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See detailA Multipotential Beta -1,6-N-Acetylglucosaminyl-Transferase Is Encoded by Bovine Herpesvirus Type 4
Vanderplasschen, Alain ULg; Markine-Goriaynoff, N.; Lomonte, P. et al

in Proceedings of the National Academy of Sciences of the United States of America (2000), 97(11), 5756-5761

The beta-1,6-N-acetylglucosaminyltransferase (beta1,6GnT) gene family encodes enzymes playing crucial roles in glycan synthesis. Important changes in beta1,6GnT expression are observed during development ... [more ▼]

The beta-1,6-N-acetylglucosaminyltransferase (beta1,6GnT) gene family encodes enzymes playing crucial roles in glycan synthesis. Important changes in beta1,6GnT expression are observed during development, oncogenesis, and immunodeficiency. The most characterized beta1,6GnTs in this gene family are the human (h) C2GnT-L and h-IGnT, which have core 2 [Galbeta1-->3(GlcNAcbeta1-->6)GalNAc] and I branching [GlcNAcbeta1-->3(GlcNAcbeta1-->6)Gal] activities, respectively. Recently, h-C2GnT-M was shown to be unique in forming core 2, core 4 [GlcNAcbeta1-->3(GlcNAcbeta1-->6)GalNAc], and I structures. To date, the beta1,6GnT gene family has been characterized only in mammals. Here, we describe that bovine herpesvirus type 4 (BHV-4) encodes a beta1,6GnT expressed during viral replication and exhibiting all of the core 2, core 4, and I branching activities. Sequencing of the BHV-4 genome revealed an ORF, hereafter called BORFF3-4, encoding a protein (pBORFF3-4) exhibiting 81.1%, 50.7%, and 36.6% amino acid identity with h-C2GnT-M, h-C2GnT-L, and h-IGnT, respectively. Reverse transcriptase-PCR analysis revealed that BORFF3-4 is expressed during BHV-4 replication. Expression of BORFF3-4 in Chinese hamster ovary cells directed the expression of core 2 branched oligosaccharides and I antigenic structures on the cell surface. Moreover, a soluble form of pBORFF3-4 had core 4 branching activity in addition to core 2 and I branching activities. Finally, infection of a C2GnT-negative cell line with BHV-4 induced expression of core 2 branched oligosaccharides. This study extends the beta1,6GnT gene family to a viral gene and provides a model to study the biological functions of a beta1,6GnT in the context of viral infection. [less ▲]

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See detailL'herpèsvirus bovin de type 4 : virus pathogène ou passager?
Thiry, Etienne ULg; Markine-Goriaynoff, Nicolas; Minner, Frédéric et al

in Point Vétérinaire (2000), 31

Bovine herpesvirus type 4 (BHV-4) is a ubiquitous herpesvirus in cattle. It has been isolated from animals showing a wide variety of clinical signs but few of the isolates have proven experimental ... [more ▼]

Bovine herpesvirus type 4 (BHV-4) is a ubiquitous herpesvirus in cattle. It has been isolated from animals showing a wide variety of clinical signs but few of the isolates have proven experimental pathogenicity. Species that are susceptible to BHV-4 include animals other than ruminants, notably cats and, surprisingly, a primate the owl monkey (Aotus trivirgatus). BHV-4 exists as a latent infection in mononuclear cells. In cattle, BHV-4 has been isolated in animals exhibiting ocular and respiratory conditions and it has been found in females with genital tract conditions such as post-partum metritis and vulvovaginitis. There is some epidemiological and experimental evidence that BHV-4 may be a cause of bovine abortion. There is no vaccine available in Europe and prevention is exclusively by hygiene measures. [less ▲]

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See detailSusceptibility of Bovine Antigen-Presenting Cells to Infection by Bovine Herpesvirus 1 and In Vitro Presentation to T Cells: Two Independent Events
Renjifo, Xiména; Letellier, Carine; Keil, Günther M. et al

in Journal of Virology (1999), 73

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See detailVaccinia virus intracellular mature virions contain only one lipid membrane
Hollinshead, M.; Vanderplasschen, Alain ULg; Smith, G. L. et al

in Journal of Virology (1999), 73

Vaccinia virus (VV) morphogenesis commences with the formation of lipid crescents that grow into spherical immature virus (IV) and then infectious intracellular mature virus (IMV) particles. Early studies ... [more ▼]

