Papers by Monique Van Oers
Figure S3. GpSGHV haplotype network in Glossina species: The haplotype network generated based on... more Figure S3. GpSGHV haplotype network in Glossina species: The haplotype network generated based on the ML tree generated based on GpSGHV concatenated sequence of VNTR-1, VNTR-2, ORF9, ORF10 and ORF38. The black dots on the lines represent mutations events between the haplotypes. The different colours represent the Glossina species. Abbreviations; KEN (Kenya), TAN (Tanzania), ETH (Ethiopia), ZIM (Zimbabwe), ZAM (Zambia), UGA (Uganda), SA (South Africa), DRC (Democratic republic of Congo), Ruck. (Ruckomechi), BRI-KALRO (Biotechnology Research institute- Kenya agricultural and livestock research organization), GR (Game reserve). (TIF 1390Â kb)
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Figure S2. Sequence alignment of GpSGHV haplotypes in Glossina species: A). Partial alignment of ... more Figure S2. Sequence alignment of GpSGHV haplotypes in Glossina species: A). Partial alignment of SGHV010 sequences showing the single nucleotide mutations that lead to non-synonymous mutations. Multiple alignment of VNTR-1 (B), VNTR-2 (C), SGHV009 (D), SGHV010 (E) and SGHV038 (F), showing different GpSGHV haplotypes and their phylogenetic relatedness. The positions are based on the reference sequence (GpSGHV-Uga). Abbreviations; Hâ =â Haplotype. (TIF 4096Â kb)
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Figure S1. Analysis of PCR-amplification products on 1.5% agarose gels: Panels A and B show the d... more Figure S1. Analysis of PCR-amplification products on 1.5% agarose gels: Panels A and B show the different band sizes of ITS1 sequences and Wolbachia diagnosis for different Glossina species, respectively. Four samples were analysed to represent each species from different geographical regions. The positive controls were Glossina species from the IPCL laboratory colonies whose identities are known. The sizes of the DNA ladder bands used for both gel images are indicated. Abbreviations: Gp (G. pallidipes), Gmm (G. m. morsitans), Gswy (G. swynnertoni), Gff (G. f. fuscipes), Gbr (G. brevipalpis), Gpp (G. p. palpalis), and Gaus (G. austeni). (TIF 7039Â kb)
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Baculovirus occlusion-derived virus (ODV) infects insect midgut cells under alkaline conditions, ... more Baculovirus occlusion-derived virus (ODV) infects insect midgut cells under alkaline conditions, a process mediated by highly conserved per os infectivity factors (PIFs), P74 (PIF0), PIF1, PIF2, PIF3, PIF4, and PIF5 (ODV-E56). Previously, a multimolecu-lar complex composed of PIF1, PIF2, PIF3, and P74 was identified which was proposed to play an essential role during ODV en-try. Recently, more proteins have been identified that play important roles in ODV oral infectivity, including PIF4, PIF5, and SF58, which might work in concert with previously known PIFs to facilitate ODV infection. In order to understand the ODV entry mechanism, the identification of all components of the PIF complex is crucial. Hence, the aim of this study was to identify additional components of the PIF complex. Coimmunoprecipitation (CoIP) combined with proteomic analysis was used to iden-tify the components of the Autographa californicamultiple nucleopolyhedrovirus (AcMNPV) PIF complex. PIF4 and P95 (AC83) ...
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Viruses
Tsetse flies cause major health and economic problems as they transmit trypanosomes causing sleep... more Tsetse flies cause major health and economic problems as they transmit trypanosomes causing sleeping sickness in humans (Human African Trypanosomosis, HAT) and nagana in animals (African Animal Trypanosomosis, AAT). A solution to control the spread of these flies and their associated diseases is the implementation of the Sterile Insect Technique (SIT). For successful application of SIT, it is important to establish and maintain healthy insect colonies and produce flies with competitive fitness. However, mass production of tsetse is threatened by covert virus infections, such as the Glossina pallidipes salivary gland hypertrophy virus (GpSGHV). This virus infection can switch from a covert asymptomatic to an overt symptomatic state and cause the collapse of an entire fly colony. Although the effects of GpSGHV infections can be mitigated, the presence of other covert viruses threaten tsetse mass production. Here we demonstrated the presence of two single-stranded RNA viruses isolated ...
