RESUMO
BACKGROUND AND AIMS: Being the most common cause of acute viral hepatitis with >20 million cases per year and 70,000 deaths annually, HEV presents a long-neglected and underinvestigated health burden. Although the entry process of viral particles is an attractive target for pharmacological intervention, druggable host factors to restrict HEV entry have not been identified so far. APPROACH AND RESULTS: Here we identify the EGF receptor (EGFR) as a novel host factor for HEV and reveal the significance of EGFR for the HEV entry process. By utilizing RNAi, chemical modulation with Food and Drug Administration-approved drugs, and ectopic expression of EGFR, we revealed that EGFR is critical for HEV infection without affecting HEV RNA replication or assembly of progeny virus. We further unveiled that EGFR itself and its ligand-binding domain, rather than its signaling function, is responsible for the proviral effect. Modulation of EGF expression in HepaRG cells and primary human hepatocytes affected HEV infection. CONCLUSIONS: Taken together, our study provides novel insights into the life cycle of HEV and identified EGFR as a possible target for future antiviral strategies against HEV.
Assuntos
Vírus da Hepatite E , Hepatócitos , Humanos , Hepatócitos/metabolismo , Antivirais/farmacologia , Receptores ErbB/metabolismo , Interferência de RNA , Transdução de Sinais , Vírus da Hepatite E/genética , Replicação ViralRESUMO
BACKGROUND AND AIMS: Severe acute respiratory syndrome coronavirus (SARS-CoV-2) preferentially infects the respiratory tract; however, several studies have implicated a multi-organ involvement. Hepatic dysfunctions caused by SARS-CoV-2 infection have been increasingly recognized and described to correlate with disease severity. To elucidate molecular factors that could contribute towards hepatic infection, we concentrated on microRNAs (miRNAs), a class of small non-coding RNAs that modulate various cellular processes and which are reported to be differentially regulated during liver injury. We aimed to study the infection of primary human hepatocytes (PHH) with SARS-CoV-2 and to evaluate the potential of miRNAs for modulating viral infection. METHODS: We analysed liver autopsies from a coronavirus disease 19 (COVID-19)-positive cohort for the presence of viral RNA using Nanopore sequencing. PHH were used for the infection with SARS-CoV-2. The candidate miRNAs targeting angiotensin converting enzyme 2 (ACE2) and transmembrane serine protease 2 (TMPRSS2) were identified using in silico approaches. To discover the potential regulatory mechanism, transfection experiments, qRT-PCRs, western blots and luciferase reporter assays were performed. RESULTS: We could detect SARS-CoV-2 RNA in COVID-19-positive liver autopsies. We show that PHH express ACE2 and TMPRSS2 and can be readily infected with SARS-CoV-2, resulting in robust replication. Transfection of selected miRNA mimics reduced SARS-CoV-2 receptor expression and SARS-CoV-2 burden in PHH. In silico and biochemical analyses supported a potential direct binding of miR-141-3p to the SARS-CoV-2 genome. CONCLUSION: We confirm that PHH are susceptible to SARS-CoV-2 infection and demonstrate selected miRNAs targeting SARS-CoV-2 entry factors and/or the viral genome reduce viral loads. These data provide novel insights into hepatic susceptibility to SARS-CoV-2 and associated dysfunctions in COVID-19.
RESUMO
AIMS: We aimed to develop a method to assess the virucidal performance of domestic laundry in a lab-scale washing machine (Rotawash) based on EN 17658. METHODS AND RESULTS: For method development, virus recovery was investigated after drying on cotton carriers for three test viruses murine norovirus (MNV), modified vaccinia virus Ankara (MVA), and bovine coronavirus (BCoV), followed by washing simulations in flasks and Rotawash. MNV and MVA demonstrated sufficient recovery from carriers after drying and washing (up to 40°C and 60 min). BCoV exhibited lower recovery, indicating less relevance as a test virus. Rotawash efficacy tests conducted with MNV, a resistant, non-enveloped virus, showed limited efficacy of a bleach-free detergent, aligning with results from a domestic washing machine. Rotawash washes achieved higher reductions in infectious virus titers than suspension tests, indicating the role of washing mechanics in virus removal. CONCLUSIONS: This study established a practical method to test the virucidal efficacy of laundry detergents in Rotawash, simulating domestic washing.
