Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 444
Filtrar
1.
Nat Commun ; 12(1): 5772, 2021 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-34599178

RESUMO

ISG15 is an interferon-stimulated, ubiquitin-like protein that can conjugate to substrate proteins (ISGylation) to counteract microbial infection, but the underlying mechanisms remain elusive. Here, we use a virus-like particle trapping technology to identify ISG15-binding proteins and discover Ring Finger Protein 213 (RNF213) as an ISG15 interactor and cellular sensor of ISGylated proteins. RNF213 is a poorly characterized, interferon-induced megaprotein that is frequently mutated in Moyamoya disease, a rare cerebrovascular disorder. We report that interferon induces ISGylation and oligomerization of RNF213 on lipid droplets, where it acts as a sensor for ISGylated proteins. We show that RNF213 has broad antimicrobial activity in vitro and in vivo, counteracting infection with Listeria monocytogenes, herpes simplex virus 1, human respiratory syncytial virus and coxsackievirus B3, and we observe a striking co-localization of RNF213 with intracellular bacteria. Together, our findings provide molecular insights into the ISGylation pathway and reveal RNF213 as a key antimicrobial effector.


Assuntos
Adenosina Trifosfatases/metabolismo , Anti-Infecciosos/metabolismo , Citocinas/metabolismo , Ubiquitina-Proteína Ligases/metabolismo , Ubiquitinas/metabolismo , Células A549 , Animais , Enterovirus/fisiologia , Células HEK293 , Células HeLa , Herpesvirus Humano 1/fisiologia , Humanos , Interferon Tipo I/metabolismo , Gotículas Lipídicas/metabolismo , Listeria monocytogenes/fisiologia , Masculino , Camundongos Endogâmicos C57BL , Ligação Proteica , Multimerização Proteica , Proteínas Modificadoras Pequenas Relacionadas à Ubiquitina/metabolismo , Células THP-1 , Ubiquitina/metabolismo
2.
Expert Opin Ther Targets ; 25(6): 479-489, 2021 06.
Artigo em Inglês | MEDLINE | ID: mdl-34253126

RESUMO

Introduction: Enteroviruses are common viruses causing a huge number of acute and chronic infections and producing towering economic costs. Similarly, coronaviruses cause seasonal mild infections, epidemics, and even pandemics and can lead to severe respiratory symptoms. It is important to develop broadly acting antiviral molecules to efficiently tackle the infections caused by thes.Areas covered: This review illuminates the differences and similarities between enteroviruses and coronaviruses and examines the most appealing therapeutic targets to combat both virus groups. Publications of both virus groups and deposited structures discovered through PubMed to March 2021 for viral proteases have been evaluated.Expert opinion: The main protease of coronaviruses and enteroviruses share similarities in their structure and function. These proteases process their viral polyproteins and thus drugs that bind to the active site have potential to target both virus groups. It is important to develop drugs that target more evolutionarily conserved processes and proteins. Moreover, it is a wise strategy to concentrate on processes that are similar between several virus families.


Assuntos
Antivirais/farmacologia , Coronavirus/fisiologia , Enterovirus/fisiologia , Animais , Coronavirus/efeitos dos fármacos , Coronavirus/enzimologia , Cisteína Endopeptidases/metabolismo , Enterovirus/efeitos dos fármacos , Enterovirus/enzimologia , Humanos , Especificidade por Substrato
3.
Int J Mol Sci ; 22(6)2021 Mar 17.
Artigo em Inglês | MEDLINE | ID: mdl-33802680

RESUMO

Coxsackievirus and adenovirus receptor (CAR) is present in epithelial and vascular endothelial cell junctions. We have previously shown a hemorrhagic phenotype in germ-line CAR knock-out mouse embryos; we have also found that CAR interacts with ZO-1 and ß-catenin. However, the role of CAR in vascular endothelial junction permeability has not been proven. To understand the roles of CAR in the vascular endothelial junctions, we generated endothelium-specific CAR knockout (CAR-eKO) mice. In the absence of CAR, the endothelial cell layer showed an increase in transmembrane electrical resistance (TER, Ω) and coxsackievirus permeability. Evans blue dye and 70 kDa dextran-FITC were delivered by tail vein injection. We observed increased vascular permeability in the hearts of adult CAR-eKO mice compare with wild-type (WT) mice. There was a marked increase in monocyte and macrophage penetration into the peritoneal cavity caused by thioglycolate-induced peritonitis. We found that CAR ablation in endothelial cells was not significantly increased coxsackievirus B3 (CVB3) induced myocarditis in murine model. However, tissue virus titers were significantly higher in CAR-eKO mice compared with WT. Moreover, CVB3 was detected in the brain of CAR-eKO mice. Endothelial CAR deletion affects the expression of major endothelial junction proteins, such as cadherin and platelet endothelial cell adhesion molecule-1 (PECAM-1) in the cultured endothelial cells as well as liver vessel. We suggest that CAR expression is required for normal vascular permeability and endothelial tight junction homeostasis. Furthermore, CVB3 organ penetration and myocarditis severities were dependent on the endothelial CAR level.


