pinecone/core-2020-05-10-deduplication huggingface dataset:
Desialylation of Peripheral Blood Mononuclear Cells Promotes Growth of HIV-1

AbstractActivation of peripheral blood CD4+helper T lymphocytes establishes a permissive state for growth of HIV-1. Activated T lymphocytes expressed increased sialidase (neuraminidase) activity and were hyposialylated. Treatment of freshly isolated peripheral blood mononuclear cells (PBMCs) with microbial neuraminidase (NANase) or phytohemagglutinin (PHA) prior to infection at low multiplicity with T cell line-adapted HIV-1IIIBresulted in production of large amounts of p24 antigen and reverse transcriptase. In contrast, neither viral component was detected in the medium of mock-treated cells infected at a similar multiplicity through 21 days in culture. The titer of a stock solution of HIV-1IIIBwas 1.4 ± 0.18 log10greater in NANase-treated PBMCs than in mock-treated cells; the titer was similarly raised 1.5 to 1.76 ± 0.18 log10in PHA-treated cells. Growth of the primary isolate HIV-191/US/056was also enhanced in NANase-treated PBMCs; the titer of a stock solution of HIV-191/US/056was 1.0 ± 0.16 log10greater in NANase-treated PBMCs than in mock-treated cells 7 days after infection. No enhancement of viral growth in PBMCs was detected when NANase was heat-inactivated or specifically inhibited with 2,3-dehydro-2-desoxy-N-acetyl-neuraminic acid prior to use. Treatment of PBMCs with NANase did not alter the distribution of lymphocyte subsets nor change the density of CD4 antigen per cell after 7 days in culture. Whereas PHA treatment of PBMCs was mitogenic, pretreatment with NANase was not; the amount of [3H]thymidine incorporated into DNA and culture growth characteristics were similar for NANase- and mock-treated cells. Thus, desialylation of PBMCs promoted a permissive state for growth of HIV-1 without affecting the rate of DNA synthesis or relative number of target CD4+cells
Characterization of Rev Function Using Subgenomic and Genomic Constructs in T and COS Cells

AbstractThe effect of the human immunodeficiency type 1 (HIV-1) Rev protein on the splicing and cytoplasmic accumulation of HIV-1 RNAs was investigated in COS and T cells. Subgenomic and genomic constructs were used which expressed varying levels of complexity in their potential RNA constituents. Using all constructs, in both cell types, an inhibitory effect of Rev on the level of fully spliced HIV-1 RNAs could be demonstrated. An increase in the nuclear level of unspliced pre-mRNA was seen in the presence of Rev with genomic constructs. Thus, the inhibitory effect on splicing was not merely due to enhancement of nuclear export of the pre-mRNA with these constructs. In both cell types, a positive effect of Rev on the cytoplasmic accumulation of HIV-1 RNAs could also be seen. However, in T cells, the Rev-dependent RNAs were still capable of accumulating at a reduced level in the cytoplasmic fraction in the absence of Rev. The identity of the cell type, construct, and RNA species impacted on the phenotypic manifestation of Rev function
Modulation of Bovine Papillomavirus DNA Replication by Phosphorylation of the Viral E1 Protein

AbstractE1 is the DNA replication origin recognition protein for bovine papillomavirus (BPV), and it carries out enzymatic functions required for initiation of viral DNA replication. Cellular mechanisms likely play a role in regulating BPV DNA replication. We are investigating the role of phosphorylation of E1 on viral replicationin vivoand on E1 activityin vitro.Serine 109 is a phosphoacceptorin vivoand is targeted by protein kinase A and protein kinase Cin vitro.A viral genome carrying a serine 109 to alanine mutation replicates more efficiently than wild-typein vivoin a transient replication assay. Furthermore, purified mutant protein, while having wild-type levels of ATPase activity, is able to bind more origin-containing DNA than wild-type E1. Phosphorylation therefore appears to play a selective role in modulating a specific E1 function during viral DNA replication
pH-Independent Uptake of Hepatitis B Virus in Primary Human Hepatocytes

