pinecone/core-2020-05-10-deduplication huggingface dataset:
Inflammatory Cytokines and the Reactivation of Kaposi's Sarcoma-Associated Herpesvirus Lytic Replication

AbstractKaposi's sarcoma (KS) is a complex proliferative lesion long suspected of being dependent on exogenous paracrine signaling molecules to stimulate its proliferative, angiogenic, and inflammatory components. In particular, both clinical and experimental observations have pointed to a potential role for inflammatory cytokines as permissive factors for KS development, but KS pathogenesis is also critically dependent on infection by an exogenous herpesvirus, the KS-associated herpesvirus (KSHV). To examine the possible links between inflammatory cytokines and KSHV replication, we tested for the ability of such cytokines to induce lytic viral reactivation in the latently infected BCBL-1 cell line. Interferon-γ consistently activated KSHV replication, whereas tumor necrosis factor, interleukin-1, interleukin-2, interleukin-6, granulocyte-macrophage colony stimulating factor, and basic fibroblast growth factor did not. Glucocorticoids also failed to induce lytic KSHV growth in these cells, but ionomycin, a calcium ionophore, induced replication and strongly augmented the known inductive effects of phorbol esters. Interferon-α had a dose-dependent inhibitory effect on KSHV induction by ionomycin. The identification of interferon-γ as an activator and interferon-α as an inhibitor of KSHV induction in vitro correlates well with in vivo observations and demonstrates for the first time that inflammatory cytokines can directly modulate KSHV replication
Simian Immunodeficiency Virus-Specific Cytotoxic T Lymphocytes and Protection Against Challenge in Rhesus Macaques Immunized with a Live Attenuated Simian Immunodeficiency Virus Vaccine

AbstractIn this study, we examined the role of simian immunodeficiency virus (SIV)-specific cytotoxic T lymphocytes (CTLs) in macaques immunized with an attenuated strain of simian immunodeficiency virus (SIVmac239Δnef) in protection against pathogenic challenge with SIVmac251. Our results indicate that attenuated SIVmac239Δnef can elicit specific CTL precursor cells (CTLp), but no correlation was observed between breadth or strength of CTLp response to structural proteins SIV-Env, -Gamg or -Pol (as measured by limiting dilution assay) and protection against infection. In one animal, we longitudinally followed the SIV-Gag-specific response to an MHC class I Mamu-A*01-restricted epitope p11C, C-M using a tetrameric MHC/peptide complex reagent. A low frequency of SIV p11C, C-M peptide-specific tetramer-reactive cells was present at the time of challenge but could be expanded in vitro. Surprisingly, the low level of Mamu-A*01/p11C, C-M-specific CTLs induced through attenuated SIVmac239Δnef vaccination increased in the absence of detectable SIVmac251 or SIVmac239Δnef proviral DNA. Overall, our results suggest that protection against infection in this model can be achieved through more than one mechanism, with SIV-specific CTLs being important in controlling SIVmac239Δnef viral replication postchallenge
Replication Rate of Feline Immunodeficiency Virus in Astrocytes Is Envelope Dependent: Implications for Glutamate Uptake

AbstractFeline immunodeficiency virus (FIV) induces neurological abnormalities in domestic cats. Previously, we demonstrated that two disparate strains of FIV (FIV-34TF10 and FIV-PPR) varied greatly in the ability to replicate in feline cortical astrocytes. To investigate the impact of the env region on the replication efficiency of these strains, we constructed two env chimera viruses, FIV-34TF10-PPRenv and FIV-PPR-34TF10env, to infect feline cortical astrocytes in vitro. Although all of these viruses infected cortical astrocytes, the efficiency of replication depended on strain, and the env region played an essential role. The viruses containing the env of 34TF10, FIV-34TF10, and FIV-PPR-34TF10env had the greatest replication rate, whereas the viruses containing the env of PPR replicated at a lower level. Other viral regions had modulatory effects on the replication rate, with the FIV-PPR genome providing a slight replication advantage over the FIV-34TF10 genome. We also monitored the effects of these viruses on an important astrocyte function, glutamate uptake; all viruses significantly decreased this activity, but only the viruses containing the env of PPR significantly impaired glutamate uptake without altering the culture viability. These results may be particularly relevant in the context of lentivirus-induced central nervous system disease in which a selective breakdown of astroglial function may contribute to neurodegeneration
African Swine Fever Virus EP153R Open Reading Frame Encodes a Glycoprotein Involved in the Hemadsorption of Infected Cells