Assuntos
Detergentes , Norovirus , Bovinos , Animais , Camundongos , Detergentes/farmacologia , TêxteisRESUMO
Increasing nonzoonotic human monkeypox virus (MPXV) infections urge reevaluation of inactivation strategies. We demonstrate efficient inactivation of MPXV by 2 World Health Organizationârecommended alcohol-based hand rub solutions. When compared with other (re)emerging enveloped viruses, MPXV displayed the greatest stability. Our results support rigorous adherence to use of alcohol-based disinfectants.
Assuntos
Desinfetantes , Mpox , Vírus , Humanos , Monkeypox virus , Desinfetantes/farmacologia , Etanol , Mpox/epidemiologia , Mpox/prevenção & controle , 2-Propanol , Organização Mundial da SaúdeRESUMO
BACKGROUND: A proportion of the convalescent SARS-CoV-2 pediatric population presents nonspecific symptoms, mental health problems, and a reduction in quality of life similar to myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) and long COVID-19 symptomatic. However, data regarding its clinical manifestation and immune mechanisms are currently scarce. METHODS: In this study, we perform a comprehensive clinical and immunological profiling of 17 convalescent COVID-19 children with post-acute COVID-19 sequelae (PASC) manifestation and 13 convalescent children without PASC manifestation. A detailed medical history, blood and instrumental tests, and physical examination were obtained from all patients. SARS-CoV-2 reactive T-cell response was analyzed via multiparametric flow cytometry and the humoral immunity was addressed via pseudovirus neutralization and ELISA assay. RESULTS: The most common PASC symptoms were shortness of breath/exercise intolerance, paresthesia, smell/taste disturbance, chest pain, dyspnea, headache, and lack of concentration. Blood count and clinical chemistry showed no statistical differences among the study groups. We detected higher frequencies of spike (S) reactive CD4+ and CD8+ T cells among the PASC study group, characterized by TNFα and IFNγ production and low functional avidity. CRP levels are positively correlated with IFNγ producing reactive CD8+ T cells. CONCLUSIONS: Our data might indicate a possible involvement of a persistent cellular inflammatory response triggered by SARS-CoV-2 in the development of the observed sequelae in pediatric PASC. These results may have implications on future therapeutic and prevention strategies.
Assuntos
COVID-19 , Síndrome de COVID-19 Pós-Aguda , Humanos , Criança , SARS-CoV-2 , Citocinas , Linfócitos T CD8-Positivos , Qualidade de Vida , Progressão da Doença , DispneiaRESUMO
Cross-reactive cellular and humoral immunity can substantially contribute to antiviral defense against SARS-CoV-2 variants of concern (VOC). While the adult SARS-CoV-2 cellular and humoral immunity and its cross-recognition potential against VOC is broadly analyzed, similar data regarding the pediatric population are missing. In this study, we perform an analysis of the humoral and cellular SARS-CoV-2 response immune of 32 convalescent COVID-19 children (children), 27 convalescent vaccinated adults(C + V+) and 7 unvaccinated convalescent adults (C + V-). Similarly to adults, a significant reduction of cross-reactive neutralizing capacity against delta and omicron VOC was observed 6 months after SARS-CoV-2 infection. While SAR-CoV-2 neutralizing capacity was comparable among children and C + V- against all VOC, children demonstrated as expected an inferior humoral response when compared to C + V+. Nevertheless, children generated SARS-CoV-2 reactive T cells with broad cross-recognition potential. When compared to V + C+, children presented even comparable frequencies of WT-reactive CD4 + and CD8 + T cells with high avidity and functionality. Taking into consideration the limitations of study - unknown disease onset for 53% of the asymptomatic pediatric subjects, serological detection of SARS-CoV-2 infection-, our results suggest that following SARS-CoV-2 infection children generate a humoral SARS-CoV-2 response with neutralizing potential comparable to unvaccinated COVID-19 convalescent adults as well a sustained SARS-CoV-2 cellular response cross-reactive to VOC.