Assuntos
Cardiomiopatias/patologia , Cardiomiopatias/virologia , Endotélio Vascular/patologia , Endotélio Vascular/virologia , Enterovirus/fisiologia , Índice de Gravidade de Doença , Animais , Antígenos CD/metabolismo , Caderinas/metabolismo , Permeabilidade Capilar , Células Cultivadas , Proteína de Membrana Semelhante a Receptor de Coxsackie e Adenovirus/metabolismo , Células Endoteliais/metabolismo , Células Endoteliais/patologia , Células Endoteliais/virologia , Enterovirus Humano B , Deleção de Genes , Inflamação/patologia , Fígado/metabolismo , Camundongos Knockout , Miocardite/complicações , Miocardite/virologia , Cavidade Peritoneal/patologia , Molécula-1 de Adesão Celular Endotelial a Plaquetas/metabolismo , Estabilidade Proteica , Replicação Viral
4.
Emerg Microbes Infect ; 10(1): 713-724, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-33745413

RESUMO

Hand, Foot and Mouth Disease (HFMD) is usually a self-limiting, mild childhood disease that is caused mainly by Coxsackie virus A16 (CVA16) and Enterovirus A71 (EV-A71), both members of the Picornaviridae family. However, recurring HFMD outbreaks and epidemics due to EV-A71 infection in the Western Pacific region, and the propensity of EV-A71 strains to cause severe neurological complications have made this neurotropic virus a serious public health concern in afflicted countries. High mutation rate leading to viral quasispecies combined with frequent intra- and inter-typic recombination events amongst co-circulating EV-A71 strains have contributed to the great diversity and fast evolution of EV-A71 genomes, making impossible any accurate prediction of the next epidemic strain. Comparative genome sequence analyses and mutagenesis approaches have led to the identification of a number of viral determinants involved in EV-A71 fitness and virulence. These viral determinants include amino acid residues located in the structural proteins of the virus, affecting attachment to the host cell surface, receptor binding, and uncoating events. Critical residues in non-structural proteins have also been identified, including 2C, 3A, 3C proteases and the RNA-dependent RNA polymerase. Finally, mutations altering key secondary structures in the 5' untranslated region were also found to influence EV-A71 fitness and virulence. While our current understanding of EV-A71 pathogenesis remains fragmented, these studies may help in the rational design of effective treatments and broadly protective vaccine candidates.


Assuntos
Infecções por Enterovirus/virologia , Enterovirus/fisiologia , Enterovirus/patogenicidade , Animais , Enterovirus/genética , Infecções por Enterovirus/microbiologia , Genoma Viral , Humanos , Mutação , Proteínas Virais/genética , Proteínas Virais/metabolismo , Virulência , Ligação Viral
5.
Int J Infect Dis ; 106: 36-40, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33771675

RESUMO

BACKGROUND: Acute Flaccid Paralyses Surveillance (AFPS) monitors the emergence of polioviruses and can track Non-Polio Enteroviruses (NPEVs). We report AFPS activity in the Lombardy region (Northern Italy) from 2016 to 2018. METHODS: Fecal and respiratory samples were collected from children <15 years who met the WHO definition of an AFP case, analyzed by virus isolation in cell cultures (RD/L20B) and by a one-step real-time RT-PCR assay specific for the 5'-noncoding-region of NPEV. NPEV-positive specimens were further analyzed by sequencing a fragment of the VP1 gene. RESULTS: 36 AFP cases (89 stool and 32 respiratory samples) were reported with an incidence of 1.1/100'000, 0.9/100'000, 0.6/100'000 children <15 years in 2016, 2017, 2018, respectively. Poliovirus was not identified, whereas NPEVs were detected in 19.4% (7/36) of AFP cases. The presence of one Echovirus-25 (2016), two EV- and D68 (2016 and 2018), one EV-A71 (2016), and one Echovirus-30 (2016) sharing high nucleotide identity with NPEVs detected in Europe was identified. CONCLUSION: The absence of polio was confirmed. The unpredicted detection of emerging EV-D68, EV-A71, and E-30 sharing high sequence nucleotide similarity with viruses involved in the latest outbreaks, provided valuable and up-to-date information, emphasizing the importance of monitoring NPEVs through AFPS.


Assuntos
Viroses do Sistema Nervoso Central/epidemiologia , Viroses do Sistema Nervoso Central/virologia , Enterovirus/isolamento & purificação , Enterovirus/fisiologia , Monitoramento Epidemiológico , Mielite/epidemiologia , Mielite/virologia , Doenças Neuromusculares/epidemiologia , Doenças Neuromusculares/virologia , Criança , Pré-Escolar , Fezes/virologia , Feminino , Humanos , Incidência , Lactente , Itália/epidemiologia , Masculino
6.
Arch Virol ; 166(4): 1203-1211, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33606112

RESUMO

Lactoferrin is part of the innate immune system, with antiviral activity against numerous DNA and RNA viruses. Rhinoviruses, the leading cause of the common cold, are associated with exacerbation of respiratory illnesses such as asthma. Here, we explored the effect of bovine lactoferrin (BLf) on RV-B14 infectivity. Using different assays, we show that the effect of BLf is strongest during adhesion of the virus to the cell and entry. Tracking the internalisation of BLf and virus revealed a degree of colocalisation, although their interaction was only confirmed in vitro using empty viral particles, indicating a possible additional influence of BLf on other infection steps.


