20 results on '"Berends, Alinda J."'
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2. Infectious bronchitis virus Mass-type (GI-1) and QX-like (GI-19) genotyping and vaccine differentiation using SYBR green RT-qPCR paired with melting curve analysis
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Laconi, Andrea, Berends, Alinda J., de Laat, Esther C.H., Urselmann, Tara A.P.M.P., and Verheije, Hélène M.
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- 2020
- Full Text
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3. miRNA repertoire and host immune factor regulation upon avian coronavirus infection in eggs
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Kemp, Vera, Laconi, Andrea, Cocciolo, Giulio, Berends, Alinda J., Breit, Timo M., and Verheije, M. Hélène
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- 2020
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4. Recombinant live attenuated avian coronavirus vaccines with deletions in the accessory genes 3ab and/or 5ab protect against infectious bronchitis in chickens
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van Beurden, Steven J., Berends, Alinda J., Krämer-Kühl, Annika, Spekreijse, Dieuwertje, Chenard, Gilles, Philipp, Hans-Christian, Mundt, Egbert, Rottier, Peter J.M., and Verheije, M. Hélène
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- 2018
- Full Text
- View/download PDF
5. Epithelial-derived galectin-9 containing exosomes contribute to the immunomodulatory effects promoted by 2’-fucosyllactose and short-chain galacto- and long-chain fructo-oligosaccharides
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Ayechu-Muruzabal, Veronica, de Boer, Merel, Blokhuis, Bart, Berends, Alinda J., Garssen, Johan, Kraneveld, Aletta D., van’t Land, Belinda, Willemsen, Linette E.M., Afd Pharmacology, Pharmacology, Afd Pharmacology, and Pharmacology
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galectins ,Immunology ,exosome ,mucosal immunity ,non-digestible oligosaccharide ,Immunology and Allergy ,intestinal-epithelial cells - Abstract
IntroductionEarly life exposure to non-digestible oligosaccharides (NDO) or microbial components is known to affect immune development. NDO in combination with a TLR9 agonist mimicking bacterial triggers (CpG) promoted the secretion of galectins through unknown pathways. We aimed to study the contribution of exosomes in epithelial galectin secretion and subsequent immunoregulation upon exposure to a mixture of NDO by inhibiting exosome biogenesis.MethodsHuman intestinal epithelial cells (IEC) (FHs 74 Int or HT-29) were apically exposed to 2’-fucosyllactose (2’FL) and short-chain galacto- and long-chain fructo-oligosaccharides (GF), alone or with CpG. Basolaterally, non-activated or αCD3/CD28-activated peripheral blood mononuclear cells (PBMC) were added. After 24 h incubation, IEC were washed and incubated in fresh medium to analyze epithelial-derived galectin secretion. Additionally, before exposure to NDO and CpG, IEC were exposed to GW4869 to inhibit exosome biogenesis. After 24 h of incubation, IEC were washed and incubated for additional 24 h in the presence of GW4869, after which epithelial-derived galectin secretion was studied. Also, epithelial-derived exosomes were isolated to study the presence of galectins within the exosomes.ResultsCompared to CpG alone, exposure to 2’FL/GF mixture and CpG, significantly enhanced Th1-type IFNγ, and regulatory IEC-derived galectin-9 secretion in the HT-29/PBMC model. Similarly, in the FHs 74 Int/PBMC co-culture, 2’FL/GF induced immunomodulatory effects in the absence of CpG. Interestingly, galectin-9 and -4 were present in CD63-expressing exosomes isolated from HT-29 supernatants after IEC/PBMC co-culture. Exposure to GW4869 suppressed 2’FL/GF and CpG induced epithelial-derived galectin-9 secretion, which subsequently prevented the rise in IL-10 and reduction in IL-13 secretion observed in the HT-29/PBMC co-culture model upon exposure to 2’FL/GF and CpG.DiscussionExposure to 2’FL/GF and CpG or 2’FL/GF promoted Th1-type regulatory effects in HT-29/PBMC or FHs 74 Int/PBMC co-culture respectively, while Th2-type IL-13 was reduced in association with increased galectin-9 release. Galectin-9 and -4 were present in exosomes from HT-29 and the inhibition of exosome biogenesis inhibited epithelial-derived galectin secretion. This, also affected immunomodulatory effects in IEC/PBMC co-culture suggesting a key role of galectin expressing IEC-derived exosomes in the mucosal immune regulation induced by NDO.
