winter season of 2021-2022. From 19 European countries, 58 institutes reported 10 481 (6.8%) EV-positive samples of which 1004 (9.6%) were identified as EV-D68 (including 852 respiratory samples). Clinical data were reported for 969 cases; 78.9% of infections were reported in children (0-5 years); and 37.9% of cases were hospitalized. Acute respiratory distress was commonly noted (93.1%) followed by fever (49.4%). Neurological problems were observed in 6.4% of cases including 6 diagnosed with AFM. Phylodynamic/Nextstrain and phylogenetic analyses based on 694 sequences showed the emergence of 2 novel B3-derived lineages, with no regional clustering. In conclusion, we describe a large-scale European EV-D68 upsurge with severe clinical impact and the emergence of B3-derived lineages.


Assuntos
Enterovirus Humano D , Infecções por Enterovirus , Filogenia , Humanos , Infecções por Enterovirus/epidemiologia , Infecções por Enterovirus/virologia , Enterovirus Humano D/genética , Enterovirus Humano D/classificação , Enterovirus Humano D/isolamento & purificação , Europa (Continente)/epidemiologia , Pré-Escolar , Masculino , Lactente , Feminino , Criança , Adolescente , Mielite/epidemiologia , Mielite/virologia , Infecções Respiratórias/virologia , Infecções Respiratórias/epidemiologia , Adulto , Viroses do Sistema Nervoso Central/epidemiologia , Viroses do Sistema Nervoso Central/virologia , Recém-Nascido , Adulto Jovem , Pessoa de Meia-Idade , Doenças Neuromusculares/epidemiologia , Doenças Neuromusculares/virologia , Idoso
8.
Emerg Infect Dis ; 30(1): 141-145, 2024 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-38147067

RESUMO

In a 2-year study in Leuven, Belgium, we investigated the use of wastewater sampling to assess community spread of respiratory viruses. Comparison with the number of positive clinical samples demonstrated that wastewater data reflected circulation levels of typical seasonal respiratory viruses, such as influenza, respiratory syncytial virus, and enterovirus D68.


Assuntos
Enterovirus Humano D , Influenza Humana , Vírus Sincicial Respiratório Humano , Humanos , Bélgica/epidemiologia , Águas Residuárias , Vírus Sincicial Respiratório Humano/genética
9.
Emerg Infect Dis ; 30(8): 1687-1691, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-39043450

RESUMO

In December 2023, we observed through hospital-based surveillance a severe outbreak of enterovirus D68 infection in pediatric inpatients in Dakar, Senegal. Molecular characterization revealed that subclade B3, the dominant lineage in outbreaks worldwide, was responsible for the outbreak. Enhanced surveillance in inpatient settings, including among patients with neurologic illnesses, is needed.


Assuntos
Surtos de Doenças , Enterovirus Humano D , Infecções por Enterovirus , Infecções Respiratórias , Humanos , Senegal/epidemiologia , Enterovirus Humano D/genética , Enterovirus Humano D/classificação , Enterovirus Humano D/isolamento & purificação , Infecções por Enterovirus/epidemiologia , Infecções por Enterovirus/virologia , Infecções por Enterovirus/diagnóstico , Infecções Respiratórias/epidemiologia , Infecções Respiratórias/virologia , Pré-Escolar , Lactente , Criança , Filogenia , Masculino , Feminino , Doença Aguda/epidemiologia , Adolescente , Hospitais , História do Século XXI
10.
Emerg Infect Dis ; 30(11): 2250-2260, 2024 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-39447143

RESUMO

Congregate homeless shelters are disproportionately affected by infectious disease outbreaks. We describe enterovirus epidemiology across 23 adult and family shelters in King County, Washington, USA, during October 2019-May 2021, by using repeated cross-sectional respiratory illness and environmental surveillance and viral genome sequencing. Among 3,281 participants >3 months of age, we identified coxsackievirus A21 (CVA21) in 39 adult residents (3.0% [95% CI 1.9%-4.8%] detection) across 7 shelters during October 2019-February 2020. We identified enterovirus D68 (EV-D68) in 5 adult residents in 2 shelters during October-November 2019. Of 812 environmental samples, 1 was EV-D68-positive and 5 were CVA21-positive. Other enteroviruses detected among residents, but not in environmental samples, included coxsackievirus A6/A4 in 3 children. No enteroviruses were detected during April 2020-May 2021. Phylogenetically clustered CVA21 and EV-D68 cases occurred in some shelters. Some shelters also hosted multiple CVA21 lineages.