AbstractThe replication cycle of the hepatitis B virus (HBV) is still incompletely understood. In particular, the early steps of the viral life cycle, such as absorption, penetration, uncoating, and nuclear translocation require further clarification. In this study we performed infection experiments with HBV in primary human hepatocyte cultures. To further elucidate the possible mechanism of virus uptake, infection experiments were performed at different pH levels, after pretreatment of viral particles with acidic buffers and in the presence of lysosomotropic agents (chloroquine and ammonium chloride, respectively). Using a selective PCR technique which discriminates between input virus DNA and the earliest replicative form, we could demonstrate viral replication 36 hr after inoculation. HBV was taken up most efficiently at a pH of 7.4. Infection was still successful after pretreatment of viral particles at low pH and was unaffected by the presence of lysosomotropic agents. In conclusion, this suggests HBV to be a pH-independent virus
Effect of 5′ and 3′ Terminal Sequences, Overall Length, and Coding Capacity on the Accumulation of Defective RNAs Associated with Broad Bean Mottle Bromovirusin Planta

AbstractBroad bean mottle bromovirus (BBMV) was shown to accumulate RNA2-derived defective interfering (DI) RNAs [Romeroet al., Virology194, 576–584 (1993); Poganyet al., Virology212, 574–586 (1995)]. In this work, we utilize three sets of BBMV RNA2-derived artificial DI RNA constructs to determine factors that affect the accumulation of defective RNAsin planta.One set of deletion constructs was used to localize sequences required for efficient accumulation within the 5′ 883 nt and the 3′ 387 nt of the DI RNAs. The second set had a gradually increasing size of 3′ nested deletions to determine the minimal length required for efficient DI RNA accumulation. The smallest DI RNA still accumulating in plants was found to be 1712 nt long. The third set consisted of frameshift mutants which showed that at least 64.4% of BBMV DI RNA sequences must have the 5′ portion of the 2a open reading frame to ensure efficient accumulation. The importance of these factors in the selection of DI RNAs is discussed
Genetic Characterization ofvif, vpr,andvpuSequences from Long-Term Survivors of Human Immunodeficiency Virus Type 1 Infection

AbstractThe identification of HIV-1-infected individuals who remain asymptomatic despite prolonged infection presents a unique opportunity to understand virologic and host factors involved in the pathogenesis of AIDS. We have previously identified 10 long-term survivors (LTS) who are clinically healthy and immunologically normal despite 13 to 15 years of HIV-1 infection. In this study, we examined three accessory genes of HIV-1,vif, vpr,andvpu,in these LTS. A total of 52vif,54vpr,and 55vpunucleotide sequences were obtained from the peripheral blood mononuclear cells of these patients. Analysis of these sequences revealed no gross deletions or insertions. Most of the clones were full-length with an intact open reading frame. Phylogenetic analyses of thevif, vpr,andvpusequences from the LTS suggested that the HIV-1 strains found in the study subjects are not significantly different from those found in patients with AIDS and that the viruses in the LTS are unlikely to share a common genetic origin. Furthermore, a similar degree of overall genetic diversity between viruses from the LTS and AIDS patients suggests that there is unlikely a significant correlation between the degree of genetic diversity and the rate of disease development. Factors other than genetic divergence, such as viral load and phenotype, are likely to impact more on disease status
Protein Composition and Morphology of Human Foamy Virus Intracellular Cores and Extracellular Particles

AbstractCharacterization of human foamy virus (HFV) gag-encoded precursors and the search for a Gag-Pol polyprotein and mature proteins derived from proteolytic processing were carried out in HFV-infected cells and with purified preassembled cores and extracellular virus by Western blotting and radioimmunoprecipitation using antisera against synthetic peptides corresponding to putative Gag and protease proteins. Precursor proteins, Pr78gag/74gagand Pr135pol, were found in the nucleus of epithelial and fibroblast cells 3–4 days after HFV infection. Kinetic analysis of HFV Pr78gagand Pr74gagindicated that Pr78gagis a precursor to Pr74gag. South-Western blot analysis indicated that Pr78gagand Pr74gaghave properties associated with nucleic acid binding protein although they lack the typical zinc-finger motifs found in retroviral nucleocapsid proteins. Western blot analyses of preassembled HFV cores isolated from the cytoplasm of infected cells and purified by sucrose gradient centrifugation demonstrated the presence of Pr78gag/74gagand Pr135pol, but no proteolytically processed Gag proteins were observed. The majority of extracellular HFV particles were found to have pentagon-shaped cores, as observed intracellularly, and are believed to be the immature extracellular form of the virus. The highest concentration of extracellular particles, estimated by EM, Western blot, and reverse transcriptase assays were found in sucrose gradient fractions having a density of 1.21–1.24 g/cm3. Western blot analysis revealed that Pr78gag/74gagand Pr135polwere the major viral proteins associated with these extracellular particles, as only small amounts of putative proteolytically cleaved capsid (p32) were observed. Our results support the notion that Pol is translated independent of Gag in HFV-infected cells
High Concentration of the EBV Latent Membrane Protein 1 in Glycosphingolipid-Rich Complexes from both Epithelial and Lymphoid Cells