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- 2022
6. Human Milk Oligosaccharide 2′-Fucosyllactose Modulates Local Viral Immune Defense by Supporting the Regulatory Functions of Intestinal Epithelial and Immune Cells
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Ayechu-Muruzabal, Veronica, Poelmann, Bente, Berends, Alinda J., Kettelarij, Nienke, Garssen, Johan, van’t Land, Belinda, Willemsen, Linette E.M., Afd Pharmacology, Sub General Pharmacology, Pharmacology, Afd Pharmacology, Sub General Pharmacology, and Pharmacology
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Milk, Human ,Galectin 4 ,Interleukin-8 ,Organic Chemistry ,Oligosaccharides ,Epithelial Cells ,C [poly I] ,Dendritic Cells ,General Medicine ,viral infection ,poly I:C ,2′-fucosyllactose ,host-defense ,moDC ,T-cell ,Catalysis ,Computer Science Applications ,Inorganic Chemistry ,Poly I ,Humans ,Physical and Theoretical Chemistry ,Trisaccharides ,Molecular Biology ,Spectroscopy - Abstract
Human milk contains bioactive components that provide protection against viral infections in early life. In particular, intestinal epithelial cells (IEC) have key regulatory roles in the prevention of enteric viral infections. Here we established an in vitro model to study the modulation of host responses against enteric viruses mimicked by poly I:C (pIC). The effects of 2′-fucosyllactose (2′FL), abundantly present in human milk, were studied on IEC and/or innate immune cells, and the subsequent functional response of the adaptive immune cells. IEC were pre-incubated with 2′FL and stimulated with naked or Lyovec™-complexed pIC (LV-pIC). Additionally, monocyte-derived dendritic cells (moDC) alone or in co-culture with IEC were stimulated with LV-pIC. Then, conditioned-moDC were co-cultured with naïve CD4+ T helper (Th)-cells. IEC stimulation with naked or LV-pIC promoted pro-inflammatory IL-8, CCL20, GROα and CXCL10 cytokine secretion. However, only exposure to LV-pIC additionally induced IFNβ, IFNλ1 and CCL5 secretion. Pre-incubation with 2′FL further increased pIC induced CCL20 secretion and LV-pIC induced CXCL10 secretion. LV-pIC-exposed IEC/moDC and moDC cultures showed increased secretion of IL-8, GROα, IFNλ1 and CXCL10, and in the presence of 2′FL galectin-4 and -9 were increased. The LV-pIC-exposed moDC showed a more pronounced secretion of CCL20, CXCL10 and CCL5. The moDC from IEC/moDC cultures did not drive T-cell development in moDC/T-cell cultures, while moDC directly exposed to LV-pIC secreted Th1 driving IL-12p70 and promoted IFNγ secretion by Th-cells. Hereby, a novel intestinal model was established to study mucosal host-defense upon a viral trigger. IEC may support intestinal homeostasis, regulating local viral defense which may be modulated by 2′FL. These results provide insights regarding the protective capacity of human milk components in early life.
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- 2022
7. Wild and domestic animals variably display Neu5Ac and Neu5Gc sialic acids
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Nemanichvili, Nikoloz, Spruit, Cindy M, Berends, Alinda J, Gröne, Andrea, Rijks, Jolianne M, Verheije, Monique H, de Vries, Robert P, Pharmacology, dPB I&I, Afd Chemical Biology and Drug Discovery, Chemical Biology and Drug Discovery, Afd Pharmacology, dI&I I&I-1, VPDC pathologie, dPB CR, Pharmacology, dPB I&I, Afd Chemical Biology and Drug Discovery, Chemical Biology and Drug Discovery, Afd Pharmacology, dI&I I&I-1, VPDC pathologie, and dPB CR
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tissue microarray ,Swine ,wildlife ,Ferrets ,host tropism ,Biochemistry ,N-Acetylneuraminic Acid ,Dogs ,Polysaccharides ,sialic acid ,Animals, Domestic ,Lectins ,Viruses ,Sialic Acids ,Animals ,Humans ,Neuraminic Acids ,Horses ,Glycolipids ,influenza A - Abstract
Sialic acids are used as a receptor by several viruses and variations in the linkage type or C-5 modifications affect the binding properties. A species barrier for multiple viruses is present due to α2,3- or α2,6-linked sialic acids. The C-5 position of the sialic acid can be modified to form N-acetylneuraminic acid (Neu5Ac) or N-glycolylneuraminic acid (Neu5Gc), which acts as a determinant for host susceptibility for pathogens such as influenza A virus, rotavirus, and transmissible gastroenteritis coronavirus. Neu5Gc is present in most mammals such as pigs and horses but is absent in humans, ferrets, and dogs. However, little is known about C-5 content in wildlife species or how many C-5 modified sialic acids are present on N-linked glycans or glycolipids. Using our previously developed tissue microarray system, we investigated how 2 different lectins specific for Neu5Gc can result in varying detection levels of Neu5Gc glycans. We used these lectins to map Neu5Gc content in wild Suidae, Cervidae, tigers, and European hedgehogs. We show that Neu5Gc content is highly variable among different species. Furthermore, the removal of N-linked glycans reduces the binding of both Neu5Gc lectins while retention of glycolipids by omitting methanol treatment of tissues increases lectin binding. These findings highlight the importance of using multiple Neu5Gc lectins as the rich variety in which Neu5Gc is displayed can hardly be detected by a single lectin.