Assuntos
Enterovirus Humano D , Infecções por Enterovirus , Pessoas Mal Alojadas , Filogenia , Humanos , Washington/epidemiologia , Pessoas Mal Alojadas/estatística & dados numéricos , Masculino , Adulto , Feminino , Infecções por Enterovirus/epidemiologia , Infecções por Enterovirus/virologia , Enterovirus Humano D/genética , Enterovirus Humano D/classificação , Pessoa de Meia-Idade , Genoma Viral , Pré-Escolar , Criança , Enterovirus/genética , Enterovirus/classificação , Adolescente , Lactente , Surtos de Doenças , Adulto Jovem , Infecções por Coxsackievirus/epidemiologia , Infecções por Coxsackievirus/virologia , Estudos Transversais , Idoso , Habitação
11.
Emerg Infect Dis ; 30(3): 423-431, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38407198

RESUMO

Surveillance for emerging pathogens is critical for developing early warning systems to guide preparedness efforts for future outbreaks of associated disease. To better define the epidemiology and burden of associated respiratory disease and acute flaccid myelitis (AFM), as well as to provide actionable data for public health interventions, we developed a multimodal surveillance program in Colorado, USA, for enterovirus D68 (EV-D68). Timely local, state, and national public health outreach was possible because prospective syndromic surveillance for AFM and asthma-like respiratory illness, prospective clinical laboratory surveillance for EV-D68 among children hospitalized with respiratory illness, and retrospective wastewater surveillance led to early detection of the 2022 outbreak of EV-D68 among Colorado children. The lessons learned from developing the individual layers of this multimodal surveillance program and how they complemented and informed the other layers of surveillance for EV-D68 and AFM could be applied to other emerging pathogens and their associated diseases.


Assuntos
Viroses do Sistema Nervoso Central , Enterovirus Humano D , Mielite , Doenças Neuromusculares , Doenças Respiratórias , Criança , Humanos , Colorado/epidemiologia , Estudos Prospectivos , Estudos Retrospectivos , Águas Residuárias , Vigilância Epidemiológica Baseada em Águas Residuárias
12.
J Virol ; 97(12): e0160023, 2023 Dec 21.
Artigo em Inglês | MEDLINE | ID: mdl-38047678

RESUMO

IMPORTANCE: Enterovirus D68 (EV-D68) is an emerging respiratory pathogen associated with acute flaccid myelitis. Currently, no approved vaccines or antiviral drugs are available. Here, we report four functionally independent neutralizing antigenic sites (I to IV) by analyses of neutralizing monoclonal antibody (MAb)-resistant mutants. Site I is located in the VP1 BC loop near the fivefold axis. Site II resides in the VP2 EF loop, and site III is situated in VP1 C-terminus; both sites are located at the south rim of the canyon. Site IV is composed of residue in VP2 ßB strand and residues in the VP3 BC loop and resides around the threefold axis. The developed MAbs targeting the antigenic sites can inhibit viral binding to cells. These findings advance the understanding of the recognition of EV-D68 by neutralizing antibodies and viral evolution and immune escape and also have important implications for the development of novel EV-D68 vaccines.


Assuntos
Anticorpos Neutralizantes , Proteínas do Capsídeo , Enterovirus Humano D , Infecções por Enterovirus , Humanos , Capsídeo , Proteínas do Capsídeo/química , Enterovirus Humano D/genética , Infecções por Enterovirus/imunologia , Infecções por Enterovirus/virologia
13.
J Virol ; 97(5): e0015623, 2023 05 31.
Artigo em Inglês | MEDLINE | ID: mdl-37154751

