Virology Commons^™

T-Cell Responses To The M3 Immune Evasion Protein Of Murid Gammaherpesvirus 68 Are Partially Protective And Induced With Lytic Antigen Kinetics, Joshua J. Obar, Douglas C. Donovan, Sarah G. Crist, Ondine Silvia, James P. Stewart, Edward J. Usherwood

Dartmouth Scholarship

DNA vaccination with the M3 gene, encoding an immune evasion molecule expressed during both the acute lytic and persistent phases of murid gammaherpesvirus 68 infection, yielded a significantly lower titer of virus in the lung than controls. The protection seen was dependent on T cells, and we mapped an epitope recognized by CD8 T cells. The immune response to this epitope follows the same kinetics as lytic cycle antigens, despite the fact that this gene is expressed in both lytic and persistent stages of infection. This has important implications for our understanding of T-cell responses to putative latency-associated gammaherpesvirus proteins …

Detection And Quantitation Of Bovine Respiratory Syncytial Virus Using Real-Time Quantitative Rt-Pcr And Quantitative Competitive Rt-Pcr Assays, Jenna Achenbaum, Christina L. Topliff, Ventzislav B. Vassilev, Ruben O. Donis, Kent M. Eskridge, Clayton L. Kelling

Nebraska Center for Virology: Faculty Publications

A single tube, fluorogenic probe-based, real-time quantitative reverse transcription-polymerase chain reaction (Q-RT-PCR) assay was developed for detection and quantitation of bovine respiratory syncytial virus (BRSV) using BioRad’s iCycler iQ™. Real-time Q-RT-PCR was compared with quantitative competitive RT-PCR (QC-RT-PCR) and viral titers. Viral mRNA levels were measured in BRSV-infected bovine turbinate cell lysate harvested at eight time points (1.5, 6, 12, 24, 36, 48, 60, 72 h) post-infection. A homologous BRSV cRNA standard was used for quantitation of the mRNA by plotting a standard curve of cycle threshold (Ct) values versus standard 10-fold dilutions of cRNA of known concentrations. Detection as …

Cd8+ T Cells Responding To Influenza Infection Reach And Persist At Higher Numbers Than Cd4+ T Cells Independently Of Precursor Frequency, Timothy J. Powell, Deborah M. Brown, Joseph A. Hollenbaugh, Tina Charbonneau, Roslyn A. Kemp, Susan L. Swain, Richard Dutton

Nebraska Center for Virology: Faculty Publications

The activation, localization, phenotypic changes, and function of CFSE-labeled naive influenza-specific CD8⁺ and CD4⁺ T cells following influenza infection were examined. Response of adoptively transferred CD8⁺ T cells was seen earliest in draining lymph node. Highly activated cells were found later in the lung, airways, and spleen, were cytolytic, and expressed IFN-γ upon restimulation. Similar amounts of division at early time points, but higher numbers of CD8⁺ T cells, were detected at 9 and 30 days postinfection after cotransfer of CD4⁺ and CD8⁺ T cells followed by infection. Transfer of much smaller numbers of …

Preserved Antigenicity Of Hiv-1 P24 Produced And Purified In High Yields From Plants Inoculated With A Tobacco Mosaic Virus (Tmv)-Derived Vector, D. M. Pérez-Filgueira, B. P. Brayfield, S. Phiri, M. V. Borca, Charles Wood, Thomas Jack Morris

Papers in Virology

Production of structural proteins from foot-and-mouth disease virus (FMDV) and bovine herpes virus (BHV-1) in Nicotiana benthamiana through the use of a tobacco mosaic virus-based vector (TMV-30B) has been reported previously. The development of the TMV-30B-HISc vector, a new version that adds a C-terminal histidine (His) sequence to the foreign protein expressed is described. Coding sequences from the FMDV VPl protein and the core protein, p24, from a clade C HIV-1 isolate from Zambia were cloned into the new vector and infective RNAs were generated for each construct to inoculate N. benthamiana plants. His-tagged proteins were purified from inoculated leaves …

In Vitro Expression Of Full-Length And Truncated Bovine Respiratory Syncytial Virus G Proteins And Their Antibody Responses In Balb/C Mice, Ryan P. Brady, Christina L. Topliff, Clayton L. Kelling