Assuntos
Antivirais/farmacologia , Enterovirus/efeitos dos fármacos , Lactoferrina/farmacologia , Internalização do Vírus/efeitos dos fármacos , Antivirais/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Efeito Citopatogênico Viral/efeitos dos fármacos , Enterovirus/fisiologia , Infecções por Enterovirus/virologia , Células HeLa , Humanos , Lactoferrina/metabolismo , Ligação Viral/efeitos dos fármacos
7.
Food Environ Virol ; 13(1): 84-92, 2021 03.
Artigo em Inglês | MEDLINE | ID: mdl-33392927

RESUMO

This study investigated the influence of viral interference on the detection of enteric viruses using the integrated cell culture (ICC)-PCR with a BGM cell line. It was possible to detect 102 plaque-forming units (PFU)/flask of enterovirus 71 (EV71) in spite of the presence of 104 PFU/flask of adenovirus 40 (AdV40). Meanwhile, 104 PFU/flask of AdV40 was not detected in the presence of 102 PFU/flask of EV71. This inhibition of AdV40 detection using ICC-PCR was attributable to the growth of EV71, because the addition of a growth inhibitor of EV71 (rupintrivir) neutralized the detection inhibition of AdV40. The growth inhibition of AdV40 under co-infection with EV71 is probably caused by the immune responses of EV71-infected cells. AdV is frequently used as a fecal contamination indicator of environmental water, but this study demonstrated that false-negative detection of infectious AdV using ICC-PCR could be caused by the co-existence of infectious EV in a water sample. The addition of rupintrivir could prevent false-negative detection of AdV using ICC-PCR. This study, therefore, emphasizes the importance of confirming the presence of multiple enteric viruses in a sample derived from environmental water prior to the application of ICC-PCR because the viral interference phenomenon may lead to the false-negative detection of target viruses.


Assuntos
Infecções por Adenoviridae/virologia , Adenoviridae/fisiologia , Interferência Viral , Adenoviridae/classificação , Adenoviridae/genética , Adenoviridae/isolamento & purificação , Técnicas de Cultura de Células , Linhagem Celular , Técnicas de Cocultura , Enterovirus/genética , Enterovirus/crescimento & desenvolvimento , Enterovirus/fisiologia , Humanos , Reação em Cadeia da Polimerase
8.
Viruses ; 13(1)2021 Jan 13.
Artigo em Inglês | MEDLINE | ID: mdl-33451106

RESUMO

Inflammatory bowel diseases (IBD), including ulcerative colitis (UC) and Crohn's disease (CD), is a multifactorial disease in which dietary, genetic, immunological, and microbial factors are at play. The role of enteric viruses in IBD remains only partially explored. To date, epidemiological studies have not fully described the role of enteric viruses in inflammatory flare-ups, especially that of human noroviruses and rotaviruses, which are the main causative agents of viral gastroenteritis. Genome-wide association studies have demonstrated the association between IBD, polymorphisms of the FUT2 and FUT3 genes (which drive the synthesis of histo-blood group antigens), and ligands for norovirus and rotavirus in the intestine. The role of autophagy in defensin-deficient Paneth cells and the perturbations of cytokine secretion in T-helper 1 and T-helper 17 inflammatory pathways following enteric virus infections have been demonstrated as well. Enteric virus interactions with commensal bacteria could play a significant role in the modulation of enteric virus infections in IBD. Based on the currently incomplete knowledge of the complex phenomena underlying IBD pathogenesis, future studies using multi-sampling and data integration combined with new techniques such as human intestinal enteroids could help to decipher the role of enteric viruses in IBD.


Assuntos
Suscetibilidade a Doenças , Infecções por Enterovirus/complicações , Infecções por Enterovirus/virologia , Enterovirus/fisiologia , Interações Hospedeiro-Patógeno , Doenças Inflamatórias Intestinais/etiologia , Animais , Autofagia , Biomarcadores , Antígenos de Grupos Sanguíneos/genética , Antígenos de Grupos Sanguíneos/imunologia , Gerenciamento Clínico , Infecções por Enterovirus/epidemiologia , Microbioma Gastrointestinal , Humanos , Doenças Inflamatórias Intestinais/diagnóstico , Doenças Inflamatórias Intestinais/epidemiologia , Doenças Inflamatórias Intestinais/terapia , Interações Microbianas , Transdução de Sinais , Viroma
9.
Viruses ; 13(2)2021 01 22.
Artigo em Inglês | MEDLINE | ID: mdl-33499355

