Virology Commons^™

Pseudomonas Bacteriophage Phi6 As A Model For Virus Emergence, Brian Elwood Ford

Dissertations, Theses, and Capstone Projects

The Pseudomonas bacteriophage Φ6 has a long and well-established history as a model organism. Here we describe a set of experiments to extend this model system to concepts previously unclaimed. Chapter 1 presents a brief background of the ecology of viruses that infect microorganisms. Chapter 2 examines genetic mutations allowing for host range expansion. Chapter 3 presents a novel paired strain assay to study how a non genetic host-acquired factor affects fitness of these enveloped viruses on subsequent hosts. Chapter 4 is an extension of this system to include how the bacteria host is affected in virus-host coevolution.

The Effect Of Environmental Selection Pressure On The Rate Of Recombination To An Advantageous Receptor Mutation In Bovine Coronavirus, Gavin Schroter, Sally Hall, Mona Hwang, Monica Borucki

STAR Program Research Presentations

Bovine Coronavirus (BCoV) is an important analogue in understanding the effectiveness of zoonotic, single-stranded, positive sense RNA viruses. Many of the most recent viral outbreaks have been attributed to RNA viruses that have one, or more, animal reservoirs [1]. BCoV is such a great candidate for studying these types of viruses because they are from the family Coronaviridae, which also contains the viruses that cause severe acute respiratory syndrome (SARS) and Middle East respiratory syndrome (MERS). The goal of this study was to observe changes in genetic makeup of the virus’ outer membrane Spike protein via recombination between two …

Kaposi’S Sarcoma-Associated Herpesvirus Transmission And Infection Among Young Zambian Children, Landon Olp

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

Kaposi’s sarcoma-associated herpesvirus (KSHV) is the etiologic agent for all forms of Kaposi’s sarcoma (KS)—one of the most common pediatric cancers in sub-Saharan Africa during the AIDS epidemic. KS was endemic in sub-Saharan Africa prior to the HIV/AIDS epidemic, but KS cases drastically increased thereafter. Our laboratory previously observed that KSHV infection is common among Zambian children and saliva is the major route of transmission. However, additional factors associated with the transmission of KSHV to children are poorly understood. Since a vaccine against KSHV is not currently available, it is paramount to understand factors associated with transmission so that alternative …

Chronic Inflammation As A Result Of Hepatitis C Virus Infection: A Review Of The Literature, Samantha L. Lane

DePaul Discoveries

Approximately 170 million people are infected with Hepatitis C virus (HCV) worldwide^5,6. It is estimated that roughly 80% of those infected suffer from persistent infection with the virus; this persistence of infection is progressive, and over time can lead to fibrosis, cirrhosis, and hepatocellular carcinoma⁷. Chronic inflammation and apoptotic deregulation are both hallmarks of chronic HCV infection, and many molecular pathways are initiated in both the innate and adaptive immune responses during infection with this viral pathogen. The aim of this review was to survey some of the major molecular mechanisms responsible for the induction of …

Detection Of Hcmv Viral Il-10 (Vil-10) In Healthy Blood Donors, Vivian P. Young

Master's Theses

Human cytomegalovirus (HCMV) is widespread in the general population and can establish lifelong latency with expression of a limited subset of viral genes. The UL111A gene is expressed during both lytic and latent infection, and at least two protein isoforms are produced. During lytic infection, the full length transcript yields cmvIL-10, a potent immunosuppressive viral ortholog of human IL-10 (hIL-10). Alternative splicing of the UL111A transcript yields a truncated protein, LAcmvIL-10, which is expressed during both lytic and latent infection but with a limited range of immunosuppressive functions. The two viral cytokines, collectively termed viral IL-10 (vIL-10), are identical in …

Infection And Dissemination Of Tav-Gfp Tagged Sindbis In Aedine Mosquitoes And Cell Lines, Jason J. Saredy

