Presymptomatic viral shedding and infective ability of SARS-CoV-2; a case report

Nissen, Karolina; Hagbom, Marie; Krambrich, Janina; Akaberi, Dario; Sharma, Sumit; Ling, Jiaxin; Hoffman, Tove; Svensson, Lennart; Salaneck, Erik

doi:10.1016/j.heliyon.2021.e06328

Cited by 4 publications

(4 citation statements)

References 25 publications

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“…For in vitro infection experiments, SARS-CoV-2 virus previously isolated in a Biosafety level 3 lab according to local safety regulations from the nasopharyngeal aspirate of a COVID-19 patient (early April 2020) was used[ 29 ]. The isolated virus was passaged five times in Vero E6 cells and for cell infection experiments, freeze-thawed medium supernatants of 4–5 days infected cells or mock supernatants were used.…”

Section: Methodsmentioning

confidence: 99%

Epigenetic rewiring of pathways related to odour perception in immune cells exposed to SARS-CoV-2in vivoandin vitro

et al. 2022

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A majority of SARS-CoV-2 recoverees develop only mild-to-moderate symptoms, while some remain completely asymptomatic. Although viruses, including SARS-CoV-2, may evade host immune responses by epigenetic mechanisms including DNA methylation, little is known about whether these modifications are important in defence against and healthy recovery from COVID-19 in the host. To this end, epigenome-wide DNA methylation patterns from COVID-19 convalescents were compared to uninfected controls from before and after the pandemic. Peripheral blood mononuclear cell (PBMC) DNA was extracted from uninfected controls, COVID-19 convalescents, and symptom-free individuals with SARS-CoV-2-specific T cell-responses, as well as from PBMCs stimulated in vitro with SARS-CoV-2. Subsequently, the Illumina MethylationEPIC 850K array was performed, and statistical/bioinformatic analyses comprised differential DNA methylation, pathway over-representation, and module identification analyses. Differential DNA methylation patterns distinguished COVID-19 convalescents from uninfected controls, with similar results in an experimental SARS-CoV-2 infection model. A SARS-CoV-2-induced module was identified in vivo , comprising 66 genes of which six ( TP53, INS, HSPA4, SP1, ESR1, and FAS ) were present in corresponding in vitro analyses. Over-representation analyses revealed involvement in Wnt, muscarinic acetylcholine receptor signalling, and gonadotropin-releasing hormone receptor pathways. Furthermore, numerous differentially methylated and network genes from both settings interacted with the SARS-CoV-2 interactome. Altered DNA methylation patterns of COVID-19 convalescents suggest recovery from mild-to-moderate SARS-CoV-2 infection leaves longstanding epigenetic traces. Both in vitro and in vivo exposure caused epigenetic modulation of pathways thataffect odour perception. Future studies should determine whether this reflects host-induced protective antiviral defense or targeted viral hijacking to evade host defence.

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Section: Methodsmentioning

confidence: 99%

Epigenetic rewiring of pathways related to odour perception in immune cells exposed to SARS-CoV-2in vivoandin vitro

et al. 2022

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“…Vero E6 cells were grown in DMEM (Gibco, 41966029) supplemented with 10% FBS (Gibco, 10500064) and 1× penicillin-streptomycin (Sigma-Aldrich, PA333) and incubated at 37 °C, 5% CO 2 atmosphere. SARS-CoV-2 was isolated at the Zoonosis Science Center (Uppsala University) from naso-oropharyngeal swab collected from a Swedish patient as described in Nissen et al 65 All infection experiments were performed in a biosafety level 3 laboratory (BSL3) at the Zoonosis Science Center (Uppsala University).…”

Section: Methodsmentioning

confidence: 99%

“…The genotype of this virus stock shows four nucleotide changes as compared with the mother Journal of the American Chemical Society ) and incubated at 37 °C, 5% CO 2 atmosphere. SARS-CoV-2 was isolated at the Zoonosis Science Center (Uppsala University) from naso-oropharyngeal swab collected from a Swedish patient as described in Nissen et al 67 All infection experiments were performed in a biosafety level 3 laboratory (BSL3) at the Zoonosis Science Center (Uppsala University).…”