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Viruses
Multiple enveloped viruses with rod-shaped nucleocapsids have been described, infecting the epith... more Multiple enveloped viruses with rod-shaped nucleocapsids have been described, infecting the epithelial cell nuclei within the hepatopancreas tubules of crustaceans. These bacilliform viruses share the ultrastructural characteristics of nudiviruses, a specific clade of viruses infecting arthropods. Using histology, electron microscopy and high throughput sequencing, we characterise two further bacilliform viruses from aquatic hosts, the brown shrimp (Crangon crangon) and the European shore crab (Carcinus maenas). We assembled the full double stranded, circular DNA genome sequences of these viruses (~113 and 132 kbp, respectively). Comparative genomics and phylogenetic analyses confirm that both belong within the family Nudiviridae but in separate clades representing nudiviruses found in freshwater and marine environments. We show that the three thymidine kinase (tk) genes present in all sequenced nudivirus genomes, thus far, were absent in the Crangon crangon nudivirus, suggesting th...
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Viruses
The genus Flavivirus contains pathogenic vertebrate-infecting flaviviruses (VIFs) and insect-spec... more The genus Flavivirus contains pathogenic vertebrate-infecting flaviviruses (VIFs) and insect-specific flaviviruses (ISF). ISF transmission to vertebrates is inhibited at multiple stages of the cellular infection cycle, via yet to be elucidated specific antiviral responses. The zinc-finger antiviral protein (ZAP) in vertebrate cells can bind CpG dinucleotides in viral RNA, limiting virus replication. Interestingly, the genomes of ISFs contain more CpG dinucleotides compared to VIFs. In this study, we investigated whether ZAP prevents two recently discovered lineage II ISFs, Binjari (BinJV) and Hidden Valley viruses (HVV) from replicating in vertebrate cells. BinJV protein and dsRNA replication intermediates were readily observed in human ZAP knockout cells when cultured at 34 °C. In ZAP-expressing cells, inhibition of the interferon response via interferon response factors 3/7 did not improve BinJV protein expression, whereas treatment with kinase inhibitor C16, known to reduce ZAP’s...
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Viruses
The mitochondrial antiviral-signaling protein (MAVS, also known as VISA, IPS-1, or CARDIF) plays ... more The mitochondrial antiviral-signaling protein (MAVS, also known as VISA, IPS-1, or CARDIF) plays an essential role in the type I interferon (IFN) response and in retinoic acid-inducible gene I (RIG-I) mediated antiviral innate immunity in mammals. In this study, the caprine MAVS gene (caMAVS, 1566 bp) was identified and cloned. The caMAVS shares the highest amino acid similarity (98.1%) with the predicted sheep MAVS. Confocal microscopy analysis of partial deletion mutants of caMAVS revealed that the transmembrane and the so-called Non-Characterized domains are indispensable for intracellular localization to mitochondria. Overexpression of caMAVS in caprine endometrial epithelial cells up-regulated the mRNA levels of caprine interferon-stimulated genes. We concluded that caprine MAVS mediates the activation of the type I IFN pathway. We further demonstrated that both the CARD-like domain and the transmembrane domain of caMAVS were essential for the activation of the IFN-β promotor. ...
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Viruses
Baculovirus expression vectors are successfully used for the commercial production of complex (gl... more Baculovirus expression vectors are successfully used for the commercial production of complex (glyco)proteins in eukaryotic cells. The genome engineering of single-copy baculovirus infectious clones (bacmids) in E. coli has been valuable in the study of baculovirus biology, but bacmids are not yet widely applied as expression vectors. An important limitation of first-generation bacmids for large-scale protein production is the rapid loss of gene of interest (GOI) expression. The instability is caused by the mini-F replicon in the bacmid backbone, which is non-essential for baculovirus replication in insect cells, and carries the adjacent GOI in between attTn7 transposition sites. In this paper, we test the hypothesis that relocation of the attTn7 transgene insertion site away from the mini-F replicon prevents deletion of the GOI, thereby resulting in higher and prolonged recombinant protein expression levels. We applied lambda red genome engineering combined with SacB counterselecti...
