An Immunoinformatics Approach to Design Novel and Potent Multi-Epitope-Based Vaccine to Target Lumpy Skin Disease

The lumpy skin disease (LSD) virus of the Poxviridae family is a serious threat that mostly affects cattle and causes significant economic loss. LSD has the potential to spread widely and its rapidly across borders. Despite the availability of information, there is still no competitive vaccine available for LSD. Therefore, the current study was conducted to develop an epitope-based LSD vaccine that is efficient, secure, and biocompatible and stimulates both innate and adaptive immune responses using immunoinformatics techniques. Initially, putative virion core proteins were manipulated;B-cell and T-cell epitopes have been predicted and connected with the help of adjuvants and linkers. Numerous bioinformatics methods, including antigenicity testing, transmembrane topology screening, allergenicity assessment, conservancy analysis, and toxicity evaluation, were employed to find superior epitopes. Based on promising vaccine candidates and immunogenic potential, the vaccine design was selected. Strong interactions between TLR4 and TLR9 and the anticipated vaccine design were revealed by molecular docking. Finally, based on the high docking score, computer simulations were performed in order to assess the stability, efficacy, and compactness of the constructed vaccine. The simulation outcomes showed that the polypeptide vaccine design was remarkably stable, with high expression, stability, immunogenic qualities, and considerable solubility. Additionally, computer-based research shows that the constructed vaccine provides adequate population coverage, making it a promising candidate for use in the design of vaccines against other viruses within the Poxviridae family and potentially other virus families as well. These outcomes suggest that the epitope-based vaccine developed in this study will be a significant candidate against LSD to control and prevent LSDV-related disorders if further investigated experimentally.

A computational two-photon fluorescence approach for revealing label-free the 3D image of viruses and bacteria.

Current solutions for bacteria and viruses identification are based on time-consuming technics with complex preparation procedures. In the present work, we revealed label-free the presence of free viral particles and bacteria with a computational two-photon fluorescence (C-TPF) strategy. Six bacteria were tested: Escherichia coli, Staphylococcus epidermidis, Proteus vulgaris, Pseudomonas fluorescens, Bacillus subtilis, and Clostridium perfringens. The two families of viral particles were the herpes virus with the cytomegalovirus (CMV, 300 nm of diameter) and the coronavirus with the SARS-CoV-2 (100 nm of diameter). The instrumental and computational pipeline FAMOUS optimized the produced 3D images. The origin of the fluorescence emission was discussed for bacteria regarding to their two-photon excitation spectra and attributed to the metabolic indicators (FAD and NADH). The optical and computational strategy constitute a new approach for imaging label-free viral particles and bacteria and paves the way to a new understanding of viral or bacterial ways of infection.

Membrane attachment and fusion of HIV-1, influenza A, and SARS-CoV-2: resolving the mechanisms with biophysical methods.

Attachment to and fusion with cell membranes are two major steps in the replication cycle of many human viruses. We focus on these steps for three enveloped viruses, i.e., HIV-1, IAVs, and SARS-CoV-2. Viral spike proteins drive the membrane attachment and fusion of these viruses. Dynamic interactions between the spike proteins and membrane receptors trigger their specific attachment to the plasma membrane of host cells. A single virion on cell membranes can engage in binding with multiple receptors of the same or different types. Such dynamic and multivalent binding of these viruses result in an optimal attachment strength which in turn leads to their cellular entry and membrane fusion. The latter process is driven by conformational changes of the spike proteins which are also class I fusion proteins, providing the energetics of membrane tethering, bending, and fusion. These viruses exploit cellular and membrane factors in regulating the conformation changes and membrane processes. Herein, we describe the major structural and functional features of spike proteins of the enveloped viruses including highlights on their structural dynamics. The review delves into some of the case studies in the literature discussing the findings on multivalent binding, membrane hemifusion, and fusion of these viruses. The focus is on applications of biophysical tools with an emphasis on single-particle methods for evaluating mechanisms of these processes at the molecular and cellular levels.

Immunomolecular assay based on selective virion capture by spike antibody and viral nucleic acid amplification for detecting intact SARS-CoV-2 particles.

