Viruses Commons^™

Overcoming Vaccine-Associated Enhanced Respiratory Disease In Swine Flu With Lipid Nanoparticle-Dna Technology, The Nguyen

School of Veterinary and Biomedical Sciences: Dissertations, Theses, and Student Research

The two primary challenges in developing vaccines against swine influenza A virus (IAV-S) are the continuous viral evolution and vaccine-associated enhanced respiratory disease (VAERD). Current vaccine strategies share a common limitation: the significant time required for licensing updated vaccines. Moreover, the strain selection for vaccine updates depends entirely on predictions, raising the risk that chosen strains may not match circulating viruses. Such mismatches not only reduce vaccine efficacy but also risk exacerbating disease severity through VAERD. Here, in this thesis, we developed and evaluated a novel DNA-based vaccine platform designed to address the need for rapid updates in response to …

Host Genetics Role In Viral Disease Susceptibility, Christian Willis Eaton

Dissertations and Doctoral Documents, University of Nebraska-Lincoln, 2023–

Viruses and their hosts exist in an evolutionary arms race, with evolutionary promotion of host genetic-based defense mechanisms and evolution of virus genetic-based mechanisms to overcome host defenses. Viruses are capable of much more rapid evolution and adaptation than their hosts, often leading to the use of host cellular mechanisms to promote viral infection and replication. In this study, we describe the generation of a detection assay for atypical porcine pestivirus (APPV), designed to target the viral 5’ UTR, a region of the viral genome which sees less mutations than the protein coding regions. This assay was generated to be …

Identification Of Benzothiophene-Derived Inhibitors Of Flaviviruses By Targeting Rna-Dependent Rna Polymerase, Leah Liu Wang, Shazeed-Ul Karim, Aidan Hand, Ryan Brunkhorst, Mackenna Petersen, Sarah Altam, Yi Liu, Luwen Zhang, Fengwei Bai, Shi-Hua Xiang

School of Biological Sciences: Faculty Publications

Flaviviruses such as Dengue, West Nile, and Zika viruses are mosquito-borne RNA viruses that can cause serious diseases in humans. To develop effective drugs for treating these viruses’ infections, we create a new approach for developing common or shared drugs that may work for several different viral species of flaviviruses. It is based on the conserved RNA-dependent RNA polymerase (RdRp), which is the key enzyme for viral replication. We built up a common structure of RdRps (POLcon) from their consensus sequence. A conserved Triple-D structural motif was identified at the active site of POLcon that has been used for virtual …

Triticale As A Source Of Resistance To Wheat Streak Mosaic Virus And Triticum Mosaic Virus, Marcos Winicius Goncalves De Souza

Department of Agronomy and Horticulture: Dissertations, Theses, and Student Research

Wheat streak mosaic virus (WSMV) and Triticum mosaic virus (TriMV) present a great threat to wheat production in the Great Plains area of the United States due to the loss of photosynthetic area causing reductions in crop yields. Effective control of these viruses is limited to a few available strategies including controlling volunteer wheat and using currently identified resistant genes to both the vector and viruses. While effective, these genes show temperature sensitivity and yield drag, and evidence that the vectors and viruses can overcome them has been found. Thus, new methods of genetic resistance are urgently needed. Field observations …

Exploring Subtype C Hiv-1 Tissue Reservoirs And The Impact Of Cannabis Use In Virally Suppressed African Individuals, Zhou Liu

Dissertations and Doctoral Documents, University of Nebraska-Lincoln, 2023–

Antiretroviral therapy (ART) has dramatically prolonged the lives of people living with HIV (PLWH), but it is not curative as it cannot eliminate integrated HIV-1 proviruses in different tissue reservoirs. Even though subtype C HIV-1 accounts for almost half of global HIV-1 infections, few studies had evaluated HIV-1 reservoirs for this subtype. Many co-factors, including the lifestyle factor cannabis use, have been investigated for their potential effects in HIV-1 infection. However, only limited information exists regarding cannabis’s effects on HIV-1 infections in vivo, and the impact of cannabis use on HIV-1 parenchymal tissue reservoirs is unexplored. This dissertation aims to …

Novel Insights Into The Autoimmune Mechanisms In Coxsackievirus B Infections And Their Prevention By Vaccination, Mahima Theresa Rasquinha

