References of "Vanderplasschen, Alain"
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See detailCharacterization of caprine herpesvirus 1 glycoprotein D gene and its translation product
Keuser, Véronique; Detry, Bruno; Thiry, Julien ULg et al

in Virus Research (2006), 115(2), 112-121

Caprine herpesvirus 1 (CpHV- 1) is responsible of systemic infection in neonatal kids as well as abortion and fertility disorders in adult goats. This virus is closely related to bovine herpesvirus 1 ... [more ▼]

Caprine herpesvirus 1 (CpHV- 1) is responsible of systemic infection in neonatal kids as well as abortion and fertility disorders in adult goats. This virus is closely related to bovine herpesvirus 1 (BoHV-1) which causes infectious bovine rhinotracheitis. Glycoprotein D (gD) mediates important functions in alphaherpesviruses and is also a main inummogen. The sequence of CpHV-1 gD gene and the biochemical properties of its translation product were analyzed and compared to those of BoHV-1 and other alphaherpesviruses. A relatively high homology was found between CpHV-1 and BoHV-1 glycoproteins D amino acid sequences (similarity of 68.8%). Moreover, six cysteine residues are conserved by CpHV-1 gD and the other studied alphaherpesviruses. CpHV-1 gD has a molecular mass similar to BoHV-1 gD and contains complex N-linked oligosaccharides. In contrast to the BoHV-1 gD, CpHV-1 gD is expressed as a late protein. In spite of the observed differences which could influence its biological functions, CpHV-1 gD, shares most characteristics with other alphaherpesviruses and especially BoHV-1. (c) 2005 Elsevier B.V. All rights reserved. [less ▲]

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See detailCloning of the genome of Alcelaphine herpesvirus 1 as an infectious and pathogenic bacterial artificial chromosome.
Dewals, Benjamin G ULg; Boudry, Christel ULg; Gillet, Laurent ULg et al

in Journal of General Virology (The) (2006), 87(Pt 3), 509-17

Alcelaphine herpesvirus 1 (AlHV-1), carried asymptomatically by wildebeest, causes malignant catarrhal fever (MCF) following cross-species transmission to a variety of susceptible species of the order ... [more ▼]

Alcelaphine herpesvirus 1 (AlHV-1), carried asymptomatically by wildebeest, causes malignant catarrhal fever (MCF) following cross-species transmission to a variety of susceptible species of the order Artiodactyla. The study of MCF pathogenesis has been impeded by an inability to produce recombinant virus, mainly due to the fact that AlHV-1 becomes attenuated during passage in culture. In this study, these difficulties were overcome by cloning the entire AlHV-1 genome as a stable, infectious and pathogenic bacterial artificial chromosome (BAC). A modified loxP-flanked BAC cassette was inserted in one of the two large non-coding regions of the AlHV-1 genome. This insertion allowed the production of an AlHV-1 BAC clone stably maintained in bacteria and able to regenerate virions when transfected into permissive cells. The loxP-flanked BAC cassette was excised from the genome of reconstituted virions by growing them in permissive cells stably expressing Cre recombinase. Importantly, BAC-derived AlHV-1 virions replicated comparably to the virulent (low-passage) AlHV-1 parental strain and induced MCF in rabbits that was indistinguishable from that of the virulent parental strain. The availability of the AlHV-1 BAC is an important advance for the study of MCF that will allow the identification of viral genes involved in MCF pathogenesis, as well as the production of attenuated recombinant candidate vaccines. [less ▲]

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See detailFelid herpesvirus 1 glycoprotein G is a structural protein that mediates the binding of chemokines on the viral envelope.
Costes, Bérénice ULg; Thirion, Muriel ULg; Dewals, Benjamin G ULg et al

in Microbes & Infection (2006), 8(11), 2657-67

Glycoprotein G (gG) orthologues have been described in several alphaherpesviruses. gG is expressed both as a membrane-anchored form on infected cells and as a secreted form. Recently, we reported that ... [more ▼]