BACKGROUND: Effective therapeutics and vaccines for coronavirus disease 2019 (COVID-19) are currently lacking because of the mutation and immune escape of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Based on the propagation characteristics of SARS-CoV-2, rapid and accurate detection of complete virions from clinical samples and the environment is critical for assessing infection risk and containing further COVID-19 outbreaks. However, currently applicable methods cannot achieve large-scale clinical application due to factors such as the high viral load, cumbersome virus isolation steps, demanding environmental conditions, and long experimental periods. In this study, we developed an immuno molecular detection method combining capture of the viral spike glycoprotein with monoclonal antibodies and nucleic acid amplification via quantitative reverse transcription PCR to rapidly and accurately detect complete virions. RESULTS: After constructing a novel pseudovirus, screening for specific antibodies, and optimizing the detection parameters, the assay achieved a limit of detection of 9 × 102 transduction units/mL of viral titer with high confidence (~ 95%) and excellent stability against human serum and common virus/pseudovirus. The coefficients of variation were 1.0 ~ 2.0% for intra-assay and inter-assay analyses, respectively. Compared with reverse transcription-PCR, the immunomolecular method more accurately quantified complete virions. SARS-CoV-2/pseudovirus was more stable on plastic and paper compared with aluminum and copper in the detection of SARS-CoV-2 pseudovirus under different conditions. Complete virions were detected up to 96 h after they were applied to these surfaces (except for copper), although the titer of the virions was greatly reduced. CONCLUSION: Convenient, inexpensive, and accurate complete virus detection can be applied to many fields, including monitoring the infectivity of convalescent and post-discharge patients and assessing high-risk environments (isolation rooms, operating rooms, patient living environments, and cold chain logistics). This method can also be used to detect intact virions, including Hepatitis B and C viruses, human immunodeficiency virus, influenza, and the partial pulmonary virus, which may further improve the accuracy of diagnoses and facilitate individualized and precise treatments.

An optical visualization of free virions for revealing the first public enemy

Viruses are unseen enemies which tend to disarmingly spread from person-to-person, therefore causing health damages and weakens the immunity system. Their invisibility give rise to various representations, projections and imaginations that allow laypeople to tame the unseen and intangible nature of viruses. For the first time to our knowledge, we are proposing a novel and uncommon scientific approach resting on the synergy between two scientific field: Technical Sciences & Medicine (TSM) with Social Sciences and Humanities (SSH). Therefore, we present the results of our investigations concerning the evaluation of the social impact of the scientific image of free virions on a specific population, particularly affected by the Covid-19 pandemic: people over 60 years old. For this research, we have implemented two scientific imaging solutions to visualize the free viral particles of SARS-CoV-2. The first one is a standard solution of electron microscopy and the second one is an optical and computational solution of microscopy. The scientific representations of SARS-CoV-2 that we have proposed is in fact highly different from the mass media image that we can see everywhere. Concerning the targeted population, we have demonstrated that the scientific image has a negative impact on the population. Thus, the socially constructed representations of these invisible enemies have a preponderant role in driving laypeople’s emotional reactions and health-related behaviors. Therefore, imaging viruses remains a critical scientific effort that contributes irrevocably to alleviate laypeople’s misrepresentations of these invisible enemies. © 2022 SPIE

An instrumental and computational pipeline for visualizing free viral particles

We are presenting the application of an optical and computational pipeline FAMOUS for revealing the presence of free viral particles named “virions”. The idea of such a protocol is to give rise to images of virions in their environment with a soft solution for recording the native image, contrary to the standard solution of imaging virions with electron microscopy (EM) for visualizing viral particles. The final aim of the current work is to observe free viral particles of SARS-CoV-2, the virions responsible for the worldwide pandemic of Covid-19. But such particles have diameters between 80 and 120 nm, a dimension smaller than the resolution limit of optical-only microscopy solutions. We have chosen to start with the biggest free virions, cytomegalovirus (CMV), a virus from the herpesvirus family also named “Human Herpes Virus 5”. Two kinds of cultures were involved: a fluorescent culture (BAD) and a label-free one (VHLE), both being collected from infected cell culture. VHLE virions were first observed after secondary immunostaining and concentrated with magnetic nanoparticles and then without labelling. The optical protocol rests on a standard solution of multiphoton microscopy combined with a computational strategy based on the point-spread-function (PSF) recordings, its mathematical modeling and the restauration of the image resting on the PSF model. A test with free viral particles of SARS-CoV-2 is led, delivering an optical visualization of the free-viral particles. The visualization of objects aggregates obtained in both situations confirm the relevance of the pipeline FAMOUS for imaging free virions. © 2022 SPIE.

