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Nebraska Center for Virology: Faculty Publications

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Mutagenic Stabilization And/Or Disruption Of A Cd4-Bound State Reveals Distinct Conformations Of The Human Immunodeficiency Virus Type 1 Gp120 Envelope Glycoprotein, Shi-Hua Xiang, Peter D. Kwong, Rishi Gupta, Carlo D. Rizzuto, David J. Casper, Richard Wyatt, Liping Wang, Wayne A. Hendrickson, Michael L. Doyle, Joseph Sodroski Jan 2002

Mutagenic Stabilization And/Or Disruption Of A Cd4-Bound State Reveals Distinct Conformations Of The Human Immunodeficiency Virus Type 1 Gp120 Envelope Glycoprotein, Shi-Hua Xiang, Peter D. Kwong, Rishi Gupta, Carlo D. Rizzuto, David J. Casper, Richard Wyatt, Liping Wang, Wayne A. Hendrickson, Michael L. Doyle, Joseph Sodroski

Nebraska Center for Virology: Faculty Publications

The human immunodeficiency virus type 1 (HIV-1) gp120 exterior envelope glycoprotein is conformationally flexible. Upon binding to the host cell receptor CD4, gp120 assumes a conformation that is recognized by the second receptor, CCR5 and/or CXCR4, and by the CD4-induced (CD4i) antibodies. Guided by the X-ray crystal structure of a gp120-CD4-CD4i antibody complex, we introduced changes into gp120 that were designed to stabilize or disrupt this conformation. One mutant, 375 S/W, in which the tryptophan indole group is predicted to occupy the Phe 43 cavity in the gp120 interior, apparently favors a gp120 conformation closer to that of the CD4-bound …

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Phylogenetic And Phenotypic Analysis Of Hiv Type 1 Env Gp120 In Cases Of Subtype C Mother-To-Child Transmission, Hong Zhang, Guillermo Ortí, Qiujiang Du, Jun He, Chipepo Kankasa, Ganapati Bhat, Charles Wood Jan 2002

Phylogenetic And Phenotypic Analysis Of Hiv Type 1 Env Gp120 In Cases Of Subtype C Mother-To-Child Transmission, Hong Zhang, Guillermo Ortí, Qiujiang Du, Jun He, Chipepo Kankasa, Ganapati Bhat, Charles Wood

Nebraska Center for Virology: Faculty Publications

Human immunodeficiency virus type 1 (HIV-1) subtype C constitutes a majority of the newly transmitted cases of HIV-1 in many developing countries. There is growing evidence suggesting that subtype C viruses may display characteristics that make them distinct from subtype B and other subtypes, and such differences may affect its transmission and pathogenesis in infected individuals. In this study, HIV-1 sequences from the C2–V4 region of the envelope gene were analyzed from four infected mother–infant pairs (MIPs) after perinatal transmission in Zambia. We found that all viral isolates from four Zambian MIPs were of the subtype C HIV-1 subgroup. All …

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Small Glutamine-Rich Protein/Viral Protein U–Binding Protein Is A Novel Cochaperone That Affects Heat Shock Protein 70 Activity, Peter C. Angeletti, Doriann Walker, Antonito T. Panganiban Jan 2002

Small Glutamine-Rich Protein/Viral Protein U–Binding Protein Is A Novel Cochaperone That Affects Heat Shock Protein 70 Activity, Peter C. Angeletti, Doriann Walker, Antonito T. Panganiban

Nebraska Center for Virology: Faculty Publications

Molecular chaperone complexes containing heat shock protein (Hsp) 70 and Hsp90 are regulated by cochaperones, including a subclass of regulators, such as Hsp70 interacting protein (Hip), C-terminus of Hsp70 interacting protein (CHIP), and Hsp70-Hsp90 organizing factor (Hop), that contain tetratricopeptide repeats (TPRs), where Hsp70 refers to Hsp70 and its nearly identical constitutive counterpart, Hsc70, together. These proteins interact with the Hsp70 to regulate adenosine triphosphatase (ATPase) and folding activities or to generate the chaperone complex. Here we provide evidence that small glutamine-rich protein/viral protein U–binding protein (SGT/UBP) is a cochaperone that negatively regulates Hsp70. By “Far-Western” and pull-down assays, SGT/UBP …