Glycoprotein G (gG) orthologues have been described in several alphaherpesviruses. gG is expressed both as a membrane-anchored form on infected cells and as a secreted form. Recently, we reported that both forms of gG encoded by alphaherpesviruses infecting large herbivores and by Felid herpesvirus 1 (FeHV-1) bind with high affinity to a broad range of CXC, CC and C-chemokines. Based on the viral species, gG has been reported either as a structural or a non-structural protein. To date, the incorporation of FeHV-1 gG into virions has never been tested, nor the property of alphaherpesvirus structural gG to bind chemokines on the virion surface. In the present study, to address these questions, various FeHV-1 gG recombinant strains were produced using an original technique based on an infectious FeHV-1 BAC clone and restriction endonuclease mediated recombination. Using the recombinants produced, we were able to determine that FeHV-1 gG is a structural protein that acts as a chemokine-binding protein on the virion surface. In the light of these results, putative roles of gG in alphaherpesvirus infections are discussed, and an evolutionary scenario is proposed to explain the structural versus non-structural property of gG amongst alphaherpesviruses. [less ▲]

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See detailRuminant alphaherpesviruses related to bovine herpesvirus 1.
Thiry, Julien ULg; Keuser, Veronique; Muylkens, Benoît ULg et al

in Veterinary Research (2006), 37(2), 169-90

Herpesviruses have mainly co-evolved with their hosts for millions of years. Consequently, different related host species may have been infected by various genetically related herpesviruses. Illustrating ... [more ▼]

Herpesviruses have mainly co-evolved with their hosts for millions of years. Consequently, different related host species may have been infected by various genetically related herpesviruses. Illustrating this concept, several ruminant alphaherpesviruses have been shown to form a cluster of viruses closely related to bovine herpesvirus 1 (BoHV-1): namely bovine herpesvirus 5, bubaline herpesvirus 1, caprine herpesvirus 1, cervid herpesviruses 1 and 2 and elk herpesvirus 1. These viruses share common antigenic properties and the serological relationships between them can be considered as a threat to BoHV-1 eradication programmes. BoHV-1 is a herpesvirus responsible for infectious bovine rhinotracheitis, which is a disease of major economic concern. In this article, the genetic properties of these ruminant alphaherpesviruses are reviewed on a comparative basis and the issue of interspecific recombination is assessed. The pathogenesis of these infections is described with emphasis on the host range and crossing of the host species barrier. Indeed, the non bovine ruminant species susceptible to these ruminant alphaherpesviruses may be potential BoHV-1 reservoirs. The differential diagnosis of these related infections is also discussed. In addition, available epidemiological data are used to assess the potential of cross-infection in ruminant populations. A better knowledge of these ruminant alphaherpesvirus infections is essential to successfully control infectious bovine rhinotracheitis. [less ▲]

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See detailEvolution of Bovine herpesvirus 4: recombination and transmission between African buffalo and cattle
Dewals, Benjamin G ULg; Thirion, Muriel ULg; Markine-Goriaynoff, N. et al

in Journal of General Virology (2006), 87(Pt 6), 1509-1519

Bovine herpesvirus 4 (BoHV-4) has been isolated from cattle throughout the world, but virological and serological studies have suggested that the African buffalo is also a natural host for this virus. It ... [more ▼]

Bovine herpesvirus 4 (BoHV-4) has been isolated from cattle throughout the world, but virological and serological studies have suggested that the African buffalo is also a natural host for this virus. It has previously been found that the Bo17 gene of BoHV-4 was acquired from an ancestor of the African buffalo, probably around 1.5 million years ago. Analysis of the variation of the Bo17 gene sequence among BoHV-4 strains suggested a relatively ancient transmission of BoHV-4 from the buffalo to the Bos primigenius lineage, followed by a host-dependent split between zebu and taurine BoHV-4 strains. In the present study, the evolutionary history of BoHV-4 was investigated by analysis of five gene sequences from each of nine strains representative of the viral species: three isolated from African buffalo in Kenya and six from cattle from Europe, North America and India. No two gene sequences had the same evolutionary tree, indicating that recombination has occurred between divergent lineages; six recombination events were delineated for these sequences. Nevertheless, exchange has been infrequent enough that a clonal evolutionary history of the strains could be discerned, upon which the recombination events were superimposed. The dates of divergence among BoHV-4 lineages were estimated from synonymous nucleotide-substitution rates. The inferred evolutionary history suggests that African buffalo were the original natural reservoir of BoHV-4 and that there have been at least three independent transmissions from buffalo to cattle, probably via intermediate hosts and - at least in the case of North American strains - within the last 500 years. [less ▲]