RESUMO

In 2014, 2016, and 2018, the United States experienced unprecedented spikes in pediatric cases of acute flaccid myelitis (AFM), which is a poliomyelitis-like paralytic illness. Accumulating clinical, immunological, and epidemiological evidence has identified enterovirus D68 (EV-D68) as a major causative agent of these biennial AFM outbreaks. There are currently no available FDA-approved antivirals that are effective against EV-D68, and the treatment for EV-D68-associated AFM is primarily supportive. Telaprevir is an food and drug administration (FDA)-approved protease inhibitor that irreversibly binds the EV-D68 2A protease and inhibits EV-D68 replication in vitro. Here, we utilize a murine model of EV-D68 associated AFM to show that early telaprevir treatment improves paralysis outcomes in Swiss Webster (SW) mice. Telaprevir reduces both viral titer and apoptotic activity in both muscles and spinal cords at early disease time points, which results in improved AFM outcomes in infected mice. Following intramuscular inoculation in mice, EV-D68 infection results in a stereotypic pattern of weakness that is reflected by the loss of the innervating motor neuron population, in sequential order, of the ipsilateral (injected) hindlimb, the contralateral hindlimb, and then the forelimbs. Telaprevir treatment preserved motor neuron populations and reduced weakness in limbs beyond the injected hindlimb. The effects of telaprevir were not seen when the treatment was delayed, and toxicity limited doses beyond 35 mg/kg. These studies are a proof of principle, provide the first evidence of benefit of an FDA-approved antiviral drug with which to treat AFM, and emphasize both the need to develop better tolerated therapies that remain efficacious when administered after viral infections and the development of clinical symptoms. IMPORTANCE Recent outbreaks of EV-D68 in 2014, 2016, and 2018 have resulted in over 600 cases of a paralytic illness that is known as AFM. AFM is a predominantly pediatric disease with no FDA-approved treatment, and many patients show minimal recovery from limb weakness. Telaprevir is an FDA-approved antiviral that has been shown to inhibit EV-D68 in vitro. Here, we demonstrate that a telaprevir treatment that is given concurrently with an EV-D68 infection improves AFM outcomes in mice by reducing apoptosis and viral titers at early time points. Telaprevir also protected motor neurons and improved paralysis outcomes in limbs beyond the site of viral inoculation. This study improves understanding of EV-D68 pathogenesis in the mouse model of AFM. This study serves as a proof of principle for the first FDA-approved drug that has been shown to improve AFM outcomes and have in vivo efficacy against EV-D68 as well as underlines the importance of the continued development of EV-D68 antivirals.


Assuntos
Viroses do Sistema Nervoso Central , Enterovirus Humano D , Infecções por Enterovirus , Animais , Estados Unidos , Camundongos , Enterovirus Humano D/fisiologia , Modelos Animais de Doenças , Paralisia/tratamento farmacológico , Paralisia/etiologia , Infecções por Enterovirus/patologia , Antivirais/farmacologia , Antivirais/uso terapêutico
14.
J Virol ; 97(4): e0042523, 2023 04 27.
Artigo em Inglês | MEDLINE | ID: mdl-37039659

RESUMO

Enterovirus D68 (EV-D68), which causes severe respiratory diseases and irreversible central nervous system damage, has become a serious public health problem worldwide. However, the mechanisms by which EV-D68 exerts neurotoxicity remain unclear. Thus, we aimed to analyze the effects of EV-D68 infection on the cleavage, subcellular translocation, and pathogenic aggregation of TAR DNA-binding protein 43 kDa (TDP-43) in respiratory or neural cells. The results showed that EV-D68-encoded proteases 2A and 3C induced TDP-43 translocation and cleavage, respectively. Specifically, 3C cleaved residue 327Q of TDP-43. The 3C-mediated cleaved TDP-43 fragments had substantially decreased protein solubility compared with the wild-type TDP-43. Hence, 3C activity promoted TDP-43 aggregation, which exerted cytotoxicity to diverse human cells, including glioblastoma T98G cells. The effects of commercially available antiviral drugs on 3C-mediated TDP-43 cleavage were screened, and the results revealed lopinavir as a potent inhibitor of EV-D68 3C protease. Overall, these results suggested TDP-43 as a conserved host target of EV-D68 3C. This study is the first to provide evidence on the involvement of TDP-43 dysregulation in EV-D68 pathogenesis. IMPORTANCE Over the past decade, the incidence of enterovirus D68 (EV-D68) infection has increased worldwide. EV-D68 infection can cause different respiratory symptoms and severe neurological complications, including acute flaccid myelitis. Thus, elucidating the mechanisms underlying EV-D68 toxicity is important to develop novel methods to prevent EV-D68 infection-associated diseases. This study shows that EV-D68 infection triggers the translocalization, cleavage, and aggregation of TDP-43, an intracellular protein closely related to degenerative neurological disorders. The viral protease 3C decreased TDP-43 solubility, thereby exerting cytotoxicity to host cells, including human glioblastoma cells. Thus, counteracting 3C activity is an effective strategy to relieve EV-D68-triggered cell death. Cytoplasmic aggregation of TDP-43 is a hallmark of degenerative diseases, contributing to neural cell damage and central nervous system (CNS) disorders. The findings of this study on EV-D68-induced TDP-43 formation extend our understanding of virus-mediated cytotoxicity and the potential risks of TDP-43 dysfunction-related cognitive impairment and neurological symptoms in infected patients.


