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Viruses
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BSL4 Viruses: Understanding and Controlling Highly Infectious Pathogens
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BSL4 Viruses: Understanding and Controlling Highly Infectious Pathogens

A special issue of Viruses (ISSN 1999-4915). This special issue belongs to the section "Human Virology and Viral Diseases".

Deadline for manuscript submissions: closed (15 March 2025) | Viewed by 4250

Special Issue Editor

Prof. Dr. Slobodan Paessler

E-Mail Website
Guest Editor
Department of Pathology, University of Texas Medical Branch, Galveston, TX, USA
Interests: venezuelan equine encephalitis virus; molecular biology; viral pathogenesis; vaccine; antivirals; diagnostics development

Special Issue Information

Dear Colleagues,

The study of Biosafety Level 4 (BSL4) viruses represents a critical frontier in infectious disease research, demanding a comprehensive understanding of the molecular mechanisms underlying viral pathogenesis, transmission, and host immune responses. BSL4 viruses, classified as highly infectious and posing significant public health threats, include notorious pathogens such as Ebola virus, Marburg virus, Lassa virus, Junin virus, Machupo virus, Nipah, and Crimean-Congo hemorrhagic fever virus, among others.

This call for papers aims to bring together cutting-edge research focused on the interdisciplinary study of BSL4 viruses, encompassing virology, immunology, epidemiology, molecular biology, bioinformatics, and translational medicine. By elucidating viral life cycles and the complex interactions between BSL4 viruses and their hosts, researchers seek to uncover novel strategies for the prevention, detection, and treatment of these deadly pathogens.

Contributions to this Special Issue may cover a wide range of topics, including:

  1. Molecular mechanisms of BSL4 virus replication, transcription, and translation;
  2. Host–pathogen interactions and immune evasion strategies employed by BSL4 viruses;
  3. Epidemiology and surveillance of BSL4 virus outbreaks, including spatiotemporal dynamics and transmission routes;
  4. Development and evaluation of vaccines, therapeutics, and diagnostic tools for BSL4 virus infections;
  5. Animal models for studying BSL4 virus pathogenesis and evaluating candidate interventions;
  6. Genomic, transcriptomic, and proteomic analyses of BSL4 virus-host interactions;
  7. Evolutionary dynamics and emergence of novel BSL4 virus strains;
  8. Public health strategies for BSL4 virus containment, biosafety, and biosecurity.

We invite researchers from around the globe to submit original research articles, reviews, and perspectives that advance our biological understanding of BSL4 viruses and contribute to the development of effective countermeasures against these pathogens. Together, we wish to share the collective knowledge and expertise of the scientific community to address the urgent global health challenges posed by BSL4 viruses.

Prof. Dr. Slobodan Paessler
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

  • hemorrhagic virus
  • viral encephalitis
  • vaccines
  • viral therapies
  • animal and tissue models
  • diagnostics
  • virus transmission
  • systemic infections

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Further information on MDPI's Special Issue policies can be found here.

Published Papers (4 papers)