RESUMO

Enteroviruses are among the most common human infectious agents. While infections are often mild, the severe neuropathogenesis associated with recent outbreaks of emerging non-polio enteroviruses, such as EV-A71 and EV-D68, highlights their continuing threat to public health. In recent years, our understanding of how non-polio enteroviruses co-opt cellular pathways has greatly increased, revealing intricate host-virus relationships. In this review, we focus on newly identified mechanisms by which enteroviruses hijack the cellular machinery to promote their replication and spread, and address their potential for the development of host-directed therapeutics. Specifically, we discuss newly identified cellular receptors and their contribution to neurotropism and spread, host factors required for viral entry and replication, and recent insights into lipid acquisition and replication organelle biogenesis. The comprehensive knowledge of common cellular pathways required by enteroviruses could expose vulnerabilities amenable for host-directed therapeutics against a broad spectrum of enteroviruses. Since this will likely include newly arising strains, it will better prepare us for future epidemics. Moreover, identifying host proteins specific to neurovirulent strains may allow us to better understand factors contributing to the neurotropism of these viruses.


Assuntos
Viroses do Sistema Nervoso Central/virologia , Sistema Nervoso Central/virologia , Infecções por Enterovirus/virologia , Enterovirus/patogenicidade , Tropismo Viral , Animais , Autofagia , Enterovirus/genética , Enterovirus/fisiologia , Genoma Viral , Interações Hospedeiro-Patógeno , Humanos , Sítios Internos de Entrada Ribossomal , Fosfolipídeos/biossíntese , Biossíntese de Proteínas , RNA Viral/biossíntese , Receptores Virais/metabolismo , Compartimentos de Replicação Viral/fisiologia , Compartimentos de Replicação Viral/ultraestrutura , Internalização do Vírus , Replicação Viral
10.
Vet Microbiol ; 252: 108921, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33191001

RESUMO

Coxsackievirus A16 (CA16) is one of predominant Enterovirus that possesses high pathogenicity. Lipid rafts, as cholesterol - and sphingolipid - enriched membrane nanodomains, are involved into many aspects of the virus life cycle. Our previous study found that lipid rafts integrity was essential for CA16 replication, but how lipid rafts regulate CA16 replication through activating downstream signaling remains largely unknown. Thus, in this study, we revealed that lipid rafts were required for activation of PI3K/Akt signaling at early stage of CA16 infection. Treatment with wortmannin significantly reduced the expression of virus protein, indicating PI3K/Akt signaling was beneficial for early stage of virus infection. In addition, lipid rafts integrity was also indispensable for PI3K/Akt activation during the late stage of CA16 infection, which played critical functions in mediating sterol regulatory element-binding proteins 1 (SREBP1) maturation. Whereas, over-expression of SREBP1 exhibited inhibition on virus replication, suggesting that PI3K/Akt signaling and SREBP1 might positively and negatively influence virus replication in two different stages of infection, respectively. Taken together, our study demonstrates an important role of the lipid raft-associated PI3K/Akt/SREBP1 signaling in modulating CA16 replication, which will deepen our understanding mechanism of CA16 infection.


Assuntos
Infecções por Coxsackievirus/veterinária , Enterovirus/fisiologia , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de Sinais , Proteína de Ligação a Elemento Regulador de Esterol 1/metabolismo , Replicação Viral , Infecções por Coxsackievirus/virologia , Microdomínios da Membrana/metabolismo , Fosfatidilinositol 3-Quinases/genética , Proteína de Ligação a Elemento Regulador de Esterol 1/genética
11.
Emerg Infect Dis ; 27(1)2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-33261719

RESUMO

In Taiwan, lower nonpolio enterovirus activity during the coronavirus disease pandemic in 2020 compared with 2014-2019 might be attributable to adherence to nonpharmaceutical interventions. The preventable fraction among unexposed persons indicated that 90% of nonpolio enterovirus activity might have been prevented during 2014-2019 by adopting the same measures enforced in 2020.


Assuntos
COVID-19/epidemiologia , Infecções por Enterovirus/epidemiologia , Enterovirus/fisiologia , SARS-CoV-2 , Adolescente , Criança , Pré-Escolar , Humanos , Lactente , Taiwan/epidemiologia
12.
Biomed Res Int ; 2020: 9850351, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33274234

RESUMO

Background: Scarce information exists about immunity to hand, foot, and mouth disease (HFMD) among household contacts of index cases in Vietnam and what that means for reducing ongoing HFMD transmission in the community. Methods: We analyzed neutralizing antibodies (NT) and the incidence of enterovirus (EVs) infection among household contacts of index cases in a province where HFMD remains endemic. Throat swab and 2 mL blood samples from household contacts were collected at enrollment, during and after 2 weeks follow-up. Results: The incidence of EV-A71 infection among household contacts was 40/84 (47.6%, 95% Cl: 36.9-58.3%), compared with 106/336 (31.5%, 95% Cl: 26.6-36.5%) for CV-A6 and 36/107 (33.6%, 95% Cl: 24.7-42.6%) for CV-A16. The incidence of CV-A6 infection was fairly constant across ages; in contrast, CV-A71 and CV-A16 had some variation across ages. At baseline, higher geometric mean titer (GMT) of EV-A71, CV-A6, and CV-A16 antibody titers was found for 25-34-year groups (range 216.3 to 305.0) compared to the other age groups. There was a statistically significant difference in GMT values of CV-A6 and CV-A16 between those who had an infection or did not have infection among households with an index case of these serotypes. Conclusions: Our results indicated that adults were becoming infected with HFMD and could be contributing to the transmission. There is, therefore, a need for considering the household setting as an additional target for intervention programs for HFMD.


