Virology Commons^™

Investigating The Viral Ecology Of Global Bee Communities With High-Throughput Metagenomics, David A. Galbraith, Zachary L. Fuller, Allyson M. Ray, Axel Brockmann, Maryann Frazier, Mary W. Gikungu, J. Francisco Iturralde Martinez, Karen M. Kapheim, Jeffrey T. Kerby, Sarah D. Kocher, Oleksiy Losyev, Elliud Muli, Harland M. Patch, Cristina Rosa, Joyce M. Sakamoto, Scott Stanley, Anthony D. Vaudo, Christina M. Grozinger

Biology Faculty Publications

Bee viral ecology is a fascinating emerging area of research: viruses exert a range of effects on their hosts, exacerbate the impacts of other environmental stressors, and, importantly, are readily shared across multiple bee species in a community. However, our understanding of bee viral communities is limited, as it is primarily derived from studies of North American and European Apis mellifera populations. Here, we examined viruses in populations of A. mellifera and 11 other bee species from 9 countries, across 5 continents and Oceania. We developed a novel pipeline to rapidly, inexpensively, and robustly screen for bee viruses. This pipeline …

Use Of Pre-S Protein-Containing Hepatitis B Virus Surface Antigens And A Powerful Adjuvant To Develop An Immune Therapy For Chronic Hepatitis B Virus Infection, J S. Yum, B C. Ahn, H J. Jo, D Y. Kim, K H. Kim, H S. Kim, Y C. Sung, J Yoon, John D. Morrey, H M. Moon

John D. Morrey

A hepatitis B virus (HBV) vaccine has been developed using a new adjuvant and HBV surface antigens produced from a CHO cell line. The purified HBV surface antigens are composed of L protein, M protein, and S protein in a mixture of 20- and 40-nm diameter particles and filamentous forms. This HBV surface antigen, formulated with L-pampo, a proprietary adjuvant, induced 10 times more antibody than the same antigen with alum and was capable of inducing strong immune responses in three different HBV transgenic mice. In spite of the presence of a large amount of HBV antigen in the blood, …

Respiratory Insufficiency Correlated Strongly With Mortality Of Rodents Infected With West Nile Virus, John D. Morrey, V Siddharthan, H Wang, J O. Hall

John D. Morrey

West Nile virus (WNV) disease can be fatal for high-risk patients. Since WNV or its antigens have been identified in multiple anatomical locations of the central nervous system of persons or rodent models, one cannot know where to investigate the actual mechanism of mortality without careful studies in animal models. In this study, depressed respiratory functions measured by plethysmography correlated strongly with mortality. This respiratory distress, as well as reduced oxygen saturation, occurred beginning as early as 4 days before mortality. Affected medullary respiratory control cells may have contributed to the animals' respiratory insufficiency, because WNV antigen staining was present …

Autonomic Nervous Dysfunction In Hamsters Infected With West Nile Virus, H Wang, V Siddharthan, J O. Hall, John D. Morrey

John D. Morrey

Clinical studies and case reports clearly document that West Nile virus (WNV) can cause respiratory and gastrointestinal (GI) complications. Other functions controlled by the autonomic nervous system may also be directly affected by WNV, such as bladder and cardiac functions. To investigate how WNV can cause autonomic dysfunctions, we focused on the cardiac and GI dysfunctions of rodents infected with WNV. Infected hamsters had distension of the stomach and intestines at day 9 after viral challenge. GI motility was detected by a dye retention assay; phenol red dye was retained more in the stomachs of infected hamsters as compared to …

Maporal Hantavirus Β-Integrin Utilization And Sensitivity To Favipiravir, Kristin K. Buys

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Hantaviruses are members of the Bunyaviridae family of viruses. Pathogenic hantaviruses are the etiologic agents of hemorrhagic fever with renal syndrome (HFRS), a disease principally endemic in the Old World, and hantavirus pulmonary syndrome (HPS), a disease primarily restricted to the Americas. Maporal virus (MPRLV), a recently isolated hantavirus, has been found to cause disease in hamsters that resembles HPS in humans. However, the virus has not been linked to human cases of HPS. Considerable evidence suggests that β-integrin usage mediating infection may serve to distinguish hantaviruses pathogenic to humans from nonpathogenic, but this receptor usage pattern information is not …

