Virology Commons^™

The Nuclear Localization Of The Arabidopsis Transcription Factor Tip Is Blocked By Its Interaction With The Coat Protein Of Turnip Crinkle Virus, Tao Ren, Feng Qu, Thomas Jack Morris

Nebraska Center for Virology: Faculty Publications

We have previously reported that TIP, an Arabidopsis protein, interacts with the coat protein (CP) of Turnip crinkle virus (TCV) in yeast cells and that this interaction correlated with the resistance response in the TCV-resistant Arabidopsis ecotype Dijon-17. TIP was also able to activate transcription of reporter genes in yeast cells, suggesting that it is likely a transcription factor. We have now verified the physical interaction between TIP and TCV CP in vitro and showed that CP mutants unable to interact with TIP in yeast cells bind TIP with much lower affinity in vitro. Secondly, we have performed gel shift …

In Vitro And In Vivo Translational Efficiencies Of The 5′ Untranslated Region From Eight Genotype 2 Bovine Viral Diarrhea Virus Field Isolates, Christina L. Topliff, Seung K. Chon, Ruben O. Donis, Kent M. Eskridge, Clayton L. Kelling

Nebraska Center for Virology: Faculty Publications

We determined the in vitro and in vivo translational efficiency mediated by the internal ribosomal entry site (IRES) from eight BVDV2 field isolates varying in virulence using a bicistronic reporter vector in rabbit reticulocyte lysates (RRL), and in primate and bovine cell lines. Using a T7-promoter system, the high virulence isolates had greater translational efficiencies in bovine lymphocytes (BL-3 cells), than did the low virulence isolates. The low virulence isolates translated with greater efficiencies than the high virulence isolates in RRL, African green monkey kidney (CV-1) and bovine turbinate (BT) cells. Our results demonstrate that despite a high degree of …

Proteomic Fingerprints Distinguish Microglia, Bone Marrow, And Spleen Macrophage Populations, Yoshimi Enose, Christopher J. Destache, Andrea L. Mack, James R. Anderson, Fred Ullrich, Pawel Ciborowski, Howard E. Gendelman

Nebraska Center for Virology: Faculty Publications

Mononuclear phagocytes (MP; dendritic cells, monocytes, tissue macrophages, and microglia) maintain tissue homeostasis and provide a first line of defense against invading pathogens. In specific circumstances, MPs also induce inflammatory responses and as such affect disease onset and progression. Despite intensive research into MP biology, little is known of the functional and molecular properties of individual MP subtypes. Using a novel proteomics platform, unique protein patterns and protein identities were observed among populations of spleen and bone marrow macrophages and microglia. Cells were obtained from C57BL/6 mice and were cultivated in macrophage colony-stimulating factor. MP subtypes were indistinguishable by morphological …

Locatization Of Human Herpesvirus 8 In Human Sperms By In Situ Pcr, Omar Bagasra, Deepa Patel, Lisa Bobroski, Jamil A. Abbasi, Alex U. Bagasra, Hasna Baidouri, Twaina Harris, Albert El-Roeiy, Zsolt Lengvarszky, Homayoon Farzadegan, Charles Wood

Nebraska Center for Virology: Faculty Publications

Objectives: Defining the mechanism of infection with human herpesvirus-8 (HHV-8) or Kaposi’s sarcoma associated herpesvirus (KSHV) is an important clinical issue. HHV-8 has been linked to Kaposi’s sarcoma (KS) development in HIV-1-infected individuals, and KS develops in 40% of those infected with both viruses. A series of epidemiological data suggest that sexual transmission is one of the routes of transmission for HHV-8. In our studies, we sought to assess the cellular reservoirs of HHV-8 DNA in the semen of HIV-1-infected men and the potential role of HHV-8 infected spermatozoa in horizontal transmission. Design and methods: A nested polymerase chain reaction …

Guidelines For The Management Of Hiv Infection In Pregnant Women And The Prevention Of Mother-To-Child Transmission Of Hiv, David Hawkins, M. Blott, P. Clayden, A, De Ruiter, G. Foster, C. Gilling-Smith, B. Gosrani, H. Lyall, D. Mercey, M.-L. Newell, S. O'Shea, R. Smith, J. Sunderland, Charles Wood, G. Taylor

Nebraska Center for Virology: Faculty Publications

The prevalence of HIV infection amongst women giving birth in England and Wales has increased every year since 1990. Results from the Unlinked Anonymous Surveys of infection in pregnancy, show that in 2003, the prevalence reached one in 180 (0.56%) in inner London, one in 271 in outer London (0.37%) and one in 1,282 (0.08%) in the rest of England [1]. The majority of these women are from sub-Saharan Africa. The Department of Health policy of recommending an HIV test to every pregnant woman [2] has resulted in an increase in the proportion of these women who are aware of …