Dissertations and Doctoral Documents, University of Nebraska-Lincoln, 2023–

Coxsackievirus Bs (CVBs) are RNA viruses of the genus Enterovirus belonging to the Picornaviridae family. CVBs commonly infect humans, and multiple CVB types can induce similar diseases such as myocarditis, pancreatitis, and type 1 diabetes (T1D). Although CVB infections can lead to the generation of autoantibodies, their role in disease is poorly defined, and vaccines are not available to prevent or minimize these infections. To fill these gaps, we first analyzed the autoantibody repertoire in mice infected with CVB3 by Phage ImmunoPrecipitation Sequencing (PhIP-Seq) and identified autoantibodies for several novel antigens not previously reported. The data were validated using enzyme-linked …

The Developmental Process For Adenoviral Vectored Canine Influenza Vaccines, Nicholas Jeanjaquet

School of Biological Sciences: Dissertations, Theses, and Student Research

Canine Influenza Virus (CIV) is a recently emerged branch of Influenza A virus that is highly infectious in dogs. The first strain was isolated in 2003, and it has quickly become endemic in areas with dense dog populations, most notably in Asia. The proximity of dogs to humans, along with their potential to serve as mixing vessels for reassortment, raises concern for possible zoonotic transmission and a potential human pandemic. Available vaccines are not frequently updated and struggle to prevent the spread of currently circulating strains. This highlights the need for a new and more effective vaccine. We outline the …

Evaluation Of Protective Efficacy Of Viral Vector Based Swine Influenza Vaccine And A Method For B Cell Culture For Monoclonal Antibody Generation, Sushmita Kumari

School of Veterinary and Biomedical Sciences: Dissertations, Theses, and Student Research

Influenza A virus of the swine (IAV-S) is an economically important swine pathogen that has the potential to spread to humans, thus posing an ongoing public health concern due to its zoonotic potential. Hemagglutinin (HA), the most abundant viral envelope protein, is known to be the key protective antigen. Anti-HA antibodies alone, have been shown to prevent IAV infection. In this study we evaluated the feasibility of a recombinant tri segmented Pichinde virus (PICV) as a viral vector to deliver IAV-S hemagglutinin antigen in pigs. Four groups of weaned pigs (T01-04) were immunized twice with PBS, rPICV-GFP as a vector …

Host Cell Responses To Zika Virus Infection, Bikash Ranjan Sahoo

School of Veterinary and Biomedical Sciences: Dissertations, Theses, and Student Research

The re-emergence of Zika virus (ZIKV) in 2015 as a significant human pathogen causing neurological diseases in infants as well as adults is a serious global health concern. A clear understanding of the mechanisms involved in ZIKV replication in infected host cells as well as the host responses to virus infection are keys to the development of therapeutic strategies against ZIKV. Studies conducted in this dissertation demonstrate that ZIKV infection induces the activation of mTOR signaling cascade that promotes viral protein accumulation and infectious progeny production. While both mTORC1 and mTORC2 are essential for ZIKV replication, the observation that depletion …

Development Of Vaccines And Antivirals Against Zika Virus, Aryamav Pattnaik

School of Veterinary and Biomedical Sciences: Dissertations, Theses, and Student Research

The re-emergence of Zika virus (ZIKV), an arbovirus, poses a major global human health concern because of its ability to cause congenital abnormalities and neurological diseases. While many candidate vaccines and antiviral drugs are in the developmental pipeline, none have been approved for use against ZIKV infection. This dissertation describes the characterization of one vaccine and two antiviral drug candidates against ZIKV infection. A bacterial ferritin-based nanoparticle vaccine, termed zDIII-F, is designed to display multiple copies of ZIKV E protein domain III on its surface. These stable nanoparticles are shown to induce robust antibody-mediated protection against lethal ZIKV infection in …

Covid19 Disease Map, A Computational Knowledge Repository Of Virus–Host Interaction Mechanisms, Marek Ostaszewski, Tomáš Helikar, Bhanwar Lal Puniya, A Host Of Co-Authors, Covid-19 Disease Map Community

