Virology Commons^™

Intracellular Signaling Molecules Activated By Epstein-Barr Virus For Induction Of Interferon Regulatory Factor 7, Luwen Zhang, Lihong Wu, Ke Hong, Joseph S. Pagano

Nebraska Center for Virology: Faculty Publications

Epstein-Barr virus (EBV) latent membrane protein 1 (LMP-1) is the principal oncogenic protein in the EBV transformation process. LMP-1 induces the expression of interferon regulatory factor 7 (IRF-7) and activates IRF-7 protein by phosphorylation and nuclear translocation. LMP-1 is an integral membrane protein with two regions in its C terminus that initiate signaling processes, the C-terminal activator regions 1 (CTAR-1) and CTAR-2. Here, genetic analysis of LMP-1 has determined that the PXQXT motif that governs the interaction between LMP-1 CTAR-1 and tumor necrosis factor receptor-associated factors (TRAFs) is needed to induce the expression of IRF-7. Mutations in the PXQXT motif …

Interferon Regulatory Factor 7: A Key Cellular Mediator Of Lmp-1 In Ebv Latency And Transformation, Luwen Zhang, Joseph S. Pagano

Nebraska Center for Virology: Faculty Publications

Interferon regulatory factor 7 (IRF-7) was cloned within the biological context of Epstein–Barr virus (EBV) latency, and has an intimate relation with EBV. EBV latent membrane protein 1 (LMP-1) regulates IRF-7 both by inducing the expression of IRF-7 and by activating IRF-7 protein through phosphorylation and nuclear translocation in a post-translational manner. The activated IRF-7 then functions to regulate both EBV and cellular target genes involved in latency, transformation and immune regulation. IRF-7 appears to be a key cellular latency protein involved in both the pathogenesis and persistence of EBV infection.

Identification Of A Cellular Protein That Interacts And Synergizes With The Rta (Orf50) Protein Of Kaposi’S Sarcoma-Associated Herpesvirus In Transcriptional Activation, Shizhen Wang, Shuhong Liu, Ming-Hoi Wu, Yunqi Geng, Charles Wood

Nebraska Center for Virology: Faculty Publications

Lytic reactivation of Kaposi’s sarcoma-associated herpesvirus (KSHV), or human herpesvirus 8, from latency requires transcriptional transactivation by the viral protein RTA encoded by the ORF50 gene. Very little is known about how RTA functions and the cellular factors that may be involved in its transactivation function. Using the yeast two-hybrid system, we have identified a human cellular protein that can interact with KSHV RTA. The cellular protein, referred to as the human hypothetical protein MGC2663 by GenBank, is encoded by human chromosome 19. This protein is 554 amino acids (aa) in size and displays sequence similarity with members of the …

Mononuclear Phagocyte Differentiation, Activation, And Viral Infection Regulate Matrix Metalloproteinase Expression: Implications For Human Immunodeficiency Virus Type 1-Associated Dementia, Anuja Ghorpade, Raisa Persidskaia, Radhika Suryadevara, Myhanh Che, Xiao Juan Liu, Yuri Persidsky, Howard Gendelman

Nebraska Center for Virology: Faculty Publications

The pathogenesis of human immunodeficiency virus type 1 (HIV-1)-associated dementia (HAD) is mediated mainly by mononuclear phagocyte (MP) secretory products and their interactions with neural cells. Viral infection and MP immune activation may affect leukocyte entry into the brain. One factor that influences central nervous system (CNS) monocyte migration is matrix metalloproteinases (MMPs). In the CNS, MMPs are synthesized by resident glial cells and affect the integrity of the neuropil extracellular matrix (ECM). To ascertain how MMPs influence HAD pathogenesis, we studied their secretion following MP differentiation, viral infection, and cellular activation. HIV-1-infected and/or immune-activated monocyte-derived macrophages (MDM) and human …

Interactions Among Murine Cytomegalovirus Us22 Family Gene Products That Influence Viral Pathogenesis, Zaruhi Karabekian

Theses and Dissertations in Biomedical Sciences

Cytomegalovirus (CMV) is a complex, ubiquitous herpesvirus that is characterized by acute, chronic, and latent infections. Monocytes-macrophages are the key target cell type involved in pathogenesis, which is most effectively studied using the murine model of CMV infection. Previously three murine CMV (MCMV) genes (M139, M140, and M141) were identified to regulate viral expression in cultured macrophages and in mice. These genes are members of the US22 gene family with respect to HCMV homology. There is no function assigned to the proteins encoded by these genes. However, deletion of M139, M140, and M141 significantly curtails growth of MCMV in macrophages …