AbstractThe open reading frame EP153R, located within the EcoRI E′ fragment of the African swine fever (ASF) virus genome, is predicted to encode a membrane protein of 153 amino acids that presents significant homology to the N-terminal region of several CD44 molecules. EP153R contains multiple putative sites for N-glycosylation, phosphorylation, and myristoylation, a central transmembrane region, a C-type animal lectin-like domain, and a cell attachment sequence. Transcription of EP153R takes place at both early and late times during the virus infection. The disruption of the gene, achieved by insertion of the marker gene LacZ within EP153R, did not change either the in vitro virus growth rate or the virus-sensitive/resistant condition of up to 17 established cell lines, but abrogated the hemadsorption phenomenon induced in ASF virus-infected cells. As the sequence and expression of the ASF virus protein pEP402R, a CD2 homolog responsible for the adhesion of erythrocytes to susceptible cells, was unaffected in cultures infected with the EP153R deletion mutant, we conclude that the gene EP153R is needed to induce and/or maintain the interaction between the viral CD2 homolog and its corresponding cell receptor
Identification of a Novel Expressed Open Reading Frame Situated between Genes UL20 and UL21 of the Herpes Simplex Virus 1 Genome

AbstractAn open reading frame (ORF) situated between the UL20 and UL21 genes encodes a protein designated as UL20.5. The UL20.5 ORF lies 5′ and in the same orientation as the UL20 ORF. The expression of the UL20.5 ORF was verified by RNase protection assays and by in-frame insertion of an amino acid sequence encoding an epitope of an available monoclonal antibody. The tagged UL20.5 protein colocalized in small dense nuclear structures with products of the α22/US1.5, UL3, and UL4 genes. Expression of the UL20.5 gene was blocked in cells infected and maintained in the presence of phosphonoacetate, indicating that it belongs to the late, or γ2, kinetic class. UL20.5 is not essential for viral replication inasmuch as a recombinant virus made by insertion of the thymidine kinase gene into the UL20.5 ORF replicates in all cell lines tested [J. D. Baines, P. L. Ward, G. Campadelli-Fiume, and B. Roizman (1991) J. Virol. 65, 6414–6424]. The genomic location of the recently discovered genes illustrates the compact nature of the viral genome
Patterned Entry and Egress by Epstein–Barr Virus in Polarized CR2-Positive Epithelial Cells

AbstractIn polarized epithelium direction of viral entry and release correlates with proclivity of a virus to establish local versus systemic infection. The Epstein–Barr virus (EBV), whose principal tissue reservoir is B lymphocytes, also has disease manifestations in epithelium, suggesting intertissue spread potentially influenced by epithelial cell polarity. We stably transfected the B lymphocyte EBV receptor (CR2/CD21) into Madin–Darby canine kidney (MDCK) epithelial cells used extensively to study effects of cell polarity on infection by both DNA and RNA viruses. CR2/CD21 was detected on both apical and basolateral surfaces of polarized MDCK cells, with predominant expression basolaterally. However, infectivity was up to four-fold greater apically, suggesting that endogenous cell surface molecules, sorted asymmetrically onto polarized plasma membranes, may be involved in EBV entry into MDCK cells. EBV gp350/220, a replicative cycle glycoprotein added to the virus envelope on egress through the cell membrane, was immunolocalized by confocal microscopy to basolateral cell surfaces only. Apical entry of EBV with subsequent basolateral release of newly replicated virus favors systemic infection by viral dissemination to underlying lymphocytic aggregations. Under conditions of long-term culture, latent EBV was not stably maintained in these cells, suggesting that the epithelial phase of acute EBV infection may be transient
HPV11 Mutant Virus-like Particles Elicit Immune Responses That Neutralize Virus and Delineate a Novel Neutralizing Domain