Assuntos
Infecções por Coxsackievirus/epidemiologia , Infecções por Coxsackievirus/virologia , Enterovirus Humano A/fisiologia , Infecções por Enterovirus/epidemiologia , Infecções por Enterovirus/virologia , Enterovirus/fisiologia , Características da Família , Adolescente , Adulto , Fatores Etários , Anticorpos Neutralizantes , Criança , Pré-Escolar , Infecções por Coxsackievirus/imunologia , Enterovirus/imunologia , Enterovirus Humano A/imunologia , Infecções por Enterovirus/imunologia , Seguimentos , Humanos , Incidência , Pessoa de Meia-Idade , Estudos Soroepidemiológicos , Sorogrupo , Vietnã/epidemiologia , Carga Viral , Adulto Jovem
13.
PLoS Pathog ; 16(12): e1009146, 2020 12.
Artigo em Inglês | MEDLINE | ID: mdl-33370422

RESUMO

Picornaviruses are important viral pathogens, but despite extensive study, the assembly process of their infectious virions is still incompletely understood, preventing the development of anti-viral strategies targeting this essential part of the life cycle. We report the identification, via RNA SELEX and bioinformatics, of multiple RNA sites across the genome of a typical enterovirus, enterovirus-E (EV-E), that each have affinity for the cognate viral capsid protein (CP) capsomer. Many of these sites are evolutionarily conserved across known EV-E variants, suggesting they play essential functional roles. Cryo-electron microscopy was used to reconstruct the EV-E particle at ~2.2 Å resolution, revealing extensive density for the genomic RNA. Relaxing the imposed symmetry within the reconstructed particles reveals multiple RNA-CP contacts, a first for any picornavirus. Conservative mutagenesis of the individual RNA-contacting amino acid side chains in EV-E, many of which are conserved across the enterovirus family including poliovirus, is lethal but does not interfere with replication or translation. Anti-EV-E and anti-poliovirus aptamers share sequence similarities with sites distributed across the poliovirus genome. These data are consistent with the hypothesis that these RNA-CP contacts are RNA Packaging Signals (PSs) that play vital roles in assembly and suggest that the RNA PSs are evolutionarily conserved between pathogens within the family, augmenting the current protein-only assembly paradigm for this family of viruses.


Assuntos
Proteínas do Capsídeo/metabolismo , Enterovirus/fisiologia , RNA Viral/genética , Montagem de Vírus/fisiologia , Sequência de Aminoácidos , Proteínas do Capsídeo/ultraestrutura , Microscopia Crioeletrônica , Enterovirus/ultraestrutura , RNA Viral/ultraestrutura
14.
mBio ; 11(6)2020 11 17.
Artigo em Inglês | MEDLINE | ID: mdl-33203755

RESUMO

Enteroviruses infect gastrointestinal epithelium cells, cause multiple human diseases, and present public health risks worldwide. However, the mechanisms underlying host immune responses in intestinal mucosa against the early enterovirus infections remain elusive. Here, we showed that human enteroviruses including enterovirus 71 (EV71), coxsackievirus B3 (CVB3), and poliovirus 1 (PV1) predominantly induce type III interferons (IFN-λ1 and IFN-λ2/3), rather than type I interferons (IFN-α and IFN-ß), in cultured human normal and cancerous intestine epithelial cells (IECs), mouse intestine tissues, and human clinical intestine specimens. Mechanistic studies demonstrated that IFN-λ production is induced upon enterovirus infection through the Toll-like receptor 3/interferon regulatory factor 1 (TLR3/IRF1) signaling pathway in IECs. In turn, the supplementation of IFN-λ subsequently induces intrinsically antiviral responses against enterovirus replication. Notably, intraperitoneal injection in neonatal C57BL/6J mice with mouse recombinant IFN-λ2 protein represses EV71 replication and protects mice from viral lethal effects. Altogether, these results revealed a distinct mechanism by which the host elicited immune responses against enterovirus infections in intestine through activating the TLR3/IRF1/type III IFN axis. The new findings would provide an antiviral strategy for the prevention and treatment of enterovirus infections and associated diseases.IMPORTANCE Enterovirus infections are significant sources of human diseases and public health risks worldwide, but little is known about the mechanism of innate immune response in host intestine epithelial surface during the viral replication. We reported the epithelial immune response in cultured human normal and cancerous cells (IECs), mouse tissues, and human clinical intestine specimens following infection with enterovirus 71. The results mechanistically revealed type III interferons (IFN-λ1 and IFN-λ2/3), rather than type I interferons (IFN-α and IFN-ß), as the dominant production through TLR3/IRF1 signaling upon multiple human enterovirus infection, including enterovirus 71 (EV71), coxsackievirus B3 (CVB3), and poliovirus 1 (PV1). IFN-λ subsequently induced antiviral activity against enterovirus replication in vitro and in vivo. These studies uncovered the role of the novel process of type III IFN production involved in the TLR3/IRF1 pathway in host intestine upon enterovirus infection, which highlighted a regulatory manner of antiviral defense in intestine during enterovirus infection.