Human Papillomavirus Virus-Like Particles Are Efficient Oral Immunogens When Coadministered With Escherichia Coli Heat-Labile Enterotoxin Mutant R192g Or Cpg Dna, S. Gerber, C. Lane, Deborah M. Brown, E. Lord, M. Dilorenzo, J. D. Clements, E. Rybicki, A.-L. Williamson, R. C. Rose

Nebraska Center for Virology: Faculty Publications

Certain human papillomaviruses (HPVs) cause most cervical cancer, which remains a significant source of morbidity and mortality among women worldwide. HPV recombinant virus-like particles (VLPs) are promising vaccine candidates for controlling anogenital HPV disease and are now being evaluated as a parenteral vaccine modality in human subjects. Vaccines formulated for injection generally are more costly, more difficult to administer, and less acceptable to recipients than are mucosally administered vaccines. Since oral delivery represents an attractive alternative to parenteral injection for large-scale human vaccination, the oral immunogenicity of HPV type 11 (HPV-11) VLPs in mice was previously investigated; it was found …

Regulation Of Human Immunodeficiency Virus Type 1 Infection, Β-Chemokine Production, And Ccr5 Expression In Cd40l-Stimulated Macrophages: Immune Control Of Viral Entry, Robin L. Cutter, Jialin Zheng, Myhanh Che, Douglas Niemann, Ying Liu, Johnny He, Elaine Thomas, Howard Gendelman

Nebraska Center for Virology: Faculty Publications

Mononuclear phagocytes (MP) and T lymphocytes play a pivotal role in the host immune response to human immunodeficiency virus type 1 (HIV-1) infection. Regulation of such immune responses can be mediated, in part, through the interaction of the T-lymphocyte-expressed molecule CD40 ligand (CD40L) with its receptor on MP, CD40. Upregulation of CD40L on CD4⁺ peripheral blood mononuclear cells during advanced HIV-1 disease has previously been reported. Based on this observation, we studied the influence of CD40L-CD40 interactions on MP effector function and viral regulation in vitro. We monitored productive viral infection, cytokine and β -chemokine production, and β-chemokine receptor …

Canine And Feline Parvoviruses Can Use Human Or Feline Transferrin Receptors To Bind, Enter, And Infect Cells, John S. L. Parker, William J. Murphy, Dai Wang, Stephen J. O'Brien, Colin R. Parrish

Biology Faculty Articles

Canine parvovirus (CPV) enters and infects cells by a dynamin-dependent, clathrin-mediated endocytic pathway, and viral capsids colocalize with transferrin in perinuclear vesicles of cells shortly after entry (J. S. L. Parker and C. R. Parrish, J. Virol. 74:1919–1930, 2000). Here we report that CPV and feline panleukopenia virus (FPV), a closely related parvovirus, bind to the human and feline transferrin receptors (TfRs) and use these receptors to enter and infect cells. Capsids did not detectably bind or enter quail QT35 cells or a Chinese hamster ovary (CHO) cell-derived cell line that lacks any TfR (TRVb cells). However, capsids bound and …

Differentiation Of Two Bovine Lentiviruses By A Monoclonal Antibody On The Basis Of Epitope Specificity, Ling Zheng, Shucheng Zhang, Charles Wood, Sanjay Kapil, Graham E. Wilcox, Thomas A. Loughlin, H. C. Minocha

Nebraska Center for Virology: Faculty Publications

Bovine immunodeficiency virus (BIV) and Jembrana disease virus (JDV) are bovine lentiviruses that are closely related genetically. A recombinant fusion protein containing the capsid protein of BIV expressed in Escherichia coli was used to immunize mice and produce monoclonal antibodies. Six hybridomas specific for BIV capsid protein were identified, and one antibody, designated 10H1, was characterized further. Competitive binding assays were performed to analyze the topography of antigenic determinants by enzyme-linked immunosorbent assay and demonstrated the existence of at least three distinct antigenic determinants on capsid protein. The monoclonal antibody reacted specifically with both BIV capsid and the recombinant fusion …

Genetic And Biological Variation In Equine Infectious Anemia, Michael Belshan, Prasith Baccum, J. Lindsay Oaks, Brett A. Sponseller, Sean C. Murphy, James Cornette, Susan Carpenter