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Research

15 pages, 1455 KiB  
Article
Successful Inactivation of High-Consequence Pathogens in PrimeStore Molecular Transport Media
by Briana Spruill-Harrell, Gregory Kocher, Maurice Boda, Kristen Akers, Denise Freeburger, Nicole Murphy, Jens H. Kuhn, Gerald Fischer, Irina Maljkovic Berry, Prabha Chandrasekaran and Jerry Torrison
Viruses 2025, 17(5), 639; https://doi.org/10.3390/v17050639 (registering DOI) - 29 Apr 2025
Abstract
Handling cultured isolates and clinical, environmental, or wildlife surveillance samples containing Risk Group 3 and 4 pathogens presents considerable biosafety challenges in minimizing human exposure during processing and transport. Safe handling typically requires high- or maximum-containment facilities, demanding substantial logistical planning and resources. [...] Read more.
Handling cultured isolates and clinical, environmental, or wildlife surveillance samples containing Risk Group 3 and 4 pathogens presents considerable biosafety challenges in minimizing human exposure during processing and transport. Safe handling typically requires high- or maximum-containment facilities, demanding substantial logistical planning and resources. We evaluated PrimeStore Molecular Transport Medium (PS-MTM), a guanidine-based solution created to kill pathogens and preserve nucleic acids at ambient temperatures, for inactivating Crimean-Congo hemorrhagic fever, eastern equine encephalitis, Ebola, Hendra, Japanese encephalitis, Lassa, Marburg, Nipah, Rift Valley fever, and West Nile viruses. To mimic diagnostic conditions, human whole blood spiked with any of these viruses was incubated with PS-MTM for 20-, 30-, or 60-min. Samples with titers up to 107 PFU/mL exposed to PS-MTM at all time points resulted in complete loss of infectivity judged by plaque assays. A 30-min incubation provided a 50% safety margin over the minimum inactivation time and was used for quantification with the tissue culture infectious dose (TCID50) assay, enabling evaluation of PS-MTM’s activity for viruses that do or do not produce well-defined plaques. Results confirmed that PS-MTM inactivated all tested viruses at titers up to 107 TCID50/mL, underscoring its reliability for enhancing biosafety in diagnostics, outbreak management, and surveillance. Full article
(This article belongs to the Special Issue BSL4 Viruses: Understanding and Controlling Highly Infectious Pathogens)
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13 pages, 2415 KiB  
Article
Development of a Luciferase Immunosorbent Assay for Detecting Crimean–Congo Hemorrhagic Fever Virus IgG Antibodies Based on Nucleoprotein
by Qi Chen, Yuting Fang, Ning Zhang and Chengsong Wan
Viruses 2025, 17(1), 32; https://doi.org/10.3390/v17010032 - 28 Dec 2024
Viewed by 901
Abstract
Crimean–Congo hemorrhagic fever (CCHF) is a serious tick-borne disease with a wide geographical distribution. Classified as a level 4 biosecurity risk pathogen, CCHF can be transmitted cross-species due to its aerosol infectivity and ability to cause severe hemorrhagic fever outbreaks with high morbidity [...] Read more.
Crimean–Congo hemorrhagic fever (CCHF) is a serious tick-borne disease with a wide geographical distribution. Classified as a level 4 biosecurity risk pathogen, CCHF can be transmitted cross-species due to its aerosol infectivity and ability to cause severe hemorrhagic fever outbreaks with high morbidity and mortality. However, current methods for detecting anti-CCHFV antibodies are limited. This study aimed to develop a novel luciferase immunosorbent assay (LISA) for the detection of CCHFV-specific IgG antibodies. We designed specific antigenic fragments of the nucleoprotein and evaluated their sensitivity and specificity in detecting IgG in serum samples from mice and horses. In addition, we compared the efficacy of our LISA to a commercial enzyme-linked immunosorbent assay (ELISA). Our results demonstrated that the optimal antigen for detecting anti-CCHFV IgG was located within the stalk cut-off domain of the nucleoprotein. The LISA exhibited high specificity for serum samples from indicated species and significantly higher sensitivity (at least 128 times) compared with the commercial ELISA. The proposed CCHFV-LISA has the potential to facilitate serological diagnosis and epidemiological investigation of CCHFV in natural foci, providing valuable technical support for surveillance and early warning of this disease. Full article
(This article belongs to the Special Issue BSL4 Viruses: Understanding and Controlling Highly Infectious Pathogens)
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10 pages, 2355 KiB  
Communication