AbstractLatent Membrane Protein 1 (LMP1) is an EBV-transforming protein which is detected both in lymphoblastoid cell lines—resulting from EBV-immortalizationin vitro—and in undifferentiated nasopharyngeal carcinoma (NPC), an EBV-associated malignancy of epithelial origin. To better define LMP1 subcellular targets, LMP1 distribution was analyzed in cellular glycosphingolipid-rich complexes (GSL-complexes) derived from epithelial and lymphoid cells. These complexes are obtained by extraction of glycosphingolipid-rich membrane domains (GSL-domains), which are clustering sites for heterotrimeric G-proteins and G-protein-associated receptors. LMP1 concentration was enriched 50-fold in GSL-complexes extracted from a NPC tumor line, C15. High concentrations of LMP1 were also observed in GSL-complexes derived from cultured lymphoid and epithelial cells. These data suggest that association with GSL-domains is an important step in LMP1 trafficking and is probably required for some aspects of its biological activity
Mucosal Immunity Induced by Parenteral Immunization with a Live Attenuated Venezuelan Equine Encephalitis Virus Vaccine Candidate

AbstractInduction of a mucosal immune response is generally thought to require introduction of an immunogen directly onto the mucosal surface. It has been observed, however, that live, attenuated mutants of the alphavirus, Venezuelan equine encephalitis virus (VEE), induce protection from virulent challenge at the respiratory mucosa even after parenteral inoculation. In this report, we propose a mechanism by which subcutaneous immunization with a molecularly cloned, attenuated double mutant of VEE is able to stimulate the production of mucosal anti-VEE IgA. Our results showed that the immunizing virus spread to, and replicated within, lymphoid tissues throughout the mouse. Several tissues known to be inductive sites of the mucosal immune system were found to be positive for the presence of VEE RNA by 48 hr postimmunization. Moreover, this mucosal lymphotropism resulted in the production of virus-specific IgA antibody detectable in vaginal secretions of immunized mice
Apoptosis in Acute and Chronic Central Nervous System Disease Induced by Theiler's Murine Encephalomyelitis Virus

AbstractApoptosis has been observed in neural development and in various neurological diseases, including viral infection and multiple sclerosis. Theiler's murine encephalomyelitis virus is divided into two subgroups based on neurovirulence: the highly neurovirulent GDVII strain produces an acute fatal polioencephalomyelitis in mice, whereas the attenuated DA strain produces demyelination with virus persistence preceded by an acute infection. TUNEL combined with immunocytochemistry was used to detect apoptosis in the central nervous system and to characterize which cell types were involved during the acute stage in both GDVII and DA virus infection and during the chronic stage in DA virus infection. We found that during the acute stage, apoptosis was induced in neurons in both virus infections. However, the number of apoptotic neurons was much greater in GDVII virus-infected mice than in DA virus-infected mice (P< 0.01). During the chronic stage of DA virus infection, apoptotic cells were detected only in the spinal cord white matter. Some of these cells were dual labeled for fragmented DNA and carbonic anhydrase II, an oligodendrocyte marker. Our results indicate that apoptosis of neurons could be responsible for the fatal outcome in GDVII virus infection. In contrast, apoptosis of oligodendrocytes can contribute to the chronic demyelinating DA virus infection
Long-Term Persistent Infection of Domestic Rabbits by the Human Foamy Virus