Assuntos
COVID-19 , SARS-CoV-2 , Adulto , Criança , Adolescente , Humanos , Imunidade Celular , Linfócitos T CD8-Positivos , Imunidade Humoral , Anticorpos Antivirais , Anticorpos NeutralizantesRESUMO
Preventing the progression to acute respiratory distress syndrome (ARDS) in COVID-19 is an unsolved challenge. The involvement of T cell immunity in this exacerbation remains unclear. To identify predictive markers of COVID-19 progress and outcome, we analyzed peripheral blood of 10 COVID-19-associated ARDS patients and 35 mild/moderate COVID-19 patients, not requiring intensive care. Using multi-parametric flow cytometry, we compared quantitative, phenotypic, and functional characteristics of circulating bulk immune cells, as well as SARS-CoV-2 S-protein-reactive T cells between the two groups. ARDS patients demonstrated significantly higher S-protein-reactive CD4+ and CD8+ T cells compared to non-ARDS patients. Of interest, comparison of circulating bulk T cells in ARDS patients to non-ARDS patients demonstrated decreased frequencies of CD4+ and CD8+ T cell subsets, with activated memory/effector T cells expressing tissue migration molecule CD11a++. Importantly, survival from ARDS (4/10) was accompanied by a recovery of the CD11a++ T cell subsets in peripheral blood. Conclusively, data on S-protein-reactive polyfunctional T cells indicate the ability of ARDS patients to generate antiviral protection. Furthermore, decreased frequencies of activated memory/effector T cells expressing tissue migratory molecule CD11a++ observed in circulation of ARDS patients might suggest their involvement in ARDS development and propose the CD11a-based immune signature as a possible prognostic marker.
Assuntos
COVID-19/imunologia , Memória Imunológica/imunologia , Pandemias , Síndrome do Desconforto Respiratório/imunologia , Adulto , Idoso , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , COVID-19/virologia , Feminino , Humanos , Masculino , Glicoproteínas de Membrana/imunologia , Pessoa de Meia-Idade , Síndrome do Desconforto Respiratório/patologia , Síndrome do Desconforto Respiratório/virologia , SARS-CoV-2/imunologia , SARS-CoV-2/patogenicidade , Subpopulações de Linfócitos T/imunologia , VitronectinaRESUMO
The optimal management in transplant recipients with coronavirus disease 2019 (COVID-19) remains uncertain. The main concern is the ability of immunosuppressed patients to generate sufficient immunity for antiviral protection. Here, we report on immune monitoring facilitating a successful outcome of severe severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-associated pneumonia, meningoencephalitis, gastroenteritis, and acute kidney and pancreas graft failure in a pancreas-kidney transplant recipient. Despite the very low numbers of circulating B, NK, and T cells identified in follow-up, a strong SARS-CoV-2 reactive T cell response was observed. Importantly, we detected T cells reactive to Spike, Membrane, and Nucleocapsid proteins of SARS-CoV-2 with majority of T cells showing polyfunctional proinflammatory Th1 phenotype at all analyzed time points. Antibodies against Spike protein were also detected with increasing titers in follow-up. Neutralization tests confirmed their antiviral protection. A correlation between cellular and humoral immunity was observed underscoring the specificity of demonstrated data. We conclude that analyzing the kinetics of nonspecific and SARS-CoV-2-reactive cellular and humoral immunity can facilitate the clinical decision on immunosuppression adjustment and allow successful outcome as demonstrated in the current clinical case. Although the antiviral protection of the detected SARS-CoV-2-reactive T cells requires further evaluation, our data prove an ability mounting a strong SARS-CoV-2-reactive T cell response with functional capacity in immunosuppressed patients.