Assuntos
Infecções por Enterovirus/imunologia , Enterovirus/imunologia , Imunidade Inata , Fator Regulador 1 de Interferon/metabolismo , Interferons/metabolismo , Receptor 3 Toll-Like/metabolismo , Animais , Enterovirus/genética , Enterovirus/fisiologia , Infecções por Enterovirus/virologia , Feminino , Humanos , Fator Regulador 1 de Interferon/genética , Interferons/genética , Intestinos/imunologia , Intestinos/virologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Transdução de Sinais , Receptor 3 Toll-Like/genética , Replicação Viral
15.
Commun Biol ; 3(1): 580, 2020 10 16.
Artigo em Inglês | MEDLINE | ID: mdl-33067530

RESUMO

Previous research suggests that hepatocytes catabolize chemical toxins but do not remove microbial agents, which are filtered out by other liver cells (Kupffer cells and endothelial cells). Here we show that, contrary to current understanding, hepatocytes trap and rapidly silence type B coxsackieviruses (CVBs). In genetically wildtype mice, this activity causes hepatocyte damage, which is alleviated in mice carrying a hepatocyte-specific deletion of the coxsackievirus-adenovirus receptor. However, in these mutant mice, there is a dramatic early rise in blood-borne virus, followed by accelerated systemic disease and increased mortality. Thus, wild type hepatocytes act similarly to a sponge for CVBs, protecting against systemic illness at the expense of their own survival. We speculate that hepatocytes may play a similar role in other viral infections as well, thereby explaining why hepatocytes have evolved their remarkable regenerative capacity. Our data also suggest that, in addition to their many other functions, hepatocytes might be considered an integral part of the innate immune system.


Assuntos
Infecções por Coxsackievirus/virologia , Resistência à Doença , Enterovirus/fisiologia , Hepatócitos/metabolismo , Hepatócitos/virologia , Interações Hospedeiro-Patógeno , Animais , Proteína de Membrana Semelhante a Receptor de Coxsackie e Adenovirus/deficiência , Infecções por Coxsackievirus/genética , Infecções por Coxsackievirus/imunologia , Infecções por Coxsackievirus/patologia , Células Dendríticas/imunologia , Células Dendríticas/metabolismo , Modelos Animais de Doenças , Resistência à Doença/genética , Resistência à Doença/imunologia , Expressão Gênica , Interações Hospedeiro-Patógeno/genética , Interações Hospedeiro-Patógeno/imunologia , Imunidade Inata , Interferon-alfa/metabolismo , Fígado/metabolismo , Fígado/patologia , Fígado/virologia , Camundongos , Camundongos Knockout , Mortalidade , Carga Viral , Viremia
16.
J Virol ; 95(2)2020 12 22.
Artigo em Inglês | MEDLINE | ID: mdl-33087467

RESUMO

Enterovirus replication requires the cellular protein GBF1, a guanine nucleotide exchange factor for small Arf GTPases. When activated, Arfs associate with membranes, where they regulate numerous steps of membrane homeostasis. The requirement for GBF1 implies that Arfs are important for replication, but which of the different Arfs function(s) during replication remains poorly understood. Here, we established cell lines expressing each of the human Arfs fused to a fluorescent tag and investigated their behavior during enterovirus infection. Arf1 was the first to be recruited to the replication organelles, where it strongly colocalized with the viral antigen 2B and mature virions but not double-stranded RNA. By the end of the infectious cycle, Arf3, Arf4, Arf5, and Arf6 were also concentrated on the replication organelles. Once on the replication membranes, all Arfs except Arf3 were no longer sensitive to inhibition of GBF1, suggesting that in infected cells they do not actively cycle between GTP- and GDP-bound states. Only the depletion of Arf1, but not other class 1 and 2 Arfs, significantly increased the sensitivity of replication to GBF1 inhibition. Surprisingly, depletion of Arf6, a class 3 Arf, normally implicated in plasma membrane events, also increased the sensitivity to GBF1 inhibition. Together, our results suggest that GBF1-dependent Arf1 activation directly supports the development and/or functioning of the replication complexes and that Arf6 plays a previously unappreciated role in viral replication. Our data reveal a complex pattern of Arf activation in enterovirus-infected cells that may contribute to the resilience of viral replication in different cellular environments.IMPORTANCE Enteroviruses include many known and emerging pathogens, such as poliovirus, enteroviruses 71 and D68, and others. However, licensed vaccines are available only against poliovirus and enterovirus 71, and specific anti-enterovirus therapeutics are lacking. Enterovirus infection induces the massive remodeling of intracellular membranes and the development of specialized domains harboring viral replication complexes, replication organelles. Here, we investigated the roles of small Arf GTPases during enterovirus infection. Arfs control distinct steps in intracellular membrane traffic, and one of the Arf-activating proteins, GBF1, is a cellular factor required for enterovirus replication. We found that all Arfs expressed in human cells, including Arf6, normally associated with the plasma membrane, are recruited to the replication organelles and that Arf1 appears to be the most important Arf for enterovirus replication. These results document the rewiring of the cellular membrane pathways in infected cells and may provide new ways of controlling enterovirus infections.