Assuntos
Proteínas de Ligação a DNA , Infecções por Enterovirus , Humanos , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/metabolismo , Proteínas de Ligação a DNA/farmacologia , Enterovirus Humano D , Infecções por Enterovirus/fisiopatologia , Infecções por Enterovirus/virologia , Linhagem Celular Tumoral , Proteases Virais 3C/metabolismo , Agregação Patológica de Proteínas/genética , Lopinavir/farmacologia , Proteólise/efeitos dos fármacos , Inativação Gênica , Inibidores de Proteases/farmacologia
15.
PLoS Pathog ; 18(7): e1010632, 2022 07.
Artigo em Inglês | MEDLINE | ID: mdl-35789345

RESUMO

Enteroviruses (EVs) are among the most prevalent viruses worldwide. They are characterized by a high genetic and phenotypic diversity, being able to cause a plethora of symptoms. EV-D68, a respiratory EV, and EV-D94, an enteric EV, represent an interesting paradigm of EV tropism heterogeneity. They belong to the same species, but display distinct phenotypic characteristics and in vivo tropism. Here, we used these two viruses as well as relevant 3D respiratory, intestinal and neural tissue culture models, to highlight key distinctive features of enteric and respiratory EVs. We emphasize the critical role of temperature in restricting EV-D68 tissue tropism. Using transcriptomic analysis, we underscore fundamental differences between intestinal and respiratory tissues, both in the steady-state and in response to infection. Intestinal tissues present higher cell proliferation rate and are more immunotolerant than respiratory tissues. Importantly, we highlight the different strategies applied by EV-D94 and EV-D68 towards the host antiviral response of intestinal and respiratory tissues. EV-D68 strongly activates antiviral pathways while EV-D94, on the contrary, barely induces any host defense mechanisms. In summary, our study provides an insightful characterization of the differential pathogenesis of EV-D68 and EV-D94 and the interplay with their main target tissues.


Assuntos
Enterovirus Humano D , Infecções por Enterovirus , Enterovirus , Infecções Respiratórias , Antígenos Virais , Antivirais , Enterovirus Humano D/fisiologia , Humanos , Tropismo
16.
PLoS Pathog ; 18(5): e1010515, 2022 05.
Artigo em Inglês | MEDLINE | ID: mdl-35639811