Efficacy of Polyphenylene Carboxymethylene (PPCM) Gel at Protecting Type I Interferon Receptors Knockout Mice from Intravaginal Ebola Virus Challenge
by Olivier Escaffre, Terry L. Juelich, Jennifer K. Smith, Lihong Zhang, Madison Pearson, Nigel Bourne and Alexander N. Freiberg
Viruses 2024, 16(11), 1693; https://doi.org/10.3390/v16111693 - 30 Oct 2024
Viewed by 979
Abstract
Ebola virus (EBOV) is one of three filovirus members of the Orthoebolavirus genus that can cause severe Ebola disease (EBOD) in humans. Transmission predominantly occurs from spillover events from wildlife but has also happened between humans with infected bodily fluids. Specifically, the sexual [...] Read more.
Ebola virus (EBOV) is one of three filovirus members of the Orthoebolavirus genus that can cause severe Ebola disease (EBOD) in humans. Transmission predominantly occurs from spillover events from wildlife but has also happened between humans with infected bodily fluids. Specifically, the sexual route through infectious male survivors could be the origin of flare up events leading to the deaths of multiple women. More studies are needed to comprehend this route of infection which has recently received more focus. The use of microbicides prior to intercourse is of interest if neither of the Ebola vaccines are an option. These experimental products have been used against sexually transmitted diseases, and recently polyphenylene carboxymethylene (PPCM) showed efficacy against EBOV in vitro. Shortly after, the first animal model of EBOV sexual transmission was established using type I interferon receptors (IFNAR−/−) knockout female mice in which mortality endpoint could be achieved. Here, we investigated PPCM efficacy against a mouse-adapted (ma)EBOV isolate in IFNAR−/− mice and demonstrated that 4% PPCM gel caused a 20% reduction in mortality in two distinct groups compared to control groups when inoculated prior to virus challenge. Among animals that succumbed to disease despite PPCM treatment, we report an increase in median survival time as well as a less infectious virus, and fewer virus positive vaginal swabs compared to those from vehicle-treated animals, altogether indicating the beneficial effect of using PPCM prior to exposure. A post-study analysis of the different gel formulations tested indicated that buffering the gels would have prevented an increase in acidity seen only in vehicles, suggesting that PPCM antiviral efficacy against EBOV was suboptimal in our experimental set-up. These results are encouraging and warrant further studies using optimized stable formulations with the goal of providing additional safe protective countermeasures from sexual transmission of EBOV in humans. Full article
(This article belongs to the Special Issue BSL4 Viruses: Understanding and Controlling Highly Infectious Pathogens)
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11 pages, 564 KiB  
Article
Inactivation Validation of Ebola, Marburg, and Lassa Viruses in AVL and Ethanol-Treated Viral Cultures
by Todd Cutts, Anders Leung, Logan Banadyga and Jay Krishnan
Viruses 2024, 16(9), 1354; https://doi.org/10.3390/v16091354 - 24 Aug 2024
Cited by 1 | Viewed by 1548
Abstract
High-consequence pathogens such as the Ebola, Marburg, and Lassa viruses are handled in maximum-containment biosafety level 4 (BSL-4) laboratories. Genetic material is often isolated from such viruses and subsequently removed from BSL-4 laboratories for a multitude of downstream analyses using readily accessible technologies [...] Read more.
High-consequence pathogens such as the Ebola, Marburg, and Lassa viruses are handled in maximum-containment biosafety level 4 (BSL-4) laboratories. Genetic material is often isolated from such viruses and subsequently removed from BSL-4 laboratories for a multitude of downstream analyses using readily accessible technologies and equipment available at lower-biosafety level laboratories. However, it is essential to ensure that these materials are free of viable viruses before removal from BSL-4 laboratories to guarantee sample safety. This study details the in-house procedure used for validating the inactivation of Ebola, Marburg, and Lassa virus cultures after incubation with AVL lysis buffer (Qiagen) and ethanol. This study’s findings show that no viable virus was detectable when high-titer cultures of Ebola, Marburg, and Lassa viruses were incubated with AVL lysis buffer for 10 min, followed by an equal volume of 95% ethanol for 3 min, using a method with a sensitivity of ≤0.8 log10 TCID50 as the limit of detection. Full article
(This article belongs to the Special Issue BSL4 Viruses: Understanding and Controlling Highly Infectious Pathogens)
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