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Cellular Tropisms And Co-Receptor Usage Of Hiv-1 Isolates From Vertically Infected Children With Neurological Abnormalities And Rapid Disease Progression, Micheline Mccarthy, Jun He, Denise Auger, Rebeca Geffin, Cecilia Hutto, Charles Wood, Gwendolyn Scott Jan 2002

Cellular Tropisms And Co-Receptor Usage Of Hiv-1 Isolates From Vertically Infected Children With Neurological Abnormalities And Rapid Disease Progression, Micheline Mccarthy, Jun He, Denise Auger, Rebeca Geffin, Cecilia Hutto, Charles Wood, Gwendolyn Scott

Nebraska Center for Virology: Faculty Publications

The longitudinal evolution of HIV-1 phenotypes was studied in a cohort of six vertically infected children with early onset and rapid progression of clinical disease. Among 30 viral isolates obtained from peripheral blood, tropisms for both human blood-derived cells (macrophages, T-lymphocytes), and for human neural (brain-derived) cells (microglia, astrocytes) were determined, as was chemokine co-receptor usage. All children harbored from birth macrophage-tropic isolates using the CCR5 co-receptor. Two children later developed T-cell tropic isolates with CXCR4 and CCR3 usage. While all six patients developed neurological abnormalities, only three produced neural cell tropic isolates, which used CCR5. However, early and persistent …

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Intracellular Signaling Molecules Activated By Epstein-Barr Virus For Induction Of Interferon Regulatory Factor 7, Luwen Zhang, Lihong Wu, Ke Hong, Joseph S. Pagano Dec 2001

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 …

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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 Jul 2001

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 …

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Interferon Regulatory Factor 7 Mediates Activation Of Tap-2 By Epstein-Barr Virus Latent Membrane Protein 1, Luwen Zhang, Joseph S. Pagano Jan 2001

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 …

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Binding Of Equine Infectious Anemia Virus Rev To An Exon Splicing Enhancer Mediates Alternative Splicing And Nuclear Export Of Viral Mrnas, Michael Belshan, Gregory S. Park, Patricia Bilodeau, C. Martin Stoltzfus, Susan Carpenter May 2000

Binding Of Equine Infectious Anemia Virus Rev To An Exon Splicing Enhancer Mediates Alternative Splicing And Nuclear Export Of Viral Mrnas, Michael Belshan, Gregory S. Park, Patricia Bilodeau, C. Martin Stoltzfus, Susan Carpenter

Nebraska Center for Virology: Faculty Publications

In addition to facilitating the nuclear export of incompletely spliced viral mRNAs, equine infectious anemia virus (EIAV) Rev regulates alternative splicing of the third exon of the tat/rev mRNA. In the presence of Rev, this exon of the bicistronic RNA is skipped in a fraction of the spliced mRNAs. In this report, the cis-acting requirements for exon 3 usage were correlated with sequences necessary for Rev binding and transport of incompletely spliced RNA. The presence of a purine-rich exon splicing enhancer (ESE) was required for exon 3 recognition, and the addition of Rev inhibited exon 3 splicing. Glutathione-S-transferase (GST)-Rev bound …

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Jembrana Disease Virus Tat Can Regulate Human Immunodeficiency Virus (Hiv) Long Terminal Repeat-Directed Gene Expression And Can Substitute For Hiv Tat In Viral Replication, Hexin Chen, Jun He, Steven Fong, Graham Wilcox, Charles Wood Mar 2000

Jembrana Disease Virus Tat Can Regulate Human Immunodeficiency Virus (Hiv) Long Terminal Repeat-Directed Gene Expression And Can Substitute For Hiv Tat In Viral Replication, Hexin Chen, Jun He, Steven Fong, Graham Wilcox, Charles Wood

Nebraska Center for Virology: Faculty Publications

Jembrana disease virus (JDV) is a bovine lentivirus genetically similar to bovine immunodeficiency virus; it causes an acute and sometimes fatal disease in infected animals. This virus carries a very potent Tat that can strongly activate not only its own long terminal repeat (LTR) but also the human immunodeficiency virus (HIV) LTR. In contrast, HIV Tat cannot reciprocally activate the JDV LTR (H. Chen, G. E. Wilcox, G. Kertayadnya, and C. Wood, J. Virol. 73:658–666, 1999). This indicates that in transactivation JDV Tat may utilize a mechanism similar to but not the same as that of the HIV Tat. To …