AbstractCharacterization of the regions of human papillomaviruses (HPVs) that elicit neutralizing immune responses supports studies on viral infectivity and provides insight for the development and evaluation of prophylactic vaccines. HPV11 is a major etiologic agent of genital warts and a likely vaccine candidate. A conformationally dependent epitope for the binding of three neutralizing monoclonal antibodies (mAbs) has been mapped to residues G131T132 of the L1 major capsid protein. The mAbs bind L1 only when it is assembled into virions or into virus-like particles (VLPs) that mimic the capsid structure. We were interested in identifying other domains of L1 that elicit neutralizing responses. To this end, we have generated a panel of mAbs against VLPs derived from HPV11 L1 harboring a G131S substitution. The new mAbs are unlike the neutralizing mAbs previously mapped to residues G131T132 in that they bind both prototype and HPV11:G131S mutant VLPs. Some of the new mAbs neutralized virus in vitro. We have mapped epitopes for three of these new mAbs, as well as a neutralizing mAb generated against HPV11 virions, by measuring binding to HPV6 VLPs substituted with HPV11-like amino acids. Two regions are critical: one defined by HPV11 L1 residues 263–290 and the other by residues 346–349. mAbs H11.H3 and H11.G131S.G3 bind HPV6 VLPs with substitutions derived from the 346–349 region; in addition, H11.G131S.G3 binds HPV6 VLPs with substitutions derived only from the 263–290 region. Although H11.H3 does not bind HPV6 VLPs with substitutions derived from the 263–290 region, binding to HPV6 VLPs is enhanced when both sets of substitutions are present. mAbs H11.G131S.I1 and H11.G131S.K5 bind HPV6 VLPs with the 263–290 substitutions, but show little binding to HPV6 VLPs with the 346–349 substitutions. However, binding to HPV6 VLPs is enhanced when substitutions at both regions are present. The 346–349 region has not previously been described as eliciting a neutralizing response for any HPV type. In addition, the work demonstrates a complex binding site contributed by two distinct regions of L1
Cell Growth Regulatory and Antiviral Effects of the P69 Isozyme of 2−5 (A) Synthetase

AbstractAmong the many interferon-induced proteins that carry out multiple cellular functions of interferons is the family of enzymes called 2′-5′ oligoadenylate synthetases. We examined the anticellular and antiviral activities of a specific member of that family, the P69 isozyme. P69 was expressed in human cells by transfection and shown to be localized primarily in the endoplasmic reticulum. For further studies, permanent cell lines expressing different levels of P69 or an enzymatically inactive mutant were isolated. Constitutive P69 expression caused inhibition of replication of encephalomyocarditis virus but not of vesicular stomatitis virus, Sendai virus, or reovirus. Increasing levels of P69 expression also caused increasing perturbations in cell growth properties. There was increasing accumulations of the P69-expressing cells in the G1 phase of the cell cycle; cell-doubling time was increased by P69 expression; and there were many multinucleated cells in the P69-expressing line, indicating a defect in cytokinesis
Alastrim Smallpox Variola Minor Virus Genome DNA Sequences

AbstractAlastrim variola minor virus, which causes mild smallpox, was first recognized in Florida and South America in the late 19th century. Genome linear double-stranded DNA sequences (186,986 bp) of the alastrim virus Garcia-1966, a laboratory reference strain from an outbreak associated with 0.8% case fatalities in Brazil in 1966, were determined except for a 530-bp fragment of hairpin-loop sequences at each terminus. The DNA sequences (EMBL Accession No. Y16780) showed 206 potential open reading frames for proteins containing ≥60 amino acids. The amino acid sequences of the putative proteins were compared with those reported for vaccinia virus strain Copenhagen and the Asian variola major strains India-1967 and Bangladesh-1975. About one-third of the alastrim viral proteins were 100% identical to correlates in the variola major strains and the remainder were ≥95% identical. Compared with variola major virus DNA, alastrim virus DNA has additional segments of 898 and 627 bp, respectively, within the left and right terminal regions. The former segment aligns well with sequences in other orthopoxviruses, particularly cowpox and vaccinia viruses, and the latter is apparently alastrim-specific
Mutational Analysis of the Cowpea Mosaic Virus Movement Protein

AbstractCowpea mosaic virus moves from cell-to-cell in a virion form through tubular structures that are assembled in modified plasmodesmata. Similar tubular structures are formed on the surface of protoplasts inoculated with cowpea mosaic virus. The RNA 2-encoded movement protein (MP) is responsible for the induction and formation of these structures. To define functional domains of the MP, an alanine-substitution mutagenesis was performed on eight positions in the MP, including two conserved sequence motifs, the LPL and D motifs. Results show that these two conserved motifs as well as the central region of the MP are essential for cell-to-cell movement. Several viruses carrying mutations in the N- or C-terminal parts of their MP retained infectivity on cowpea plants. Coexpression studies revealed that mutant MPs did not interfere with the activity of wild-type MP and could not mutually complement their defects
Identification of Two Major Virion Protein Genes of White Spot Syndrome Virus of Shrimp