Antigenicity And Immunogenicity Of A Synthetic Human Immunodeficiency Virus Type 1 Group M Consensus Envelope Glycoprotein, Feng Gao, Eric A. Weaver, Zhongjing Lu, Yingying Li, Hua-Xin Liao, Benjiang Ma, S. Munir Alam, Richard M. Scearce, Laura L. Sutherland, Jae-Sung Yu, Julie M. Decker, George M. Shaw, David C. Montefiori, Bette T. Korber, Beatrice H. Hahn, Barton F. Haynes

Nebraska Center for Virology: Faculty Publications

Genetic variation of human immunodeficiency virus (HIV-1) represents a major obstacle for AIDS vaccine development. To decrease the genetic distances between candidate immunogens and field virus strains, we have designed and synthesized an artificial group M consensus env gene (CON6 gene) to be equidistant from contemporary HIV-1 subtypes and recombinants. This novel envelope gene expresses a glycoprotein that binds soluble CD4, utilizes CCR5 but not CXCR4 as a coreceptor, and mediates HIV-1 entry. Key linear, conformational, and glycan-dependent monoclonal antibody epitopes are preserved in CON6, and the glycoprotein is recognized equally well by sera from individuals infected with different HIV-1 …

Antigenicity And Immunogenicity Of A Synthetic Human Immunodeficiency Virus Type 1 Group M Consensus Envelope Glycoprotein, Feng Gao, Eric A. Weaver, Zhongjing Lu, Yingying Li, Hua-Xin Liao, Benjiang Ma, S. Munir Alam, Richard M. Scearce, Laura L. Sutherland, Jae-Sung Yu, Julie M. Decker, George M. Shaw, David C. Montefiori, Bette T. Korber, Beatrice H. Hahn, Barton F. Haynes

Nebraska Center for Virology: Faculty Publications

Genetic variation of human immunodeficiency virus (HIV-1) represents a major obstacle for AIDS vaccine development. To decrease the genetic distances between candidate immunogens and field virus strains, we have designed and synthesized an artificial group M consensus env gene (CON6 gene) to be equidistant from contemporary HIV-1 subtypes and recombinants. This novel envelope gene expresses a glycoprotein that binds soluble CD4, utilizes CCR5 but not CXCR4 as a coreceptor, and mediates HIV-1 entry. Key linear, conformational, and glycan-dependent monoclonal antibody epitopes are preserved in CON6, and the glycoprotein is recognized equally well by sera from individuals infected with different HIV-1 …

Genome Of Deerpox Virus, C. L. Afonso, Gustavo A. Delhon, E. R. Tuhlman, Z. Lu, A. Zsak, V. M. Becerra, L. Zsak, G. F. Kutish, D. L. Rock

Nebraska Center for Virology: Faculty Publications

Deerpox virus (DPV), an uncharacterized and unclassified member of the Poxviridae, has been isolated from North American free-ranging mule deer (Odocoileus hemionus) exhibiting mucocutaneous disease. Here we report the genomic sequence and comparative analysis of two pathogenic DPV isolates, W-848-83 (W83) and W-1170-84 (W84). The W83 and W84 genomes are 166 and 170 kbp, containing 169 and 170 putative genes, respectively. Nucleotide identity between DPVs is 95% over the central 157 kbp. W83 and W84 share similar gene orders and code for similar replicative, structural, virulence, and host range functions. DPV open reading frames (ORFs) with putative …

Epidemiological Characteristics Of Human Herpesvirus-8 Infection In A Large Population Of Antenatal Women In Zambia, W. Klaskala, B. P. Brayfield, C. Kankasa, G. Bhat, J. T. West, C. D. Mitchell, Charles Wood

Nebraska Center for Virology: Faculty Publications

Comprehensive data describing epidemiological characteristics of the human herpesvirus-8 or Kaposi’s sarcoma-associated herpesvirus (HHV-8 or KSHV) infection among pregnant women in a central sub-Saharan Africa are not available. This study determined virus prevalence estimates and the risk factors associated with HHV-8 infection. Cross-sectional, enrollment visit data were analyzed from a prospective cohort study of perinatal transmission of HHV-8 in Lusaka, Zambia. Exposure data were obtained via structured interview, physical examination, medical chart review, and laboratory testing. Among 3,160 antenatal women serologically screened for HHV-8 between September 1998 and October 2000, 40.2% were seropositive. The HHV-8 positive women were more likely …