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Interferon Regulatory Factor 7 Is Induced By Epstein-Barr Virus Latent Membrane Protein 1, Luwen Zhang, Joseph S. Pagono Feb 2000

Interferon Regulatory Factor 7 Is Induced By Epstein-Barr Virus Latent Membrane Protein 1, Luwen Zhang, Joseph S. Pagono

Nebraska Center for Virology: Faculty Publications

Infection by Epstein-Barr virus (EBV) generates several types of latency with different profiles of gene expression but with expression of Epstein-Barr nuclear antigen 1 (EBNA-1) in common. The BamHI Q promoter (Qp) is used for the transcription of EBNA-1 mRNA in type I latency, which is an EBV infection state exemplified by Burkitt’s lymphoma (BL). However, Qp is inactive in type III latency, and other promoters (C/Wp) are used for transcription of EBNA-1, which raises the question of how usage of these promoters is governed. Interferon (IFN) regulatory factor 7 (IRF-7) was identified first as a negative regulator of …

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Identification Of The Transactivation Domain Of The Transcription Factor Sox-2 And An Associated Co-Activator, Tamara K. Nowling, Lance R. Johnson, Matthew S. Wiebe, Angie Rizzino Jan 2000

Identification Of The Transactivation Domain Of The Transcription Factor Sox-2 And An Associated Co-Activator, Tamara K. Nowling, Lance R. Johnson, Matthew S. Wiebe, Angie Rizzino

Nebraska Center for Virology: Faculty Publications

The importance of interactions between Sox and POU transcription factors in the regulation of gene expression is becoming increasingly apparent. Recently, many examples of the involvement of Sox-POU partnerships in transcription have been discovered, including a partnership between Sox-2 and Oct-3. Little is known about the mechanisms by which these factors modulate transcription. To better understand the molecular interactions involved, we mapped the location of the transactivation do-main of Sox-2. This was done in the context of its interaction with Oct-3, as well as its ability to transactivate as a fusion protein linked to the DNA-binding domain of Gal4. Both …

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Interferon Regulatory Factor 2 Represses The Epstein-Barr Virus BamHi Q Latency Promoter In Type Iii Latency, Luwen Zhang, Joseph S. Pagano Apr 1999

Interferon Regulatory Factor 2 Represses The Epstein-Barr Virus BamHi Q Latency Promoter In Type Iii Latency, Luwen Zhang, Joseph S. Pagano

Nebraska Center for Virology: Faculty Publications

Epstein-Barr virus (EBV) nuclear antigen 1 (EBNA-1) is the essential protein for maintenance of the EBV episome and establishment of latency. The BamHI Q promoter (Qp) is used for the transcription of EBNA-1 mRNA in type I and type II latency, which are EBV infection states exemplified by Burkitt’s lymphoma and nasopharyngeal carcinoma. However, Qp is inactive in type III latency, and other promoters (the BamHI C promoter and/or the BamHI W promoter) are used for EBNA-1. The involvement of interferon regulatory factors (IRFs) in the regulation of Qp is suggested by the presence of an essential interferon-stimulated response element …

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Characterization Of The Jembrana Disease Virus Tat Gene And The Cis- And Trans-Regulatory Elements In Its Long Terminal Repeats, Hexin Chen, Graham Wilcox, Gde Kertayadnya, Charles Wood Jan 1999

Characterization Of The Jembrana Disease Virus Tat Gene And The Cis- And Trans-Regulatory Elements In Its Long Terminal Repeats, Hexin Chen, Graham Wilcox, Gde Kertayadnya, Charles Wood

Nebraska Center for Virology: Faculty Publications

Jembrana disease virus (JDV) is a newly identified bovine lentivirus that is closely related to the bovine immunodeficiency virus (BIV). JDV contains a tat gene, encoded by two exons, which has potent transactivation activity. Cotransfection of the JDV tat expression plasmid with the JDV promoter chloramphenicol acetyltransferase (CAT) construct pJDV-U3R resulted in a substantial increase in the level of CAT mRNA transcribed from the JDV long terminal repeat (LTR) and a dramatic increase in the CAT protein level. Deletion analysis of the LTR sequences showed that sequences spanning nucleotides 268 to 153, including the TATA box and the predicted first …

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Biological Characterization Of Rev Variation In Equine Infectious Anemia Virus, Michael Belshan, Matthew E. Harris, Anne E. Shoemaker, Thomas J. Hope, Susan Carpenter May 1998

