References of "Vanderplasschen, Alain"
     in
Bookmark and Share    
Full Text
Peer Reviewed
See detailReplacement of Glycoprotein B in Alcelaphine Herpesvirus 1 by Its Ovine Herpesvirus 2 Homolog : Implications in Vaccine Development for Sheep-Associated Malignant Catarrhal Fever.
Cunha, Cristina W.; Taus, Naomi S.; Dewals, Benjamin G ULg et al

in mSphere (2016), 1(4), 00108-16

Vaccine development is a top priority in malignant catarrhal fever (MCF) research. In the case of sheep-associated MCF (SA-MCF) caused by ovine herpesvirus 2 (OvHV-2), progress toward this objective has ... [more ▼]

Vaccine development is a top priority in malignant catarrhal fever (MCF) research. In the case of sheep-associated MCF (SA-MCF) caused by ovine herpesvirus 2 (OvHV-2), progress toward this objective has been hindered by the absence of methods to attenuate or modify the virus, since it cannot be propagated in vitro. As an alternative for vaccine development, in this study, we tested the hypothesis that one of the SA-MCF vaccine candidate targets, OvHV-2 glycoprotein B (gB), could be expressed by a nonpathogenic alcelaphine herpesvirus 1 (AlHV-1) and then evaluated the potential of the AlHV-1/OvHV-2 chimera to be used as a vaccine and a diagnostic tool. The construction and characterization of an AlHV-1/OvHV-2 chimeric virus that is nonpathogenic and expresses an OvHV-2 vaccine target are significant steps toward the development of an SA-MCF vaccine and also provide a valuable means to study OvHV-2 biology. [less ▲]

Detailed reference viewed: 14 (1 ULg)
Full Text
Peer Reviewed
See detailLong term-cultured and cryopreserved primordial germ cells from various chicken breeds retain high proliferative potential and gonadal colonisation competency
Tonus, Céline ULg; Cloquette, Karine; Ectors, Fabien ULg et al

in Reproduction, Fertility and Development (2016), 28(5), 628-639

Detailed reference viewed: 135 (74 ULg)
Full Text
Peer Reviewed
See detailDeletion of Murid Herpesvirus 4 ORF63 Affects the Trafficking of Incoming Capsids toward the Nucleus.
Latif, Muhammad Bilal ULg; Machiels, Bénédicte ULg; Xiao, Xue ULg et al

in Journal of virology (2016), 90(5), 2455-72

Gammaherpesviruses are important human and animal pathogens. Despite the fact that they display the classical architecture of herpesviruses, the function of most of their structural proteins is still ... [more ▼]

Gammaherpesviruses are important human and animal pathogens. Despite the fact that they display the classical architecture of herpesviruses, the function of most of their structural proteins is still poorly defined. This is especially true for tegument proteins. Interestingly, a potential role in immune evasion has recently been proposed for the tegument protein encoded by Kaposi's sarcoma-associated herpesvirus open reading frame 63 (ORF63). To gain insight about the roles of ORF63 in the life cycle of a gammaherpesvirus, we generated null mutations in the ORF63 gene of murid herpesvirus 4 (MuHV-4). We showed that disruption of ORF63 was associated with a severe MuHV-4 growth deficit both in vitro and in vivo. The latter deficit was mainly associated with a defect of replication in the lung but did not affect the establishment of latency in the spleen. From a functional point of view, inhibition of caspase-1 or the inflammasome did not restore the growth of the ORF63-deficient mutant, suggesting that the observed deficit was not associated with the immune evasion mechanism identified previously. Moreover, this growth deficit was also not associated with a defect in virion egress from the infected cells. In contrast, it appeared that MuHV-4 ORF63-deficient mutants failed to address most of their capsids to the nucleus during entry into the host cell, suggesting that ORF63 plays a role in capsid movement. In the future, ORF63 could therefore be considered a target to block gammaherpesvirus infection at a very early stage of the infection. IMPORTANCE: The important diseases caused by gammaherpesviruses in human and animal populations justify a better understanding of their life cycle. In particular, the role of most of their tegument proteins is still largely unknown. In this study, we used murid herpesvirus 4, a gammaherpesvirus infecting mice, to decipher the role of the protein encoded by the viral ORF63 gene. We showed that the absence of this protein is associated with a severe growth deficit both in vitro and in vivo that was mainly due to impaired migration of viral capsids toward the nucleus during entry. Together, our results provide new insights about the life cycle of gammaherpesviruses and could allow the development of new antiviral strategies aimed at blocking gammaherpesvirus infection at the very early stages. [less ▲]