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See detailRecombinant bovine herpesvirus 4 (BoHV-4) expressing glycoprotein D of BoHV-1 is immunogenic and elicits serum-neutralizing antibodies against BoHV-1 in a rabbit model.
Donofrio, Gaetano; Cavirani, Sandro; Vanderplasschen, Alain ULg et al

in Clinical and Vaccine Immunology (2006), 13(11), 1246-54

Several biological characteristics of bovine herpesvirus 4 (BoHV-4) make it a good candidate as a gene delivery vector for vaccination purposes. These characteristics include little or no pathogenicity ... [more ▼]

Several biological characteristics of bovine herpesvirus 4 (BoHV-4) make it a good candidate as a gene delivery vector for vaccination purposes. These characteristics include little or no pathogenicity, unlikely oncogenicity, the capability to accommodate large amounts of foreign genetic material, the ability to infect several cell types coming from different animal species, and the ability to maintain transgene expression in both undifferentiated and differentiated cells. Starting from BoHV-4 cloned as a bacterial artificial chromosome (BAC), we used MuA transposase-mediated in vitro transposition to generate recombinant BoHV-4 expressing the immunodominant glycoprotein D (gD) of BoHV-1, one of the most important pathogens of cattle. Although a cis-acting element from woodchuck hepatitis virus (the woodchuck hepatitis virus posttranscriptional regulatory element [WPRE]) in the 3' end of the gD expression cassette was required for maximal gD expression from plasmids in transient transfection assays, this element was not necessary for efficient expression of gD from recombinant BoHV-4 genomes. BoHV-4 recombinants containing gD expression cassettes with or without the WPRE expressed gD at similarly high levels. Several cell lines originating from different animal species expressed gD when infected with BoHV-4 recombinants. When rabbits were immunized with one of the recombinants, high levels of serum neutralizing antibodies against BoHV-1 were generated. This work is one of the first demonstrations of the use BoHV-4 as a vector for vaccine purposes and may provide the basis for BoHV-1 vaccination of cattle with recombinant BoHV-4. [less ▲]

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See detailDemonstration by flow cytometry that CD5+CD8+ cells carry alcelaphine herpesvirus 1 in inoculated rabbits developing malignant catarrhal fever
Dewals, Benjamin G ULg; Gillet, Laurent ULg; Vanderplasschen, Alain ULg

Poster (2005, November 18)

Alcelaphine herpesvirus 1 (AlHV 1), carried by wildebeest (Connochaetes taurinus) asymptomatically, causes malignant catarrhal fever (MCF) when cross species transmitted to a variety of susceptible ... [more ▼]

Alcelaphine herpesvirus 1 (AlHV 1), carried by wildebeest (Connochaetes taurinus) asymptomatically, causes malignant catarrhal fever (MCF) when cross species transmitted to a variety of susceptible species of the Artiodactyla order. MCF is a fascinating disease described as a combination of lymphoproliferative and degenerative lesions. The study of MCF pathogenesis has been impeded by an inability to produce recombinant virus, due mainly to the fact that AlHV 1 becomes attenuated during passage in culture. Here, we have overcome these difficulties by (i) cloning the entire AlHV 1 genome as a stable, infectious and pathogenic bacterial artificial chromosome (BAC), and (ii) by using prokaryotic recombination technology for the production of an AlHV 1 recombinant. Firstly, the AlHV 1 genome was BAC cloned using one insertion site in a region containing no open reading frame. This insertion allowed the production of an AlHV 1 BAC clone stably maintained in bacteria and able to regenerate virions when transfected into permissive cells. BAC derived AlHV 1 virions induced MCF in rabbits comparably to the AlHV 1 wild type (WT) strain. Secondly, a two step mutagenesis procedure in E. coli was used to generate a recombinant strain expressing enhanced-green fluorescent protein (EGFP) as a reporter gene. After reconstitution of recombinant virions into permissive cells and excision of the BAC cassette, flow cytometry analyses were performed to validate the recombinant strain and to investigate the pathogenesis of MCF. The results of these analyses can be summarized as follows: (i) the validity of the EGFP expression cassette as a reporter gene has been demonstrated by in vitro infections; (ii) inoculation of rabbits revealed that the recombinant strain has retained the pathogenicity of its parental strain and that the cell types carrying AlHV-1 in peripheral blood mononuclear cells, lymph nodes and the spleen are mainly CD5+ CD8+ cells. [less ▲]