Department of Biochemistry: Faculty Publications

We need to effectively combine the knowledge from surging literature with complex datasets to propose mechanistic models of SARS-CoV-2 infection, improving data interpretation and predicting key targets of intervention. Here, we describe a large-scale community effort to build an open access, interoperable and computable repository of COVID-19 molecular mechanisms. The COVID-19 Disease Map (C19DMap) is a graphical, interactive representation of disease-relevant molecular mechanisms linking many knowledge sources. Notably, it is a computational resource for graph-based analyses and disease modelling. To this end, we established a framework of tools, platforms and guidelines necessary for a multifaceted community of biocurators, domain experts, …

Smrt Sequencing Of Paramecium Bursaria Chlorella Virus-1 Reveals Diverse Methylation Stability In Adenines Targeted By Restriction Modification Systems, Samantha R. Coy, Eric R. Gann, Spiridon E. Papoulis, Michael E. Holder, Nadim J. Ajami, Joseph F. Petrosino, Erik R. Zinser, James L. Van Etten, Steven W. Wilhelm

James Van Etten Publications

Chloroviruses (family Phycodnaviridae) infect eukaryotic, freshwater, unicellular green algae. A unique feature of these viruses is an abundance of DNA methyltransferases, with isolates dedicating up to 4.5% of their protein coding potential to these genes. This diversity highlights just one of the long-standing values of the chlorovirus model system; where group-wide epigenomic characterization might begin to elucidate the function(s) of DNA methylation in large dsDNA viruses. We characterized DNA modifications in the prototype chlorovirus, PBCV-1, using single-molecule real time (SMRT) sequencing (aka PacBio). Results were compared to total available sites predicted in silico based on DNA sequence alone. SMRT-software detected …

On The Inadequacy Of Species Distribution Models For Modelling The Spread Of Sars-Cov-2: Response To Araújo And Naimi, Joseph D. Chipperfield, Blas M. Benito, Robert B. O'Hara, Richard J. Telford, Colin J. Carlson

United States Public Health Resources

The ongoing pandemic of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is causing significant damage to public health and economic livelihoods, and is putting significant strains on healthcare services globally. This unfolding emergency has prompted the preparation and dissemination of the article “Spread of SARS-CoV-2 Coronavirus likely to be constrained by climate” by Araújo and Naimi (2020). The authors present the results of an ensemble forecast made from a suite of species distribution models (SDMs), where they attempt to predict the suitability of the climate for the spread of SARS-CoV-2 over the coming months. They argue that climate is …

Environmental Sampling Techniques For Herd-Level Surveillance Of Bovine Viral Diarrhea Virus, Jaden Carlson

School of Veterinary and Biomedical Sciences: Dissertations, Theses, and Student Research

Control of Bovine Viral Diarrhea Virus (BVDV) relies on resource-intensive individual animal sampling to detect and remove persistently infected (PI) cattle. Herd-level surveillance tools would be useful for herds with unknown BVDV status and for monitoring herds with BVDV-free status. The overall objective of this thesis is to explore the viability of BVDV surveillance at a herd-level using samples collected without handling individual animals. The first objective was to determine the feasibility of using stable flies as a sampling tool to detect BVDV. The second objective was to determine the feasibility of using drinking water to detect BVDV. To accomplish …

Regulation Of Vaccinia Virus Replication: A Story Of Viral Mimicry And A Novel Antagonistic Relationship Between Vaccinia Kinase And Pseudokinase, Annabel T. Olson

School of Biological Sciences: Dissertations, Theses, and Student Research

Poxviruses employ sophisticated signaling pathways that thwart cellular defense mechanisms and simultaneously ensure viral factors are modulated properly. Yet, our understanding of these complex signaling networks are incomplete. For example, the vaccinia B1 kinase plays a vital role in inactivating the cellular antiviral factor BAF, and is suggested to orchestrate other pathways. B1 is highly conserved among poxviruses and exhibits a remarkable degree of similarity to VRKs, a family of cellular kinases, suggesting that the viral enzyme has evolved to mimic VRK activity. Indeed, B1 and VRKs have been demonstrated to target a shared substrate, the DNA binding protein BAF, …

