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Viruses
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Advanced Microscopy and Imaging Approaches to Dissect Virus–Host Interactions
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Advanced Microscopy and Imaging Approaches to Dissect Virus–Host Interactions

A special issue of Viruses (ISSN 1999-4915). This special issue belongs to the section "General Virology".

Deadline for manuscript submissions: 30 July 2024 | Viewed by 4520

Special Issue Editor

Dr. Nicholas Eyre

E-Mail Website
Guest Editor
College of Medicine & Public Health, Flinders University, Bedford Park, SA 5042, Australia
Interests: flaviviruses; virus-host interactions; viral replication; innate immunity; high-resolution imaging
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

As obligate intracellular parasites, viruses interact with, and exploit, a myriad of host factors during their replication cycles. Developing a detailed understanding of these interactions is critical to understanding how viruses cause disease, and how therapeutic or preventative interventions may disrupt these virus–host interactions to limit viral pathogenesis and spread. Imaging is a long-standing and increasingly important component of the approaches employed to understand virus–host interactions and the validity and mechanisms of antiviral interventions. Electron microscopy techniques, such as cryo-electron microscopy (cryo-EM), cryo-electron tomography (cryo-ET), transmission electron microscopy (TEM), electron tomography (ET), scanning electron microscopy (SEM), focused ion beam (FIB)-SEM, correlative light and electron microscopy (CLEM), and hybrid approaches have revealed critical details about viruses and virus–host interactions at remarkable resolution in fixed samples. In addition to reliable and evolving confocal and widefield fluorescence microscopy approaches, super-resolution fluorescence microscopy techniques such as structured illumination microscope (SIM), stimulated emission depletion (STED) microscopy, and single molecule localization microscopy (SMLM) have the power to unveil important details about virus–host interactions at high spatial resolutions in live or minimally processed single cells and organoids. Concurrently, technological advances in small animal imaging and improvements in associated reporter and labelling systems have enabled the detailed examination of viral spread and host responses at a whole-organism level.

This Special Issue of Viruses will cover the application of a variety of advanced imaging techniques to study virus–host interactions. Researchers are invited to contribute primary research articles, brief reports, technical notes, and reviews that focus on advanced imaging approaches to better understand viral replication cycles and virus–host interactions and facilitate the development of safe and effective approaches to combat viral infections.

Dr. Nicholas Eyre
Guest Editor

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Viruses is an international peer-reviewed open access monthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2600 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • high-resolution imaging
  • virus–host
  • correlative microscopy
  • viral replication cycle

Published Papers (3 papers)