Effect Of Dipterinyl Calcium Pentahydrate On Hepatitis B Virus Replication In Transgenic Mice, P Moheno, John D. Morrey, D Fuchs

John D. Morrey

Dipterinyl calcium pentahydrate (DCP) has previously been shown to inhibit MDA-MB-231 human breast cancer xenographs in nude mice in a manner correlated with increases in plasma IL-12 and IL-4 concentrations, and decreases in plasma IL-6 levels. DCP also inhibits indoleamine 2,3-dioxygenase (IDO), an immuno-inhibitory enzyme, in human PBMCs (Peripheral Blood Mononuclear Cells).

Assessing Changes In Vascular Permeability In A Hamster Model Of Viral Hemorrhagic Fever, B B. Gowen, J G. Julander, N R. London, M H. Wong, D Larson, John D. Morrey, D Y. Li, M Bray

John D. Morrey

A number of RNA viruses cause viral hemorrhagic fever (VHF), in which proinflammatory mediators released from infected cells induce increased permeability of the endothelial lining of blood vessels, leading to loss of plasma volume, hypotension, multi-organ failure, shock and death. The optimal treatment of VHF should therefore include both the use of antiviral drugs to inhibit viral replication and measures to prevent or correct changes in vascular function. Although rodent models have been used to evaluate treatments for increased vascular permeability (VP) in bacterial sepsis, such studies have not been performed for VHF.

Development Of A New Tacaribe Arenavirus Infection Model And Its Use To Explore Antiviral Activity Of A Novel Aristeromycin Analog, B B. Gowen, M H. Wong, D Larson, W Ye, K H. Jung, E J. Sefing, R Skirpstunas, D F. Smee, John D. Morrey, S W. Schneller

John D. Morrey

A growing number of arenaviruses can cause a devastating viral hemorrhagic fever (VHF) syndrome. They pose a public health threat as emerging viruses and because of their potential use as bioterror agents. All of the highly pathogenic New World arenaviruses (NWA) phylogenetically segregate into clade B and require maximum biosafety containment facilities for their study. Tacaribe virus (TCRV) is a nonpathogenic member of clade B that is closely related to the VHF arenaviruses at the amino acid level. Despite this relatedness, TCRV lacks the ability to antagonize the host interferon (IFN) response, which likely contributes to its inability to cause …

Effects Of The Combination Of Favipiravir (T-705) And Oseltamivir On Influenza A Virus Infections In Mice, D F. Smee, B L. Hurst, M H. Wong, K W. Bailey, E B. Tarbet, John D. Morrey, Y Furuta

John D. Morrey

Favipiravir (T-705 [6-fluoro-3-hydroxy-2-pyrazinecarboxamide]) and oseltamivir were combined to treat influenza virus A/NWS/33 (H1N1), A/Victoria/3/75 (H3N2), and A/Duck/MN/1525/81 (H5N1) infections. T-705 alone inhibited viruses in cell culture at 1.4 to 4.3 µM. Oseltamivir inhibited these three viruses in cells at 3.7, 0.02, and 0.16 µM and in neuraminidase assays at 0.94, 0.46, and 2.31 nM, respectively. Oral treatments were given twice daily to mice for 5 to 7 days starting, generally, 24 h after infection. Survival resulting from 5 days of oseltamivir treatment (0.1 and 0.3 mg/kg/day) was significantly better in combination with 20 mg/kg of body weight/day of T-705 against …

Effects Of Combinations Of Favipiravir (T-705) And Oseltamivir On Influenza A (H1n1, H3n2, And H5n1) Virus Infections In Mice, D F. Smee, B L. Hurst, M H. Wong, K W. Bailey, E B. Tarbet, John D. Morrey, Y Furuta