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Viruses
Zika virus (ZIKV) is a mosquito-borne pathogen that caused a large outbreak in the Americas in 20... more Zika virus (ZIKV) is a mosquito-borne pathogen that caused a large outbreak in the Americas in 2015 and 2016. The virus is currently present in tropical areas around the globe and can cause severe disease in humans, including Guillain-Barré syndrome and congenital microcephaly. The tropical yellow fever mosquito, Aedes aegypti, is the main vector in the urban transmission cycles of ZIKV. The discovery of ZIKV in wild-caught Culex mosquitoes and the ability of Culex quinquefasciatus mosquitoes to transmit ZIKV in the laboratory raised the question of whether the common house mosquito Culex pipiens, which is abundantly present in temperate regions in North America, Asia and Europe, could also be involved in ZIKV transmission. In this study, we investigated the vector competence of Cx. pipiens (biotypes molestus and pipiens) from the Netherlands for ZIKV, using Usutu virus as a control. After an infectious blood meal containing ZIKV, none of the tested mosquitoes accumulated ZIKV in th...
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Viruses
The genus Orthobunyavirus (family Peribunyaviridae, order Bunyavirales) comprises over 170 named ... more The genus Orthobunyavirus (family Peribunyaviridae, order Bunyavirales) comprises over 170 named mosquito- and midge-borne viruses, several of which cause severe disease in animals or humans. Their three-segmented genomes enable reassortment with related viruses, which may result in novel viruses with altered host or tissue tropism and virulence. One such reassortant, Schmallenberg virus (SBV), emerged in north-western Europe in 2011. Shuni virus (SHUV) is an orthobunyavirus related to SBV that is associated with neurological disease in horses in southern Africa and recently caused an outbreak manifesting with neurological disease and birth defects among ruminants in Israel. The zoonotic potential of SHUV was recently underscored by its association with neurological disease in humans. We here report a reverse genetics system for SHUV and provide first evidence that the non-structural (NSs) protein of SHUV functions as an antagonist of host innate immune responses. We furthermore rep...
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Insects
In this special issue the focus is on the factors and (molecular) mechanisms that determine the t... more In this special issue the focus is on the factors and (molecular) mechanisms that determine the transmission efficiency of a variety of insect pathogens in a number of insect hosts. In this editorial, we summarize the main findings of the twelve papers in this special issue and conclude that much more needs to be learned for an in-depth understanding of pathogen transmission in field and cultured insect populations. Analyses of mutual interactions between pathogens or between endosymbionts and pathogens, aspects rather under-represented in the scientific literature, are described in a number of contributions to this special issue.
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Viruses
Rabbit haemorrhagic disease virus (RHDV) type 2 (GI.2/RHDV2/b) is an emerging pathogen in wild ra... more Rabbit haemorrhagic disease virus (RHDV) type 2 (GI.2/RHDV2/b) is an emerging pathogen in wild rabbits and in domestic rabbits vaccinated against RHDV (GI.1). Here we report the genome sequence of a contemporary RHDV2 isolate from the Netherlands and investigate the immunogenicity of virus-like particles (VLPs) produced in insect cells. RHDV2 RNA was isolated from the liver of a naturally infected wild rabbit and the complete viral genome sequence was assembled from sequenced RT-PCR products. Phylogenetic analysis based on the VP60 capsid gene demonstrated that the RHDV2 NL2016 isolate clustered with other contemporary RHDV2 strains. The VP60 gene was cloned in a baculovirus expression vector to produce VLPs in Sf9 insect cells. Density-gradient purified RHDV2 VLPs were visualized by transmission electron microscopy as spherical particles of around 30 nm in diameter with a morphology resembling authentic RHDV. Immunization of rabbits with RHDV2 VLPs resulted in high production of se...