RESUMO

Worldwide outbreaks of enterovirus D68 (EV-D68) in 2014 and 2016 have caused serious respiratory and neurological disease. We collected samples from several European countries during the 2018 outbreak and determined 53 near full-length genome ('whole genome') sequences. These sequences were combined with 718 whole genome and 1,987 VP1-gene publicly available sequences. In 2018, circulating strains clustered into multiple subgroups in the B3 and A2 subclades, with different phylogenetic origins. Clusters in subclade B3 emerged from strains circulating primarily in the US and Europe in 2016, though some had deeper roots linking to Asian strains, while clusters in A2 traced back to strains detected in East Asia in 2015-2016. In 2018, all sequences from the USA formed a distinct subgroup, containing only three non-US samples. Alongside the varied origins of seasonal strains, we found that diversification of these variants begins up to 18 months prior to the first diagnostic detection during a EV-D68 season. EV-D68 displays strong signs of continuous antigenic evolution and all 2018 A2 strains had novel patterns in the putative neutralizing epitopes in the BC- and DE-loops. The pattern in the BC-loop of the USA B3 subgroup had not been detected on that continent before. Patients with EV-D68 in subclade A2 were significantly older than patients with a B3 subclade virus. In contrast to other subclades, the age distribution of A2 is distinctly bimodal and was found primarily among children and in the elderly. We hypothesize that EV-D68's rapid evolution of surface proteins, extensive diversity, and high rate of geographic mixing could be explained by substantial reinfection of adults. Better understanding of evolution and immunity across diverse viral pathogens, including EV-D68 and SARS-CoV-2, is critical to pandemic preparedness in the future.


Assuntos
COVID-19 , Enterovirus Humano D , Infecções por Enterovirus , Infecções Respiratórias , Adulto , Idoso , Criança , Demografia , Surtos de Doenças , Enterovirus Humano D/genética , Infecções por Enterovirus/epidemiologia , Humanos , Filogenia , SARS-CoV-2
17.
J Med Virol ; 96(7): e29810, 2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-39049549

RESUMO

Enterovirus D68 (EV-D68) is an emerging agent for which data on the susceptible adult population is scarce. We performed a 6-year analysis of respiratory samples from influenza-like illness (ILI) admitted during 2014-2020 in 4-10 hospitals in the Valencia Region, Spain. EV-D68 was identified in 68 (3.1%) among 2210 Enterovirus (EV)/Rhinovirus (HRV) positive samples. Phylogeny of 59 VP1 sequences showed isolates from 2014 clustering in B2 (6/12), B1 (5/12), and A2/D1 (1/12) subclades; those from 2015 (n = 1) and 2016 (n = 1) in B3 and A2/D1, respectively; and isolates from 2018 in A2/D3 (42/45), and B3 (3/45). B1 and B2 viruses were mainly detected in children (80% and 67%, respectively); B3 were equally distributed between children and adults; whereas A2/D1 and A2/D3 were observed only in adults. B3 viruses showed up to 16 amino acid changes at predicted antigenic sites. In conclusion, two EV-D68 epidemics linked to ILI hospitalized cases occurred in the Valencia Region in 2014 and 2018, with three fatal outcomes and one ICU admission. A2/D3 strains from 2018 were associated with severe respiratory infection in adults. Because of the significant impact of non-polio enteroviruses in ILI and the potential neurotropism, year-round surveillance in respiratory samples should be pursued.


Assuntos
Enterovirus Humano D , Infecções por Enterovirus , Hospitalização , Influenza Humana , Filogenia , Humanos , Espanha/epidemiologia , Infecções por Enterovirus/epidemiologia , Infecções por Enterovirus/virologia , Enterovirus Humano D/genética , Enterovirus Humano D/classificação , Enterovirus Humano D/isolamento & purificação , Criança , Adulto , Pré-Escolar , Masculino , Adolescente , Feminino , Pessoa de Meia-Idade , Lactente , Idoso , Adulto Jovem , Hospitalização/estatística & dados numéricos , Influenza Humana/epidemiologia , Influenza Humana/virologia , Infecções Respiratórias/epidemiologia , Infecções Respiratórias/virologia , Estações do Ano , Idoso de 80 Anos ou mais , Efeitos Psicossociais da Doença , Recém-Nascido
18.
J Med Virol ; 96(10): e29947, 2024 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-39370858

RESUMO

To clarify the epidemiology of enterovirus D68 (EV-D68), an enterovirus rarely identified in the 20th century, we performed seroepidemiological analysis against EV-D68 using sera collected in 1976, 1985, 1990, 1999, 2009, and 2019, as well as Yamagata isolate (EVD68/Yamagata.JPN/2023-89), in Yamagata, Japan. The neutralizing antibody (Ab)-positive rates for those under 20 years old were 61.0%, 82.5%, 84.3%, 46.7%, 50.5%, and 67.9%, in each year, whereas the rates for those above 20 years old were between 93.4% and 99.1%. Generally, geometric mean titers (GMTs)increased with age among children and the total GMT in each year was 25.4, 49.2, 37.2, 30.8, 29.5, and 33.9, from 1976 to 2019, respectively. The findings in this Yamagata-based study showed that the seroprevalence of EV-D68 over the last four decades has increased with age among children, as a susceptible group, and then reaches a plateau of over approximately 80% among adults. This study clearly revealed that EV-D68 was stably transmitted among children in the 20th century, when EV-D68 detection was quite rare.