The N-Glycan Structures Of The Antigenic Variants Of Chlorovirus Pbcv-1 Major Capsid Protein Help To Identify The Virus-Encoded Glycosyltransferases, Immacolata Speciale, Garry A. Duncan, Luca Unione, Irina Agarkova, Domenico Garozzo, Jesus Jimenez-Barbero, Sicheng Lin, Todd L. Lowary, Antonio Molinaro, Eric Noel, Maria Elena Laugieri, Michela Tonetti, James L. Van Etten, Cristina De Castro

James Van Etten Publications

The chlorovirus Paramecium bursaria chlorella virus 1 (PBCV-1) is a large dsDNA virus that infects the microalga Chlorella variabilis NC64A. Unlike most other viruses, PBCV-1 encodes most, if not all, of the machinery required to glycosylate its major capsid protein (MCP). The structures of the four N-linked glycans from the PBCV-1 MCP consist of nonasaccharides, and similar glycans are not found elsewhere in the three domains of life. Here, we identified the roles of three virus-encoded glycosyltransferases (GTs) that have four distinct GT activities in glycan synthesis. Two of the three GTs were previously annotated as GTs but the third …

Chloroviruses Lure Hosts Through Long-Distance Chemical Signaling, David Dunigan, Maitham Ahmed Al-Sammak, Zeina Al-Ameeli, Irina Agarkova, John Delong, James L. Van Etten

James Van Etten Publications

Chloroviruses exist in aquatic systems around the planet where they infect certain eukaryotic green algae that are mutualistic endosymbionts in a variety of protists and metazoans. Natural chlorovirus populations are seasonally dynamic but the precise temporal changes in these populations and the mechanisms that underlie them have, heretofore, been unclear. We recently reported the novel concept that predator/prey-mediated virus activation regulates chlorovirus population dynamics, and in the current manuscript demonstrate virus packaged chemotactic modulation of prey behavior.

Viruses have not previously been reported to act as chemotactic/chemo-attractive agents. Rather, viruses as extracellular entities are generally viewed as non-metabolically active spore-like …

Host Factors Against Plant Viruses, Hernan García-Ruiz

Department of Plant Pathology: Faculty Publications

Plant virus genome replication and movement is dependent on host resources and factors. However, plants respond to virus infection through several mechanisms, such as autophagy, ubiquitination, mRNA decay and gene silencing, that target viral components. Viral factors work in synchrony with pro-viral host factors during the infection cycle and are targeted by antiviral responses. Accordingly, establishment of virus infection is genetically determined by the availability of the pro-viral factors necessary for genome replication and movement, and by the balance between plant defence and viral suppression of defence responses. Sequential requirement of pro-viral factors and the antagonistic activity of antiviral factors …

Contributions Of The Gp120 Variable Loops To Envelope Glycoprotein Trimer Stability In Primate Lentiviruses, Dane M. Bowder

School of Biological Sciences: Dissertations, Theses, and Student Research

Human Immunodeficiency Virus (HIV) is the etiological agent of AIDS and is responsible for the AIDS pandemic worldwide. According to UNAIDS, as of 2016, there were approximately 36.7 million people living with HIV globally, and 1.8 million new infections that year. While antiretroviral therapies and education continue to reduce these numbers, a preventative vaccine is still required to curb this epidemic. The envelope glycoprotein trimer of HIV, which is the sole protein on the surface of the virus and facilitates entry of the virus into host cells, is of keen interest to the HIV vaccine and drug-development field. Recently, the …

Evaluation Of A Human Papillomavirus Genotyping Assay For Cervical Cancer Screening In Tanzania, Kandali S. Kapie

School of Biological Sciences: Dissertations, Theses, and Student Research

Abstract: Mucosal high-risk Human papillomavirus (HR-HPV) has been identified as the primary factor in causing cervical cancer, the most common cancer in women of low and middle-income countries (LMICs). Cervical cancer screening, incorporating the HPV test shown to be more advantageous than screening by visual inspection with acetic acid (VIA) alone. However, due to resources constrain these countries could not afford the available HPV tests. We, therefore, validated a cost-effective Multiplex PCR HPV (mPCR) assay, a recently developed assay for detecting sixteen genital HPV, fourteen HR-HPV and 2 low-risk genotypes. We postulate that this HPV multiplex assay will be at …