Biological Characterization Of Rev Variation In Equine Infectious Anemia Virus, Michael Belshan, Matthew E. Harris, Anne E. Shoemaker, Thomas J. Hope, Susan Carpenter

Nebraska Center for Virology: Faculty Publications

Sequence analysis identified significant variation in the second exon of equine infectious anemia virus (EIAV) rev. Functional analysis indicated that limited amino acid variation in Rev significantly altered the export activity of the protein but did not affect Rev-dependent alternative splicing. EIAV Rev can mediate export through two independent cis -acting Rev-responsive elements (RREs), and differences among Rev variants were more pronounced when both RREs were present. Variation in Rev may be an important mechanism for regulation of virus replication in vivo and may contribute to changes in clinical disease.

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Human Immunodeficiency Virus Neurotropism: An Analysis Of Viral Replication And Cytopathicity For Divergent Strains In Monocytes And Microglia, Anuja Ghorpade, Adeline Nukuna, Myhanh Che, Sheryl Haggerty, Yuri Persidsky, Eboni Carter, Leeroy Carhart, Laura Shafer, Howard Gendelman Apr 1998

Human Immunodeficiency Virus Neurotropism: An Analysis Of Viral Replication And Cytopathicity For Divergent Strains In Monocytes And Microglia, Anuja Ghorpade, Adeline Nukuna, Myhanh Che, Sheryl Haggerty, Yuri Persidsky, Eboni Carter, Leeroy Carhart, Laura Shafer, Howard Gendelman

Nebraska Center for Virology: Faculty Publications

Productive replication of human immunodeficiency virus type 1 (HIV-1) in brain macrophages and microglia is a critical component of viral neuropathogenesis. However, how virus-macrophage interactions lead to neurological disease remains incompletely understood. Possibly, a differential ability of virus to replicate in brain tissue macrophages versus macrophages in other tissues underlies HIV-1 neurovirulence. To these ends, we established systems for the isolation and propagation of pure populations of human microglia and then analyzed the viral life cycles of divergent HIV-1 strains in these cells and in cultured monocytes by using identical viral inocula and indicator systems. The HIV-1 isolates included those …

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Human Immunodeficiency Virus Neurotropism: An Analysis Of Viral Replication And Cytopathicity For Divergent Strains In Monocytes And Microglia, Anuja Ghorpade, Adeline Nukuna, Myhanh Che, Sherryl Haggerty, Yuri Persidsky, Eboni Carter, Leeroy Carhart, Laura Shafer, Howard Gendelman Apr 1998

Human Immunodeficiency Virus Neurotropism: An Analysis Of Viral Replication And Cytopathicity For Divergent Strains In Monocytes And Microglia, Anuja Ghorpade, Adeline Nukuna, Myhanh Che, Sherryl Haggerty, Yuri Persidsky, Eboni Carter, Leeroy Carhart, Laura Shafer, Howard Gendelman

Nebraska Center for Virology: Faculty Publications

Productive replication of human immunodeficiency virus type 1 (HIV-1) in brain macrophages and microglia is a critical component of viral neuropathogenesis. However, how virus-macrophage interactions lead to neurological disease remains incompletely understood. Possibly, a differential ability of virus to replicate in brain tissue macrophages versus macrophages in other tissues underlies HIV-1 neurovirulence. To these ends, we established systems for the isolation and propagation of pure populations of human microglia and then analyzed the viral life cycles of divergent HIV-1 strains in these cells and in cultured monocytes by using identical viral inocula and indicator systems. The HIV-1 isolates included those …

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Human Herpesvirus 8 As A Potential Sexually Transmitted Agent In Honduras, C. Sosa, W. Klaskala, B. Chandran, R. Soto, L. Sieczkowski, M. H. Wu, M. Baum, Charles Wood Jan 1998

Human Herpesvirus 8 As A Potential Sexually Transmitted Agent In Honduras, C. Sosa, W. Klaskala, B. Chandran, R. Soto, L. Sieczkowski, M. H. Wu, M. Baum, Charles Wood