UNF Graduate Theses and Dissertations

Arthropod-borne-viruses (arboviruses) pose a global threat due to their ability to be transmitted by hematophagous insects to vertebrate hosts resulting in a range of serious infectious diseases. Sindbis virus (SINV) is the prototype arbovirus of the genus Alphavirus in the family Togaviridae. The purpose of this study was to investigate the use of a fluorescent tagged reporter virus in both in vitro and in vivo environments. The fluorescent protein GFP was inserted between the Capsid and PE2 in the genome of TR339; SINV TaV-GFP (Wm. Klimstra Lab). This virus construct should have the same infectivity and virulence as wild type …

Enhanced Expression Of Codon Optimized Mycobacterium Avium Subsp. Paratuberculosis Antigens In Lactobacillus Salivarius, Christopher D. Johnston, John P. Bannatine, Rodney Govender, Lorraine Endersen, Daniel Pletzer, Helge Weingart, Aidan Coffey, Jim O'Mahony, Roy D. Sleator

Department of Biological Sciences Publications

It is well documented that open reading frames containing high GC content show poor expression in A+T rich hosts. Specifically, G+C-rich codon usage is a limiting factor in heterologous expression of Mycobacterium avium subsp. paratuberculosis (MAP) proteins using Lactobacillus salivarius. However, re-engineering opening reading frames through synonymous substitutions can offset codon bias and greatly enhance MAP protein production in this host. In this report, we demonstrate that codon-usage manipulation of MAP2121c can enhance the heterologous expression of the major membrane protein (MMP), analogous to the form in which it is produced natively by MAP bacilli. When heterologously over-expressed, antigenic determinants …

Cell Kinase Inhibitor Panel Reveals Multiple Targets To Prevent Replication Of Varicella-Zoster Virus, Bryan Egan Bunnell

Renée Crown University Honors Thesis Projects - All

The alphaherpesvirus that causes chicken pox and shingles, varicella-zoster virus (VZV), infects skin fibroblasts and keratinocytes. These cells are typically quiescent and it is known that VZV manipulates their intracellular environment to activate MAPK signaling cascades, cell cycle regulators, and many transcription factors for its replication. We hypothesized that inhibition of cell kinases would prevent VZV replication and also elucidate which pathways are most important. We evaluated 80 kinase inhibitors for cytotoxicity and anti-proliferative effects on human foreskin fibroblasts, and then determined their antiviral efficacy against VZV-ORF9-Luc strain. Ten well-tolerated, potent kinase inhibitors whose targets are critical mediators of VZV …

The Sumoylation Of The Non-Structural Protein 1 Of The Influenza A Virus Plays A Dual Role During Viral Infection, Jason Michael Chacon

Open Access Theses & Dissertations

The potential for a highly pathogenic influenza pandemic remains a concern for global health. The ability of the influenza virus to undergo antigenic shift and genetic drift give circulating strains of influenza the high probability of developing resistance to current antiviral therapies. Emerging strains of higher virulence, to which the infected person has not had any previous exposure to viral antigens, pose a greater threat of serious illness and death. Currently, there is no antiviral therapy that is effective against all strains of influenza, emphasizing the need to develop new strategies that target cellular systems required for viral replication to …

Different Meal, Same Flavor: Cospeciation And Host Switching Of Haemosporidian Parasites In Some Non-Passerine Birds, Diego Santiago-Alarcon, Adriana Rodríguez-Ferraro, Patricia Parker, Robert Ricklefs

Biology Department Faculty Works

BackgroundPrevious studies have shown that haemosporidian parasites (Haemoproteus(Parahaemoproteus) and Plasmodium) infecting passerine birds have an evolutionary history of host switching with little cospeciation, in particular at low taxonomic levels (e.g., below the family level), which is suggested as the main speciation mechanism of this group of parasites. Recent studies have characterized diverse clades of haemosporidian parasites (H. (Haemoproteus) and H. (Parahaemoproteus)) infecting non-passerine birds (e.g., Columbiformes, Pelecaniiformes). Here, we explore the cospeciation history of H. (Haemoproteus) and H. (Parahaemoproteus) parasites with their non-passerine hosts.MethodsWe sequenced the mtDNA cyt b gene of both haemosporidian parasites and their avian non-passerine hosts. We …