Nebraska Center for Virology: Faculty Publications

Bovine respiratory syncytial virus (BRSV) is a primary cause of lower respiratory tract disease in calves. Protection is incomplete following vaccination or natural infection, as re-infections are common. The objectives of this study were to create plasmid DNA constructs encoding the full-length, secreted, or conserved region of the BRSV G glycoprotein, and to compare and evaluate their expression in cell culture and potential to induce antibody responses in BALB/c mice. Transfection of COS-7 cells with plasmid DNA resulted in expression of the BRSV G region from each of the plasmid DNA constructs. Following inoculation of BALB/c mice with plasmid DNA, …

Heat Shock Protein 70 Protects Cells From Cell Cycle Arrest And Apoptosis Induced By Human Immunodeficiency Virus Type 1 Viral Protein R, Yuqi Zhao, Larisa Dubrovsky, Tatiana Iordanskaya, Mongzhong Chen, Dong Liang, Michael Bukrinsky, Sergey Iordanskiy

Nebraska Center for Virology: Faculty Publications

Viral protein R (Vpr) of human immunodeficiency virus type 1 (HIV-1) is an accessory protein that plays an important role in viral pathogenesis. This pathogenic activity of Vpr is related in part to its capacity to induce cell cycle G₂ arrest and apoptosis of target T cells. A screening for multicopy suppressors of these Vpr activities in fission yeast identified heat shock protein 70 (Hsp70) as a suppressor of Vpr-induced cell cycle arrest. Hsp70 is a member of a family of molecular chaperones involved in innate immunity and protection from environmental stress. In this report, we demonstrate that HIV-1 …

Prion Infection Of Skeletal Muscle Cells And Papillae In The Tongue, Ellyn R. Mulcahy, Jason C. Bartz, Anthony E. Kincaid, Richard Bessen

Nebraska Center for Virology: Faculty Publications

The presence of the prion agent in skeletal muscle is thought to be due to the infection of nerve fibers located within the muscle. We report here that the pathological isoform of the prion protein, PrP^Sc, accumulates within skeletal muscle cells, in addition to axons, in the tongue of hamsters following intralingual and intracerebral inoculation of the HY strain of the transmissible mink encephalopathy agent. Localization of PrP^Sc to the neuromuscular junction suggests that this synapse is a site for prion agent spread between motor axon terminals and muscle cells. Following intracerebral inoculation, the majority of PrP …

Distribution Of Kaposi Sarcoma–Associated Herpesvirus/Human Herpesvirus 8 In Maternal Saliva And Breast Milk In Zambia: Implications For Transmission, Brad P. Brayfield, Chipepo Kankasa, John T. West, Jubra Muyanga, Ganapati Bhat, Winslow Klaskala, Charles D. Mitchell, Charles Wood

Nebraska Center for Virology: Faculty Publications

Background. The seroprevalence of Kaposi sarcoma–associated herpesvirus (KSHV)/human herpesvirus 8 (HHV-8) in sub-Saharan Africa suggests that multiple routes of transmission exist. In the present study,we examined 2 possible routes of mother-to-child transmission, through breast milk and saliva, during the first 6 months after delivery. Methods. The prevalence of HHV-8 DNA in the breast-milk cells (n=75), milk supernatant (n=56), colostrum (n=2), and saliva cells (n=65) of HHV-8–seropositive mothers who recently gave birth was examined. Polymerase chain reaction analysis was performed for the detection of HHV-8 in cross-sectional samples isolated at 2, 4, and 6 months after delivery.

Results. None of the …

Cd4 T Cell Responses To Influenza Infection, Deborah M. Brown, Eulogia Roman, Susan L. Swain

Nebraska Center for Virology: Faculty Publications

Immune responses to viral infections involve a complex orchestration between innate signals and adaptive responses of specific T and B cells. Anti-viral CD4 cells can direct CD8 responses by secreting a Type 1 panel of cytokines including IFN-γ, IL-2 and TNF- α and can drive B cell production of IgG2a to neutralize infective viral particles. This review will focus specifically on the role of CD4 cells in the immune response to influenza, an acute, localized respiratory viral infection. We suggest that CD4 cells act as direct effectors in protection against influenza, may contribute to immunopathology and generate functionally distinct memory …

Multiple Signal Transducers And Activators Of Transcription Are Induced By Ebv Lmp-1, Luwen Zhang, Ke Hong, Jun Zhang, Joseph S. Pagano

Nebraska Center for Virology: Faculty Publications

Epstein-Barr virus (EBV) latent membrane protein 1 (LMP-1) is required for EBV immortalization of primary B cells in vitro. Signal transducers and activators of transcription (STATs) play a pivotal role in the initiation and maintenance of certain cancers. STAT proteins, especially STAT-1, -3, and -5, are persistently tyrosine phosphorylated or activated in many cancers. We show here that EBV-infected type III latency cells, in which the EBV oncoprotein, LMP-1 is expressed, express high levels of four STATs (STAT-1, -2, -3, and -5A) and that LMP-1 is responsible for the induction of three (STAT-1, -2, and -3). In addition, the C-terminal …