Detailed reference viewed: 22 (6 ULg)
Full Text
Peer Reviewed
See detailStructural Proteomics of Herpesviruses.
Leroy, Baptiste; Gillet, Laurent ULg; Vanderplasschen, Alain ULg et al

in Viruses (2016), 8(2),

Herpesviruses are highly prevalent viruses associated with numerous pathologies both in animal and human populations. Until now, most of the strategies used to prevent or to cure these infections have ... [more ▼]

Herpesviruses are highly prevalent viruses associated with numerous pathologies both in animal and human populations. Until now, most of the strategies used to prevent or to cure these infections have been unsuccessful because these viruses have developed numerous immune evasion mechanisms. Therefore, a better understanding of their complex lifecycle is needed. In particular, while the genome of numerous herpesviruses has been sequenced, the exact composition of virions remains unknown for most of them. Mass spectrometry has recently emerged as a central method and has permitted fundamental discoveries in virology. Here, we review mass spectrometry-based approaches that have recently allowed a better understanding of the composition of the herpesvirus virion. In particular, we describe strategies commonly used for proper sample preparation and fractionation to allow protein localization inside the particle but also to avoid contamination by nonstructural proteins. A collection of other important data regarding post-translational modifications or the relative abundance of structural proteins is also described. This review also discusses the poorly studied importance of host proteins in herpesvirus structural proteins and the necessity to develop a quantitative workflow to better understand the dynamics of the structural proteome. In the future, we hope that this collaborative effort will assist in the development of new strategies to fight these infections. [less ▲]

Detailed reference viewed: 32 (6 ULg)
Full Text
Peer Reviewed
See detailBovine Herpesvirus 4 Modulates Its beta-1,6-N-Acetylglucosaminyltransferase Activity through Alternative Splicing.
Lété, Céline ULg; Markine-Goriaynoff, Nicolas; Machiels, Bénédicte ULg et al

in Journal of virology (2016), 90(4), 2039-51

Carbohydrates play major roles in host-virus interactions. It is therefore not surprising that, during coevolution with their hosts, viruses have developed sophisticated mechanisms to hijack for their ... [more ▼]

Carbohydrates play major roles in host-virus interactions. It is therefore not surprising that, during coevolution with their hosts, viruses have developed sophisticated mechanisms to hijack for their profit different pathways of glycan synthesis. Thus, the Bo17 gene of Bovine herpesvirus 4 (BoHV-4) encodes a homologue of the cellular core 2 protein beta-1,6-N-acetylglucosaminyltransferase-mucin type (C2GnT-M), which is a key player for the synthesis of complex O-glycans. Surprisingly, we show in this study that, as opposed to what is observed for the cellular enzyme, two different mRNAs are encoded by the Bo17 gene of all available BoHV-4 strains. While the first one corresponds to the entire coding sequence of the Bo17 gene, the second results from the splicing of a 138-bp intron encoding critical residues of the enzyme. Antibodies generated against the Bo17 C terminus showed that the two forms of Bo17 are expressed in BoHV-4 infected cells, but enzymatic assays revealed that the spliced form is not active. In order to reveal the function of these two forms, we then generated recombinant strains expressing only the long or the short form of Bo17. Although we did not highlight replication differences between these strains, glycomic analyses and lectin neutralization assays confirmed that the splicing of the Bo17 gene gives the potential to BoHV-4 to fine-tune the global level of core 2 branching activity in the infected cell. Altogether, these results suggest the existence of new mechanisms to regulate the activity of glycosyltransferases from the Golgi apparatus. IMPORTANCE: Viruses are masters of adaptation that hijack cellular pathways to allow their growth. Glycans play a central role in many biological processes, and several studies have highlighted mechanisms by which viruses can affect glycosylation. Glycan synthesis is a nontemplate process regulated by the availability of key glycosyltransferases. Interestingly, bovine herpesvirus 4 encodes one such enzyme which is a key enzyme for the synthesis of complex O-glycans. In this study, we show that, in contrast to cellular homologues, this virus has evolved to alternatively express two proteins from this gene. While the first one is enzymatically active, the second results from the alternative splicing of the region encoding the catalytic site of the enzyme. We postulate that this regulatory mechanism could allow the virus to modulate the synthesis of some particular glycans for function at the location and/or the moment of infection. [less ▲]