Assuntos
Anticorpos Antivirais/imunologia , COVID-19/epidemiologia , Imunidade Humoral , Transplante de Rim , Monitorização Imunológica/métodos , Transplante de Pâncreas/métodos , SARS-CoV-2/imunologia , COVID-19/virologia , Tomada de Decisão Clínica , Comorbidade , Rejeição de Enxerto/epidemiologia , Rejeição de Enxerto/imunologia , Humanos , Hospedeiro Imunocomprometido , PandemiasAssuntos
COVID-19 , Infecções por HIV , Humanos , SARS-CoV-2 , Imunidade Celular , Vacinação , Anticorpos Antivirais , Imunidade HumoralAssuntos
Vacinas contra COVID-19 , Imunidade Celular/fisiologia , Imunidade Humoral/fisiologia , Diálise Renal , Insuficiência Renal/imunologia , SARS-CoV-2/imunologia , Anticorpos Antivirais , Vacina BNT162 , COVID-19/prevenção & controle , Estudos de Casos e Controles , Humanos , Insuficiência Renal/terapiaAssuntos
Imunidade Humoral , Falência Renal Crônica , Anticorpos Antivirais , Humanos , Imunidade Celular , VacinaçãoAssuntos
COVID-19 , SARS-CoV-2 , Linfócitos B , Humanos , Memória Imunológica , Glicoproteína da Espícula de CoronavírusRESUMO
In recent years, multiple coronaviruses have emerged, with the latest one, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), causing a global pandemic. Besides respiratory symptoms, some patients experienced extrapulmonary effects, such as cardiac damage or renal injury, indicating the broad tropism of SARS-CoV-2. The ability of the virus to effectively invade the renal cellular environment can eventually cause tissue-specific damage and disease. Indeed, patients with severe coronavirus disease 2019 exhibited a variety of symptoms such as acute proximal tubular injury, ischemic collapse, and severe acute tubular necrosis resulting in irreversible kidney failure. This review summarizes the current knowledge on how it is believed that SARS-CoV-2 influences the renal environment and induces kidney disease, as well as current therapy approaches.
Assuntos
COVID-19 , Rim , SARS-CoV-2 , Tropismo Viral , Humanos , COVID-19/virologia , SARS-CoV-2/fisiologia , SARS-CoV-2/patogenicidade , Rim/virologia , Rim/fisiopatologia , Rim/patologia , Nefropatias/virologia , AnimaisRESUMO
Equine parvovirus-hepatitis (EqPV-H) was first reported from the serum and liver tissue of a horse diagnosed with Theiler's disease in the United States in 2018. Theiler's disease, also known as equine serum hepatitis, is a severe hepatitis with fulminant hepatic necrosis. The disease has most frequently been reported following the administration of equine-origin biological products; however, it has also been reported in in-contact horses with no prior biologic administration. EqPV-H has been detected in clinically healthy horses in North America (USA, Canada), Europe (Germany, Austria, Slovenia), Asia (China, South Korea), and South America (Brazil). Previous prevalence studies conducted worldwide have shown the presence of EqPV-H DNA in serum or plasma ranging from 3.2 to 19.8%. This study investigated the prevalence of EqPV-H DNA in 170 healthy broodmares of various breeds located on 37 farms in southern Ontario, Canada. The occurrence of EqPV-H infection was determined by quantitative PCR for EqPV-H DNA in serum samples. The effects of age, breed, season, pregnancy status, and equine herpesvirus-1 (EHV-1) vaccination history on EqPV-H status were also investigated. There was a prevalence of 15.9% (27/170) with viral loads of EqPV-H ranging from detectable to 2900 copies/mL. Statistical analysis showed that increasing age was a significant factor in the detection of EqPV-H DNA. Neither breed, season, pregnancy status, nor EHV-1 vaccination history was significant in predicting EqPV-H infection status.