John D. Morrey

Favipiravir (T-705 [6-fluoro-3-hydroxy-2-pyrazinecarboxamide]) and oseltamivir were combined to treat influenza virus A/NWS/33 (H1N1), A/Victoria/3/75 (H3N2), and A/Duck/MN/1525/81 (H5N1) infections. T-705 alone inhibited viruses in cell culture at 1.4 to 4.3 µM. Oseltamivir inhibited these three viruses in cells at 3.7, 0.02, and 0.16 µM and in neuraminidase assays at 0.94, 0.46, and 2.31 nM, respectively. Oral treatments were given twice daily to mice for 5 to 7 days starting, generally, 24 h after infection. Survival resulting from 5 days of oseltamivir treatment (0.1 and 0.3 mg/kg/day) was significantly better in combination with 20 mg/kg of body weight/day of T-705 against …

Persistent West Nile Virus Associated With A Neurological Sequela In Hamsters Identified By Motor Unit Number Estimation, V Siddharthan, H Wang, N E. Motter, J O. Hall, R D. Skinner, R T. Skirpstunas, John D. Morrey

John D. Morrey

To investigate the hypothesis that neurological sequelae are associated with persistent West Nile virus (WNV) and neuropathology, we developed an electrophysiological motor unit number estimation (MUNE) assay to measure the health of motor neurons temporally in hamsters. The MUNE assay was successful in identifying chronic neuropathology in the spinal cords of infected hamsters. MUNE was suppressed at days 9 to 92 in hamsters injected subcutaneously with WNV, thereby establishing that a long-term neurological sequela does occur in the hamster model. MUNE suppression at day 10 correlated with the loss of neuronal function as indicated by reduced choline acetyltransferase staining (R2 …

Alkoxyalkyl Esters Of 9-(S)-(3-Hydroxy-2-Phosphonomethoxypropyl) Adenine Are Potent And Selective Inhibitors Of Hepatitis B Virus (Hbv) Replication In Vitro And In Hbv Transgenic Mice In Vivo, John D. Morrey, B E. Korba, J R. Beadle, D L. Wyles, K Y. Hostetler

John D. Morrey

Alkoxyalkyl esters of acyclic nucleoside phosphonates have previously been shown to have increased antiviral activity when they are administered orally in animal models of viral diseases, including lethal infections with vaccinia virus, cowpox virus, ectromelia virus, murine cytomegalovirus, and adenovirus. 9-(S)-(3-Hydroxy-2-phosphonomethoxypropyl)adenine [(S)-HPMPA] was previously shown to have activity against hepatitis B virus (HBV) in vitro. To assess the effect of alkoxyalkyl esterification of (S)-HPMPA, we prepared the hexadecyloxypropyl (HDP), 15-methyl-hexadecyloxypropyl (15M-HDP), and octadecyloxyethyl (ODE) esters and compared their activities with the activity of adefovir dipivoxil in vitro and in vivo. Alkoxyalkyl esters of (S)-HPMPA were 6 to 20 times more …

Activity Of T-705 In A Hamster Model Of Yellow Fever Virus Infection In Comparison With That Of A Chemically Related Compound, T-1106, J G. Julander, K Shafer, D F. Smee, John D. Morrey, Y Furuta

John D. Morrey

Treatment with the nucleoside analog T-1106 was previously shown to be effective in a hamster model of yellow fever virus (YFV) disease, even though it had only slight activity in cell culture. In the study described in this report, the activity of T-705, a chemically related compound currently undergoing clinical trials for the treatment of influenza (FDANews 4:1, 2007), was tested against YFV in cell culture and in the hamster model. The antiviral efficacy of T-705 in cell culture occurred at a concentration of 330 µM, which was more than threefold lower than the concentration at which T-1106 had antiviral …

Effects Of Double Combinations Of Amantadine, Oseltamivir, And Ribavirin On Influenza A (H5n1) Virus Infections In Cell Culture And In Mice, D F. Smee, B L. Hurst, M H. Wong, K W. Bailey, John D. Morrey