Detailed reference viewed: 28 (8 ULg)
Full Text
Peer Reviewed
See detailSmall RNA deep sequencing identifies viral microRNAs during malignant catarrhal fever induced by alcelaphine herpesvirus 1
Sorel, Océane ULg; Tuddenham, Lee; Myster, Françoise ULg et al

in Journal of General Virology (The) (2015), 96(11), 3360-3372

Alcelaphine herpesvirus 1 (AlHV-1) is a c-herpesvirus (c-HV) carried asymptomatically by wildebeest. Upon cross-species transmission, AlHV-1 induces a fatal lymphoproliferative disease named malignant ... [more ▼]

Alcelaphine herpesvirus 1 (AlHV-1) is a c-herpesvirus (c-HV) carried asymptomatically by wildebeest. Upon cross-species transmission, AlHV-1 induces a fatal lymphoproliferative disease named malignant catarrhal fever (MCF) in many ruminants, including cattle, and the rabbit model. Latency has been shown to be essential for MCF induction. However, the mechanisms causing the activation and proliferation of infected CD8+T cells are unknown. Many c-HVs express microRNAs (miRNAs). These small non-coding RNAs can regulate expression of host or viral target genes involved in various pathways and are thought to facilitate viral infection and/or mediate activation and proliferation of infected lymphocytes. The AlHV-1 genome has been predicted to encode a large number of miRNAs. However, their precise contribution in viral infection and pathogenesis in vivo remains unknown. Here, using cloning and sequencing of small RNAs we identified 36 potential miRNAs expressed in a lymphoblastoid cell line propagated from a calf infected with AlHV-1 and developing MCF. Among the sequenced candidate miRNAs, 32 were expressed on the reverse strand of the genome in two main clusters. The expression of these 32 viral miRNAs was further validated using Northern blot and quantitative reverse transcription PCR in lymphoid organs of MCF- developing calves or rabbits. To determine the concerted contribution in MCF of 28 viral miRNAs clustered in the non-protein-coding region of the AlHV-1 genome, a recombinant virus was produced. The absence of these 28 miRNAs did not affect viral growth in vitro or MCF induction in rabbits, indicating that the AlHV-1 miRNAs clustered in this non-protein-coding genomic region are dispensable for MCF induction. [less ▲]

Detailed reference viewed: 28 (2 ULg)
Full Text
Peer Reviewed
See detailThe genome of a tortoise herpesvirus (testudinid herpesvirus 3) has a novel structure and contains a large region that is not required for replication in vitro or virulence in vivo.
Gandar, Frederic ULg; Wilkie, Gavin S.; Gatherer, Derek et al

in Journal of Virology (2015), 89(22), 11438-11456

Testudinid herpesvirus 3 (TeHV-3) is the causative agent of a lethal disease affecting several tortoise species. The threat that this virus poses to endangered animals is focusing efforts on ... [more ▼]