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- 2022
8. Antiviral activity of selected cathelicidins against infectious bronchitis virus
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Verheije, M. Hélène, Coorens, Maarten, Weerts, Erik A.W.S., Berends, Alinda J., van Harten, Roel M., Angel, Marloes, Kooij, Jannetje, Ordonez, Soledad R., van Beurden, Steven J., van Dijk, Albert, Haagsman, Henk P., Veldhuizen, Edwin J.A., dPB I&I, VP pathologie, LS Moleculaire Afweer, VPDC pathologie, Afd Pharmacology, dI&I I&I-1, Moleculaire afweer, dI&I I&I-3, LS Pathologie, Immunologie, dPB I&I, VP pathologie, LS Moleculaire Afweer, VPDC pathologie, Afd Pharmacology, dI&I I&I-1, Moleculaire afweer, dI&I I&I-3, LS Pathologie, and Immunologie
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Innate immune system ,Chemistry ,host defense peptides ,Organic Chemistry ,coronavirus ,Biophysics ,Infectious bronchitis virus ,medicine.disease_cause ,Virology ,Biochemistry ,Cathelicidins ,Biomaterials ,medicine ,innate immunity ,Coronavirus - Abstract
Avian infectious bronchitis (IB) is a highly contagious disease caused by infectious bronchitis virus (IBV), a coronavirus of domestic fowl. IB is a major concern in the poultry industry, causing worldwide economic losses through decreased egg production and quality and by increasing the chicken's susceptibility for secondary bacterial infections, particularly Escherichia coli. In this study, the anti-IBV activity of cathelicidins, small antimicrobial peptides of the innate immune system was investigated. The cell culture adapted (nonvirulent) IBV strain Beaudette was effectively inhibited by the human cathelicidin LL-37 in bovine hamster kidney-21 cells at nontoxic concentrations. The peptide needed to be present during virus inoculation to effectively inhibit the infection of IBV-Beaudette, indicating that LL-37 likely bound viral particles. However, no clear morphological changes in the IBV virion upon binding were observed by electron microscopy. In this cell culture model, chicken cathelicidins (CATH1-3) were inactive against IBV-Beaudette. In contrast, in multicellular infection models using the virulent IBV-M41 strain the activities of human and chicken cathelicidins were different. In particular, upon inoculation of 10-day-old embryonic eggs with IBV-M41, CATH-2 reduced the viral load to a higher extend than LL-37. Similarly, viral infection of chicken tracheal organ cultures with IBV-M41 was significantly reduced in the presence of CATH-2 but not LL-37. These results indicate a potential antiviral role for CATH-2 upon IBV infection in vivo.
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- 2022
9. Evidence of Hearing Loss and Unrelated Toxoplasmosis in a Free-Ranging Harbour Porpoise (Phocoena phocoena)
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Morell, Maria, Ijsseldijk, Lonneke L., Berends, Alinda J., Gröne, Andrea, Siebert, Ursula, Raverty, Stephen A., Shadwick, Robert E., Kik, Marja J.L., VPDC pathologie, dPB CR, Afd Pharmacology, dI&I I&I-1, dPB I&I, and VP pathologie
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inner ear ,Noise‐induced hearing loss ,Veterinary medicine ,encephalitis ,Toxoplasma gondii ,live stranding ,veterinary(all) ,hair cell ,noise-induced hearing loss ,Post‐mortem examination ,Live stranding ,QL1-991 ,Inner ear ,SF600-1100 ,otorhinolaryngologic diseases ,Encephalitis ,Animal Science and Zoology ,North Sea ,Hair cell ,post-mortem examination ,Zoology - Abstract
Evidence of hearing impairment was identified in a harbour porpoise (Phocoena phocoena) on the basis of scanning electron microscopy. In addition, based on histopathology and immunohistochemistry, there were signs of unrelated cerebral toxoplasmosis. The six-year old individual live stranded on the Dutch coast at Domburg in 2016 and died a few hours later. The most significant gross lesion was multifocal necrosis and haemorrhage of the cerebrum. Histopathology of the brain revealed extensive necrosis and haemorrhage in the cerebrum with multifocal accumulations of degenerated neutrophils, lymphocytes and macrophages, and perivascular lymphocytic cuffing. The diagnosis of cerebral toxoplasmosis was confirmed by positive staining of protozoa with anti-Toxoplasma gondii antibodies. Tachyzoites were not observed histologically in any of the examined tissues. Ultrastructural evaluation of the inner ear revealed evidence of scattered loss of outer hair cells in a 290 µm long segment of the apical turn of the cochlea, and in a focal region of ~ 1.5 mm from the apex of the cochlea, which was compatible with noise-induced hearing loss. This is the first case of concurrent presumptive noise-induced hearing loss and toxoplasmosis in a free-ranging harbour porpoise from the North Sea.