Vaccinia virus (VV) morphogenesis commences with the formation of lipid crescents that grow into spherical immature virus (IV) and then infectious intracellular mature virus (IMV) particles. Early studies proposed that the lipid crescents were synthesized de novo and matured into IMV particles that contained a single lipid bilayer (S. Dales and E. H. Mosbach, Virology 35:564-583, 1968), but a more recent study reported that the lipid crescent was derived from membranes of the intermediate compartment (IC) and contained a double lipid bilayer (B. Sodiek et al., J. Cell Biol. 121:521-541, 1993). In the present study, we used high-resolution electron microscopy to reinvestigate the structures of the lipid crescents, IV, and IMV particles in order to determine if they contain one or two membranes. Examination of thin sections of Epon-embedded, VV-infected cells by use of a high-angular-tilt series of single sections, serial-section analysis, and high-resolution digital-image analysis detected only a single, 5-nm-thick lipid bilayer in virus crescents, IV, and IMV particles that is covered by a 8-nm-thick protein coat. In contrast, it was possible to discern tightly apposed cellular membranes, each 5 nm thick, in junctions between cells and in the myelin sheath of Schwann cells around neurons. Serial-section analysis and angular tilt analysis of sections detected no continuity between virus lipid crescents or IV particles and cellular membrane cisternae. Moreover, crescents were found to form at sites remote from IC membranesnamely, within the center of virus factories and within the nucleusdemonstrating that crescent formation can occur independently of IC membranes. These data leave unexplained the mechanism of single-membrane formation, but they have important implications with regard to the mechanism of entry of IMV and extracellular enveloped virus into cells; topologically, a one-to-one membrane fusion suffices for delivery of the IMV core into the cytoplasm. Consistent with this, we have demonstrated previously by confocal microscopy that uncoated virus cores within the cytoplasm lack the IMV surface protein D8L, and we show here that intracellular cores lack the surface protein coat and lipid membrane. [less ▲]

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See detailBovine herpesvirus 1-induced apoptotic cell death: role of glycoprotein D
Hanon, Emmanuel; Keil, Günther; Van Drunen Littel-Van Den Hurk, Sylvia et al

in Virology (1999), 257

Bovine herpesvirus 1 (BHV-1) induces apoptotic cell death in peripheral blood mononuclear cells and in bovine B lymphoma (BL-3) cells. Attachment but not penetration of BHV-1 is necessary to induce ... [more ▼]

Bovine herpesvirus 1 (BHV-1) induces apoptotic cell death in peripheral blood mononuclear cells and in bovine B lymphoma (BL-3) cells. Attachment but not penetration of BHV-1 is necessary to induce apoptosis in target cells, suggesting that one or more BHV-1 envelope glycoproteins could be involved in the activation of the apoptotic process. In the present study, we demonstrate that, although BHV-1 virions devoid of glycoprotein D (BHV-1 gD-/-) still bind to BL-3 cells, they are no longer able to induce apoptosis. In contrast, virions that contain glycoprotein D (gD) in the viral envelope but do not genetically encode gD (BHV-1 gD-/+) induce a level of apoptosis comparable to that produced by wild-type (wt) BHV-1. In addition, monoclonal antibodies directed against gD, but not against gB or gC, strongly reduced the high levels of apoptosis induced by BHV-1. These observations demonstrate that the induction of apoptosis is directly due to BHV-1 viral particles harboring gD in the viral envelope. Interestingly, binding of affinity-purified gD to BL-3 cells did not induce apoptosis but inhibited the ability of wt BHV-1 to induce apoptosis. Altogether, these results provide evidence for the direct or indirect involvement of gD in the mechanism by which BHV-1 induces apoptosis. [less ▲]

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See detailComparison of intracellular and extracellular particles of the vaccinia virus
Vanderplasschen, Alain ULg

in Bulletin et Mémoires de l'Académie Royale de Médecine de Belgique (1999), 154

The construction of recombinant vaccinia viruses (VV) is definitely a promising approach to develop vaccines for pets. To produce safer VV recombinant vaccines, it is necessary to understand the ... [more ▼]

The construction of recombinant vaccinia viruses (VV) is definitely a promising approach to develop vaccines for pets. To produce safer VV recombinant vaccines, it is necessary to understand the pathogenesis of VV infection. The goal of this study was to compare the biological properties of the extracellular and intracellular infectious particles of VV. Four biological properties were investigated: (i) the binding into cellular receptor(s); (ii) the mechanism of entry; (iii) the resistance to neutralisation by antibodies; (iv) the resistance to complement lysis. The results obtained in this study emphasize the differences existing between the two infectious forms of VV and demonstrate the adaptation of the extracellular particle to its role of virus dissemination in vivo. The results of this study implicate that VV strains used as expression vector in vaccinology should be deleted for genes required for EEV morphogenesis. [less ▲]

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See detailIn vitro study of the interactions between bovine herpesvirus 4 and the bovine host cells
Vanderplasschen, Alain ULg

in Bulletin et Mémoires de l'Académie Royale de Médecine de Belgique (1999), 154

This work was devoted to the study of the interactions between bovine herpesvirus 4 (BHV-4) and bovine cells in vitro. It led to the discovery of two interesting properties of BVH-4 replication cycle ... [more ▼]