Testudinid herpesvirus 3 (TeHV-3) is the causative agent of a lethal disease affecting several tortoise species. The threat that this virus poses to endangered animals is focusing efforts on characterizing its properties, in order to enable the development of prophylactic methods. We have sequenced the genomes of the two most studied TeHV-3 strains (1976 and 4295). TeHV-3 strain 1976 has a novel genome structure and is most closely related to a turtle herpesvirus, thus supporting its classification into genus Scutavirus, subfamily Alphaherpesvirinae, family Herpesviridae. The sequence of strain 1976 also revealed viral counterparts of cellular interleukin-10 and semaphorin, which have not been described previously in members of subfamily Alphaherpesvirinae. TeHV-3 strain 4295 is a mixture of three forms (m1, m2, and M), in which, in comparison to strain 1976, the genomes exhibit large, partially overlapping deletions of 12.5 to 22.4 kb. Viral subclones representing these forms were isolated by limiting dilution assays, and each replicated in cell culture comparably to strain 1976. With the goal of testing the potential of the three forms as attenuated vaccine candidates, strain 4295 was inoculated intranasally into Hermann's tortoises (Testudo hermanni). All inoculated subjects died, and PCR analyses demonstrated the ability of the m2 and M forms to spread and invade the brain. In contrast, the m1 form was detected in none of the organs tested, suggesting its potential as the basis of an attenuated vaccine candidate. Our findings represent a major step toward characterizing TeHV-3 and developing prophylactic methods against it. [less ▲]

Detailed reference viewed: 53 (9 ULg)
Full Text
See detailLong term culture, cryopreservation and genetic modification of chicken primordial germ cells
Tonus, Céline ULg; Garcia Gil, Francisco José ULg; Cloquette, Karine et al

Poster (2015, October 16)

Avian primordial germ cells (PGCs) are precursor of gametes and appear during early stages of embryonic development. Under appropriate culture conditions, these cells can keep their germ cells properties ... [more ▼]

Avian primordial germ cells (PGCs) are precursor of gametes and appear during early stages of embryonic development. Under appropriate culture conditions, these cells can keep their germ cells properties in vitro and are foreseen as promising tools for developing efficient avian genetic engineering and preservation of germplasm. We propose original methods that allow long term expansion, efficient cryopreservation and genetic modification of primary cultures of undifferentiated PGCs. PGCs are collected from embryonic blood during their migratory period and grown in cell-culture insert in the presence of feeder cells (BRL). This physically separated co-culture system along with selective culture medium promoted emergence, selection and proliferation of PGCs lines. Forty percent of blood samples gave rise to lines originating from three commercial layer and two Belgian endangered breeds. PGCs lines were characterized for the expression of the stem cells and PGCs marker SSEA-1 by FACS. RT-PCR confirmed expression of germ-line specific markers (CVH, CDH, DAZL), pluripotency markers (cPouV, cSox2, cNanog), telomerase and CXCR4 receptor. All lines were male although isolated from pooled male and female blood samples. Two cryopreservation methods were developed based upon slow-freezing and aseptic vitrification. Both have shown a similar effectiveness in allowing storage without phenotype drift. Stably expressing lines were obtained by Lipofectamine® mediated transfection of a GFP plasmid. PGCs were subsequently injected in recipient embryos. Persistence of exogenous PGCs in the developing gonad of recipient embryos confirmed that PGCs retain their gonadal colonisation ability, both after long-term culture and after cryopreservation. [less ▲]