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Viruses
Small RNA mediated responses are essential for antiviral defence in mosquitoes, however, they app... more Small RNA mediated responses are essential for antiviral defence in mosquitoes, however, they appear to differ per virus-vector combination. To further investigate the diversity of small RNA responses against viruses in mosquitoes, we applied a small RNA deep sequencing approach on five mosquito cell lines: Culex tarsalis CT cells, Aedes albopictus U4.4 and C6/36 cells, Ae. aegypti Aag2 cells (cleared from cell fusing agent virus and Culex Y virus (CYV) by repetitive dsRNA transfections) and Ae. pseudoscutellaris AP-61 cells. De novo assembly of small RNAs revealed the presence of Phasi Charoen-like virus (PCLV), Calbertado virus, Flock House virus and a novel narnavirus in CT cells, CYV in U4.4 cells, and PCLV in Aag2 cells, whereas no insect-specific viruses (ISVs) were detected in C6/36 and AP-61 cells. Next, we investigated the small RNA responses to the identified ISVs and to acute infection with the arthropod-borne West Nile virus (WNV). We demonstrate that AP-61 and C6/36 cel...
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Journal of General Virology
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Frontiers in microbiology, 2018
The salivary gland hypertrophy virus (GpSGHV) infects tsetse flies predominantly asymptomatically... more The salivary gland hypertrophy virus (GpSGHV) infects tsetse flies predominantly asymptomatically and occasionally symptomatically. Symptomatic infections are characterized by overt salivary gland hypertrophy (SGH) in mass reared tsetse flies, which causes reproductive dysfunctions and colony collapse, thus hindering tsetse control via sterile insect technique (SIT). Asymptomatic infections have no apparent cost to the fly's fitness. Here, small RNAs were sequenced and profiles in asymptomatically and symptomatically infected flies determined. Thirty-eight host-encoded microRNAs (miRNAs) were present in both the asymptomatic and symptomatic fly profiles, while nine host miRNAs were expressed specifically in asymptomatic flies versus 10 in symptomatic flies. Of the shared 38 miRNAs, 15 were differentially expressed when comparing asymptomatic with symptomatic flies. The most up-regulated host miRNAs in symptomatic flies was predicted to target immune-related mRNAs of the host. Si...
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BMC Microbiology
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BMC Microbiology
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Insects
Baculoviruses are arthropod-specific large DNA viruses that orally infect the larvae of lepidopte... more Baculoviruses are arthropod-specific large DNA viruses that orally infect the larvae of lepidopteran, hymenopteran and dipteran insect species. These larvae become infected when they eat a food source that is contaminated with viral occlusion bodies (OBs). These OBs contain occlusion-derived viruses (ODVs), which are released upon ingestion of the OBs and infect the endothelial midgut cells. At least nine different ODV envelope proteins are essential for this oral infectivity and these are denoted per os infectivity factors (PIFs). Seven of these PIFs form a complex, consisting of PIF1, 2, 3 and 4 that form a stable core complex and PIF0 (P74), PIF6 and PIF8 (P95) that associate with this complex with lower affinity than the core components. The existence of a PIF complex and the fact that the pif genes are conserved in baculovirus genomes suggests that PIF-proteins cooperatively mediate oral infectivity rather than as individual functional entities. This review therefore discusses ...
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Frontiers in Microbiology, 2018
The Glossina pallidipes salivary gland hypertrophy virus (GpSGHV) infects tsetse flies predominan... more The Glossina pallidipes salivary gland hypertrophy virus (GpSGHV) infects tsetse flies predominantly asymptomatically and occasionally symptomatically. Symptomatic infections are characterized by overt salivary gland hypertrophy (SGH) in mass reared tsetse flies, which causes reproductive dysfunctions and colony collapse, thus hindering tsetse control via sterile insect technique (SIT). Asymptomatic infections have no apparent cost to the fly's fitness. Here, small RNAs were sequenced and profiles in asymptomatically and symptomatically infected G. pallidipes flies determined. Thirty-eight host-encoded microRNAs (miRNAs) were present in both the asymptomatic and symptomatic fly profiles, while nine host miRNAs were expressed specifically in asymptomatic flies versus 10 in symptomatic flies. Of the shared 38 miRNAs, 15 were differentially expressed when comparing asymptomatic with symptomatic flies. The most up-regulated host miRNAs in symptomatic flies was predicted to target im...
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Papers by Monique Van Oers