John D. Morrey

An amantadine-resistant influenza A/Duck/MN/1525/81 (H5N1) virus was developed from the low-pathogenic North American wild-type (amantadine-sensitive) virus for studying treatment of infections in cell culture and in mice. Double combinations of amantadine, oseltamivir (or the cell culture-active form, oseltamivir carboxylate), and ribavirin were used. Amantadine-oseltamivir carboxylate and amantadine-ribavirin combinations showed synergistic interactions over a range of doses against wild-type virus in Madin-Darby canine kidney (MDCK) cell culture, but oseltamivir carboxylate-ribavirin combinations did not. Primarily additive interactions were seen with oseltamivir carboxylate-ribavirin combinations against amantadine-resistant virus. The presence of amantadine in drug combinations against the resistant virus did not improve activity. The …

Treatment Of Late Stage Disease In A Model Of Arenaviral Hemorrhagic Fever: T-705 Efficacy And Reduced Toxicity Suggests An Alternative To Ribavirin, B B. Gowen, D F. Smee, M H. Wong, J O. Hall, K H. Jung, K W. Bailey, J R. Stevens, Y Furuta, John D. Morrey

John D. Morrey

A growing number of arenaviruses are known to cause viral hemorrhagic fever (HF), a severe and life-threatening syndrome characterized by fever, malaise, and increased vascular permeability. Ribavirin, the only licensed antiviral indicated for the treatment of certain arenaviral HFs, has had mixed success and significant toxicity. Since severe arenaviral infections initially do not present with distinguishing symptoms and are difficult to clinically diagnose at early stages, it is of utmost importance to identify antiviral therapies effective at later stages of infection. We have previously reported that T-705, a substituted pyrazine derivative currently under development as an anti-influenza drug, is highly …

Phosphorothioate Di- And Trinucleotides As A Novel Class Of Anti-Hepatitis B Virus Agents, R. P. Iyer, Y. Jin, A. Roland, John D. Morrey, S. Mounir, B. Korba

John D. Morrey

Several nucleoside analogs are under clinical development for use against hepatitis B virus (HBV). Lamivudine (3TC), a nucleoside analog, and adefovir dipivoxil (ADV), an acyclonucleotide analog, are clinically approved. However, long-term treatment can induce viral resistance, and following the cessation of therapy, viral rebound is frequently observed. There continues to be a need for new antiviral agents with novel mechanisms of action. A library of more than 600 di- and trinucleotide compounds synthesized by parallel synthesis using a combinatorial strategy was screened for potential inhibitors of HBV replication using the chronically HBV-producing cell line 2.2.15. Through an iterative process of …

In Vivo Influenza Virus-Inhibitory Effects Of The Cyclopentane Neuraminidase Inhibitor Rjw-270201, R W. Sidwell, D F. Smee, J H. Huffman, D L. Barnard, K W. Bailey, John D. Morrey, Y S. Babu

John D. Morrey

The cyclopentane influenza virus neuraminidase inhibitor RWJ-270201 was evaluated against influenza A/NWS/33 (H1N1), A/Shangdong/09/93 (H3N2), A/Victoria/3/75 (H3N2), and B/Hong Kong/05/72 virus infections in mice. Treatment was by oral gavage twice daily for 5 days beginning 4 h pre-virus exposure. The influenza virus inhibitor oseltamivir was run in parallel, and ribavirin was included in studies with the A/Shangdong and B/Hong Kong viruses. RWJ-270201 was inhibitory to all infections using doses as low as 1 mg/kg/day. Oseltamivir was generally up to 10-fold less effective than RWJ-270201. Ribavirin was also inhibitory but was less tolerated by the mice at the 75-mg/kg/day dose used. …

In Vivo Activation Of Human Immunodeficiency Virus Type 1 Long Terminal Repeat By Uv Type A (Uv-A) Light Plus Psoralen And Uv-B Light In The Skin Of Transgenic Mice, John D. Morrey, S M. Bourn, T D. Bunch, M K. Jackson, R W. Sidwell, L R. Barrows, R A. Daynes, C A. Rosen