Section: ■ Materials and Methodsmentioning

confidence: 99%

Ultralarge Virtual Screening Identifies SARS-CoV-2 Main Protease Inhibitors with Broad-Spectrum Activity against Coronaviruses

et al. 2022

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Drugs targeting SARS-CoV-2 could have saved millions of lives during the COVID-19 pandemic, and it is now crucial to develop inhibitors of coronavirus replication in preparation for future outbreaks. We explored two virtual screening strategies to find inhibitors of the SARS-CoV-2 main protease in ultralarge chemical libraries. First, structure-based docking was used to screen a diverse library of 235 million virtual compounds against the active site. One hundred top-ranked compounds were tested in binding and enzymatic assays. Second, a fragment discovered by crystallographic screening was optimized guided by docking of millions of elaborated molecules and experimental testing of 93 compounds. Three inhibitors were identified in the first library screen, and five of the selected fragment elaborations showed inhibitory effects. Crystal structures of target–inhibitor complexes confirmed docking predictions and guided hit-to-lead optimization, resulting in a noncovalent main protease inhibitor with nanomolar affinity, a promising in vitro pharmacokinetic profile, and broad-spectrum antiviral effect in infected cells.

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“…For in vitro infection experiments, SARS-CoV-2 virus previously isolated in a Biosafety level 3 lab according to local safety regulations from the nasopharyngeal aspirate of a COVID-19 patient (early April 2020) was used. 26 The isolated virus was passaged five times in Vero E6 cells and for cell infection experiments, freeze-thawed medium supernatants of 4-5 days infected cells or mock supernatants were used. Virus titers were determined using immunoperoxidase assay.…”

Section: Methodsmentioning

confidence: 99%

Epigenome-wide DNA methylation profiling of healthy COVID-19 recoverees reveals a unique signature in circulating immune cells

Huoman

Sayyab

Apostolou

et al. 2021

Preprint

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Background: Coronaviruses such as SARS-CoV-2 may circumvent host defence mechanisms by hijacking host proteins, possibly by altering DNA methylation patterns in host cells. While most epigenetic studies have been performed in severely ill COVID-19 patients, studies on individuals who have recovered from mild-to-moderate disease remain scarce. The aim of this study was to assess epigenome-wide DNA methylation patterns in COVID-19 convalescents compared to uninfected controls from before and after the pandemic outbreak began. Methods: DNA was extracted from peripheral blood mononuclear cells originating from uninfected controls before (Pre20, n=5) and after (Con, n=18) 2020, COVID-19 convalescents (CC19, n=14) and symptom-free individuals with a SARS-CoV-2-specific T cell response (SFT, n=6), as well as from Pre20 (n=4) samples stimulated in vitro with SARS-CoV-2. Subsequently, epigenome-wide DNA methylation analyses were performed using the Illumina MethylationEPIC 850K array, and statistical and bioinformatic analyses comprised differential DNA methylation, pathway over-representation and module identification network analyses. Results: DNA methylation patterns of COVID-19 convalescents were altered as compared to uninfected controls, with similar results observed in in vitro stimulations of PBMC with SARS-CoV-2. Differentially methylated genes from the in vivo comparison constituted the foundation for the identification of a possibly SARS-CoV-2-induced module, containing 66 genes of which six could also be identified in corresponding analyses of the in vitro data (TP53, INS, HSPA4, SP1, ESR1 and FAS). Pathway over-representation analyses revealed involvement of Wnt, cadherin and apoptosis signalling pathways amongst others. Furthermore, numerous interactions were found between the obtained differentially methylated genes from both settings and the network analyses when overlaying the data unto the SARS-CoV-2 interactome. Conclusions: Epigenome-wide DNA methylation patterns of individuals that have recovered from mild-to-moderate COVID-19 are different from those of non-infected controls. The observed alterations during both in vivo and in vitro exposure to SARS-CoV-2 showed involvement in interactions and pathways that are highly relevant to COVID-19. The present study provides indications that DNA methylation is one of several epigenetic mechanisms that is altered upon SARS-CoV-2 infection. Further studies on the mechanistic underpinnings should determine whether the observed effects are reflecting host-protective antiviral defence or targeted viral hijacking to evade host defence.

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Presymptomatic viral shedding and infective ability of SARS-CoV-2; a case report

Cited by 4 publications

References 25 publications

Epigenetic rewiring of pathways related to odour perception in immune cells exposed to SARS-CoV-2in vivoandin vitro

Epigenetic rewiring of pathways related to odour perception in immune cells exposed to SARS-CoV-2in vivoandin vitro

Ultralarge Virtual Screening Identifies SARS-CoV-2 Main Protease Inhibitors with Broad-Spectrum Activity against Coronaviruses

Epigenome-wide DNA methylation profiling of healthy COVID-19 recoverees reveals a unique signature in circulating immune cells

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