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- 2021
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10. Tissue Microarrays to Visualize Influenza D Attachment to Host Receptors in the Respiratory Tract of Farm Animals
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Nemanichvili, Nikoloz, Berends, Alinda J, Wubbolts, Richard W, Gröne, Andrea, Rijks, Jolianne M, de Vries, Robert P, Verheije, Monique H, VP pathologie, dI&I I&I-1, dPB I&I, Celbiologie, IOV CCB, dB&C I&I, dPB CR, VPDC pathologie, Afd Chemical Biology and Drug Discovery, Pharmacology, Chemical Biology and Drug Discovery, VP pathologie, dI&I I&I-1, dPB I&I, Celbiologie, IOV CCB, dB&C I&I, dPB CR, VPDC pathologie, Afd Chemical Biology and Drug Discovery, Pharmacology, and Chemical Biology and Drug Discovery
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0301 basic medicine ,Farm animals ,Hemagglutination ,Swine ,030106 microbiology ,Respiratory System ,lcsh:QR1-502 ,Host tropism ,Hemagglutinins, Viral ,Virus Attachment ,Biology ,Influenza D ,Virus ,lcsh:Microbiology ,Article ,Microbiology ,Tissue microarray ,03 medical and health sciences ,9-O-acetylated sialic acid ,Virology ,medicine ,Animals ,Horses ,Receptor ,Tropism ,Sheep ,Host Microbial Interactions ,Goats ,Epithelium ,Recombinant Proteins ,Viral Tropism ,030104 developmental biology ,medicine.anatomical_structure ,Infectious Diseases ,Tissue Array Analysis ,Animals, Domestic ,Tissue tropism ,biology.protein ,Sialic Acids ,Cattle ,Antibody ,Thogotovirus ,Viral Fusion Proteins - Abstract
The trimeric hemagglutinin-esterase fusion protein (HEF) of influenza D virus (IDV) binds 9-O-acetylated sialic acid receptors, which are expressed in various host species. While cattle are the main reservoir for IDV, the viral genome has also been detected in domestic pigs. In addition, antibodies against IDV have been detected in other farm animals such as sheep, goats, and horses, and even in farmers working with IDV positive animals. Viruses belonging to various IDV clades circulate, but little is known about their differences in host and tissue tropism. Here we used recombinantly produced HEF proteins (HEF S57A) from the major clades D/Oklahoma (D/OK) and D/Oklahoma/660 (D/660) to study their host and tissue tropism and receptor interactions. To this end, we developed tissue microarrays (TMA) composed of respiratory tissues from various farm animals including cattle, domestic pigs, sheep, goats, and horses. Protein histochemical staining of farm animal respiratory tissue-microarrays with HEF proteins showed that cattle have receptors present over the entire respiratory tract while receptors are only present in the nasal and pharyngeal epithelium of pigs, sheep, goats, and horses. No differences in tropism for tissues and animals were observed between clades, while hemagglutination assays showed that D/OK has a 2-fold higher binding affinity than D/660 for receptors on red blood cells. The removal of O-acetylation from receptors via saponification treatment confirmed that receptor-binding of both clades was dependent on O-acetylated sialic acids.
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- 2021
11. Three Amino Acid Changes In Avian Coronavirus Spike Protein Allows Binding To Kidney Tissue
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Bouwman, Kim M, Parsons, Lisa M, Berends, Alinda J, de Vries, Robert P, Cipollo, John F, Verheije, Monique H, Chemical Biology and Drug Discovery, Afd Chemical Biology and Drug Discovery, dPB I&I, LS Pathologie, and dI&I I&I-1
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infectious bronchitis virus ,coronavirus ,receptors ,receptor-binding domain ,spike protein ,virus-host interactions - Abstract
Infectious bronchitis virus (IBV) infects ciliated epithelial cells in the chicken respiratory tract. While some IBV strains replicate locally, others can disseminate to various organs, including the kidney. Here we elucidate the determinants for kidney tropism by studying interactions between the receptor binding domain (RBD) of the viral attachment protein spike from two IBV strains with different tropisms. Recombinantly produced RBDs from the nephropathogenic IBV strain QX and from the non-nephropathogenic strain M41 bound to the epithelial cells of the trachea. In contrast, only QX-RBD binds more extensively to cells of the digestive tract, urogenital tract, and kidneys. While removal of sialic acids from tissues prevented binding of all proteins to all tissues, binding of QX-RBD to trachea and kidney could not be blocked by pre-incubation with synthetic alpha-2,3-linked sialic acids. The lack of binding of QX-RBD to a previously identified IBV-M41 receptor was confirmed by ELISA, demonstrating that tissue binding of QX-RBD is dependent on a different sialylated glycan receptor. Using chimeric RBD proteins, we discovered that the region encompassing amino acids 99-159 of QX-RBD was required to establish kidney binding. In particular, QX-RBD amino acids 110-112 (KIP) were sufficient to render IBV-M41 with the ability to bind to kidney, while the reciprocal mutations in IBV-QX abolished kidney binding completely. Structural analysis of both RBDs suggests that the receptor binding site for QX is located at a different location on the spike than that of M41.Importance: Infectious bronchitis virus is the causative agent of Infectious bronchitis in chickens. Upon infection of chicken flocks, the poultry industry faces substantial economic losses by diminished egg quality and increased morbidity and mortality of infected animals. While all IBV strains infect the chicken respiratory tract via the ciliated epithelial layer of the trachea, some strains can also replicate in the kidneys, dividing IBV in two pathotypes: non-nephropathogenic (example IBV-M41) and nephropathogenic viruses (including IBV-QX). Here we set out to identify the determinants for the extended nephropathogenic tropism of IBV-QX. Our data reveal that each pathotype makes use of a different sialylated glycan ligand, with binding sites on opposite sides of the attachment protein. This knowledge should facilitate the design of antivirals to prevent coronavirus infections in the field.