Assuntos
Fatores de Ribosilação do ADP/metabolismo , Infecções por Enterovirus/metabolismo , Enterovirus/fisiologia , Compartimentos de Replicação Viral/metabolismo , Fatores de Ribosilação do ADP/genética , Antígenos Virais/metabolismo , Enterovirus/classificação , Infecções por Enterovirus/virologia , Fatores de Troca do Nucleotídeo Guanina/genética , Fatores de Troca do Nucleotídeo Guanina/metabolismo , Células HeLa , Humanos , Membranas Intracelulares/metabolismo , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , RNA Viral/metabolismo , Replicação Viral
17.
PLoS One ; 15(8): e0238080, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32834009

RESUMO

The purpose of this study was to evaluate potential insights into the pathogenesis of acute posterior multifocal placoid pigment epitheliopathy (APMPPE) using multimodal diagnostic imaging and laboratory evaluation in long-term follow-up. A retrospective, single-center case series was conducted on seven consecutive patients (14 eyes) who were given a diagnosis of APMPPE from March 1, 2011, through June 30, 2019 with at least three months of follow-up. Clinical characteristics (age, symptoms, visual acuity [VA]), laboratory testing including coxsackievirus titers, and multimodal imaging from fundus photography, spectral-domain optical coherence tomography (SD-OCT), fundus autofluorescence (FAF), fluorescein angiography (FA), and indocyanine green angiography (ICG) were analyzed for each patient. The initial median VA was 20/71 and final median VA was 20/22. Coxsackievirus B (CVB) titers were elevated (≥ 1:80) in six of seven patients, with a four-fold increase in convalescent titers seen in two patients suggestive of recent infection. All patients were treated with oral corticosteroids, and five patients underwent corticosteroid-sparing immunomodulatory therapy. Initially, multifocal deep choroidal lesions were observed in the posterior pole corresponding to patches of hypocyanescence on ICG. Overlying retinal pigment epithelium (RPE) disease was observed on FAF, although this finding was not universally observed, suggesting that RPE disease may occur as a sequelae to unchecked choroidal inflammation. SD-OCT architectural changes confirmed outer retina and ellipsoid zone disruption. FA of active lesions showed early hypofluorescence and late hyperfluorescence with surrounding leakage while inactive disease showed areas of staining. Long-term follow-up of multimodal diagnostic imaging in APMPPE revealed that choroidal inflammation likely precedes RPE change and photoreceptor damage. Elevation of coxsackievirus titers with seroconversion may be associated with an infectious trigger in concert with immune-mediated disease in this posterior uveitis syndrome.


Assuntos
Enterovirus/fisiologia , Exposição Ambiental/efeitos adversos , Coroidite Multifocal/diagnóstico por imagem , Coroidite Multifocal/virologia , Imagem Multimodal , Doenças Retinianas/diagnóstico por imagem , Doenças Retinianas/virologia , Doença Aguda , Adolescente , Adulto , Idoso , Angiografia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Imagem Óptica , Estudos Retrospectivos , Tomografia de Coerência Óptica , Adulto Jovem
18.
Nat Commun ; 11(1): 4332, 2020 08 28.
Artigo em Inglês | MEDLINE | ID: mdl-32859902

RESUMO

The group of enteroviruses contains many important pathogens for humans, including poliovirus, coxsackievirus, rhinovirus, as well as newly emerging global health threats such as EV-A71 and EV-D68. Here, we describe an unbiased, system-wide and time-resolved analysis of the proteome and phosphoproteome of human cells infected with coxsackievirus B3. Of the ~3,200 proteins quantified throughout the time course, a large amount (~25%) shows a significant change, with the majority being downregulated. We find ~85% of the detected phosphosites to be significantly regulated, implying that most changes occur at the post-translational level. Kinase-motif analysis reveals temporal activation patterns of certain protein kinases, with several CDKs/MAPKs immediately active upon the infection, and basophilic kinases, ATM, and ATR engaging later. Through bioinformatics analysis and dedicated experiments, we identify mTORC1 signalling as a major regulation network during enterovirus infection. We demonstrate that inhibition of mTORC1 activates TFEB, which increases expression of lysosomal and autophagosomal genes, and that TFEB activation facilitates the release of virions in extracellular vesicles via secretory autophagy. Our study provides a rich framework for a system-level understanding of enterovirus-induced perturbations at the protein and signalling pathway levels, forming a base for the development of pharmacological inhibitors to treat enterovirus infections.