Evolution Of Subtype C Hiv-1 Env In A Slowly Progressing Zambian Infant, Hong Zhang, Federico Hoffmann, Jun He, Xiang He, Chipepo Kankasa, Ruth Ruprecht, John T. West, Guillermo Orti, Charles Wood

Nebraska Center for Virology: Faculty Publications

Background: Given the high prevalence of mother to child infection, the development of a better understanding of African subtype C HIV-1 transmission and natural evolution is of significant importance. In this study, we genotypically and phenotypically characterized subtype C viruses isolated over a 67-month follow-up period from an in utero-infected Zambian infant. Changes in genotype and phenotype were correlated to alterations of the host humoral immune response.

Results: A comparison of baseline maternal and infant samples indicated that the infant sequences are monophyletic and contain a fraction of the diversity observed in the mother. This finding suggests that selective …

Compartmentalization Of Hiv-1 In The Central Nervous System: Role Of The Choroid Plexus, Evan J. Burkala, Jun He, John T. West, Charles Wood, Carol K. Petito

Nebraska Center for Virology: Faculty Publications

Objectives: To determine the genetic and phenotypic composition of HIV-1 found in the choroid plexus (CPx) and its relationship to virus in the brain and peripheral lymphoid tissue. Design: Phenotypic and molecular comparisons of HIV-1 found in autopsy brain, CPx, and spleen tissues. Methods: HIV-1 was co-cultured from matched postmortem brain (basal ganglia), CPx, and spleen tissues of AIDS patients with and without HIV-associated encephalitis and dementia. Viral phenotypes were determined by infection of monocyte-derived macrophages, MT-2 or co-receptor-specific cell lines. Viral env and pol sequences were determined from genomic DNA isolated directly from tissues or co-cultures, and phylogenetic comparisons …

Dichotomy In Cross-Clade Reactivity And Neutralization By Hiv-1 Sera: Implications For Active And Passive Immunotherapy, Lisa A. Cavacini, Mark Duval, Ajay Patil, Charles Wood, Kanneth H. Mayer, Ruth M. Ruprecht, Marshall L. Posner

Nebraska Center for Virology: Faculty Publications

The identification of broadly reactive and cross-clade neutralizing antibodies will facilitate the development of a more universally effective vaccine for human immunodeficiency virus (HIV). Antibodies in sera from individuals infected with Clade B HIV bind native primary viral isolates, and virus binding correlates with neutralization and stable clinical disease. In this study, we quantified cross-clade antibody reactivity and neutralization by Clades B and C sera. Primary viral isolates were captured by serum IgG bound to anti-human IgG and quantitated as p24 released by lysis of captured virus. Neutralization was determined using PHA-stimulated PBMC. Clade B antibodies reacted more frequently with …

Replication And Encapsidation Of Papillomaviruses In Saccharomyces Cerevisiae, Peter C. Angeletti

Nebraska Center for Virology: Faculty Publications

Improvements in methodologies to recapitulate and study particular biological functions of the

papillomavirus life cycle have led to great advances in our knowledge of these viruses. Described in

this chapter are techniques that allow low-copy and high-copy replication of full-length human

papillomavirus (HPV) genomes, as well as assembly of virus-like particles, in Saccharomyces

cerevisiae (yeast). This system has several distinct advantages that make it an attractive complement

to the well-established raft-culturing system. First, yeast are inexpensive, rapid, and simple to culture

in the lab. Second, they provide an ever-widening array of genetic tools to analyze HPV functions

—most recently notable, …

Association Of Dc-Sign Promoter Polymorphism With Increased Risk For Parenteral, But Not Mucosal, Acquisition Of Human Immunodeficiency Virus Type 1 Infection, Maureen P. Martin, Michael M. Lederman, Holli B. Hutcheson, James J. Goedert, George W. Nelson, Yvette Van Kooyk, Roger Detels, Susan Buchbinder, Keith Hoots, David Vlahov, Stephen J. O'Brien, Mary Carrington

Biology Faculty Articles

There is considerable debate about the fundamental mechanisms that underlie and restrict acquisition of human immunodeficiency virus type 1 (HIV-1) infection. In light of recent studies demonstrating the ability of C type lectins to facilitate infection with HIV-1, we explored the potential relationship between polymorphisms in the DC-SIGN promoter and risk for acquisition of HIV-1 according to route of infection. Using samples obtained from 1,611 European-American participants at risk for parenteral (n = 713) or mucosal (n = 898) infection, we identified single-nucleotide polymorphisms in the DC-SIGN promoter using single-strand conformation polymorphism. Individuals at risk for parenterally acquired …