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- 2020
12. A reverse genetics system for avian coronavirus infectious bronchitis virus based on targeted RNA recombination
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van Beurden, Steven J, Berends, Alinda J, Krämer-Kühl, Annika, Spekreijse, Dieuwertje, Chénard, Gilles, Philipp, Hans-Christian, Mundt, Egbert, Rottier, Peter J M, Verheije, M Hélène, LS Virologie, dI&I I&I-1, dPB I&I, LS Virologie, dI&I I&I-1, and dPB I&I
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0301 basic medicine ,Embryonated eggs ,animal structures ,030106 microbiology ,Infectious bronchitis virus ,medicine.disease_cause ,Recombinant virus ,Poultry ,Cell Line ,Vaccine development ,lcsh:Infectious and parasitic diseases ,03 medical and health sciences ,Mice ,Mouse hepatitis virus ,Virology ,medicine ,Animals ,lcsh:RC109-216 ,Gene ,Coronavirus ,Targeted RNA recombination ,Recombination, Genetic ,biology ,Research ,Avian coronavirus ,Embryonated ,RNA ,Avian infectious bronchitis ,biology.organism_classification ,Chicken ,Reverse genetics ,Reverse Genetics ,030104 developmental biology ,Infectious Diseases ,Reverse genetics system ,Gene Targeting ,embryonic structures ,RNA, Viral ,Chickens - Abstract
Background Avian coronavirus infectious bronchitis virus (IBV) is a respiratory pathogen of chickens that causes severe economic losses in the poultry industry worldwide. Major advances in the study of the molecular biology of IBV have resulted from the development of reverse genetics systems for the highly attenuated, cell culture-adapted, IBV strain Beaudette. However, most IBV strains, amongst them virulent field isolates, can only be propagated in embryonated chicken eggs, and not in continuous cell lines. Methods We established a reverse genetics system for the IBV strain H52, based on targeted RNA recombination in a two-step process. First, a genomic and a chimeric synthetic, modified IBV RNA were co-transfected into non-susceptible cells to generate a recombinant chimeric murinized (m) IBV intermediate (mIBV). Herein, the genomic part coding for the spike glycoprotein ectodomain was replaced by that of the coronavirus mouse hepatitis virus (MHV), allowing for the selection and propagation of recombinant mIBV in murine cells. In the second step, mIBV was used as the recipient. To this end a recombination with synthetic RNA comprising the 3′-end of the IBV genome was performed by introducing the complete IBV spike gene, allowing for the rescue and selection of candidate recombinants in embryonated chicken eggs. Results Targeted RNA recombination allowed for the modification of the 3′-end of the IBV genome, encoding all structural and accessory genes. A wild-type recombinant IBV was constructed, containing several synonymous marker mutations. The in ovo growth kinetics and in vivo characteristics of the recombinant virus were similar to those of the parental IBV strain H52. Conclusions Targeted RNA recombination allows for the generation of recombinant IBV strains that are not able to infect and propagate in continuous cell lines. The ability to introduce specific mutations holds promise for the development of rationally designed live-attenuated IBV vaccines and for studies into the biology of IBV in general. Electronic supplementary material The online version of this article (doi:10.1186/s12985-017-0775-8) contains supplementary material, which is available to authorized users.
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- 2017
13. Deletion of accessory genes 3a, 3b, 5a or 5b from avian coronavirus infectious bronchitis virus induces an attenuated phenotype both in vitro and in vivo
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Laconi, Andrea, van Beurden, Steven J, Berends, Alinda J, Krämer-Kühl, Annika, Jansen, Christine A, Spekreijse, Dieuwertje, Chénard, Gilles, Philipp, Hans-Christian, Mundt, Egbert, Rottier, Peter J M, Hélène Verheije, M, dI&I I&I-1, LS Pathologie, dPB I&I, dI&I RA-I&I I&I, LS Immunologie, LS GZ Landbouwhuisdieren, LS Virologie, dI&I I&I-1, LS Pathologie, dPB I&I, dI&I RA-I&I I&I, LS Immunologie, LS GZ Landbouwhuisdieren, and LS Virologie
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0301 basic medicine ,infectious bronchitis virus ,animal structures ,chicken ,Accessory genes ,Infectious bronchitis virus ,030106 microbiology ,accessory genes ,Biology ,In ovo ,medicine.disease_cause ,law.invention ,Accessory proteins ,03 medical and health sciences ,law ,Interferon ,Virology ,medicine ,Coronavirus ,live attenuated virus ,Attenuated vaccine ,Live attenuated virus ,Embryonated ,Avian infectious bronchitis ,biology.organism_classification ,Chicken ,030104 developmental biology ,accessory proteins ,embryonic structures ,Recombinant DNA ,medicine.drug ,Research Article - Abstract
Avian coronavirus infectious bronchitis virus (IBV) infects domestic fowl, resulting in respiratory disease and causing serious losses in unprotected birds. Its control is mainly achieved by using live attenuated vaccines. Here we explored the possibilities for rationally attenuating IBV to improve our knowledge regarding the function of IBV accessory proteins and for the development of next-generation vaccines with the recently established reverse genetic system for IBV H52 based on targeted RNA recombination and selection of recombinant viruses in embryonated eggs. To this aim, we selectively removed accessory genes 3a, 3b, 5a and 5b individually, and rescued the resulting recombinant (r) rIBV-Δ3a, rIBV-Δ3b, rIBV-Δ5a and rIBV-Δ5b. In vitro inoculation of chicken embryo kidney cells with recombinant and wild-type viruses demonstrated that the accessory protein 5b is involved in the delayed activation of the interferon response of the host after IBV infection. Embryo mortality after the inoculation of 8-day-old embryonated chicken eggs with recombinant and wild-type viruses showed that rIBV-Δ3b, rIBV-Δ5a and rIBV-Δ5b had an attenuated phenotype in ovo, with reduced titres at 6 h p.i. and 12 h p.i. for all viruses, while growing to the same titre as wild-type rIBV at 48 h p.i. When administered to 1-day-old chickens, rIBV-Δ3a, rIBV-Δ3b, rIBV-Δ5a and rIBV-Δ5b showed reduced ciliostasis in comparison to the wild-type viruses. In conclusion, individual deletion of accessory genes in IBV H52 resulted in mutant viruses with an attenuated phenotype.