Assuntos
Infecções por Coxsackievirus/metabolismo , Interações Hospedeiro-Patógeno/fisiologia , Proteoma/análise , Animais , Autofagia , Fatores de Transcrição de Zíper de Leucina e Hélice-Alça-Hélix Básicos/genética , Fatores de Transcrição de Zíper de Leucina e Hélice-Alça-Hélix Básicos/metabolismo , Linhagem Celular , Sobrevivência Celular , Enterovirus/fisiologia , Enterovirus Humano B/fisiologia , Técnicas de Inativação de Genes , Células HeLa , Humanos , Alvo Mecanístico do Complexo 1 de Rapamicina , Fosforilação , Transdução de Sinais , Proteínas Virais/metabolismo
19.
Curr Opin Virol ; 40: 55-60, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-32711392

RESUMO

Recent outbreaks of limb paralysis similar to poliomyelitis, termed acute flaccid myelitis (AFM), have prompted intense investigation into potential etiology. Peaks of AFM were seen in the United States in 2012, 2014, 2016 and 2018, coincident with peaks in enterovirus transmission, particularly EV-D68. Similar peaks of AFM and EV-D68 circulation were reported in other parts of the world. The causal relationship between EV-D68 is still not widely accepted as it is for poliovirus and EV-A71, the latter of which is endemic in the US. Recent in vitro and mouse model data as well as enhanced-sensitivity diagnostic assays have provided further evidence linking the causal relationship between EV-D68 and AFM. In addition, an outbreak of EV-A71-associated AFM was recently described, highlighting the possibility of an additional emerging non-polio enterovirus of public health concern. As AFM is a devastating disease with poor prognosis in many children, particularly those with EV-D68, recent studies call for increased surveillance, pursuit of novel therapeutics and strategies to prevent transmission before the next outbreak.


Assuntos
Viroses do Sistema Nervoso Central/virologia , Infecções por Enterovirus/virologia , Enterovirus/fisiologia , Mielite/virologia , Doenças Neuromusculares/virologia , Animais , Viroses do Sistema Nervoso Central/epidemiologia , Surtos de Doenças , Enterovirus/classificação , Enterovirus/genética , Enterovirus/isolamento & purificação , Infecções por Enterovirus/epidemiologia , Humanos , Mielite/epidemiologia , Doenças Neuromusculares/epidemiologia , Estados Unidos/epidemiologia
20.
Medicine (Baltimore) ; 99(29): e21258, 2020 Jul 17.
Artigo em Inglês | MEDLINE | ID: mdl-32702909

RESUMO

INTRODUCTION: Hand, foot, and mouth disease (HFMD) has been an important public health concern worldwide, especially in the Asia-Pacific region. Unfortunately, the effect of current measures on preventing and controlling HFMD may be limited. Isolation of infectious sources is reported as an important way to prevent and control this disease. The isolation period is determined on the basis of duration of viral shedding in patients with HFMD. However, the results of previous researches on duration of viral shedding remain controversial. Here, we present a protocol of a systematic review and single-arm meta-analysis for assessing the duration of viral shedding in patients with HFMD induced by Enterovirus 71 (EV71), Coxsackievirus A16 (CA16), or Coxsackievirus A6 (CA6). METHODS AND ANALYSIS: A comprehensive literature search will be performed in PubMed, EMBASE, the Cochrane library, Chinese National Knowledge Infrastructure (CNKI), Chinese Biomedical Literature Database (CBM), and Wanfang Database, covering the period from inception to May 1, 2019. Point estimate of positive rate with corresponding 95% confidence intervals (CIs) of EV71, CA16, or CA6 in HFMD patients' fecal or throat samples will be carried out using STATA 14.0. Subgroup analyses will be performed for mild cases, severe cases, and close contacts. Sensitive analysis will also be performed to evaluate the influences of individual studies on the final effect by exclusion of a few articles of poor quality. We will assess the risk of bias for the final studies included in our meta-analysis using previously available tools and the modified risk of bias tool. RESULTS: The results of this systematic review and meta-analysis will be published in a peer-reviewed journal. CONCLUSION: To the best of our knowledge, this paper will be the first systematic review and meta-analysis for assessing the duration of viral shedding in patients with HFMD induced by EV71, CA16, or CA6. The conclusions drawn from this review will provide the scientific basis to formulate the isolation period of HFMD. ETHICS AND DISSEMINATION: Ethical review is not required as this article is for a systematic review since there is no direct involvement of patients in the whole process. We will publish the results of this systematic review and meta-analysis of single-arm studies in a peer-reviewed journal. REGISTRATION NUMBER: Prospero CRD42020139999.


Assuntos
Enterovirus Humano A/fisiologia , Enterovirus/fisiologia , Doença de Mão, Pé e Boca/virologia , Eliminação de Partículas Virais , Humanos , Fatores de Tempo
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...