Type 2 Bovine Viral Diarrhea Virus NPro Suppresses Type I Interferon Pathway Signaling In Bovine Cells And Augments Bovine Respiratory Syncytial Virus Replication, Abdulrahman Abdulaziz A. Alkheraif

School of Veterinary and Biomedical Sciences: Dissertations, Theses, and Student Research

Bovine viral diarrhea virus (BVDV) and bovine respiratory syncytial virus (BRSV) infections contribute to the bovine respiratory disease complex (BRDC), which is a multi-factorial disorder involving co-infections of viruses and bacteria including mycoplasma. BRDC causes great economic losses to the United States feedlot industry. BVDV infection induces immunosuppression in infected animals. BVDV N^pro binds and degrades the transcription factor interferon regulatory factor-3 (IRF-3) and effectively blocks type I interferon (type I IFN) expression in host cells. BRSV nonstructural proteins, NS1 and NS2, block activation of IRF-3. In calves, concurrent infection with BVDV and BRSV resulted in more severe clinical …

Biophysical Approaches To Solve The Structures Of The Complex Glycan Shield Of Chloroviruses, Cristina De Castro, Garry Duncan, Domenico Garozzo, Antonio Molinaro, Luisa Sturiale, Michela Tonetti, James L. Van Etten

James Van Etten Publications

The capsid of Paramecium bursaria chlorella virus (PBCV-1) contains a heavily glycosylated major capsid protein, Vp54. The capsid protein contains four glycans, each N-linked to Asn. The glycan structures are unusual in many aspects: (1) they are attached by a β-glucose linkage, which is rare in nature; (2) they are highly branched and consist of 8–10 neutral monosaccharides; (3) all four glycoforms contain a dimethylated rhamnose as the capping residue of the main chain, a hyper-branched fucose residue and two rhamnose residues ''with opposite absolute configurations; (4) the four glycoforms differ by the nonstoichiometric presence of two monosaccharides, l-arabinose and …

Genes For Membrane Transport Proteins: Not So Rare In Viruses, Timo Greiner, Anna Moroni, James L. Van Etten, Gerhard Thiel

James Van Etten Publications

Some viruses have genes encoding proteins with membrane transport functions. It is unknown if these types of proteins are rare or are common in viruses. In particular, the evolutionary origin of some of the viral genes is obscure, where other viral proteins have homologs in prokaryotic and eukaryotic organisms. We searched virus genomes in databases looking for transmembrane proteins with possible transport function. This effort led to the detection of 18 different types of putative membrane transport proteins indicating that they are not a rarity in viral genomes. The most abundant proteins are K+ channels. Their predicted structures vary between …

Size-Dependent Catalysis Of Chlorovirus Population Growth By A Messy Feeding Predator, John Delong, Zeina Al-Ameeli, Shelby Lyon, James L. Van Etten, David Dunigan

James Van Etten Publications

Many chloroviruses replicate in endosymbiotic zoochlorellae that are protected from infection by their symbiotic host. To reach the high virus concentrations that often occur in natural systems, a mechanism is needed to release zoochlorellae from their hosts. We demonstrate that the ciliate predator Didinium nasutum foraging on zoochlorellae-bearing Paramecium bursaria can release live zoochlorellae from the ruptured prey cell that can then be infected by chloroviruses. The catalysis process is very effective, yielding roughly 95% of the theoretical infectious virus yield as determined by sonication of P. bursaria. Chlorovirus activation is more effective with smaller Didinia, as larger …

Cross-Reactive Immunity To Porcine Reproductive And Respiratory Syndrome Virus And Its Contribution To Protection, Ignacio Correas

School of Veterinary and Biomedical Sciences: Dissertations, Theses, and Student Research

Due to the vast geographical distribution and significant economic losses generated, porcine reproductive and respiratory syndrome virus (PRRSV) can be considered the most important swine pathogen of contemporary times. Current control and eradication strategies against PRRSV have difficulty succeeding because of their complex nature and the absence of an effective vaccine. A major obstacle for PRRSV vaccine development is the broad heterogeneity of the virus, both at the genetic and antigenic level, its rapid evolution, and an incomplete knowledge of the immune responses responsible for clearing the virus from the host. Specifically, how known correlates of protection against PRRSV—neutralizing antibodies …