Characterizing Dsrna Production In Virus-Infected Fish Cells, Amal Brek Aloufi

Theses and Dissertations (Comprehensive)

Viral dsRNA is produced by almost all viruses sometime during their replicative cycle. These viral nucleic acids are potent inducers of both innate and adaptive immune responses, and are therefore considered important immuno-modulators. Previous studies have shown that viruses produce dsRNA when replicating in mammalian cells; however, to date no one has demonstrated viral dsRNA production in virus infected fish cells. Therefore, the goal of this study is to investigate dsRNA production by fish viruses in fish cells, verifying production and performing initial characterization of the dsRNA molecules being produced. Three different rainbow trout cell lines were used in this …

Reversible Inactivation And Desiccation Tolerance Of Silicified Viruses, James R. Laidler, Jessica A. Shugart, Sherry L. Cady, Keith S. Bahjat, Kenneth M. Stedman

Biology Faculty Publications and Presentations

Long-distance host-independent virus dispersal is poorly understood, especially for viruses found in isolated ecosystems. To
demonstrate a possible dispersal mechanism, we show that bacteriophage T4, archaeal virus Sulfolobus spindle-shaped virus Kamchatka, and vaccinia virus are reversibly inactivated by mineralization in silica under conditions similar to volcanic hot
springs. In contrast, bacteriophage PRD1 is not silicified. Moreover, silicification provides viruses with remarkable desiccation
resistance, which could allow extensive aerial dispersal.

Barrier To Autointegration Factor (Baf) Inhibits Vaccinia Virus Intermediate Transcription In The Absence Of The Viral B1 Kinase, Nouhou Ibrahim, April Wicklund, Augusta Jamin, Matthew S. Wiebe

Nebraska Center for Virology: Faculty Publications

Barrier to autointegration factor (BAF/BANF1) is a cellular DNA-binding protein found in the nucleus and cytoplasm. Cytoplasmic BAF binds to foreign DNA and can act as a defense against vaccinia DNA replication. To evade BAF, vaccinia expresses the B1 kinase, which phosphorylates BAF and blocks its ability to bind DNA. Interestingly, B1 is also needed for viral intermediate gene expression via an unknown mechanism. Therefore, we evaluated the impact of B1-BAF signaling on vaccinia transcription. Strikingly, the decrease in vaccinia transcription caused by loss of B1 can be rescued by depletion of BAF. The repressive action of BAF is greatest …

Defining The Requirements For Early Gene Expression In Bacteriophage Hk639, Amanda L. Seaton

Masters Theses & Specialist Projects

Lambdoid phages suppress transcription termination to fully express their genes. Antitermination of early gene expression in most lambdoid phages is mediated by an interaction between the N protein and a number of host-encoded factors. Bacteriophage HK022 does not rely on a protein for antitermination. To promote full expression of early phage genes, the transcripts of the HK022 put sites interact directly with RNA polymerase to convert it to a termination resistant form. Bacteriophage HK639 also uses RNA-mediated antitermination. However, it only possesses a single put-like element in its left operon. Because most lambdoid phages, including HK022, have antiterminator elements in …

In Vitro Characterization Of Avian Influenza Virus Isolates With A Truncated Ns1 Gene Segment, Aaron Dick

Honors Scholar Theses

Avian Influenza Virus represents a significant threat to the world poultry population, and is a potential threat to humans due to the possibility of cross-species AIV infection. Our approach is to characterize a number of avian virus populations with respect to their content of biologically active particles that include hemagglutinating particles (HAP), plaque forming particles (PFP), interferon inducing particles (IFP), interferon induction-suppressing particles (ISP), defective-interfering particles (DIP), cell-killing particles (CKP) and non-infectious cell killing particles (niCKP) using unique in vitro assays developed for avian influenza virus in the Marcus-Sekellick Laboratory. Specifically, we will use a strain of Avian influenza virus, …

Innate Immune Responses To Respiratory Syncytial Virus: Age-Associated Changes, Terianne Maiko Wong

USF Tampa Graduate Theses and Dissertations

Respiratory syncytial virus (RSV) infection causes ~64 million cases of respiratory disease and 200,000 deaths annually worldwide, yet there is no broadly effective prophylactic or treatment regimen. RSV can produce acute respiratory illness in patients of all ages but strikes the age extremes, infants and the elderly, with highest frequency presumably due to innate immune deficiencies. A higher morbidity and mortality has been reported for the elderly above 65 years of age, which has been attributed to immune senescence. Efforts to generate an effective vaccine have thus far been unsuccessful.

