Ultrastructural analysis of SARS-CoV-2 interactions with the host cell via high resolution scanning electron microscopy

Caldas, Lúcio Ayres; Carneiro, Fabiana A.; Higa, Luiza M.; Monteiro, Fábio Luiz; Silva, Gustavo Peixoto da; Costa, Luciana Jesus da; Durigon, Edison Luiz; Tanuri, Amílcar; Souza, Wanderley de

doi:10.1038/s41598-020-73162-5

Cited by 92 publications

(87 citation statements)

References 39 publications

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“…1F ) . Recently, similar vesicles loaded with mature virions have been identified as vacuoles or deacidified lysosomal compartments used by SARS-CoV-2 for viral egress ( 37 , 38 ). These results indicate that SARS-CoV-2 is able to infect, replicate in, and rapidly propagate among human CMs.…”

Section: Resultsmentioning

confidence: 99%

SARS-CoV-2 infection of human iPSC–derived cardiac cells reflects cytopathic features in hearts of patients with COVID-19

et al. 2021

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Although coronavirus disease 2019 (COVID-19) causes cardiac dysfunction in up to 25% of patients, its pathogenesis remains unclear. Exposure of human induced pluripotent stem cell (iPSC)-derived heart cells to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) revealed productive infection and robust transcriptomic and morphological signatures of damage, particularly in cardiomyocytes. Transcriptomic disruption of structural genes corroborates adverse morphologic features, which included a distinct pattern of myofibrillar fragmentation and nuclear disruption. Human autopsy specimens from patients with COVID-19 reflected similar alterations, particularly sarcomeric fragmentation. These striking cytopathic features in cardiomyocytes provide insights into SARS-CoV-2-induced cardiac damage, offer a platform for discovery of potential therapeutics, and raise concerns about the long-term consequences of COVID-19 in asymptomatic as well as severe cases.

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Section: Resultsmentioning

confidence: 99%

SARS-CoV-2 infection of human iPSC–derived cardiac cells reflects cytopathic features in hearts of patients with COVID-19

et al. 2021

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“…We suggest that this resembles the particle clustering by host defense protein BST-2 as it was observed for human coronavirus229E and quantified in HeLa cells by Wang and co-workers [ 38 ]. However, the metal coating applied by Wang et al is clearly visible at high resolution in the SEM images as a rough layer on the cell membrane that hides the true topography [ 25 , 39 ]. In contrast, the HIM images presented here not only allow for the quantification of particles and clusters, but also enable an unveiled view on the interaction of virus particles with the cell membrane.…”

Section: Resultsmentioning

confidence: 99%

Imaging of SARS-CoV-2 infected Vero E6 cells by helium ion microscopy

Frese¹,

Schmerer²,

Wortmann³

et al. 2021

Beilstein J. Nanotechnol.

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Helium ion microscopy (HIM) offers the opportunity to obtain direct views of biological samples such as cellular structures, virus particles, and microbial interactions. Imaging with the HIM combines sub-nanometer resolution, large depth of field, and high surface sensitivity. Due to its charge compensation capability, the HIM can image insulating biological samples without additional conductive coatings. Here, we present an exploratory HIM study of SARS-CoV-2 infected Vero E6 cells, in which several areas of interaction between cells and virus particles, as well as among virus particles, were imaged. The HIM pictures show the three-dimensional appearance of SARS-CoV-2 and the surface of Vero E6 cells at a multiplicity of infection of approximately 1 with great morphological detail. The absence of a conductive coating allows for a distinction between virus particles bound to the cell membrane and virus particles lying on top of the membrane. After prolonged imaging, it was found that ion-induced deposition of hydrocarbons from the vacuum renders the sample sufficiently conductive to allow for imaging even without charge compensation. The presented images demonstrate the potential of the HIM in bioimaging, especially for the imaging of interactions between viruses and their host organisms.

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“…Interestingly, cellular extensions containing the S protein were observed for the wt and each of the mutants (Fig. S4, white arrows) [ 63 ]. Finally, luciferase reporter gene assays were performed.…”

Section: Resultsmentioning

confidence: 99%

Effect of mutations in the SARS-CoV-2 spike protein on protein stability, cleavage, and cell-cell fusion function

Barrett

Neal

Edmonds

et al. 2021

Preprint

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The SARS-CoV-2 spike protein (S) is the sole viral protein responsible for both viral binding to a host cell and the membrane fusion event needed for cell entry. In addition to facilitating fusion needed for viral entry, S can also drive cell-cell fusion, a pathogenic effect observed in the lungs of SARS-CoV-2 infected patients. While several studies have investigated S requirements involved in viral particle entry, examination of S stability and factors involved in S cell-cell fusion remain limited. We demonstrate that S must be processed at the S1/S2 border in order to mediate cell-cell fusion, and that mutations at potential cleavage sites within the S2 subunit alter S processing at the S1/S2 border, thus preventing cell-cell fusion. We also identify residues within the internal fusion peptide and the cytoplasmic tail that modulate S cell-cell fusion. Additionally, we examine S stability and protein cleavage kinetics in a variety of mammalian cell lines, including a bat cell line related to the likely reservoir species for SARS-CoV-2, and provide evidence that proteolytic processing alters the stability of the S trimer. This work therefore offers insight into S stability, proteolytic processing, and factors that mediate S cell-cell fusion, all of which help give a more comprehensive understanding of this highly sought-after therapeutic target.

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Ultrastructural analysis of SARS-CoV-2 interactions with the host cell via high resolution scanning electron microscopy

Cited by 92 publications

References 39 publications

SARS-CoV-2 infection of human iPSC–derived cardiac cells reflects cytopathic features in hearts of patients with COVID-19

SARS-CoV-2 infection of human iPSC–derived cardiac cells reflects cytopathic features in hearts of patients with COVID-19

Imaging of SARS-CoV-2 infected Vero E6 cells by helium ion microscopy

Effect of mutations in the SARS-CoV-2 spike protein on protein stability, cleavage, and cell-cell fusion function

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