Multiple Origins Of Endosymbionts In Chlorellaceae With No Reductive Effects On The Plastid Or Mitochondrial Genomes, Weishu Fan, Wenhu Guo, James L. Van Etten, Jeffrey P. Mower

James Van Etten Publications

Ancient endosymbiotic relationships have led to extreme genomic reduction in many bacterial and eukaryotic algal endosymbionts. Endosymbionts in more recent and/or facultative relationships can also experience genomic reduction to a lesser extent, but little is known about the effects of the endosymbiotic transition on the organellar genomes of eukaryotes. To understand how the endosymbiotic lifestyle has affected the organellar genomes of photosynthetic green algae, we generated the complete plastid genome (plastome) and mitochondrial genome (mitogenome) sequences from three green algal endosymbionts (Chlorella heliozoae, Chlorella variabilis and Micractinium conductrix). The mitogenomes and plastomes of the three newly sequenced endosymbionts …

Induction And Suppression Of Antiviral Rna Silencing By Tomato Spotted Wilt Virus, Sergio M. Gabriel Peralta

Department of Agronomy and Horticulture: Dissertations, Theses, and Student Research

Tomato spotted wilt virus (TSWV) is an emerging pathogen with wide host range and one of the most important viruses of plants. Information regarding processing of negative single stranded RNA viruses such as TSWV in the RNA silencing pathway remains limited. In nature TSWV is only transmitted by thrips as vectors and since infection occurs in both thrips and plants, an experimental system to transmit using thrips and the detection of TSWV were established. In order to understand the processing of TSWV in the RNA silencing pathway, Arabidopsis thaliana as a model plant was used in the genetic analysis against …

Identification Of Intrahelical Bifurcated H‑Bonds As A New Type Of Gate In K+ Channels, Oliver Rauh, Martin Urban, Leonhard M. Henkes, Tobias Winterstein, Timo Greiner, James L. Van Etten, Anna Moroni, Stefan M. Kast, Gerhard Thiel, Indra Schroeder

James Van Etten Publications

Gating of ion channels is based on structural transitions between open and closed states. To uncover the chemical basis of individual gates, we performed a comparative experimental and computational analysis between two K⁺ channels, Kcv_S and Kcv_NTS. These small viral encoded K+ channel proteins, with a monomer size of only 82 amino acids, resemble the pore module of all complex K+ channels in terms of structure and function. Even though both proteins share about 90% amino acid sequence identity, they exhibit different open probabilities with ca. 90% in Kcv_NTS and 40% in Kcv_S. …

Structure Of The Chlorovirus Pbcv-1 Major Capsid Glycoprotein Determined By Combining Crystallographic And Carbohydrate Molecular Modeling Approaches, Cristina De Castro, Thomas Klose, Immacolata Speciale, Rosa Lanzetta, Antonio Molinaro, James L. Van Etten, Michael G. Rossmann

James Van Etten Publications

The glycans of the major capsid protein (Vp54) of Paramecium bursaria chlorella virus (PBCV-1) were recently described and found to be unusual. This prompted a reexamination of the previously reported Vp54 X-ray structure. A detailed description of the complete glycoprotein was achieved by combining crystallographic data with molecular modeling. The crystallographic data identified most of the monosaccharides located close to the protein backbone, but failed to detect those further from the glycosylation sites. Molecular modeling complemented this model by adding the missing monosaccharides and examined the conformational preference of the whole molecule, alone or within the crystallographic environment. Thus, combining …

Conversion Of An Instantaneous Activating K+ Channel Into A Slow Activating Inward Rectifier, Dirk Baumeister, Brigitte Hertel, Indra Schroeder, Sabrina Gazzarrini, Stefan M. Kast, James L. Van Etten, Anna Moroni, Gerhard Thiel

James Van Etten Publications

The miniature channel, Kcv, is a structural equivalent of the pore of all K⁺ channels. Here, we follow up on a previous observation that a largely voltage-insensitive channel can be converted into a slow activating inward rectifier after extending the outer transmembrane domain by one Ala. This gain of rectification can be rationalized by dynamic salt bridges at the cytosolic entrance to the channel; opening is favored by voltage-sensitive formation of salt bridges and counteracted by their disruption. Such latent voltage sensitivity in the pore could be relevant for the understanding of voltage gating in complex Kv channels.