The innate immune system provides the first line of defense …

Arbovirus Persistence And Selection Of Persistent Variants Following Chronic Infection In Aedine Mosquitoes: A Comparative Study Between Ae. Aegypti And Ae. Albopictus 30 Days Post Infection With Sindbis Virus, Zoe Leah Lyski

UNF Graduate Theses and Dissertations

In the present investigation two container breeding mosquitoes, Aedes aegypti and Aedes albopictus were exposed to Sindbis virus (SINV). Mosquitoes were proffered a viremic bloodmeal using a vertical membrane feeder in an attempt to infect females while preserving virus-gut interactions that are speculated to be responsible for amplification and selective roles encountered in nature. Cohorts of 50 mosquitoes of each species were exposed to SINV strain TR339 or EMEM in a bloodmeal and incubated in insectary conditions for 30 days. Once infected, the mosquito remains persistently infected for life, accumulating mutations in the virus RNA genome. Plaque size variants may …

Role Of Post-Translational Modifications In The Hiv Cofactor Activity Of Ledgf/P75, Denisse Adriana Gutierrez

Open Access Theses & Dissertations

see document.

Differential Virus Host-Ranges Of The Fuselloviridae Of Hyperthermophilic Archaea: Implications For Evolution In Extreme Environments, Ruben M. Ceballos, Caleb D. Marceau, Joshua O. Marceau, Steven Morris, Adam J. Clore, Kenneth M. Stedman

Biology Faculty Publications and Presentations

An emerging model for investigating virus-host interactions in hyperthermophilic Archaea is the Fusellovirus-Sulfolobus system. The host, Sulfolobus, is a hyperthermophilic acidophile endemic to sulfuric hot springs worldwide. The Fuselloviruses, also known as Sulfolobus Spindle-shaped Viruses (SSVs), are "lemon" or "spindle"-shaped double-stranded DNA viruses, which are also found worldwide. Although a few studies have addressed the host-range for the type virus, Sulfolobus Spindle-shaped Virus 1 (SSV1), using common Sulfolobus strains, a comprehensive host-range study for SSV-Sulfolobus systems has not been performed. Herein, we examine six bona fide SSV strains (SSV1, SSV2, SSV3, SSVL1, SSVK1, SSVRH) and their respective infection characteristics on …

A Plant Virus Substrate Induces Early Upregulation Of Bmp2 For Rapid Bone Formation, Pongkwan Sitasuwan, L. Andrew Lee, Peng Bo, Erin N. Davis, Yuan Lin, Qian Wang

Faculty Publications

Many nanoscale materials have been developed to investigate the effects on stem cell differentiations via topographical and chemical cues for applications in tissue engineering and regenerative medicine. The use of plant viruses as cell supporting substrates has been of particular interest due to the rapid induction of bone marrow derived mesenchymal stem cells (BMSCs) towards osteogenic cells. In this study, the role of Tobacco mosaic virus (TMV) and its early effects on osteoinduction with particular emphasis on the regulation of bone morphogenetic protein-2 (BMP2) was examined. We observed that the cells on the virus substrate immediately aggregated and formed bone-like …

The Us27 Protein Of Human Cytomegalovirus Enhances Signaling By Human Chemokine Receptor Cxcr4, Kathleen Arnolds, Juliet V. Spencer

Creative Activity and Research Day - CARD

Human cytomegalovirus (HCMV) is a prevalent pathogen that causes little disease except in immune-compromised hosts. HCMV establishes life-long latency by manipulating host immune responses. We have found that the US27 protein profoundly alters the activity of host chemokine receptor CXCR4. Cells expressing US27 and CXCR4 exhibited greater migration and calcium signaling than cells expressing CXCR4 alone. The mechanism for this enhanced signaling appears to be increased expression of CXCR4, and we are continuing to investigate how US27 causes up-regulation of CXCR4. This work is expected to clarify the role of US27 in immune evasion and provide insights into treatment strategies.