John D. Morrey

UV irradiation has been shown to activate the human immunodeficiency virus type 1 (HIV-1) long terminal repeat (LTR) in cell culture; however, only limited studies have been described in vivo. UV light has been categorized as UV-A (400 to 315 nm), -B (315 to 280 nm), or -C (<280 nm); the longer wavelengths are less harmful but more penetrative. Highly penetrative UV-A radiation constitutes the vast majority of UV sunlight reaching the earth's surface but is normally harmless. UV-B irradiation is more harmful but less prevalent than UV-A. In this report, the HIV-1 LTR-luciferase gene in the skin of transgenic mice was markedly activated when exposed to UV-B irradiation. The LTR in the skin of transgenic mice pretreated topically with a photosensitizing agent (psoralen) was also activated to similar levels when exposed to UV-A light. A 2-h exposure to sunlight activated the LTR in skin treated with psoralen, whereas the LTR in skin not treated with psoralen was activated after 7 h of sunlight exposure. The HIV-1 LTR-β-galactosidase reporter genes have been used to demonstrate the in vivo UV-induced activation of the LTR and might be used to evaluate other environmental factors or pharmacologic substances that might potential activate the HIV-1 LTR in vivo

Bluetongue Virus Evolution: Sequence Analyses Of The Gene Coding For The Major Serogroup Antigen, Timothy F. Kowalik

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

A study was undertaken to better understand the genetic relationship of five United States prototype bluetongue virus serotypes. Genomic double-stranded RNA segment S1, which encodes the major core protein and serogroup antigen, VP7, was used as a marker gene for the sequence analyses. The S1 segments from BTV-2, 11, 13, and 17 were cloned and sequenced by methods developed during the course of this investigation. These results were compared with previously published sequence data from segment S1 of BTV-10. The Sl segments are 1156 base pairs long and contain a common open reading frame capable of coding for a protein …

Tissue-Specific Replication Of Friend And Moloney Murine Leukemia Viruses In Infected Mice, L H. Evans, John D. Morrey

John D. Morrey

We have studied the replication of ecotropic murine leukemia viruses (MuLV) in the spleens and thymuses of mice infected with the lymphocytic leukemia-inducing virus Moloney MuLV (M-MuLV), with the erythroleukemia-inducing virus Friend MuLV (F-MuLV), or with in vitro-constructed recombinants between these viruses in which the long terminal repeat (LTR) sequences have been exchanged. At 1 week after infection both the parents and the LTR recombinants replicated predominantly in the spleens with only low levels of replication in the thymus. At 2 weeks after infection, the patterns of replication in the spleens and thymuses were strongly influenced by the type of …

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 …

The Effect Of Urea On The Sedimentation Coefficient Of The Curly Top Virus Dimer, Allen H. Smith

Undergraduate Honors Capstone Projects

The purpose of this project is to investigate the effect of increasing concentrations of urea on the sedimentation rate of the curly top virus dimer as measured by ultracentrifugation techniques. In general, urea causes the breaking of hydrogen bonds in macromolecules--in the case of the virus, increased concentrations of urea should cause changes in the configuration of the virus and may possibly cause a separation of the dimer into monomers. From previous centrifugation studies, we know that the dimer has a characteristic sedimentation coefficient(S) of about 80 Svedbergs; the monomer has an S value of about 55 Svedberg units. Increased …

Detection Of Neonatal Calf Diarrhea Virus (Ncdv) And Human Infant Reolike Diarrhea Virus, Martin Wyatt Peterson

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

The purpose of this study was to develop a diagnostic test and conduct a survey for the neonatal calf diarrhea virus (NCDV) and human infant reolike diarrhea virus. Two immunologic methods were developed in this investigation.

Immune electron microscopy (IEM) and the fluorescent viral precipitin test (FVPT) are the methods used to detect NCDV and the human virus. Both methods are based upon the principle that viral aggregates form when the virus is reacted with anti-NCDV antibody. Aggregates in the IEM method are negatively stained and observed with the use of an electron microscope. Fluorescein labeled antibody is used in …