Nebraska Center for Virology: Faculty Publications

The seroprevalence of human herpesvirus 8 (HHV-8) was studied in 326 human immunodeficiency virus (HIV)–positive and –negative persons from Honduras; women constituted 77% (n = 251) of the subjects. Sera were tested for lytic HHV-8 antibodies by an IFA, and positive samples were confirmed by a radioimmunoprecipitation assay. Of the 326 persons tested, 58 (17.8%) had HHV- 8 antibodies. Among the HIV-infected women, 22.7% were seropositive; 11.3% of the HIV-negative women were seropositive. HHV-8 seroprevalence was almost four times higher in HIV-positive female commercial sex workers (36%) than in HIV-negative female non–commercial sex workers (9.9%; odds ratio = 3.8, 95% …

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Seroprevalence Of Human Herpesvirus 8 Among Zambian Women Of Childbearing Age Without Kaposi’S Sarcoma (Ks) And Mother-Child Pairs With Ks, Jun He, Ganapati Bhat, Chipepo Kankasa, Chifumbe Chintu, Charles Mitchell, Wenjing Duan, Charles Wood Jan 1998

Seroprevalence Of Human Herpesvirus 8 Among Zambian Women Of Childbearing Age Without Kaposi’S Sarcoma (Ks) And Mother-Child Pairs With Ks, Jun He, Ganapati Bhat, Chipepo Kankasa, Chifumbe Chintu, Charles Mitchell, Wenjing Duan, Charles Wood

Nebraska Center for Virology: Faculty Publications

The seroprevalence of human herpesvirus 8 (HHV-8) among a group of Zambian women of reproductive age and among mother-child pairs in which either one of them has Kaposi’s sarcoma (KS) was determined. A cross-sectional group of 378 pregnant women was randomly recruited into the study, and 183 (48.4%) had HHV-8 antibodies. Among the human immunodeficiency virus (HIV)-1–infected women, 51.1% were HHV-8–seropositive, whereas of HIV- 1–negative women, 47.3% were HHV-8–seropositive. In addition, 21 women index patients with KS and 5 young children index patients with KS were studied. All children with KS had mothers who were HHV-8–seropositive, while not all children …

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Porcine Reproductive And Respiratory Syndrome Virus Replicates In Testicular Germ Cells, Alters Spermatogenesis, And Induces Germ Cell Death By Apoptosis, Jung-Hyang Sur, Alan R. Doster, James S. Christian, Judith A. Galeota, Robert W. Wills, Jeff J. Zimmerman, Fernando A. Osorio Dec 1997

Porcine Reproductive And Respiratory Syndrome Virus Replicates In Testicular Germ Cells, Alters Spermatogenesis, And Induces Germ Cell Death By Apoptosis, Jung-Hyang Sur, Alan R. Doster, James S. Christian, Judith A. Galeota, Robert W. Wills, Jeff J. Zimmerman, Fernando A. Osorio

Nebraska Center for Virology: Faculty Publications

Like other arteriviruses, porcine reproductive and respiratory syndrome virus (PRRSV) is shed in semen, a feature that is critical for the venereal transmission of this group of viruses. In spite of its epidemiological importance, little is known of the association of PRRSV or other arteriviruses with gonadal tissues. We experimentally infected a group of boars with PRRSV 12068-96, a virulent field strain. By combined use of in situ hybridization and immunohistochemistry, we detected infection by PRRSV in the testes of these boars. The PRRSV testicular replication in testis centers on two types of cells: (i) epithelial germ cells of the …

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Human Herpesvirus Type 8 Dna Sequences In Cell-Free Plasma And Mononuclear Cells Of Kaposi's Sarcoma Patients, William J. Harrington, Omar Bagasra, Carlos E. Sosa, Lisa E. Bobroski, Marianna Baum, Xue Lan Wen, Lisa Cabral, Gerald E. Byrne, Roger J. Pomerantz, Charles Wood Nov 1996

Human Herpesvirus Type 8 Dna Sequences In Cell-Free Plasma And Mononuclear Cells Of Kaposi's Sarcoma Patients, William J. Harrington, Omar Bagasra, Carlos E. Sosa, Lisa E. Bobroski, Marianna Baum, Xue Lan Wen, Lisa Cabral, Gerald E. Byrne, Roger J. Pomerantz, Charles Wood