Intra And Interhost Dynamics Shaping Arbovirus Adaptation And Evolution, Alexander T. Ciota

Legacy Theses & Dissertations (2009 - 2024)

Arthropod-borne viruses (arboviruses), which are predominately mosquito-borne and almost exclusively RNA viruses, are maintained in nature in complex transmission cycles involving blood sucking invertebrates and vertebrate hosts. Although over 120 arboviruses are human pathogens responsible for causing a significant and expanding global health burden, a detailed understanding of the complex interactions between these pathogens and their hosts, particularly invertebrate hosts, is lacking. Defining these interactions is necessary if we are to understand the selective pressures and, therefore, evolutionary, adaptive, and epidemiological potential of arboviruses. This requires experimental infection and evolution studies, particularly in vivo, with natural hosts. The results presented …

Development And Validation Of A Novel Reporter Assay For Human Papillomavirus Type 16 Late Gene Expression, Beatrice Orru, Ciaran Cunniffe, Fergus Ryan, Stefan Schwartz

Articles

To facilitate the investigations of HPV-16 late gene expression HPV-16 reporter plasmids were generated using previously described sub-genomic HPV-16 plasmids, named pBEL and pBELM, that, similar to the full viral genome, produce primarily HPV-16 early mRNAs and very little, if any, late mRNAs in cervical cancer cells. The HPV-16 late L1 gene was replaced by the chloramphenicol acetyltransferase (CAT) reporter gene, or green fluorescent protein (GFP), preceded by the poliovirus internal ribosome entry site (IRES). Results show that the reporter genes mimic the expression of L1 from these plasmids. For example, overexpression of adenovirus E4orf4 protein (E4orf4), polypyrimidine tract binding …

Role Of Porcine Reproductive And Respiratory Syndrome Virus Non-Structural Protein 1 In Viral Replications And Pathogenesis, Lalit Kumar Beura

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

Porcine reproductive and respiratory syndrome virus (PRRSV) is an infectious agent of significant concern to the global swine industry. PRRSV infection of pigs is initiated by a long viremia after which the virus enters in an extended persistent phase of 3-4 months that ultimately is resolved by the immune system. The delayed and weak host adaptive response is responsible for the protracted convalescence period. An initial sub-optimal innate immune response is postulated to be the reason behind such meager adaptive immune response. A major focus of the studies undertaken in this dissertation was to identify the viral non-structural proteins (nsps) …

Upregulation Of Reactive Oxygen Species During The Retrovirus Life Cycle And Their Roles In A Mutant Of Moloney Murine Leukemia Virus, Ts1-Mediated Neurodegeneration, Soo Jin Kim

Dissertations and Theses (Open Access)

Viral invasion of the central nervous system (CNS) and development of neurological symptoms is a characteristic of many retroviruses. The mechanism by which retrovirus infection causes neurological dysfunction has yet to be fully elucidated. Given the complexity of the retrovirus-mediated neuropathogenesis, studies using small animal models are extremely valuable. Our laboratory has used a mutant moloney murine leukemia retrovirus, ts1-mediated neurodegneration. We hypothesize that astrocytes play an important role in ts1-induced neurodegeneration since they are retroviral reservoirs and supporting cells for neurons. It has been shown that ts1 is able to infect astrocytes in vivo and in …

Molecular Coevolution Of Pacific Northwest Hantaviruses And Their Host, The Deer Mouse, Peromyscus Maniculatus, Philip Darren Jones

Dissertations and Theses

Sin Nombre virus (SNV, family Bunyaviridae, genus Hantavirus), hosted by the deer mouse, Peromyscus maniculatus (family Cricetidae, Subfamily Neotominae), is the primary etiological agent of Hantavirus pulmonary syndrome (HPS) in the western United States. HPS, with known pathogenicity only to humans and for which there is no cure or prophylaxis, affects the epithelium of the lungs by making the capillaries leaky, thereby resulting in bilateral infiltrates, and eventually leading to respiratory failure and death by drowning in approximately 38% of hospitalized patients.