This work was devoted to the study of the interactions between bovine herpesvirus 4 (BHV-4) and bovine cells in vitro. It led to the discovery of two interesting properties of BVH-4 replication cycle: first, the cellular receptor heparan sulfate was proven to mediate BVH-4 binding to target cells. This is the first description of the implication of heparan sulfate in the binding process of a gammaherpesvirus. Second, using synchronised cells, the replication of BVH-4 DNA was proven to be dependent on the S phase of the cell cycle. This dependence could explain some properties of BVH-4 infection in vitro and could play an important role in the biology of the infection in vivo. Finally, in order to produce monoclonal antibodies against BVH-4 IE1 and IE2 proteins, the genes coding for these proteins were cloned and expressed in prokaryotic cells. [less ▲]

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See detailUsing Confocal Microscopy to Study Virus Binding and Entry into Cells
Vanderplasschen, Alain ULg; Smith, G. L.

in Methods in Enzymology (1999), 307

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See detailAttachment but Not Penetration of Bovine Herpesvirus 1 Is Necessary to Induce Apoptosis in Target Cells
Hanon, Emmanuel; Meyer, Gilles; Vanderplasschen, Alain ULg et al

in Journal of Virology (1998), 72(9), 7638-41

Bovine herpesvirus 1 (BHV-1) induces apoptotic cell death in bovine peripheral blood mononuclear cells and B-lymphoma cells. Using a BHV-1 glycoprotein H null mutant, we have demonstrated that although ... [more ▼]

Bovine herpesvirus 1 (BHV-1) induces apoptotic cell death in bovine peripheral blood mononuclear cells and B-lymphoma cells. Using a BHV-1 glycoprotein H null mutant, we have demonstrated that although penetration of BHV-1 is not required, attachment of BHV-1 viral particles is essential for the induction of apoptosis. [less ▲]

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See detailIntracellular and extracellular vaccinia virions enter cells by different mechanisms
Vanderplasschen, Alain ULg; Hollinshead, M.; Smith, G. L.

in Journal of General Virology (The) (1998), 79(Pt 4), 877-887

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See detailExtracellular enveloped vaccinia virus is resistant to complement because of incorporation of host complement control proteins into its envelope
Vanderplasschen, Alain ULg; Mathew, E.; Hollinshead, M. et al

in Proceedings of the National Academy of Sciences of the United States of America (1998), 95

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See detailExtracellular enveloped vaccinia virus. Entry, egress, and evasion
Smith, G. L.; Vanderplasschen, Alain ULg

in Advances in Experimental Medicine and Biology (1998), 440

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See detailStudy of vaccinia virus binding and entry using confocal microscopy, an original approach
Vanderplasschen, Alain ULg; Smith, G. L.

in Biology of the Cell (1998), 90

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See detailOptimization of murine CD8+ cytotoxic T-lymphocyte responses to pseudorabies virus
Depierreux, C.; Graff, I.; Lancelot, V. et al

in Journal of Immunological Methods (1997), 203

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See detailAntibodies against vaccinia virus do not neutralize extracellular enveloped virus but prevent virus release from infected cells and comet formation
Vanderplasschen, Alain ULg; Hollinshead, M.; Smith, G. L.

in Journal of General Virology (The) (1997), 78(Pt 8), 2041-2048

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See detailA novel virus blinding assay using confocal microscopy: demonstration that the intracellular and extracellular vaccinia virions bind to different cellular receptors
Vanderplasschen, Alain ULg; Smith, G. L.

in Journal of Virology (1997), 71

Vaccinia virus (VV) produces two antigenically and structurally distinct infectious virions, intracellular mature virus (IMV) and extracellular enveloped virus (EEV), which bind to unidentified and ... [more ▼]

Vaccinia virus (VV) produces two antigenically and structurally distinct infectious virions, intracellular mature virus (IMV) and extracellular enveloped virus (EEV), which bind to unidentified and possibly different cellular receptors. Studies of VV binding have been hampered by having two infectious virions and by the rupture of the EEV outer membrane in the majority of EEV virions during purification. To overcome these problems, we have developed a novel approach to study VV binding that is based on confocal microscopy and does not require EEV purification. In this assay, individual virus particles adsorbed to the cell are simultaneously distinguished and quantified by double immunofluorescence labelling with antibody markers for EEV and IMV. By this method, we show unequivocally that IMV and EEV bind to different cellular receptors. Three independent observations allow this conclusion. First, the efficiencies with which IMV and EEV bind to different cell lines are unrelated; second, cell surface digestion with some enzymes affects IMV and EEV binding differently; and third, the binding of a monoclonal antibody to cells prevents IMV binding but not EEV binding. This technique may be widely applicable for studying the binding of different viruses. [less ▲]

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