Detailed reference viewed: 109 (11 ULg)
Full Text
Peer Reviewed
See detailThe site of administration influences both the type and the magnitude of the immune response induced by DNA vaccine electroporation
Vandermeulen, G; Vanvarenberg, K; De Beuckelaer, A et al

in Vaccine (2015), 33

Detailed reference viewed: 27 (2 ULg)
Full Text
Peer Reviewed
See detailRational Development of an Attenuated Recombinant Cyprinid Herpesvirus 3 Vaccine Using Prokaryotic Mutagenesis and In Vivo Bioluminescent Imaging
Boutier, Maxime ULg; Ronsmans, Maygane ULg; Ouyang, Ping et al

in PLoS Pathogens (2015), 11(2), 1004690

Cyprinid herpesvirus 3 (CyHV 3) is causing severe economic losses worldwide in common and koi carp industries, and a safe and efficacious attenuated vaccine compatible with mass vaccination is needed. We ... [more ▼]

Cyprinid herpesvirus 3 (CyHV 3) is causing severe economic losses worldwide in common and koi carp industries, and a safe and efficacious attenuated vaccine compatible with mass vaccination is needed. We produced single deleted recombinants using prokaryotic mutagenesis. When producing a recombinant lacking open reading frame 134 (ORF134), we unexpectedly obtained a clone with additional deletion of ORF56 and ORF57. This triple deleted recombinant replicated efficiently in vitro and expressed an in vivo safety/efficacy profile compatible with use as an attenuated vaccine. To determine the role of the double ORF56-57 deletion in the phenotype and to improve further the quality of the vaccine candidate, a series of deleted recombinants was produced and tested in vivo. These experiments led to the selection of a double deleted recombinant lacking ORF56 and ORF57 as a vaccine candidate. The safety and efficacy of this strain were studied using an in vivo bioluminescent imaging system (IVIS), qPCR, and histopathological examination, which demonstrated that it enters fish via skin infection similar to the wild type strain. However, compared to the parental wild type strain, the vaccine candidate replicated at lower levels and spread less efficiently to secondary sites of infection. Transmission experiments allowing water contamination with or without additional physical contact between fish demonstrated that the vaccine candidate has a reduced ability to spread from vaccinated fish to naïve sentinel cohabitants. Finally, IVIS analyses demonstrated that the vaccine candidate induces a protective mucosal immune response at the portal of entry. Thus, the present study is the first to report the rational development of a recombinant attenuated vaccine against CyHV 3 for mass vaccination of carp. We also demonstrated the relevance of the CyHV 3 carp model for studying alloherpesvirus transmission and mucosal immunity in teleost skin. [less ▲]

Detailed reference viewed: 57 (11 ULg)
Full Text
Peer Reviewed
See detailCyprinid herpesvirus 3 : an archetype of fish alloherpesviruses
Boutier, Maxime ULg; Ronsmans, Maygane ULg; Rakus, Krzysztof ULg et al

in Advances in Virus Research (2015), 93

Detailed reference viewed: 54 (11 ULg)
Full Text
Peer Reviewed
See detailThe alpha2,3-Sialyltransferase Encoded by Myxoma Virus Is a Virulence Factor that Contributes to Immunosuppression.
Boutard, Berengere; Vankerckhove, Sophie; Markine-Goriaynoff, Nicolas et al

in PloS one (2015), 10(2), 0118806

Myxoma virus (MYXV) induces a lethal disease called Myxomatosis in European rabbits. MYXV is one of the rare viruses that encodes an alpha2,3-sialyltransferase through its M138L gene. In this study, we ... [more ▼]

Myxoma virus (MYXV) induces a lethal disease called Myxomatosis in European rabbits. MYXV is one of the rare viruses that encodes an alpha2,3-sialyltransferase through its M138L gene. In this study, we showed that although the absence of the enzyme was not associated with any in vitro deficit, the M138L deficient strains are highly attenuated in vivo. Indeed, while all rabbits infected with the parental and the revertant strains died within 9 days post-infection from severe myxomatosis, all but one rabbit inoculated with the M138L deficient strains survived the infection. In primary lesions, this resistance to the infection was associated with an increased ability of innate immune cells, mostly neutrophils, to migrate to the site of virus replication at 4 days post-infection. This was followed by the development of a better specific immune response against MYXV. Indeed, at day 9 post-infection, we observed an important proliferation of lymphocytes and an intense congestion of blood vessels in lymph nodes after M138L knockouts infection. Accordingly, in these rabbits, we observed an intense mononuclear cell infiltration throughout the dermis in primary lesions and higher titers of neutralizing antibodies. Finally, this adaptive immune response provided protection to these surviving rabbits against a challenge with the MYXV WT strain. Altogether, these results show that expression of the M138L gene contributes directly or indirectly to immune evasion by MYXV. In the future, these results could help us to better understand the pathogenesis of myxomatosis but also the importance of glycans in regulation of immune responses. [less ▲]