AbstractWhite Spot Syndrome Virus (WSSV) is an invertebrate virus, causing considerable mortality in shrimp. Two structural proteins of WSSV were identified. WSSV virions are enveloped nucleocapsids with a bacilliform morphology with an approximate size of 275 × 120 nm, and a tail-like extension at one end. The double-stranded viral DNA has an approximate size 290 kb. WSSV virions, isolated from infected shrimps, contained four major proteins: 28 kDa (VP28), 26 kDa (VP26), 24 kDa (VP24), and 19 kDa (VP19) in size, respectively. VP26 and VP24 were found associated with nucleocapsids; the others were associated with the envelope. N-terminal amino acid sequences of nucleocapsid protein VP26 and the envelope protein VP28 were obtained by protein sequencing and used to identify the respective genes (vp26 and vp28) in the WSSV genome. To confirm that the open reading frames of WSSV vp26 (612) and vp28 (612) are coding for the putative major virion proteins, they were expressed in insect cells using baculovirus vectors and analyzed by Western analysis. A polyclonal antiserum against total WSSV virions confirmed the virion origin of VP26 and VP28. Both proteins contained a putative transmembrane domain at their N terminus and many putative N- and O-glycosylation sites. These major viral proteins showed no homology to baculovirus structural proteins, suggesting, together with the lack of DNA sequence homology to other viruses, that WSSV may be a representative of a new virus family, Whispoviridae
Mutagenesis of Murine Cytomegalovirus Using a Tn3-Based Transposon

AbstractA transposon derived from Escherichia coli Tn3 was introduced into the genome of murine cytomegalovirus (MCMV) to generate a pool of viral mutants. We analyzed three of the constructed recombinant viruses that contained the transposon within the M25, M27, and m155 open reading frames. Our studies provide the first direct evidence to suggest that M25 and M27 are not essential for viral replication in mouse NIH 3T3 cells. Studies in cultured cells and Balb/c mice indicated that the transposon insertion is stable during viral propagation both in vitro and in vivo. Moreover the virus that contained the insertion mutation in M25 exhibited a titer similar to that of the wild-type virus in the salivary glands, lungs, livers, spleens, and kidneys of the Balb/c mice that were intraperitoneally infected with these viruses. These results suggest that M25 is dispensable for viral growth in these organs and the presence of the transposon sequence in the viral genome does not significantly affect viral replication in vivo. The Tn3-based system can be used as a mutagenesis approach for studying the function of MCMV genes in both tissue culture and in animals
Characterization of the UL33 Gene Product of Herpes Simplex Virus 1

AbstractThe UL33 protein is one of six genes (including UL6, UL15, UL17, UL28, and UL32) required for cleavage of viral concatemeric DNA into unit-length genomes and packaging of the virus genomes into preformed capsids. The UL25 gene product is dispensable for cleavage of viral DNA but essential for packaging of DNA into capsids. A polyclonal antiserum was produced against an affinity-purified protein containing the full-length UL33 gene product of herpes simplex virus 1 fused to glutathione-S-transferase. A protein of approximate Mr 19,000 that reacted with the antiserum was detected in immunoblots of herpes simplex virus 1-infected cellular lysates. This protein was not detected in lysates of mock-infected cells or cells infected with a mutant virus containing a stop codon in UL33, indicating that the 19,000 Mr protein is the product of the UL33 open reading frame. The UL33 gene product was not detected in purified virions or capsids. Accumulation of the UL33 protein to detectable levels required viral DNA synthesis, indicating that the protein was regulated as a late gene. Indirect immunofluorescence analysis demonstrated that UL33 protein accumulated predominantly within replication compartments in the central domains of infected cell nuclei and within the cytoplasm. Localization of the UL33 gene product in replication compartments was maintained in cells infected with a variety of cleavage/packaging mutants
Incorporation of Simian Virus 5 Fusion Protein into Murine Leukemia Virus Particles and Its Effect on the Co-incorporation of Retroviral Envelope Glycoproteins

AbstractWe describe the generation of murine leukemia virus (MLV) virus particles carrying the paramyxovirus fusion protein F from simian virus 5 (SV5-F). This glycoprotein was expressed in cells providing Moloney MLV (MoMLV) Gag and Pol proteins and a lacZ retroviral vector. SV5-F was correctly expressed, processed, and efficiently incorporated into retroviral particles. SV5-F-bearing retroviruses were not infectious although a weak binding to primate and rodent cells could be detected and SV5-F could mediate cell to cell fusion. We then co-expressed the SV5-F glycoprotein in retroviral particles with chimeric and wild-type MoMLV envelope glycoproteins. Our results show that F strongly inhibited infection via the retroviral envelopes although the mechanism of inhibition was different depending on the retroviral envelope used
Oral DNA Vaccination Promotes Mucosal and Systemic Immune Responses to HIV Envelope Glycoprotein