Understanding viruses and viral infections by biophotonic methods

In the last few decades outbreaks of viral infections have often challenged the world‐wide health infrastructure and caused a significant financial burden as well as human suffering despite progress in diagnostic technologies. The recent outbreaks of the Ebola virus in the African continent, the Zika virus in the American continent, severe acute respiratory syndrome (SARS), Middle East respiratory syndrome (MN1 -https://media.proquest.com/media/hms/PFT/1/9NKwM?_a=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%3D%3D&_s=kRWbjm7Gq64Rq8ZmQhDgAEQ7U9s%3D ERS), influenza A and lately severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) viral infections have repeatedly highlighted the importance of technological advancement enabling a better understanding of virions. In this review, we systematically discuss different aspects of virions and how their properties and functions can be studied using different light‐based technologies. We focus on virion classification, detection and interactions with the host's immune system. Further, the potential of advanced biophotonic methods, for example, Raman, infrared reflection, absorption and fluorescence spectroscopy, advanced microscopic techniques and biosensor‐based approaches for diagnosing viral infections, investigating therapeutics and vaccine development are described. Although significant advancements have already been made in photonic technologies, which even enable visualizing virion‐host interactions on single‐cell level, the continuous evolution of viruses demands further progress in biophotonic solutions for fast, affordable and robust health monitoring devices for screening viral infections.

(INVITED)Quantitative detection of SARS-CoV-2 virions in aqueous mediums by IoT optical fiber sensors

The COVID-19 pandemic has emphasized the need for portable, small-size, low-cost, simple to use, and highly sensitive sensors able to measure a specific substance, with the capability of the transmission over the Internet of statistical data, such as in this specific case on the spread of the SARS-CoV-2 virions. Moreover, to resolve the COVID-19 emergency, the possibility of making selective SARS-CoV-2 measurements in different aqueous matrices could be advantageous. Thus, the realization of rapid and innovative point-of-care diagnostics tests has become a global priority. In response to the current need for quick, highly sensitive and on-site detection of the SARS-CoV-2 virions in different aqueous solutions, two different nanolayer biorecognition systems separately combined with an adaptable optical fiber sensor have been reported in this work. More specifically, two SARS-CoV-2 sensors have been developed and tested by exploiting a plasmonic plastic optical fiber (POF) sensor coupled with two different receptors, both designed for the specific recognition of the SARS-CoV-2 Spike protein;one is aptamer-based and the other one Molecular Imprinted Polymer-based. The preliminary tests on SARS-CoV-2 virions, performed on samples of nasopharyngeal (NP) swabs in universal transport medium (UTM), were compared with data obtained using reverse-transcription polymerase chain reaction (RT-PCR). According to these preliminary experimental results obtained exploiting both receptors, the sensitivity of the proposed SARS-CoV-2 optical fiber sensors proved to be high enough to detect virions. Furthermore, a relatively fast response time (a few minutes) to detect virions was obtained without additional reagents, with the capability to transmit the data via the Internet automatically. © 2021 The Authors

Numerical simulation of particle transport in the urban boundary layer with implications for SARS-CoV-2 virion distribution

This paper presents a description, verification, and application of a model simulating the transport of aerosol particles with different properties in the urban boundary layer by using a Lagrangian approach. The model takes input fields of air flow characteristics from arbitrary external models or analytical solutions and allows estimating the movement, sedimentation, and decay of particles. In this paper, the accuracy of the model is successfully estimated on the basis of exact analytical solutions. Simulations are made for a series of urban canyons under different conditions of stratification and wind speed to assess the effects of these meteorological parameters on particle transport in urban areas. Under similar conditions, the transport of particles simulating SARS-CoV-2 coronavirus particles is calculated.

Molecular Communications in Viral Infections Research: Modeling, Experimental Data, and Future Directions.