Phylogenetic Analyses Of Texas Isolates Indicate An Evolving Subtype Of The Clade B Feline Immunodeficiency Viruses, Eric A. Weaver, Ellen W. Collisson, Margaret Slater, Guan Zhu

Nebraska Center for Virology: Faculty Publications

Rigorous phylogenetic analyses were used to compare the nucleotide sequences of feline immunodeficiency virus strains isolated from Texas and throughout the world. The envelope V3-V4 sequences and capsid gene of the Texas isolates formed a cluster between subtypes B and E. Statistical comparisons with other published sequences confirmed that the Texas group is a unique cluster, possibly a new subtype, arising from subtype B.

Feline immunodeficiency virus (FIV) was initially isolated in 1987 from a cat in California with severe immunodeficiency and has been recognized as a common worldwide feline pathogen (11, 14, 19, 20, 33). FIV-infected cats exhibit a …

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 …

Cd40-Associated Traf 6 Signaling Is Required For Disease Induction In A Retrovirus-Induced Murine Immunodeficiency, Kathy A. Green, Cory L. Ahonen, W. James Cook, William R. Green

Dartmouth Scholarship

LP-BM5 retrovirus-infected C57BL/6 mice develop splenomegaly, lymphadenopathy, hypergammaglobulinemia, and immunodeficiency; thus, this disease has been named mouse AIDS. In this syndrome, CD154/CD40 interactions are required for but do not mediate disease by upregulation of CD80 or CD86. We report here that there is nonetheless a necessity for CD40 signaling competence, specifically an intact tumor necrosis factor receptor-associated factor 6 (TRAF 6) binding site.

Cd46-Mediated Transduction Of A Species D Adenovirus Vaccine Improves Mucosal Vaccine Efficacy, Zenaido T. Camacho, Mallory A. Turner, Michael A. Barry, Eric A. Weaver

Nebraska Center for Virology: Faculty Publications

The high levels of preexisting immunity against Adenovirus type 5 (Ad5) have deemed Ad5 unusable for translation as a human vaccine vector. Low seroprevalent alternative viral vectors may be less impacted by preexisting immunity, but they may also have significantly different phenotypes from that of Ad5. In this study we compare species D Ads (26, 28, and 48) to the species C Ad5. In vitro transduction studies show striking differences between the species C and D viruses. Most notably, Ad26 transduced human dendritic cells much more effectively than Ad5. In vivo imaging studies showed strikingly different transgene expression profiles. The …

Phylogenetic Analyses Of Texas Isolates Indicate An Evolving Subtype Of The Clade B Feline Immunodeficiency Viruses, Eric A. Weaver, Ellen W. Collisson, Margaret Slater, Guan Zhu

Nebraska Center for Virology: Faculty Publications

Rigorous phylogenetic analyses were used to compare the nucleotide sequences of feline immunodeficiency virus strains isolated from Texas and throughout the world. The envelope V3-V4 sequences and capsid gene of the Texas isolates formed a cluster between subtypes B and E. Statistical comparisons with other published sequences confirmed that the Texas group is a unique cluster, possibly a new subtype, arising from subtype B.

Genomes Of The Parapoxviruses Orf Virus And Bovine Papular Stomatitis Virus, Gustavo A. Delhon, E. R. Tuhlman, C. L. Afonso, Z. Lu, A. De La Concha-Bermejillo, H. D. Lehmkuhl, M. E. Piccone, G. F. Kutish, D. L. Rock

Nebraska Center for Virology: Faculty Publications

Bovine papular stomatitis virus (BPSV) and orf virus (ORFV), members of the genus Parapoxvirus of the Poxviridae, are etiologic agents of worldwide diseases affecting cattle and small ruminants, respectively. Here we report the genomic sequences and comparative analysis of BPSV strain BV-AR02 and ORFV strains OV-SA00, isolated from a goat, and OV-IA82, isolated from a sheep. Parapoxvirus (PPV) BV-AR02, OV-SA00, and OV-IA82 genomes range in size from 134 to 139 kbp, with an average nucleotide composition of 64% G+C. BPSV and ORFV genomes contain 131 and 130 putative genes, respectively, and share colinearity over 127 genes, 88 of which …