AbstractIn this report, we described induction of HIV envelope (env)-specific systemic and mucosal immune responses by oral vaccination of BALB/c mice with env-encoded plasmid DNA encapsulated in poly(dl-lactide-co-glycolide) (PLG) microparticles. We demonstrated that intragastric administration of the encapsulated plasmid DNA resulted in transduced expression of the env glycoprotein in the intestinal epithelium. Mice immunized orally exhibited env-specific type 1 and cytotoxic T lymphocyte (CTL) responses in spleen and the inductive (Peyer's patches) and effector (lamina propria) mucosal tissues of gut. Oral administration of PLG-encapsulated plasmid DNA encoding gp160 also induced env-specific serum antibodies, and an increased level of IgA directed to gp160 was detected in fecal washes of the immunized mice. In contrast, intramuscular (i.m.) administration of naked or PLG-encapsulated DNA vaccine induced only systemic cellular and humoral responses to the env glycoprotein. Using an HIV env-expressing recombinant vaccinia viral intrarectal murine challenge system, we observed higher resistance to mucosal viral transmission in mice immunized orally than in animals injected i.m. with PLG-encapsulated plasmid DNA encoding gp160. Results of these studies demonstrate the feasibility of using orally delivered PLG microparticles containing plasmid DNA-encoded HIV gp160 for induction of env-specific systemic and mucosal immune responses and protection against recombinant HIV env vaccinia virus challenge
The N-Terminal Matrix Domain of HIV-1 Gag Is Sufficient but Not Necessary for Viral Protein U-Mediated Enhancement of Particle Release through a Membrane-Targeting Mechanism

AbstractViral protein U (Vpu) is an 81 amino acid phosphoprotein found in human immunodeficiency virus type 1 (HIV-1)-infected cells. One function of Vpu is to enhance the release of virus particles from the plasma membrane in infected cells. Using subcellular fractionation, we observed that Vpu promotes the targeting of Pr55 Gag to the plasma membrane, the site of viral assembly. Deletions of Pr55, which removed most of the N-terminal matrix domain (p39) or the C-terminal domains of nucleocapsid and p6 (p41), still allowed for virus-like particle production. Moreover, the release of these particles remained Vpu-responsive. The N-terminal matrix (MA) domain of Gag, which contains its membrane-binding domain, is sufficient for Vpu-mediated enhanced release into the supernatant. Furthermore, a MA-GFP fusion protein showed enhanced membrane binding in the presence of Vpu. This demonstrates that Vpu action may be mediated by allowing Gag, specifically the N-terminal matrix domain, to efficiently associate with the plasma membrane. Thus MA appears sufficient but not necessary for Vpu-mediated enhanced particle release
Effects of Oligomerization on the Epitopes of the Human Immunodeficiency Virus Type 1 Envelope Glycoproteins

AbstractTo understand the differential expression of epitopes on monomeric and oligomeric forms of the envelope glycoproteins, nine human monoclonal antibodies (mAbs) were derived from the cells of human immunodeficiency virus-infected subjects by selection with soluble oligomeric gp140 (o.140). These nine mAbs and 12 human mAbs selected with V3 peptides, viral lysates, and rgp120, specific for the V2, V3, C5, CD4-binding domain (CD4bd), and gp41, were tested in a binding assay to compare the exposure of these regions on monomeric gp120 or gp41 and on o.140. None of the 21 mAbs were oligomer specific. However, mAbs to V3 and CD4bd were “oligomer sensitive,” whereas mAbs to V2 and the distal epitope of C5 tended to be “monomer sensitive” (i.e., to react better with the oligomer or monomer, respectively). The majority of anti-gp41 mAbs reacted similarly with monomer and oligomer. Although the uncleaved o.140 used in this study differs from the cleaved gp120/41 oligomer found on the native virus particle, these results suggest that new epitopes are not introduced by oligomerization of viral envelope proteins, that such oligomer-specific epitopes, if they exist, are not highly immunogenic, and/or that they are not efficiently selected using soluble o.140