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See detailClonage de l’herpèsvirus alcélaphin 1 sous forme d’un chromosome artificiel bactérien infectieux
Dewals, Benjamin G ULg; Boudry, Christel; Markine-Goriaynoff, Nicolas et al

Poster (2005, April)

L’herpèsvirus alcélaphin 1 (AlHV-1) est un Gammaherpesvirinae du genre Rhadinovirus ayant pour hôte naturel le gnou (Connochaetes taurinus). Apathogène pour son hôte naturel, ce virus induit par ailleurs ... [more ▼]

L’herpèsvirus alcélaphin 1 (AlHV-1) est un Gammaherpesvirinae du genre Rhadinovirus ayant pour hôte naturel le gnou (Connochaetes taurinus). Apathogène pour son hôte naturel, ce virus induit par ailleurs une pathologie mortelle lorsqu’il est transmis à un grand nombre d’espèces de ruminants sensibles. Cette pathologie, appelée forme africaine du coryza gangreneux (FACG) est associée à une lymphoprolifération et une destruction des tissus de l’hôte infecté. Pour étudier le rôle de l’AlHV-1 dans la pathogenèse de la FACG, il est nécessaire de pouvoir manipuler le génome viral afin de générer des virus recombinants et révertants pour certains gènes. A ce jour, aucune souche d’AlHV-1 recombinante n’a pu être produite. Cette carence résulte du fait que ce virus est strictement associé aux cellules et qu’il possède la caractéristique de s’atténuer spontanément lors de sa multiplication in vitro. De ce fait, la réalisation de virus mutants se révèle impossible par une approche classique de recombinaison homologue en cellules eucaryotes. Récemment, le développement des technologies de chromosome artificiel bactérien (BAC) a permis de cloner le génome entier de plusieurs gammaherpèsvirus de manière stable en bactérie ainsi que la production rapide de nombreuses souches recombinantes. Dans la présente étude, nous avons cloné le génome entier de la souche C500 de l’AlHV-1 sous forme d’un BAC appelé ci-après BAC-AlHV-1. Les résultats obtenus peuvent se résumer comme suit : (i) le BAC-AlHV-1 permet une propagation stable du génome de l’AlHV-1 en bactérie ; (ii) la transfection du BAC-AlHV-1 en cellules eucaryotes permet de régénérer des particules virales infectieuses ; (iii) la cassette BAC insérée initialement dans le génome de l’AlHV-1 étant flanquée de séquences loxP, la multiplication des virions générés à partir du BAC-AlHV-1 en cellules EBL-Cre (embryonic bovine lung ; exprimant la Cre recombinase) permet l’excision de la cassette BAC ; (iv) enfin, la capacité des virions générés à partir du BAC à induire la FACG en modèle lapin a été démontrée. En conclusion, le clone BAC-AlHV-1 généré dans cette étude va enfin permettre l’étude des rôles des différents gènes de l’AlHV-1 dans la genèse de la FACG. [less ▲]

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See detailVaricella-zoster virus IE63 protein represses the basal transcription machinery by disorganizing the pre-initiation complex
Di Valentin, Emmanuel ULg; Bontems, Sébastien ULg; Habran, Lionel ULg et al

in Biological Chemistry (2005), 386(3), 255-267

Using transient transfection assays, regulation properties of varicella-zoster virus (VZV)-encoded IE63 protein were analyzed on several VZV immediate early (ORF4), early (ORF28) and late (ORF67 ... [more ▼]

Using transient transfection assays, regulation properties of varicella-zoster virus (VZV)-encoded IE63 protein were analyzed on several VZV immediate early (ORF4), early (ORF28) and late (ORF67) promoters. IE63 was shown to repress the basal activity of most of the promoters tested in epithelial (Vero) and neuronal (ND7) cells to various extents. Trans -repressing activities were also observed on heterologous viral and cellular promoters. Since a construct carrying only a TATA box sequence and a series of wild-type or mutated interleukin (IL)-8 promoters was also repressed by IE63, the role of upstream regulatory elements was ruled out. Importantly, the basal activity of a TATA-less promoter was not affected by IE63. Using a series of IE63 deletion constructs, amino acids 151-213 were shown to be essential to the transrepressing activity in Vero cells, while in ND7 cells the essential region extended to a much larger carboxy-terminal part of the protein. We also demonstrate that IE63 is capable of disrupting the transcriptional pre-initiation complex and of interacting with several general transcription factors. The central and carboxy-terminal domains of IE63 are important for these effects. Altogether, these results demonstrate that IE63 protein is a transcriptional repressor whose activity is directed towards general transcription factors. [less ▲]