L'hépatite à parvovirus équin (EqPV-H) a été signalée pour la première fois à partir du sérum et du tissu hépatique d'un cheval diagnostiqué avec la maladie de Theiler aux États-Unis en 2018. La maladie de Theiler, également connue sous le nom d'hépatite sérique équine, est une hépatite sévère avec nécrose hépatique fulminante. La maladie a été le plus souvent rapportée à la suite de l'administration de produits biologiques d'origine équine; cependant, il a également été signalé chez des chevaux en contact sans administration préalable de produit biologique. EqPV-H a été détecté chez des chevaux cliniquement sains en Amérique du Nord (États-Unis, Canada), en Europe (Allemagne, Autriche, Slovénie), en Asie (Chine, Corée du Sud) et en Amérique du Sud (Brésil). Des études de prévalence antérieures menées dans le monde entier ont montré la présence d'ADN EqPV-H dans le sérum ou le plasma allant de 3,2 à 19,8 %. Cette étude a examiné la prévalence de l'ADN EqPV-H chez 170 poulinières en bonne santé de différentes races situées dans 37 fermes du sud de l'Ontario, au Canada. La survenue d'une infection par EqPV-H a été déterminée par PCR quantitative pour l'ADN d'EqPV-H dans des échantillons de sérum. Les effets de l'âge, de la race, de la saison, de l'état de grossesse et des antécédents de vaccination contre l'herpèsvirus équin-1 (EHV-1) sur le statut EqPV-H ont également été étudiés. Il y avait une prévalence de 15,9 % (27/170) avec des charges virales d'EqPV-H allant de détectable à 2900 copies/mL. L'analyse statistique a montré que l'augmentation de l'âge était un facteur significatif dans la détection de l'ADN EqPV-H. Ni la race, ni la saison, ni l'état de grossesse, ni les antécédents de vaccination contre l'EHV-1 n'étaient significatifs pour prédire l'état de l'infection par l'EqPV-H.(Traduit par Docteur Serge Messier).
Assuntos
Hepatite Viral Animal , Hepatite , Infecções por Herpesviridae , Doenças dos Cavalos , Infecções por Parvoviridae , Parvovirus , Animais , Cavalos , Gravidez , Feminino , Parvovirus/genética , Infecções por Parvoviridae/epidemiologia , Infecções por Parvoviridae/veterinária , Ontário/epidemiologia , Prevalência , Hepatite Viral Animal/epidemiologia , Doenças dos Cavalos/epidemiologia , Infecções por Herpesviridae/veterináriaRESUMO
The role of adaptive SARS-CoV-2 specific immunity in post-acute sequelae of COVID-19 (PASC) is not well explored, although a growing population of convalescent COVID-19 patients with manifestation of PASC is observed. We analyzed the SARS-CoV-2-specific immune response, via pseudovirus neutralizing assay and multiparametric flow cytometry in 40 post-acute sequelae of COVID-19 patients with non-specific PASC manifestation and 15 COVID-19 convalescent healthy donors. Although frequencies of SARS-CoV-2-reactive CD4+ T cells were similar between the studied cohorts, a stronger SARS-CoV-2 reactive CD8+ T cell response, characterized by IFNγ production and predominant TEMRA phenotype but low functional TCR avidity was detected in PASC patients compared to controls. Of interest, high avidity SARS-CoV-2-reactive CD4+ and CD8+ T cells were comparable between the groups demonstrating sufficient cellular antiviral response in PASC. In line with the cellular immunity, neutralizing capacity in PASC patients was not inferior compared to controls. In conclusion, our data suggest that PASC may be driven by an inflammatory response triggered by an expanded population of low avidity SARS-CoV-2 reactive pro-inflammatory CD8+ T cells. These pro-inflammatory T cells with TEMRA phenotype are known to be activated by a low or even without TCR stimulation and lead to a tissue damage. Further studies including animal models are required for a better understanding of underlying immunopathogensis. Summary: A CD8+ driven persistent inflammatory response triggered by SARS-CoV-2 may be responsible for the observed sequelae in PASC patients.