In the Americas, Peromyscus has been co-evolving with hantaviruses for approximately 12–20 million years, since the first …

Molecular Characterisation Of A Bovine-Like Rotavirus Detected From A Giraffe, Emily Mulherin, Jill Bryan, Marijke Beltman, Luke O'Grady, Eugene Pidgeon, Lucie Garon, Andrew Lloyd, John Bainbridge, Helen O'Shea, Paul Whyte, Séamus Fanning

Department of Biological Sciences Publications

Background

Rotavirus (RV), is a member of the Reoviridae family and an important etiological agent of acute viral gastroenteritis in the young. Rotaviruses have a wide host range infecting a broad range of animal species, however little is known about rotavirus infection in exotic animals. In this paper we report the first characterisation of a RV strain from a giraffe calf.

Results

This report describes the identification and detailed molecular characterisation of a rotavirus strain detected from a 14-day-old Giraffe (Giraffa camelopardalis), presenting with acute diarrhea. The RV strain detected from the giraffe was characterized molecularly as G10P[11]. …

Comprehensive Analysis Of Class I And Class Ii Hla Antigens And Chronic Hepatitis B Virus Infection, Chloe L. Thio, David L. Thomas, Peter Karacki, Xiaojiang Gao, Darlene Marti, Richard A. Kaslow, James J. Goedert, Margaret Hilgartner, Steffanie A. Strathdee, Priya Duggal, Stephen J. O'Brien, Jacquie Astemborski, Mary Carrington

Biology Faculty Articles

Following an acute hepatitis B virus (HBV) infection, clearance or persistence is determined in part by the vigor and breadth of the host immune response. Since the human leukocyte antigen system (HLA) is an integral component of the immune response, we hypothesized that the highly polymorphic HLA genes are key determinants of viral clearance. HLA class I and II genes were molecularly typed in 194 Caucasian individuals with viral persistence and 342 matched controls who had cleared the virus. A single class I allele, A*0301 (odds ratio [OR], 0.47; 95% confidence interval [CI], 0.30 to 0.72; P = 0.0005) was …

Phylogenetic Analysis Of Segment 10 From African Horsesickness Virus And Cognate Genes From Other Orbiviruses, Rafael O. De Sá, Marla Zellner, Marvin J. Grubman

Biology Faculty Publications

Utilizing the reverse transcriptase-polymerase chain reaction (RT-PCR) procedure, we have synthesized full-length copies of segment 10 from African horsesickness virus (AHSV) serotypes 1,4 and 8. The genes were cloned, sequenced and compared with the sequence of the cognate gene from AHSV serotypes 3 and 9. Sequences were analyzed to assess evolutionary relationships among serotypes using cladistics. Based on this analysis the data support a close relationship between serotypes 4 and 9 and between serotypes 1 and 8 and a closer relationship of serotype 3 to the 4 and 9 group.

Effects Of Folic Acid Or Zinc Malnutrition On Rotaviral Infection In A Murine Model, John Douglas Morrey

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

The influences of dietary deficiencies of folic acid or zinc on rotaviral disease infant mice were studied. Preliminary studies indicated that bovine and simian rotaviruses, but not porcine rotavirus, caused diarrhea in infant mice. Bovine and porcine rotaviruses were not, however, sufficiently infectious to replace murine rotavirus in studies utilizing the murine model. It was also determined that murine rotavirus purified by a cesium chloride gradient was highly infectious and useful for subsequent studies on nutritional influences of rotaviral disease. In dietary studies, female Swiss Webster mice were fed diets containing deficient, moderately deficient, or adequate levels of folic acid …