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- 2018
14. Influenza D binding properties vary amongst the two major virus clades and wildlife species.
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Nemanichvili, Nikoloz, Berends, Alinda J., Tomris, Ilhan, Barnard, Karen N., Parrish, Colin R., Gröne, Andrea, Rijks, Jolianne M., Verheije, Monique H., and de Vries, Robert P.
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DOMESTIC animals , *WATER buffalo , *SENDAI virus , *CELL receptors , *VIRAL tropism , *WILD boar , *SWINE , *INFLUENZA - Abstract
• Influenza D has a wide host tropism in both domestic and wild animals despite a very specific receptor preference. • Differences in tissue tropism exist between the two major influenza D clades in water buffalo, Asian elephant, and hedgehog. • Despite positive serosurveillance data, wild Suidae and Cervidea do not express influenza D receptors. • Influenza D has potential to spread from domesticated animals to wildlife and vice versa. The influenza D virus (IDV) uses a trimeric hemagglutinin-esterase fusion protein (HEF) for attachment to 9-O-acetylated sialic acid receptors on the cell surface of host species. So far research has revealed that farm animals such as cattle, domestic pigs, goats, sheep and horses contain the necessary receptors on the epithelial surface of the respiratory tract to accommodate binding of the IDV HEF protein of both worldwide clades D/Oklahoma (D/OK) and D/Oklahoma/660 (D/660). More recently, seroprevalence studies have identified IDV-seropositive wildlife such as wild boar, deer, dromedaries, and small ruminants. However, no research has thus far been conducted in wildlife to reveal the distribution of acetylated sialic acid receptors that accommodate binding of IDV. Using our previously developed tissue microarray (TMA) system, we developed TMAs containing respiratory tissues of various wild and domestic species including wild boar, deer, dromedary, springbok, water buffalo, tiger, hedgehog, and Asian elephant. Protein histochemical staining of these TMAs with HEF proteins showed no receptor binding for wild Suidae, Cervidae and tiger. However, receptors were present in dromedary, springbok, water buffalo, Asian elephant, and hedgehog. In contrast to previously tested farm animals, a difference in host tropism was observed between the D/OK and D/660 clade HEF proteins in Asian elephant, and water buffalo. These results show that IDV can attach to the respiratory tract of wildlife which might facilitate transmission of IDV between wildlife and domestic animals. [ABSTRACT FROM AUTHOR]
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- 2022
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15. Infectious bronchitis virus Mass-type (GI-1) and QX-like (GI-19) genotyping and vaccine differentiation using SYBR green RT-qPCR paired with melting curve analysis
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Laconi, Andrea, Berends, Alinda J, de Laat, Esther C H, Urselmann, Tara A P M P, Verheije, Hélène M, LS Pathologie, dPB I&I, dI&I I&I-1, LS Pathologie, dPB I&I, and dI&I I&I-1
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0301 basic medicine ,infectious bronchitis virus ,Genotyping ,animal structures ,Genotype ,030106 microbiology ,Infectious bronchitis virus ,Biology ,Real-Time Polymerase Chain Reaction ,Vaccines, Attenuated ,Sensitivity and Specificity ,Melting curve analysis ,Virus ,Article ,03 medical and health sciences ,Viral Proteins ,Virology ,Animals ,Multiplex ,Poultry Diseases ,Vaccine differentiation ,Attenuated vaccine ,RT-qPCR ,Viral Vaccines ,Vaccination ,030104 developmental biology ,Melting curve ,melting curve ,Molecular Diagnostic Techniques ,genotyping ,vaccine differentiation ,embryonic structures ,Coronavirus Infections ,Chickens - Abstract
Highlights • Development of a reliable and cost-effective tool for IBV Mass-type and QX-like genotyping. • IBV QX vaccine markers enable vaccine differentiation via melting curve analysis. • SYBRgreen RT-qPCR assay supports multiplexing and enables IBV genotyping., Infectious Bronchitis Virus (IBV) is a highly contagious virus of chicken, causing huge economic losses in the poultry industry. Many genotypes circulate in a given area, and optimal protection relies on vaccination with live attenuated vaccines of the same genotype. As these live vaccines are derived from field viruses and circulate, understanding the prevalence of different IBV genotypes in any area is complex. In a recent study, the genome comparison of an IBV QX vaccine and its progenitor field strain led to the identification of vaccine markers. Here we developed a simplex SYBRgreen RT-qPCR assay for differentiation between QX-like field and vaccine strains and a multiplex SYBRgreen RT-qPCR assay for IBV genotyping with melting curve analysis, as each virus produced distinct and reliable melting peaks. Both the simplex and the multiplex assays showed excellent efficiency, sensitivity and specificity representing a low cost diagnostic tool for IBV genotyping and vaccine differentiation.
16. Three Amino Acid Changes in Avian Coronavirus Spike Protein Allow Binding to Kidney Tissue.
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Bouwman, Kim M., Parsons, Lisa M., Berends, Alinda J., de Vries, Robert P., Cipollo, John F., and Verheije, Monique H.