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See detailGynogenesis induction and sex determination in the Eurasian perch, Perca fluviatilis
Rougeot, Carole ULg; Ngingo, J. V.; Gillet, Laurent ULg et al

in Aquaculture (2005), 243(1-4), 411-415

In the present study, we used meiotic gynogenesis, widely used in studies on sex determination, to confirm female homogamety in Eurasian perch, Perca fluviatilis. Sperm irradiated with UV for 400 s was ... [more ▼]

In the present study, we used meiotic gynogenesis, widely used in studies on sex determination, to confirm female homogamety in Eurasian perch, Perca fluviatilis. Sperm irradiated with UV for 400 s was used to artificially fertilized eggs. The diploid of the resulting embryos was restored by a heat shock (30 degreesC) applied to the eggs 5 min postfertilization, for 25 min. Fertilization (ranging between 45% and 75%) and survival rates at hatching (ranging between 3.4% and 46.6%) were not significantly different (P>0.05) between the diploid control and gynogenetics. The diploid controls and two batches of gynogenetics contained 100% diploid larvae, whereas two other batches of gynogenetics contained 6.7% and 10.0% triploid larvae. The sex ratios of the diploid controls were not significantly different from 1:1, whereas all gynogenetic families were 100% female. These results confirm female homogamety in Eurasian perch, demonstrated by the use of hormonally mascilinized breeders in a previous study. (C) 2004 Elsevier B.V. All rights reserved. [less ▲]

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See detailShort communication: Pasteurization of milk abolishes bovine herpesvirus 4 infectivity.
Bona, C.; Dewals, Benjamin G ULg; Wiggers, L. et al

in Journal of Dairy Science (2005), 88(9), 3079-83

Bovine herpesvirus 4 (BoHV-4) is a gammaherpesvirus highly prevalent in the cattle population that has been isolated from the milk and the serum of healthy infected cows. Several studies reported the ... [more ▼]

Bovine herpesvirus 4 (BoHV-4) is a gammaherpesvirus highly prevalent in the cattle population that has been isolated from the milk and the serum of healthy infected cows. Several studies reported the sensitivity and the permissiveness of some human cells to BoHV-4 infection. Moreover, our recent study demonstrated that some human cells sensitive but not permissive to BoHV-4 support a persistent infection protecting them from tumor necrosis factor-alpha-induced apoptosis. Together, these observations suggested that BoHV-4 could represent a danger for public health. To evaluate the risk of human infection by BoHV-4 through milk or serum derivatives, we investigated the resistance of BoHV-4 to the mildest thermal treatments usually applied to these products. The results demonstrated that milk pasteurization and thermal decomplementation of serum abolish BoHV-4 infectivity by inactivation of its property to enter permissive cells. Consequently, our results demonstrate that these treatments drastically reduce the risk of human infection by BoHV-4 through treated milk or serum derivatives. [less ▲]

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See detailBovine herpesvirus 4 induces apoptosis of human carcinoma cell lines in vitro and in vivo
Gillet, Laurent ULg; Dewals, Benjamin G ULg; Farnir, Frédéric ULg et al

in Cancer Research (2005), 65(20), 9463-9472

The idea of using oncolytic viruses for the treatment of cancers was proposed a century ago. During the last two decades, viruses able to replicate specifically in cancer cells and to induce their lysis ... [more ▼]