Hundreds of millions of people worldwide are affected by viral infections each year, and yet, several of them neither have vaccines nor effective treatment during and post-infection. This challenge has been highlighted by the COVID-19 pandemic, showing how viruses can quickly spread and impact society as a whole. Novel interdisciplinary techniques must emerge to provide forward-looking strategies to combat viral infections, as well as possible future pandemics. In the past decade, an interdisciplinary area involving bioengineering, nanotechnology and information and communication technology (ICT) has been developed, known as Molecular Communications. This new emerging area uses elements of classical communication systems to molecular signalling and communication found inside and outside biological systems, characterizing the signalling processes between cells and viruses. In this paper, we provide an extensive and detailed discussion on how molecular communications can be integrated into the viral infectious diseases research, and how possible treatment and vaccines can be developed considering molecules as information carriers. We provide a literature review on molecular communications models for viral infection (intra-body and extra-body), a deep analysis on their effects on immune response, how experimental can be used by the molecular communications community, as well as open issues and future directions.

Airway and Alveoli Organoids as Valuable Research Tools in COVID-19.

The coronavirus disease 2019 (COVID-19), caused by the novel coronavirus, SARS-CoV-2, affects tissues from different body systems but mostly the respiratory system, and the damage evoked in the lungs may occasionally result in severe respiratory complications and eventually lead to death. Studies of human respiratory infections have been limited by the scarcity of functional models that mimic in vivo physiology and pathophysiology. In the last decades, organoid models have emerged as potential research tools due to the possibility of reproducing in vivo tissue in culture. Despite being studied for over one year, there is still no effective treatment against COVID-19, and investigations using pulmonary tissue and possible therapeutics are still very limited. Thus, human lung organoids can provide robust support to simulate SARS-CoV-2 infection and replication and aid in a better understanding of their effects in human tissue. The present review describes methodological aspects of different protocols to develop airway and alveoli organoids, which have a promising perspective to further investigate COVID-19.

Virology from the perspective of theoretical colloid and interface science.

Viral infections occur at very different length and time scales and include various processes, which can often be described using the models developed and/or employed in colloid and interface science. Bearing in mind the currently active COVID-19, I discuss herein the models aimed at viral transmission via respiratory droplets and the contact of virions with the epithelium. In a more general context, I outline the models focused on penetration of virions via the cellular membrane, initial stage of viral genome replication, and formation of viral capsids in cells. In addition, the models related to a new generation of drug delivery vehicles, for example, lipid nanoparticles with size about 100-200 nm, are discussed as well. Despite the high current interest in all these processes, their understanding is still limited, and this area is open for new theoretical studies.

The porcine deltacoronavirus accessory protein NS6 is expressed in vivo and incorporated into virions.

Porcine deltacoronavirus (PDCoV) is one of the emerged coronaviruses posing a significant threat to the swine industry. Previous work showed the presence of a viral accessory protein NS6 in PDCoV-infected cells. In this study, we detected the expression of the NS6 protein in small intestinal tissues of PDCoV-infected piglets. In addition, SDS-PAGE and Western blot analysis of sucrose gradient-purified virions showed the presence of a 13-kDa NS6 protein. Further evidences of the presence of NS6 in the PDCoV virions were obtained by immunogold staining of purified virions with anti-NS6 antiserum, and by immunoprecipitation of NS6 from purified virions. Finally, the anti-NS6 antibody was not able to neutralize PDCoV in cultured cells. These data establish for the first time that the accessory protein NS6 is expressed during infection in vivo and incorporated into PDCoV virions.

Polymers with antiviral properties: A brief review.

Viruses that are pathogenic to humans and livestock pose a serious epidemiological threat and challenge the world's population. The SARS-CoV-2/COVID-19 pandemic has made the world aware of the scale of the threat. The surfaces of various materials can be a source of viruses that remain temporarily contagious in the environment. Few polymers have antiviral effects that reduce infectivity or the presence of a virus in the human environment. Some of the effects are due to certain physical properties, e.g., high hydrophobicity. Other materials owe their antiviral activity to a modified physicochemical structure favoring the action on specific virus receptors or on their biochemistry. Current research areas include: gluten, polyvinylidene fluoride, polyimide, polylactic acid, graphene oxide, and polyurethane bound to copper oxide. The future belongs to multi-component mixtures or very thin multilayer systems. The rational direction of research work is the search for materials with a balanced specificity in relation to the most dangerous viruses and universality in relation to other viruses.