AbstractHuman foamy virus (HFV) belongs to the spumaretrovirus group of the Retroviridae taxonomic family. Attempts to associate HFV or other foamy viruses to a specific pathology still remain unsuccessful. However, viral gene expression as well as tissue-specific tropism in anin vivocontext remain poorly analyzed. To address this issue, we have infected domestic rabbits with a single dose of HFV and followed them at the biological and molecular levels for 5 years. No apparent pathology was detectable in the infected animals which have developed a strong immunological response against major viral proteins. We found that HFV provirus in blood cells and several organs persisted predominantly in its defective form, ΔHFV, suggesting thatin vivoviral persistence could be related to homologous interference as was recently shownin vitro.This animal model might be useful for studying thein vivotargets of HFV and should also be convenient for testing therapeutic effects of anti-retroviral drugs
Immune Response to Neonatal Genetic Immunization

AbstractThe effect of genetic immunization of neonatal mice was tested with a plasmid vector expressing the rabies virus glycoprotein. Mice inoculated within 24 hr after birth with the plasmid DNA developed antibodies as well as T helper cells to the rabies virus glycoprotein. The response could not be distinguished from that seen upon vaccination of adult mice. Taken together, these data clearly show that the immune system, known to be prone to induction of immunological tolerance to some antigens applied during the early neonatal period, can readily respond to rabies virus glycoprotein induced by a plasmid vector
Characterization of a Chemokine Receptor-Related Gene in Human Herpesvirus 8 and Its Expression in Kaposi's Sarcoma

AbstractHuman herpesvirus 8 (HHV-8) is a recently discovered virus that is highly associated with Kaposi's sarcoma (KS) and AIDS-associated body cavity lymphomas, although it is also found in some normal individuals. HHV-8 is related by nucleotide sequence homology to herpesvirus saimiri (HVS), which causes T cell lymphomas in some New World monkeys, and to Epstein–Barr virus (EBV), a human herpesvirus linked etiologically with Burkitt's lymphoma and nasopharyngeal carcinoma. We report that, like HVS but unlike EBV, HHV-8 contains a gene (ORF74) with significant sequence homology to the high-affinity IL-8 receptor, a member of the α (CXC) chemokine receptor family of transmembrane G protein-coupled receptors. We also show by reverse transcription PCR that the chemokine receptor-related HHV-8 gene is detectable in some RNA samples from KS tissue, and that its expression varies independently from that of ORF26, a minor capsid protein. The presence of a potential chemokine receptor in HHV-8 and its expression in KS tissue suggests that it may be important in the regulation of viral gene expression and may play a role in the etiology of KS and AIDS-related body cavity lymphomas
A Full-Length and Replication-Competent Proviral Clone of SIVAGMfrom Tantalus Monkeys

AbstractAfrican green monkeys (AGM) are classified into four distinct species (commonly termed vervet, grivet, sabaeus, and tantalus monkeys), all of which are known to be infected with simian immunodeficiency virus (SIVAGM) in the wild. Sequence analysis of partialgagandenvregions has indicated that each of the four species harbors a phylogenetically distinct SIVAGMsubtype. This species-specific diversity suggests that African green monkeys have been infected with SIVAGMfor an extended period of time, possibly even before their speciation from a common ancestor. However, our understanding of the evolutionary history of this group of viruses is still incomplete, in part because sequence information for most isolates is limited to small subgenomic regions. There are only six SIVAGMproviruses which have been sequenced in their entirety, and these represent only three of the four SIVAGMlineages (i.e., SIVAGMgri, SIVAGMver, and SIVAGMsab). In this paper, we have generated the first full-length proviral clone for SIVAGMinfecting tantalus monkeys (SIVAGMtan). Lambda phage techniques were employed to clone this provirus (TAN) as a single genomic unit from productively infected Molt 4 (clone 8) cells, and sequence analysis confirmed the integrity of all major open reading frames, exceptvprwhich contained an in-frame stop codon. The proviral clone was also biologically active since transfection yielded replication-competent virions. Amino acid sequence comparisons of all major viral proteins indicated that TAN was roughly equidistant from previously characterized sabaeus, grivet, and vervet strains, thus confirming that it represents a fourth independent SIVAGMlineage. Given the need for well-characterized reference reagents, this full-length tantalus provirus should facilitate future studies of SIVAGMmolecular biology and evolution
Mouse Mammary Tumor Virus Superantigens Require N-Linked Glycosylation for Effective Presentation to T Cells