The idea of using oncolytic viruses for the treatment of cancers was proposed a century ago. During the last two decades, viruses able to replicate specifically in cancer cells and to induce their lysis were identified and were genetically modified to improve their viro-oncolytic properties. More recently, a new approach consisting of inducing selective apoptosis in cancer cells through viral infection has been proposed; this approach has been called viro-oncoapoptosis. In the present study, we report the property of bovine herpesvirus-4 (BoHV-4) to induce, in vitro and in vivo, apoptosis of some human carcinomas. This conclusion relies on the following observations: (a) In vitro, BoHV-4 infection induced apoptosis of A549 and OVCAR carcinoma cell lines in a time- and dose-dependent manner. (b) Apoptosis was induced by the expression of an immediate-early or an early BoHV-4 gene, but did not require viral replication. (c) Cell treatment with caspase inhibitors showed that apoptosis induced by BoHV-4 relied mainly on caspase-10 activation. (d) Infection of cocultures of A549 or OVCAR cells mixed with human 293 cells (in which BoHV-4 does not induce apoptosis) showed that BoHV-4 specifically eradicated A549 or OVCAR cancer cells from the cocultures. (e) Finally, in vivo experiments done with nude mice showed that BoHV-4 intratumoral injections reduced drastically the growth of preestablished A549 xenografts. Taken together, these results suggest that BoHV-4 may have potential as a viro-oncoapoptotic agent for the treatment of some human carcinomas. Moreover, further identification of BoHV-4 proapoptotic gene(s) and the cellular pathways targeted by this or these gene(s) could lead to the design of new cancer therapeutic strategies. [less ▲]

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See detailAntibodies against bovine herpesvirus 4 are highly prevalent in wild African buffaloes throughout eastern and southern Africa
Dewals, Benjamin G ULg; Gillet, Laurent ULg; Gerdes, Truuske et al

in Veterinary Microbiology (2005), 110(3-4), 209-220

Bovine herpesvirus 4 (BoHV-4) has been isolated from cattle throughout the world. Interestingly, a survey of wild African buffaloes mainly from the Maasai Mara Game Reserve in Kenya revealed that 94% of ... [more ▼]

Bovine herpesvirus 4 (BoHV-4) has been isolated from cattle throughout the world. Interestingly, a survey of wild African buffaloes mainly from the Maasai Mara Game Reserve in Kenya revealed that 94% of the animals tested had anti-BoHV-4 antibodies [Rossiter, P.B., Gumm, I.D., Stagg, D.A., Conrad, PA., Mukolwe, S., Davies, F.G., White, H., 1989. Isolation of bovine herpesvirus-3 from African buffaloes (Syncerus caffer). Res. Vet. Sci. 46, 337-343]. These authors also proposed that the serological antigenic relationship existing between BoHV-4 and alcelaphine herpesvirus I (A1HV-1) could confer to BoHV-4 infected buffaloes a protective immune response against lethal A1HV-1 infection. In the present study, we addressed two questions related to Rossiter et al. paper. Firstly, to investigate the role of the African buffalo as a natural host species of BoHV-4, the seroprevalence of anti-BoHV-4 antibodies was analysed in wild African buffaloes throughout eastern and southern Africa. A total of 400 sera was analysed using two complementary immunofluorescent assays. These analyses revealed that independently of their geographical origin, wild African buffaloes exhibit a seroprevalence of anti-BoHV-4 antibodies higher than 68%. This result is by far above the seroprevalence generally observed in cattle. Our data are discussed in the light of our recent phylogenetic study demonstrating that the BoHV-4 Bo17 gene has been acquired from a recent ancestor of the African buffalo. Secondly, we investigated the humoral antigenic relationship existing between BoHV-4 and A1HV-1. Our results demonstrate that among the antigens expressed in A1HV-1 infected cells, epitope(s) recognised by anti-BoHV-4 antibodies are exclusively nuclear, suggesting that the putative property of BoHV-4 to confer an immune protection against A1HV-1 relies on a cellular rather than on a humoral immune response. (c) 2005 Elsevier B.V. All rights reserved. [less ▲]

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See detailDevelopment of bovine herpesvirus 4 as an expression vector using bacterial artificial chromosome cloning.
Gillet, Laurent ULg; Daix, V.; Donofrio, G. et al

in Journal of General Virology (The) (2005), 86(Pt 4), 907-17

Several features make bovine herpesvirus 4 (BoHV-4) attractive as a backbone for use as a viral expression vector and/or as a model to study gammaherpesvirus biology. However, these developments have been ... [more ▼]