Detailed reference viewed: 39 (10 ULg)
Peer Reviewed
See detailA gammaherpesvirus infection protects against allergic asthma.
Machiels, Bénédicte ULg; Dourcy, Mickael ULg; Sabatel, Catherine ULg et al

Poster (2014, December 12)

The “hygiene hypothesis” proposes that the augmentation of allergic diseases in developed countries could be linked to a reduced exposure to infections during childhood. Surprisingly, the potential ... [more ▼]

The “hygiene hypothesis” proposes that the augmentation of allergic diseases in developed countries could be linked to a reduced exposure to infections during childhood. Surprisingly, the potential protective role of herpesvirus infections against allergy development has never been addressed directly. In this study, we used the Murid herpesvirus 4 (MuHV-4) to study the impact of a persistent gammaherpesvirus infection on the development of House Dust Mites (HDM)-induced allergic asthma. Our results revealed that MuHV-4 infection affects both the sensitization and the challenging phases of HDM-induced airway allergy. In particular, we highlighted that MuHV-4 infection strongly impacts the lung innate immune response. Indeed, while the dendritic cells remained competent to uptake antigens and to migrate to the draining lymph nodes, MuHV-4 infection impaired their ability to trigger HDM sensitization. In the future, these results could allow us to develop strategies to prevent the development of TH2-skewed responses against respiratory allergens. [less ▲]

Detailed reference viewed: 50 (17 ULg)
Full Text
Peer Reviewed
See detailSchistosoma mansoni egg-induced inflammation inhibits γ-herpesvirus replication
Dougall, Annette ULg; Rolot, Marion ULg; Vanderplasschen, Alain ULg et al

Poster (2014, December)

Geographically, S. mansoni overlaps with human γ-herpesvirus infections such as Kaposi’s sarcoma-associated herpesvirus. The strongly regulated Th2-type immune response generated during infection by S ... [more ▼]

Geographically, S. mansoni overlaps with human γ-herpesvirus infections such as Kaposi’s sarcoma-associated herpesvirus. The strongly regulated Th2-type immune response generated during infection by S. mansoni may jeopardize or improve the host’s ability to generate effective immunity against co-infecting pathogens, such as viruses. Here, we have trialled two approaches using murine herpesvirus-4 (MuHV-4). The first used a S. mansoni egg model to induce lung granulomas followed by intranasal infection with the MuHV-4-luc+ recombinant virus. Alternatively, we naturally infected mice with S. mansoni cercariae to induce a systemic Th2-type response and granulomas in the liver and intestine before intranasal MuHV-4-luc+ infection. We observed in both models a significant reduction of MuHV-4 replication in the lungs at day 5 and 7 pi associated with reduced weight loss caused by MuHV-4 infection. These results indicate that helminth induce Th2-type responses could inhibit and protect against viral infection. [less ▲]

Detailed reference viewed: 33 (3 ULg)
Full Text
Peer Reviewed
See detailThe replication characteristics of infectious laryngotracheitis virus (ILTV) in the respiratory and conjunctival mucosa
Reddy, V.R.; Steukers, L.; Li, Y. et al

in Avian Pathology : Journal of the W.V.P.A (2014), 21

Detailed reference viewed: 50 (4 ULg)