Assuntos
Anticorpos Neutralizantes , Anticorpos Antivirais , Enterovirus Humano D , Infecções por Enterovirus , Humanos , Estudos Soroepidemiológicos , Japão/epidemiologia , Infecções por Enterovirus/epidemiologia , Infecções por Enterovirus/virologia , Criança , Enterovirus Humano D/imunologia , Pré-Escolar , Adolescente , Anticorpos Antivirais/sangue , Adulto Jovem , Adulto , Anticorpos Neutralizantes/sangue , Lactente , Feminino , Masculino , Pessoa de Meia-Idade , Idoso , Idoso de 80 Anos ou mais
19.
MMWR Morb Mortal Wkly Rep ; 73(4): 70-76, 2024 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-38300829

RESUMO

Acute flaccid myelitis (AFM) is a serious neurologic condition primarily affecting children; AFM can cause acute respiratory failure and permanent paralysis. AFM is a rare but known complication of various viral infections, particularly those of enteroviruses (EVs). Increases in AFM cases during 2014, 2016, and 2018 were associated with EV-D68 infection. This report examines trends in confirmed AFM cases during 2018-2022 and patients' clinical and laboratory characteristics. The number of AFM cases was low during 2019-2022 (28-47 cases per year); the number of cases remained low in 2022 despite evidence of increased EV-D68 circulation in the United States. Compared with cases during the most recent peak year (2018), fewer cases during 2019-2021 had upper limb involvement, prodromal respiratory or febrile illness, or cerebrospinal fluid pleocytosis, and more were associated with lower limb involvement. It is unclear why EV-D68 circulation in 2022 was not associated with an increase in AFM cases or when the next increase in AFM cases will occur. Nonetheless, clinicians should continue to suspect AFM in any child with acute flaccid limb weakness, especially those with a recent respiratory or febrile illness.


Assuntos
Viroses do Sistema Nervoso Central , Enterovirus Humano D , Infecções por Enterovirus , Mielite , Doenças Neuromusculares , Criança , Humanos , Estados Unidos/epidemiologia , Doenças Neuromusculares/epidemiologia , Paralisia , Mielite/epidemiologia , Viroses do Sistema Nervoso Central/epidemiologia , Infecções por Enterovirus/epidemiologia
20.
Euro Surveill ; 29(41)2024 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-39392006

RESUMO

We report a considerable increase in enterovirus D68 (EV-D68) cases since July 2024, culminating in an ongoing outbreak of acute respiratory infections in northern Italy, accounting for nearly 90% of all enterovirus infections. The outbreak was identified by community- and hospital-based surveillance systems, detecting EV-D68 in individuals with mild-to-severe respiratory infections. These strains belonged to B3 and a divergent A2 lineage. An increase in adult cases was observed. Enhanced surveillance and molecular characterisation of EV-D68 across Europe are needed.


Assuntos
Surtos de Doenças , Enterovirus Humano D , Infecções por Enterovirus , Infecções Respiratórias , Humanos , Infecções Respiratórias/epidemiologia , Infecções Respiratórias/virologia , Infecções Respiratórias/diagnóstico , Infecções por Enterovirus/epidemiologia , Infecções por Enterovirus/diagnóstico , Infecções por Enterovirus/virologia , Itália/epidemiologia , Enterovirus Humano D/isolamento & purificação , Enterovirus Humano D/genética , Adulto , Adolescente , Criança , Masculino , Pré-Escolar , Feminino , Pessoa de Meia-Idade , Lactente , Idoso , Adulto Jovem , Vigilância da População , Filogenia
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