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PROTEIN binding , *PROXIMAL kidney tubules , *AMINO acids , *AVIAN infectious bronchitis virus , *CHIMERIC proteins , *KIDNEYS - Abstract
Infectious bronchitis virus (IBV) infects ciliated epithelial cells in the chicken respiratory tract. While some IBV strains replicate locally, others can disseminate to various organs, including the kidney. Here, we elucidate the determinants for kidney tropism by studying interactions between the receptor-binding domain (RBD) of the viral attachment protein spike from two IBV strains with different tropisms. Recombinantly produced RBDs from the nephropathogenic IBV strain QX and from the nonnephropathogenic strain M41 bound to the epithelial cells of the trachea. In contrast, only QX-RBD binds more extensively to cells of the digestive tract, urogenital tract, and kidneys. While removal of sialic acids from tissues prevented binding of all proteins to all tissues, binding of QX-RBD to trachea and kidney could not be blocked by preincubation with synthetic alpha-2,3-linked sialic acids. The lack of binding of QX-RBD to a previously identified IBV-M41 receptor was confirmed by enzyme-linked immunosorbent assay (ELISA), demonstrating that tissue binding of QX-RBD is dependent on a different sialylated glycan receptor. Using chimeric RBD proteins, we discovered that the region encompassing amino acids 99 to 159 of QXRBD was required to establish kidney binding. In particular, QX-RBD amino acids 110 to 112 (KIP) were sufficient to render IBV-M41 with the ability to bind to kidney, while the reciprocal mutations in IBV-QX abolished kidney binding completely. Structural analysis of both RBDs suggests that the receptor-binding site for QX is located at a different location on the spike than that of M41. [ABSTRACT FROM AUTHOR]
- Published
- 2020
- Full Text
- View/download PDF
17. Human Milk Oligosaccharide 2'-Fucosyllactose Modulates Local Viral Immune Defense by Supporting the Regulatory Functions of Intestinal Epithelial and Immune Cells.
- Author
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Ayechu-Muruzabal V, Poelmann B, Berends AJ, Kettelarij N, Garssen J, Van't Land B, and Willemsen LEM
- Subjects
- Dendritic Cells, Epithelial Cells, Galectin 4, Humans, Oligosaccharides pharmacology, Poly I, Trisaccharides, Interleukin-8, Milk, Human
- Abstract
Human milk contains bioactive components that provide protection against viral infections in early life. In particular, intestinal epithelial cells (IEC) have key regulatory roles in the prevention of enteric viral infections. Here we established an in vitro model to study the modulation of host responses against enteric viruses mimicked by poly I:C (pIC). The effects of 2'-fucosyllactose (2'FL), abundantly present in human milk, were studied on IEC and/or innate immune cells, and the subsequent functional response of the adaptive immune cells. IEC were pre-incubated with 2'FL and stimulated with naked or Lyovec™-complexed pIC (LV-pIC). Additionally, monocyte-derived dendritic cells (moDC) alone or in co-culture with IEC were stimulated with LV-pIC. Then, conditioned-moDC were co-cultured with naïve CD4
+ T helper (Th)-cells. IEC stimulation with naked or LV-pIC promoted pro-inflammatory IL-8, CCL20, GROα and CXCL10 cytokine secretion. However, only exposure to LV-pIC additionally induced IFNβ, IFNλ1 and CCL5 secretion. Pre-incubation with 2'FL further increased pIC induced CCL20 secretion and LV-pIC induced CXCL10 secretion. LV-pIC-exposed IEC/moDC and moDC cultures showed increased secretion of IL-8, GROα, IFNλ1 and CXCL10, and in the presence of 2'FL galectin-4 and -9 were increased. The LV-pIC-exposed moDC showed a more pronounced secretion of CCL20, CXCL10 and CCL5. The moDC from IEC/moDC cultures did not drive T-cell development in moDC/T-cell cultures, while moDC directly exposed to LV-pIC secreted Th1 driving IL-12p70 and promoted IFNγ secretion by Th-cells. Hereby, a novel intestinal model was established to study mucosal host-defense upon a viral trigger. IEC may support intestinal homeostasis, regulating local viral defense which may be modulated by 2'FL. These results provide insights regarding the protective capacity of human milk components in early life.- Published
- 2022
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18. Wild and domestic animals variably display Neu5Ac and Neu5Gc sialic acids.