Nebraska Center for Virology: Faculty Publications

Human herpesvirus (HHV) type 8 bas been detected in both classical and AIDS-related Kaposi's sarcoma, body-cavity lymphomas, and other types of tumors. HHV-8 has also been detected in DNA from peripheral blood mononuclear cells (PBMC) of some Kaposi's sarcoma patients and more readily in B cell fractions derived from panned cell subpopulations. Two patients were followed using several methods: in situ hybridization, solution-based polymerase chain reaction (PCR), and in situ PCR. HHV-8 was intermittently detected in plasma and detection correlated with detection in PBMC. In situ PCR demonstrated HHV-8 sequences in both peripheral blood B lymphocytes and, to a lesser …

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Tumor Necrosis Factor Alpha-Induced Apoptosis In Human Neuronal Cells: Protection By The Antioxidant N-Acetylcysteine And The Genes Bcl-2 And Crma, Angela K. Talley, Stephen Dewhurst, Seth W. Perry, Sheila C. Dollard, Suryaram Gummuluru, Steven M. Fine, Deborah New, Leon Epstein, Howard Gendelman, Harris A. Gelbard May 1995

Tumor Necrosis Factor Alpha-Induced Apoptosis In Human Neuronal Cells: Protection By The Antioxidant N-Acetylcysteine And The Genes Bcl-2 And Crma, Angela K. Talley, Stephen Dewhurst, Seth W. Perry, Sheila C. Dollard, Suryaram Gummuluru, Steven M. Fine, Deborah New, Leon Epstein, Howard Gendelman, Harris A. Gelbard

Nebraska Center for Virology: Faculty Publications

Tumor necrosis factor alpha (TNF-α) is a candidate human immunodeficiency virus type 1-induced neurotoxin that contributes to the pathogenesis of AIDS dementia complex. We report here on the effects of exogenous TNF-α on SK-N-MC human neuroblastoma cells differentiated to a neuronal phenotype with retinoic acid. TNF-α caused a dose-dependent loss of viability and a corresponding increase in apoptosis in differentiated SK-N-MC cells but not in undifferentiated cultures. Importantly, intracellular signalling via TNF receptors, as measured by activation of the transcription factor NF-kB, was unaltered by retinoic acid treatment. Finally, overexpression of bcl-2 or crmA conferred resistance to apoptosis mediated by …

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Media Components Influence Viral Gene Expression Assays In Human Fetal Astrocyte Cultures, Micheline Mccarthy, Charles Wood, Larisa Fedoseyeva, Scott R. Whittemore Jan 1995

Media Components Influence Viral Gene Expression Assays In Human Fetal Astrocyte Cultures, Micheline Mccarthy, Charles Wood, Larisa Fedoseyeva, Scott R. Whittemore

Nebraska Center for Virology: Faculty Publications

In vitro neurovirological studies of viral infectivity or viral gene expression may be confounded by the mulHple neural cell types and/or fibrob last contamination present in early passage cultures prepared from dissociated human central nervous system (eNS) tissue. We have developed highly enriched astrocyte cultures for neurovirological study by culturing in a serum-free defined medium, 816, supplemented with basic fibroblast growth factor (FGF-2). Subculture in this medium selects against fibroblast proliferation and favors sustained proliferation of a highly enriched glial fibrillary acidic protein (GFAP)-positive cell population. These astrocytes support productive replication of cytomegalovirus (CMV) and transient expression of transfected CMVand …

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Book Review: The Baculovirus Expression System: A Laboratory Guide (1992) King, L. A. & Possee, R. D., David D. Dunigan Jan 1993

Book Review: The Baculovirus Expression System: A Laboratory Guide (1992) King, L. A. & Possee, R. D., David D. Dunigan

Nebraska Center for Virology: Faculty Publications

The power of molecular biology is unleashed with the ability to clone and sequence genes, and then express these genes in heterologous systems. This sets the stage for the full analysis of proteins that are otherwise difficult to isolate and/or purify, especially when present at very low copy number per cell or when isolated from relatively precious materials. Overexpression of protein is now possible in a number of systems including prokaryotes (e.g., E. coli) and various eukaryotes (yeast, insects, and plants). The issue then becomes, which system (1) most closely reflects the homologous expression with respect to posttranslational modifications, …

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Loss Of Infectivity By Progeny Virus From Alpha Interferon- Treated Human Immunodeficiency Virus Type 1-Infected T Cells Is Associated With Defective Assembly Of Envelope Gp120, Brian D. Hanson, Peter L. Nara, Radha K. Maheshwari, Girmel S. Sidhu, John G. Bernbaum, David Hoekzema, Monte S. Meltzer, Howard Gendelman Dec 1992