AbstractMouse mammary tumor viruses (MMTVs) encode superantigens that associate with major histocompatibility complex class II products on antigen-presenting cells and stimulate T cells in a Vβ-specific manner. This T cell activation is critical for completion of the viral life cycle and vertical transmission to the next generation. To investigate the functional significance of extensive viral superantigen (Sag) glycosylation, we disrupted the six potential sites for N-linked carbohydrate addition in the Sag encoded by proviral integrantMtv-1.Shifts in the apparent molecular mass of these mutant glycoproteins suggested that wild-typeMtv-1Sag is glycosylated on four of its six sites. Intracellular and cell surface staining of the panel of mutants indicated that any decrease in glycosylation resulted in reduced levels of intracellular protein and undetectable surface expression, suggesting that decreased glycosylation leads to rapid Sag degradation and abates trafficking to the plasma membrane. Nevertheless, several mutants with intermediate levels of glycosylation expressed enough Sag on the B cell surface to potently stimulate reactive T cell hybrids. We show there is no specific site bearing N-linked glycosylation that is essential for activity, but at least one carbohydrate addition is necessary for effective B cell presentation of MMTV superantigens to T cells
Close Phylogenetic Relationship between Egg Drop Syndrome Virus, Bovine Adenovirus Serotype 7, and Ovine Adenovirus Strain 287

AbstractA cloned egg drop syndrome (EDS) virus genomic DNA fragment containing the protease gene has been identified and the complete nucleotide sequence of the protease and partial nucleotide sequence of the hexon genes has been determined. Phylogenetic analysis of the protease gene has revealed EDS virus to be genetically more closely related to bovine adenovirus type 7 (BAV-7) and ovine adenovirus isolate 287 (OAV287) than either of these two viruses are to other members of the genusMastadenovirusor EDS virus is to an other member of theAviadenovirusgenus. The three viruses share further similarities in that they have a high percentage AT content in their genome and are characterized by having more compact genomes than other adenoviruses. The protease gene from all three viruses contained the active site residues (H55–D72–C122 triad) and C104 (providing a disulfide bond to cofactor pVIc). However, P137, found in all other members of theMastadenovirusgenus, and thought to be involved in trafficking, was missing from the protease of the EDS virus, BAV-7, and OAV287. These results suggest that EDS virus should be classified along with BAV-7 and OAV287 in a separate taxon
Neutralization Susceptibility of African Swine Fever Virus Is Dependent on the Phospholipid Composition of Viral Particles

AbstractIn this study we have investigated the generation of African swine fever (ASF) virus variants resistant to neutralizing antibodies after cell culture propagation. All highly passaged ASF viruses analyzed were resistant to neutralization by antisera from convalescent pigs or antibodies generated against individual viral proteins which neutralized low-passage viruses. A molecular analysis of neutralizable and nonneutralizable virus isolates by sequencing of the genes encoding for neutralizing proteins revealed that the absence of neutralization of high-passage viruses is not due to antigenic variability of critical epitopes. A comparative analysis of phospholipid composition of viral membranes between low- and high-passage viruses revealed differences in the relative amount of phosphatidylinositol in these two groups of viruses, independent of the cells in which the viruses were grown. Further purification of low- and high-passage viruses by Percoll sedimentation showed differences in the phospholipid composition identical to those found with the partially purified viruses and confirmed the susceptibility of these viruses to neutralization. The incorporation of phosphatidylinositol into membranes of high-passage viruses rendered a similar neutralization susceptibility to low-passage viruses, in which this is a major phospholipid. In contrast, other phospholipids did not interfere with high-passage virus neutralization, suggesting that phosphatidylinositol is essential for a correct epitope presentation to neutralizing antibodies. Additionally, the removal of phosphatidylinositol from a low-passage virus by a specific lipase transformed this virus from neutralizable to nonneutralizable. These data constitute clear evidence of the importance of the lipid composition of the viral membranes for the protein recognition by antibodies and may account in part for the past difficulties in reproducibly demonstrating ASF virus-neutralizing antibodies by using high-passage viruses