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Research

18 pages, 7858 KiB  
Article
Development of a Mouse-Adapted Reporter SARS-CoV-2 as a Tool for Two-Photon In Vivo Imaging
by Hiroshi Ueki, Maki Kiso, Yuri Furusawa, Shun Iida, Seiya Yamayoshi, Noriko Nakajima, Masaki Imai, Tadaki Suzuki and Yoshihiro Kawaoka
Viruses 2024, 16(4), 537; https://doi.org/10.3390/v16040537 - 29 Mar 2024
Viewed by 546
Abstract
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) often causes severe viral pneumonia. Although many studies using mouse models have examined the pathogenicity of SARS-CoV-2, COVID-19 pathogenesis remains poorly understood. In vivo imaging analysis using two-photon excitation microscopy (TPEM) is useful for elucidating the [...] Read more.
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) often causes severe viral pneumonia. Although many studies using mouse models have examined the pathogenicity of SARS-CoV-2, COVID-19 pathogenesis remains poorly understood. In vivo imaging analysis using two-photon excitation microscopy (TPEM) is useful for elucidating the pathology of COVID-19, providing pathological insights that are not available from conventional histological analysis. However, there is no reporter SARS-CoV-2 that demonstrates pathogenicity in C57BL/6 mice and emits sufficient light intensity for two-photon in vivo imaging. Here, we generated a mouse-adapted strain of SARS-CoV-2 (named MASCV2-p25) and demonstrated its efficient replication in the lungs of C57BL/6 mice, causing fatal pneumonia. Histopathologic analysis revealed the severe inflammation and infiltration of immune cells in the lungs of MASCV2-p25-infected C57BL/6 mice, not unlike that observed in COVID-19 patients with severe pneumonia. Subsequently, we generated a mouse-adapted reporter SARS-CoV-2 (named MASCV-Venus-p9) by inserting the fluorescent protein-encoding gene Venus into MASCV2-p25 and sequential lung-to-lung passages in C57BL/6 mice. C57BL/6 mice infected with MASCV2-Venus-p9 exhibited severe pneumonia. In addition, the TPEM of the lungs of the infected C57BL/6J mice showed that the infected cells emitted sufficient levels of fluorescence for easy observation. These findings suggest that MASCV2-Venus-p9 will be useful for two-photon in vivo imaging studies of the pathogenesis of severe COVID-19 pneumonia. Full article
(This article belongs to the Special Issue Advanced Microscopy and Imaging Approaches to Dissect Virus–Host Interactions)
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22 pages, 1922 KiB  
Article
SERINC5-Mediated Restriction of HIV-1 Infectivity Correlates with Resistance to Cholesterol Extraction but Not with Lipid Order of Viral Membrane
by Gokul Raghunath, Yen-Cheng Chen, Mariana Marin, Hui Wu and Gregory B. Melikyan
Viruses 2022, 14(8), 1636; https://doi.org/10.3390/v14081636 - 27 Jul 2022
Cited by 3 | Viewed by 1986
Abstract
Serine incorporator 5 (SER5) is a protein that upon incorporation into virions inhibits HIV-1 infectivity by interfering with the ability of the Env glycoprotein to promote viral fusion. The mechanisms by which SER5 antagonizes HIV-1 fusion are not well understood. A recent study [...] Read more.
Serine incorporator 5 (SER5) is a protein that upon incorporation into virions inhibits HIV-1 infectivity by interfering with the ability of the Env glycoprotein to promote viral fusion. The mechanisms by which SER5 antagonizes HIV-1 fusion are not well understood. A recent study of SER5’s structure revealed a lipid-binding pocket, suggesting the ability to sequester lipids. This finding, along with the well-documented modulation of HIV-1 infectivity by viral lipids, especially cholesterol, prompted our examination of SER5′s effect on the general lipid order of the HIV-1 membrane. Pseudoviruses bearing the SER5-sensitive HXB2-Env and containing SER5 or SER2, a control protein that lacks antiviral activity, were analyzed using two distinct lipid-order probes. We show that SER5 incorporation does not noticeably affect the lipid order of pseudoviruses. Although viral cholesterol extraction reduces HIV-1 infectivity, SER5+ viruses are less sensitive to cholesterol extraction than the control samples. In contrast, the virus’ sensitivity to cholesterol oxidation was not affected by SER5 incorporation. The hydrolytic release of sphingomyelin-sequestered cholesterol had a minimal impact on the apparent resistance to cholesterol extraction. Based on these results, we propose that a subpopulation of more stable Env glycoproteins responsible for the residual infectivity of SER5+ viruses is less sensitive to the cholesterol content of the viral membrane. Full article
(This article belongs to the Special Issue Advanced Microscopy and Imaging Approaches to Dissect Virus–Host Interactions)
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26 pages, 5460 KiB  
Article
Generation of Reporter-Expressing New World Arenaviruses: A Systematic Comparison
by Lucie Fénéant, Anne Leske, Karla Günther and Allison Groseth
Viruses 2022, 14(7), 1563; https://doi.org/10.3390/v14071563 - 18 Jul 2022
Cited by 3 | Viewed by 1730
Abstract
Replication-competent reporter-expressing viruses are crucial tools in molecular virology with applications that range from antiviral screening to live-cell imaging of protein spatiotemporal dynamics. However, there is currently little information available regarding viable strategies to develop reporter-expressing arenaviruses. To address this, we used Tacaribe [...] Read more.
Replication-competent reporter-expressing viruses are crucial tools in molecular virology with applications that range from antiviral screening to live-cell imaging of protein spatiotemporal dynamics. However, there is currently little information available regarding viable strategies to develop reporter-expressing arenaviruses. To address this, we used Tacaribe virus (TCRV), an apathogenic BSL2 arenavirus, to assess the feasibility of different reporter expression approaches. We first generated trisegmented TCRV viruses with either the glycoprotein (GP) or nucleoprotein (NP) replaced by a reporter (GFP, mCherry, or nanoluciferase). These viruses were all viable, but showed marked differences in brightness and attenuation. Next, we generated terminal fusions with each of the TCRV proteins (i.e., NP, GP, polymerase (L), matrix protein (Z)) either with or without a T2A self-cleavage site. We tested both the function of the reporter-fused proteins alone, and the viability of corresponding recombinant TCRVs. We successfully rescued viruses with both direct and cleavable reporter fusions at the C-terminus of Z, as well as cleavable N-terminal fusions with NP. These viruses all displayed detectable reporter activity, but were also moderately attenuated. Finally, reporter proteins were inserted into a flexible hinge region within L. These viruses were also viable and showed moderate attenuation; however, reporter expression was only detectable for the luminescent virus. These strategies provide an exciting range of new tools for research into the molecular biology of TCRV that can likely also be adapted to other arenaviruses. Full article
(This article belongs to the Special Issue Advanced Microscopy and Imaging Approaches to Dissect Virus–Host Interactions)
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