Loss Of Infectivity By Progeny Virus From Alpha Interferon- Treated Human Immunodeficiency Virus Type 1-Infected T Cells Is Associated With Defective Assembly Of Envelope Gp120, Brian D. Hanson, Peter L. Nara, Radha K. Maheshwari, Girmel S. Sidhu, John G. Bernbaum, David Hoekzema, Monte S. Meltzer, Howard Gendelman

Nebraska Center for Virology: Faculty Publications

Levels of human immunodeficiency virus (HIV) DNA, RNA, or p24 antigen and reverse transcriptase activity in T-cell cultures treated with 500 IU of recombinant alpha interferon (rIFNα) per ml were comparable to those in control cultures. Radioimmunoprecipitation analysis of proteins in lysates of IFN-treated T cells documented a marked accumulation of HlV proteins. Localization of gp120 by immunofluorescence showed a diffuse pattern in IFN-treated cells quite distinct from the ring pattern in untreated control cells. That large quantities of a120 in aberrant cell compartments might affect HlV morphogenesis was confirmed in infectivity studies: virions from IFN-treated cells were 100- to …

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Altered Expression Of Adenovirus 12 Dna-Binding Protein But Not Dna Polymerase During Abortive Infection Of Hamster Cells, Lynne A. Lucher, Benjawan Khuntirat, Jiansheng Zhao, Peter C. Angeletti Jan 1992

Altered Expression Of Adenovirus 12 Dna-Binding Protein But Not Dna Polymerase During Abortive Infection Of Hamster Cells, Lynne A. Lucher, Benjawan Khuntirat, Jiansheng Zhao, Peter C. Angeletti

Nebraska Center for Virology: Faculty Publications

Replication of human adenovirus type 12 DNA is blocked in abortively infected baby hamster kidney cells. The activity and accumulation of adenovirus 12 DNA polymerase is equivalent in infected hamster and human cell extracts. However, the accumulation of adenovirus type 12 DNA-binding protein is approximately 120-fold lower in extracts from infected hamster cells when compared to infected permissive human cells. This difference in accumulation is not because of replication of viral DNA during productive infection, since this difference is observed in the presence of hydroxyurea. The DNA-binding protein from infected hamster cells retains the ability to bind denatured DNA-cellulose. An …

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The Inability Of Human Immunodeficiency Virus To Infect Chimpanzee Monocytes Can Be Overcome By Serial Viral Passage In Vivo, Howard Gendelman, Garth D. Ehrlich, Lisa M. Baca, Shawn Conley, Jorge Ribas, D. Chester Kalter, Monte S. Meltzer, Bernard J. Poiesz, Peter Nara Jul 1991

The Inability Of Human Immunodeficiency Virus To Infect Chimpanzee Monocytes Can Be Overcome By Serial Viral Passage In Vivo, Howard Gendelman, Garth D. Ehrlich, Lisa M. Baca, Shawn Conley, Jorge Ribas, D. Chester Kalter, Monte S. Meltzer, Bernard J. Poiesz, Peter Nara

Nebraska Center for Virology: Faculty Publications

Studies of lentivirus infection in ruminants, nonhuman primates, and humans suggest that virus infection of macrophages plays a central role in the disease process. To investigate whether human immunodeficiency virus type 1 (HIV-1) can infect chimpanzee macrophages, we recovered monocytes from peripheral blood mononuclear cells of HIV-1-negative animals and inoculated these and control human monocytes with a panel of four human-passaged monocytotropic virus strains and one chimpanzee-passaged isolate. HIV-1-infected human monocytes synthesized proviral DNA, viral mRNA, p24 antigen, and progeny virions. In contrast, except for the chimpanzee-passaged HIV-1 isolate, chimpanzee monocytes failed to support HIV-1 replication when cultured under both …

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Reliable Detection Of Individuals Seropositive For The Human Immunodeficiency Virus (Hiv) By Competitive Immunoassays Using Escherichia Coli - Expressed Hiv Structural Proteins, G. J. Dawson, J. S. Heller, Charles Wood, R. A. Gutierrez, J. S. Webber, J. C. Hunt, S. A. Hojvat, D. Senn, S. G. Devare, R. H. Decker Jan 1988