Interferon Regulatory Factor 7 Is Associated With Epstein-Barr Virus-Transformed Central Nervous System Lymphoma And Has Oncogenic Properties, Luwen Zhang, Jun Zhang, Que Lambert, Channing J. Der, Luis Del Valle, Judith Miklossy, Kamel Kahlili, You Zhou, Joseph S. Pagano

Nebraska Center for Virology: Faculty Publications

EBV transforms primary B cells, and the major EBV oncoprotein, latent membrane protein 1 (LMP-1), is required for the process. LMP-1 both induces the expression of IRF-7 and activates the IRF-7 protein by phosphorylation and nuclear translocation. Here we report that the expression of IRF-7 is increased in EBV-immortalized B lymphocytes compared with that in primary B cells. IRF-7 was phosphorylated and predominantly localized in the nucleus in the immortalized cells. The expression of IRF-7 was detected in 19 of 27 specimens of primary lymphomas of the human central nervous system by immunohistochemical analysis. The association between LMP-1 and IRF-7 …

Infectious Molecular Clone Of A Recently Transmitted Pediatric Human Immunodeficiency Virus Clade C Isolate From Africa: Evidence Of Intraclade Recombination, Ricky D. Grisson, Agnes-Laurence Chenine, Lan-Yu Yeh, Jun He, Charles Wood, Ganapati J. Bhat, Weidong Zu, Chipepo Kankasa, Ruth M. Ruprecht

Nebraska Center for Virology: Faculty Publications

Although human immunodeficiency virus type 1 (HIV-1) clade C continues to dominate the pandemic, only two infectious clade C proviral DNA clones have been described (N. Mochizuki, N. Otsuka, K. Matsuo, T. Shiino, A. Kojima, T. Kurata, K. Sakai, N. Yamamoto, S. Isomura, T. N. Dhole, Y. Takebe, M. Matsuda, and M. Tatsumi, AIDS Res. Hum. Retrovir. 15:1321–1324, 1999; T. Ndung’u, B. Renjifo, and M. Essex, J. Virol. 75:4964–4972, 2001). We have generated an infectious molecular clone of a pediatric clade C strain, HIV1084i, which was isolated from a Zambian infant infected either intrapartum or through breastfeeding. HIV1084i is an …

The Latent Membrane Protein 1 Of Epstein-Barr Virus Establishes An Antiviral State Via Induction Of Interferon-Stimulated Genes, Jun Zhang, Subash C. Das, Catherine Kotalik, Asit K. Pattnaik, Luwen Zhang

Nebraska Center for Virology: Faculty Publications

Epstein-Barr virus (EBV) infection is associated with several human cancers. Latent membrane protein 1 (LMP-1) is one of the key viral proteins required for transformation of primary B cells in vitro and establishment of EBV latency. In this report, we show that LMP-1 is able to induce the expression of several interferon (IFN)-stimulated genes (ISGs) with antiviral properties such as 2’-5’ oligoadenylate synthetase (OAS), stimulated trans-acting factor of 50 kDa (STAF- 50), and ISG-15. LMP-1 inhibits vesicular stomatitis virus (VSV) replication at low multiplicity of infection (0.1 pfu/cell). The antiviral effect of LMP-1 is associated with the ability of LMP-1 …

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 …

Apobec3g Genetic Variants And Their Influence On The Progression To Aids, Ping An, Gabriela Bleiber, Priya Duggal, George Nelson, Margaret May, Bastien Mangeat, Irene Alobwede, Didier Trono, David Vlahov, Sharyne Donfield, James J. Goedert, John Phair, Susan Buchbinder, Stephen J. O'Brien, Amalio Telenti, Cheryl Winkler

Biology Faculty Articles

The cytosine deaminase APOBEC3G, in the absence of the human immunodeficiency virus type 1 (HIV-1) accessory gene HIV-1 viral infectivity factor (vif), inhibits viral replication by introducing G→A hypermutation in the newly synthesized HIV-1 DNA negative strand. We tested the hypothesis that genetic variants of APOBEC3G may modify HIV-1 transmission and disease progression. Single nucleotide polymorphisms were identified in the promoter region (three), introns (two), and exons (two). Genotypes were determined for 3,073 study participants enrolled in six HIV-AIDS prospective cohorts. One codon-changing variant, H186R in exon 4, was polymorphic in African Americans (AA) (f < 37%) and rare in European Americans (f < 3%) or Europeans (f …