Several features make bovine herpesvirus 4 (BoHV-4) attractive as a backbone for use as a viral expression vector and/or as a model to study gammaherpesvirus biology. However, these developments have been impeded by the difficulty in manipulating its large genome using classical homologous recombination in eukaryotic cells. In the present study, the feasibility of exploiting bacterial artificial chromosome (BAC) cloning and prokaryotic recombination technology for production of BoHV-4 recombinants was explored. Firstly, the BoHV-4 genome was BAC cloned using two potential insertion sites. Both sites of insertion gave rise to BoHV-4 BAC clones stably maintained in bacteria and able to regenerate virions when transfected into permissive cells. Reconstituted virus replicated comparably to wild-type parental virus and the loxP-flanked BAC cassette was excised by growing them on permissive cells stably expressing Cre recombinase. Secondly, BoHV-4 recombinants expressing Ixodes ricinus anti-complement protein I or II (IRAC I/II) were produced using a two-step mutagenesis procedure in Escherichia coli. Both recombinants induced expression of high levels of functional IRAC molecules in the supernatant of infected cells. This study demonstrates that BAC cloning and prokaryotic recombination technology are powerful tools for the development of BoHV-4 as an expression vector and for further fundamental studies of this gammaherpesvirus. [less ▲]

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See detailRecombination in alphaherpesviruses
Thiry, Etienne ULg; Meurens, F.; Muylkens, Benoît ULg et al

in Reviews in Medical Virology (2005), 15(2, Mar-Apr), 89-103

Within the Herpesviridae family, Alphaherpesvirinae is an extensive subfamily which contains numerous mammalian and avian viruses. Given the low rate of herpesvirus nucleotide substitution, recombination ... [more ▼]

Within the Herpesviridae family, Alphaherpesvirinae is an extensive subfamily which contains numerous mammalian and avian viruses. Given the low rate of herpesvirus nucleotide substitution, recombination can be seen as an essential evolutionary driving force although it is likely underestimated. Recombination in alphaherpesviruses is intimately linked to DNA replication. Both viral and cellular proteins participate in this recombination-dependent replication. The presence of inverted repeats in the alphaherpesvirus genomes allows segment inversion as a consequence of specific recombination between repeated sequences during DNA replication. High molecular weight intermediates of replication, called concatemers, are the site of early recombination events. The analysis of concatemers, from cells coinfected by two distinguishable alphaherpesviruses provides an efficient tool to study recombination without the bias introduced by invisible or non-viable recombinants, and by dominance of a virus over recombinants. Intraspecific recombination frequently occurs between strains of the same alphaherpesvirus species. Interspecific recombination depends on enough sequence similarity to enable recombination between distinct alphaherpesvirus species. The most important prerequisite for successful recombination is coinfection of the individual host by different virus strains or species. Consequently the following factors affecting the distribution of different viruses to shared target cells need to be considered: dose of inoculated virus, time interval between inoculation of the first and the second virus, distance between the marker mutations, genetic homology, virulence and latency. Recombination, by exchanging genomic segments, may modify the virulence of alphaherpesviruses. It must be carefully assessed for the biosafety of antiviral therapy, alphaherpesvirus-based vectors and live attenuated vaccines. Copyright (C) 2004 John Wiley Sons, Ltd. [less ▲]

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See detailPro-inflammatory properties for thiazolidinediones.
Desmet, Christophe ULg; Warzée, Barbara ULg; Gosset, Philippe et al

in Biochemical Pharmacology (2005), 69(2), 255-265

Thiazolidinediones (TZDs) are pharmacological ligands of the peroxisome proliferator-activated receptor (PPAR)-gamma that are extensively used in the treatment of type II diabetes. Recently, an anti ... [more ▼]

Thiazolidinediones (TZDs) are pharmacological ligands of the peroxisome proliferator-activated receptor (PPAR)-gamma that are extensively used in the treatment of type II diabetes. Recently, an anti-inflammatory potential for TZDs has been suggested, based on observations that these compounds may inhibit pro-inflammatory cytokine expression in vitro and may attenuate the inflammatory response in vivo. Here, we show that the TZDs rosiglitazone (RSG) and troglitazone (TRO) do not inhibit the inflammatory response to tumor necrosis factor (TNF)-alpha in various epithelial cell types. On the contrary, both RSG and TRO significantly potentiated TNF-alpha-induced production of granulocyte/macrophage-colony-stimulating factor, interleukin (IL)-6 and/or IL-8 in these cells. This increase in pro-inflammatory cytokine expression was functionally significant as supernatants from cells co-treated with TNF-alpha and TZDs displayed increased neutrophil pro-survival activity when compared with supernatants from cells treated with TNF-alpha alone. Additionally, it was shown that TZDs enhance cytokine expression at the transcriptional level, but that the pro-inflammatory effects of TZDs are independent on PPARgamma, nuclear factor kappaB or mitogen-activated protein kinase activation. Our study shows that TZDs may potentiate the inflammatory response in epithelial cells, a previously unappreciated effect of these compounds [less ▲]