Reliable Detection Of Individuals Seropositive For The Human Immunodeficiency Virus (Hiv) By Competitive Immunoassays Using Escherichia Coli - Expressed Hiv Structural Proteins, G. J. Dawson, J. S. Heller, Charles Wood, R. A. Gutierrez, J. S. Webber, J. C. Hunt, S. A. Hojvat, D. Senn, S. G. Devare, R. H. Decker

Nebraska Center for Virology: Faculty Publications

We molecularly cloned the gag and env genes of the human immunodeficiency virus (HIV) and expressed fragments of these genes in Escherichia coli. Using the recombinant core and envelope proteins, we developed two competitive immunoassays (CIAs). Samples that recognized either the envelope or core proteins were considered positive for antibodies to HIV. This test system was comparable with western blot in detecting antibodies in patients with AIDS or AIDS-related complex that were repeatably reactive in the HIV screening test. All 360 individuals who were positive by western blot were positive by the CIA. A total of 844 samples repeatably …

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Activation Of The Human Immunodeficiency Virus By Herpes Simplex Virus Type 1, Jeffrey M. Ostrove, John Leonard, Karen E. Weck, Arnold B. Rabson, Howard Gendelman Dec 1987

Activation Of The Human Immunodeficiency Virus By Herpes Simplex Virus Type 1, Jeffrey M. Ostrove, John Leonard, Karen E. Weck, Arnold B. Rabson, Howard Gendelman

Nebraska Center for Virology: Faculty Publications

Herpes simplex virus type 1 (HSV-1) and some of its immediate-early genes stimulate expression of the human immunodeficiency virus (HIV) long terminal repeat (LTR) sequences and the replication of HIV itself. To demonstrate this, the HIV LTR was linked to the indicator gene chloramphenicol acetyltransferase (CAT) and transfected into Vero cells with or without the trans-activating gene (tat) of HIV. Infection of these cells with HSV-1 strain KOS or temperature-sensitive mutant tsB2l or tsE6 resulted in a large increase in CAT activity in the absence of tat and further augmentation in the presence of tat. This stimulation was seen at …

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Productive, Persistent Infection Of Human Colorectal Cell Lines With Human Immunodeficiency Virus, Akio Adachi, Scott Koenig, Howard Gendelman, Daryl Daugherty, Sebastiano Gattoni-Celli, Anthony Fauci, Malcom A. Martin Jan 1987

Productive, Persistent Infection Of Human Colorectal Cell Lines With Human Immunodeficiency Virus, Akio Adachi, Scott Koenig, Howard Gendelman, Daryl Daugherty, Sebastiano Gattoni-Celli, Anthony Fauci, Malcom A. Martin

Nebraska Center for Virology: Faculty Publications

Thirteen adherent human non-lymphocyte cell lines were tested for their susceptibility to infection by human immunodeficiency virus. Productive infection could be demonstrated in three of five colorectal carcinoma cell lines examined; the other eight human non-lymphocyte cell lines were uninfectible. A susceptible colon carcinoma cell line (HT29), as well as normal colonic mucosa, was shown to contain a 3.0-kilobase species of poly(A)+ CD4 RNA, whereas uninfectible colon carcinoma and rhabdomyosarcoma cell lines synthesized no detectable T4 RNA. A persistently infected colon carcinoma cell line was established that continued to produce progeny human immunodeficiency virus for more than 10 weeks …

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Production Of Acquired Immunodeficiency Syndrome-Associated Retrovirus In Human And Nonhuman Cells Transfected With An Infectious Molecular Clone, Akio Adachi, Howard E. Gendelman, Scott Koenig, Thomas Folks, Ronald Willey, Arnold Rabson, Malcom A. Martin Aug 1986

Production Of Acquired Immunodeficiency Syndrome-Associated Retrovirus In Human And Nonhuman Cells Transfected With An Infectious Molecular Clone, Akio Adachi, Howard E. Gendelman, Scott Koenig, Thomas Folks, Ronald Willey, Arnold Rabson, Malcom A. Martin

Nebraska Center for Virology: Faculty Publications

We constructed an infectious molecular clone of acquired immunodeficiency syndrome-associated retrovirus. Upon transfection, this clone directed the production of infectious virus particles in a wide variety of cells in addition to human T4 cells. The progeny, infectious virions, were synthesized in mouse, mink, monkey, and several human non-T cell lines, indicating the absence of any intracellular obstacle to viral RNA or protein production or assembly. During the course of these studies, a human colon carcinoma cell line, exquisitely sensitive to DNA transfection, was identified.

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