- Author
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Nemanichvili N, Spruit CM, Berends AJ, Gröne A, Rijks JM, Verheije MH, and de Vries RP
- Subjects
- Animals, Animals, Domestic metabolism, Dogs, Ferrets metabolism, Glycolipids, Horses, Humans, Lectins, N-Acetylneuraminic Acid metabolism, Neuraminic Acids, Polysaccharides, Swine, Sialic Acids metabolism, Viruses
- Abstract
Sialic acids are used as a receptor by several viruses and variations in the linkage type or C-5 modifications affect the binding properties. A species barrier for multiple viruses is present due to α2,3- or α2,6-linked sialic acids. The C-5 position of the sialic acid can be modified to form N-acetylneuraminic acid (Neu5Ac) or N-glycolylneuraminic acid (Neu5Gc), which acts as a determinant for host susceptibility for pathogens such as influenza A virus, rotavirus, and transmissible gastroenteritis coronavirus. Neu5Gc is present in most mammals such as pigs and horses but is absent in humans, ferrets, and dogs. However, little is known about C-5 content in wildlife species or how many C-5 modified sialic acids are present on N-linked glycans or glycolipids. Using our previously developed tissue microarray system, we investigated how 2 different lectins specific for Neu5Gc can result in varying detection levels of Neu5Gc glycans. We used these lectins to map Neu5Gc content in wild Suidae, Cervidae, tigers, and European hedgehogs. We show that Neu5Gc content is highly variable among different species. Furthermore, the removal of N-linked glycans reduces the binding of both Neu5Gc lectins while retention of glycolipids by omitting methanol treatment of tissues increases lectin binding. These findings highlight the importance of using multiple Neu5Gc lectins as the rich variety in which Neu5Gc is displayed can hardly be detected by a single lectin., (© The Author(s) 2022. Published by Oxford University Press.)
- Published
- 2022
- Full Text
- View/download PDF
19. Tissue Microarrays to Visualize Influenza D Attachment to Host Receptors in the Respiratory Tract of Farm Animals.
- Author
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Nemanichvili N, Berends AJ, Wubbolts RW, Gröne A, Rijks JM, de Vries RP, and Verheije MH
- Subjects
- Animals, Animals, Domestic virology, Cattle, Goats, Hemagglutinins, Viral genetics, Horses, Host Microbial Interactions, Recombinant Proteins metabolism, Sheep, Sialic Acids metabolism, Swine, Thogotovirus chemistry, Thogotovirus genetics, Viral Fusion Proteins genetics, Hemagglutinins, Viral metabolism, Respiratory System virology, Thogotovirus physiology, Tissue Array Analysis, Viral Fusion Proteins metabolism, Viral Tropism, Virus Attachment
- Abstract
The trimeric hemagglutinin-esterase fusion protein (HEF) of influenza D virus (IDV) binds 9-O-acetylated sialic acid receptors, which are expressed in various host species. While cattle are the main reservoir for IDV, the viral genome has also been detected in domestic pigs. In addition, antibodies against IDV have been detected in other farm animals such as sheep, goats, and horses, and even in farmers working with IDV positive animals. Viruses belonging to various IDV clades circulate, but little is known about their differences in host and tissue tropism. Here we used recombinantly produced HEF proteins (HEF S57A) from the major clades D/Oklahoma (D/OK) and D/Oklahoma/660 (D/660) to study their host and tissue tropism and receptor interactions. To this end, we developed tissue microarrays (TMA) composed of respiratory tissues from various farm animals including cattle, domestic pigs, sheep, goats, and horses. Protein histochemical staining of farm animal respiratory tissue-microarrays with HEF proteins showed that cattle have receptors present over the entire respiratory tract while receptors are only present in the nasal and pharyngeal epithelium of pigs, sheep, goats, and horses. No differences in tropism for tissues and animals were observed between clades, while hemagglutination assays showed that D/OK has a 2-fold higher binding affinity than D/660 for receptors on red blood cells. The removal of O-acetylation from receptors via saponification treatment confirmed that receptor-binding of both clades was dependent on O-acetylated sialic acids.
- Published
- 2021
- Full Text
- View/download PDF
20. Deletion of accessory genes 3a, 3b, 5a or 5b from avian coronavirus infectious bronchitis virus induces an attenuated phenotype both in vitro and in vivo.
- Author
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Laconi A, van Beurden SJ, Berends AJ, Krämer-Kühl A, Jansen CA, Spekreijse D, Chénard G, Philipp HC, Mundt E, Rottier PJM, and Hélène Verheije M
- Abstract
Avian coronavirus infectious bronchitis virus (IBV) infects domestic fowl, resulting in respiratory disease and causing serious losses in unprotected birds. Its control is mainly achieved by using live attenuated vaccines. Here we explored the possibilities for rationally attenuating IBV to improve our knowledge regarding the function of IBV accessory proteins and for the development of next-generation vaccines with the recently established reverse genetic system for IBV H52 based on targeted RNA recombination and selection of recombinant viruses in embryonated eggs. To this aim, we selectively removed accessory genes 3a, 3b, 5a and 5b individually, and rescued the resulting recombinant (r) rIBV-Δ3a, rIBV-Δ3b, rIBV-Δ5a and rIBV-Δ5b. In vitro inoculation of chicken embryo kidney cells with recombinant and wild-type viruses demonstrated that the accessory protein 5b is involved in the delayed activation of the interferon response of the host after IBV infection. Embryo mortality after the inoculation of 8-day-old embryonated chicken eggs with recombinant and wild-type viruses showed that rIBV-Δ3b, rIBV-Δ5a and rIBV-Δ5b had an attenuated phenotype in ovo, with reduced titres at 6 h p.i. and 12 h p.i. for all viruses, while growing to the same titre as wild-type rIBV at 48 h p.i. When administered to 1-day-old chickens, rIBV-Δ3a, rIBV-Δ3b, rIBV-Δ5a and rIBV-Δ5b showed reduced ciliostasis in comparison to the wild-type viruses. In conclusion, individual deletion of accessory genes in IBV H52 resulted in mutant viruses with an attenuated phenotype.
- Published
- 2018
- Full Text
- View/download PDF
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