Nebraska Center for Virology: Faculty Publications

Genetic and biological variation in the regulatory protein Rev of equine infectious anemia virus (EIAV) were examined throughout a clinically dynamic disease course of an experimentally infected pony. Following infection with the virulent EIAV_Wyo, the pony underwent a variable disease course, including an acute fever episode at 12 days postinfection (DPI), multiple recurrent fever episodes until 135 DPI, a prolonged subclinical period, and two late fever episodes. Viral RNA was isolated from the inoculum and sequential sera samples, and the rev exon 2/gp45 overlapping ORFs were amplified, cloned, and sequenced. Novel variants were found throughout infection, and genetic …

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. …

Human Herpesvirus 8 Can Be Transmitted Through Blood In Drug Addicts, Carlos Sosa, Jorge Benetucci, Colleen Hanna, Laura Sieczkowski, Gabriel Deluchi, Ana Maria Canizal, Hamakwa Mantina, Winslow Klaskala, Marianna Baum, Charles Wood

Nebraska Center for Virology: Faculty Publications

Human Herpes virus type-8 (HHV-8) seroprevalence was studied in a population of HIV positive intravenous drug users (IVDUs) from Argentina. Analysis of this population also indirectly made it possible to study HHV-8 blood transmission, because these individuals frequently engage in needle sharing behavior and are capable of acquiring a broad array of blood borne pathogens, including Hepatitis B/C virus. The seroprevalence of HHV-8 in IVDUs was compared to a group of non-IVDUs and HIV negative individuals. Of the 223 individuals tested, 13.45% were HHV-8 positive, 16.99% in the IVDUs group, and 5.71% in the non-IVDUs. Among HIV positive IVDUs, 25/144 …

Interferon Regulatory Factor 7 Mediates Activation Of Tap-2 By Epstein-Barr Virus Latent Membrane Protein 1, Luwen Zhang, Joseph S. Pagano

Nebraska Center for Virology: Faculty Publications

Transporter associated with antigen processing 2 (Tap-2) is responsible for ATP-dependent transport of peptides from the cytosol to the endoplasmic reticulum, where peptides bind to newly synthesized human leukocyte antigen (HLA) class I molecules, which are essential for cellular immune responses. Epstein-Barr virus (EBV) latent membrane protein 1 (LMP-1) has been shown to induce the expression of Tap-2. In this study, the induction of endogenous Tap-2 by LMP-1 is shown to be associated with and requires the expression of interferon regulatory factor 7 (IRF-7). In DG75 Burkitt’s lymphoma (BL) cells, in which LMP-1 induces the expression of IRF-7, LMP-1 induced …

Vertical Transmission Of Kaposi's Sarcoma-Associated Herpesvirus, Hamakwa Mantina, Chipepo Kankasa, Winslow Klaskala, Brad Brayfield, James Campbell, Quijiang Du, Ganapati Bhat, Francis Kasolo, Charles Mitchell, Charles Wood

Nebraska Center for Virology: Faculty Publications

Little is presently known about the specific routes of transmission of Kaposi’s sarcoma-associated herpesvirus (KSHV) or human herpesvirus-8 (HHV-8). To investigate whether this agent might be transmitted vertically from mother to infant, we conducted a study on 89 KSHV seropositive mothers and their newborn infants. Thirteen mothers (14.6%) had KSHV DNA detected in their peripheral blood mononuclear cells (PBMC). Two of 89 samples drawn at birth from infants born to KSHV seropositive mothers had KSHV DNA detectable within their PBMC. These findings suggest that KSHV can be transmitted perinatally, but infrequently. Other routes of transmission such as horizontal transmission remain …

The Investigation Of Rna Viruses Using The Atomic Force Microscope, Michelle Moloney

Theses

The subject of viral entry into, and subsequent progeny virus egress, from cultured cells, has been extensively studied using numerous scientific techniques in fields ranging from cell culture to biochemistry to microscopy. The atomic force microscope (AFM) is a novel, developing instrument with unprecedented capabilities and this study was undertaken to observe enveloped Semliki Forest virus (SFV) and non-enveloped Theiler's Murine Encephalomyelitis virus (TMEV) entry into and egress from cultured cells using the AFM. Virus particle structure of both enveloped and non-enveloped viruses was also directly examined.

Cytopathic effect studies revealed that TMEV's are unable to produce progeny viruses in …