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See detailBoth soluble and membrane-anchored forms of Felid herpesvirus 1 glycoprotein G function as a broad-spectrum chemokine-binding protein
Costes, Bérénice ULg; Ruiz-Arguello, M. B.; Bryant, N. A. et al

in Journal of General Virology (2005), 86(Pt 12), 3209-3214

Recently, glycoprotein G (gG) of several alphaherpesviruses infecting large herbivores was shown to belong to a new family of chemokine-binding proteins (vCKBPs). In the present study, the function of ... [more ▼]

Recently, glycoprotein G (gG) of several alphaherpesviruses infecting large herbivores was shown to belong to a new family of chemokine-binding proteins (vCKBPs). In the present study, the function of Felid herpesvirus 1 (FeHV-1) gG as a vCKBP was investigated and the following conclusions were reached: (i) FeHV-1 secreted gG is a high-affinity broad-spectrum vCKBP that binds CC, CXC and C chemokines; (ii) gG is the only vCKBP expressed by FeHV-1 that binds CCL3 and CXCL1; (iii) secreted gG blocks chemokine activity by preventing their interaction with high-affinity cellular receptors; (iv) the membrane-anchored form of gG expressed on the surface of infected cells is also able to bind chemokines; and (v) the vCKBP activity is conserved among different field isolates of FeHV-1. Altogether, these data demonstrate that FeHV-1 gG is a new member of the vCKBP-4 family. Moreover, this study is the first to demonstrate that gG expressed at the surface of FeHV-1-infected cells can also bind chemokines. [less ▲]

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See detailSTAT5 promotes granulocyte survival during inflammation
Fievez, Laurence ULg; Desmet, Christophe ULg; Seumois, G. et al

in Proceedings: The American Thoracic Society (2005)

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See detailLes glycosyltransférases virales
Markine-Goriaynoff, N.; Vanderplasschen, Alain ULg

in Virologie (2005), 9(1), 35-48

Les glycanes constituent une classe biochimique difficile à investiguer, mais dont les rôles essentiels à tous les niveaux de la biologie se sont imposés comme une évidence ces dernières décennies. Il ... [more ▼]

Les glycanes constituent une classe biochimique difficile à investiguer, mais dont les rôles essentiels à tous les niveaux de la biologie se sont imposés comme une évidence ces dernières décennies. Il apparaît maintenant qu'ils représentent la quatrième catégorie de molécules bio-informatives après l'ADN, l'ARN et les protéines. À ce titre, après la génomique, la transcriptomique et la protéomique, l'ère de la glycomique est appelée à concentrer beaucoup d'intérêts. Des millions d'années durant, les virus ont co-évolué avec leurs hôtes; au cours de ce processus adaptatif, ils ont évolué de manière à favoriser leur multiplication par l'acquisition de moyens visant à imiter, détourner ou saboter les mécanismes les plus complexes de la physiologie de leurs hôtes, y compris les mécanismes destinés à interférer avec le glycome. La découverte de l'importance du glycome dans le contexte de la régulation des interactions entre les virus et leurs hôtes a récemment mené à la notion de « glycovirologie ». Un aspect fascinant de la glycovirologie est l'étude des mécanismes viraux visant à modifier le glycome. Les virus affectent le glycome de deux façons: en régulant l'expression des glycosyltransférases de la cellule hôte ou en exprimant leur propre glycosyltransférase. Cette revue est consacrée aux glycosyltransférases virales et à la description de leurs fonctions. La description de ces enzymes illustre différents concepts fondamentaux de virologie et démontre indirectement le rôle crucial joué par la glycomique dans les processus biologiques. [less ▲]

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See detailViral subversion of the immune system
Gillet, L.; Vanderplasschen, Alain ULg

in Applications of gene-based technologies for improving animal production and health in developing countries (2005)

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