A5044139506- Influenza Virus Research Studies
- SARS-CoV-2 and COVID-19 Research
- Respiratory viral infections research
- SARS-CoV-2 detection and testing
- Animal Virus Infections Studies
- Virus-based gene therapy research
- Diabetes and associated disorders
- Glycosylation and Glycoproteins Research
- Animal Disease Management and Epidemiology
- Immune Cell Function and Interaction
- COVID-19 Clinical Research Studies
- Cytomegalovirus and herpesvirus research
- RNA and protein synthesis mechanisms
- Protein Structure and Dynamics
- Immunotherapy and Immune Responses
- Herpesvirus Infections and Treatments
- Biosensors and Analytical Detection
- Advanced biosensing and bioanalysis techniques
- Electrochemical sensors and biosensors
- Long-Term Effects of COVID-19
- interferon and immune responses
- Virology and Viral Diseases
- Viral gastroenteritis research and epidemiology
- T-cell and B-cell Immunology
- Immune Response and Inflammation
Utrecht University
2018-2024
Pharmo Institute
2020-2024
Amsterdam University Medical Centers
2021
University of Amsterdam
2021
Emerging evidence suggests that host glycans influence severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. Here, we reveal the receptor-binding domain (RBD) of spike (S) protein on SARS-CoV-2 recognizes oligosaccharides containing sialic acid (Sia), with preference for monosialylated gangliosides. Gangliosides embedded within an artificial membrane also bind to RBD. The monomeric affinities (Kd = 100–200 μM) gangliosides RBD are similar another negatively charged glycan...
Stromal-derived follicular dendritic cells (FDCs) are essential for germinal centers (GCs), the site where B maturate their antibodies. FDCs present native antigen to and maintain a CXCL13 gradient form cell follicle. Yet despite role, transcriptome of human remains undefined. Using single-cell RNA sequencing microarray, we provided these enigmatic as comprehensive resource. Key genes were validated by flow cytometry microscopy. Surprisingly, marginal reticular (MRCs) rather than expressed...
Receptor binding studies on sarbecoviruses would benefit from an available toolkit of recombinant spike proteins, or domains thereof, that recapitulate receptor properties native viruses. We hypothesized trimeric Binding Domain (RBD) proteins be suitable candidates to study SARS-CoV-1 and -2. Here we created monomeric fluorescent RBD derived adherent HEK293T, as well in GnTI-/- mutant cells, analyze the effect complex vs high mannose glycosylation binding. The results demonstrate trimeric,...
SARS-CoV-2 viruses engage ACE2 as a functional receptor with their spike protein. The S1 domain of the protein contains C-terminal binding (RBD) and an N-terminal (NTD). NTD other coronaviruses includes glycan cleft. However, for NTD, protein–glycan was only observed weakly sialic acids highly sensitive methods. Amino acid changes in variants concern (VoC) show antigenic pressure, which can be indication NTD-mediated binding. Trimeric proteins SARS-CoV-2, alpha, beta, delta, omicron did not...
Influenza A virus carries hundreds of trimeric hemagglutinin (HA) proteins on its viral envelope that interact with various sialylated glycans a host cell. This interaction represents multivalent binding event is present in all the current receptor assays, including those employing viruses or precomplexed HA trimers. To study nature such events, we fused superfolder green fluorescent protein (sfGFP) to C-terminus allow for direct visualization HA-receptor interactions without need additional...
SARS-CoV-2 attaches to angiotensin-converting enzyme 2 (ACE2) gain entry into cells after which the spike protein is cleaved by transmembrane serine protease (TMPRSS2) facilitate viral-host membrane fusion. ACE2 and TMPRSS2 expression profiles have been analyzed at genomic, transcriptomic, single-cell RNAseq levels. However, transcriptomic data actual validation convey conflicting information regarding distribution of biologically relevant receptor in whole tissues. To describe organ-level...
The development of an effective human immunodeficiency virus (HIV-1) vaccine is a high global health priority. Soluble native-like HIV-1 envelope glycoprotein trimers (Env), including those based on the SOSIP design, have shown promise as candidates by inducing neutralizing antibody responses against autologous in animal models. However, to overcome HIV-1's extreme diversity needs induce broadly antibodies (bNAbs). Such bNAbs can protect non-human primates (NHPs) and humans from infection....
Influenza A viruses (IAV) initiate infection by binding to glycans with terminal sialic acids on the cell surface. Hosts of IAV variably express two major forms acid, N-acetylneuraminic acid (NeuAc) and N-glycolylneuraminic (NeuGc). NeuGc is produced in most mammals, including horses pigs, but absent humans, ferrets, birds. The only known naturally occurring that exclusively bind are extinct highly pathogenic equine H7N7 viruses. We determined crystal structure a representative H7...
The emergence of new SARS-CoV-2 variants and the dangers long-covid necessitate development broad-acting therapeutics that can reduce viral burden. employs heparan sulfate (HS) as an initial cellular attachment factor, therefore, there is interest in developing heparin a therapeutic for SARS-CoV-2. Its use is, however, complicated by structural heterogeneity risk causing bleeding thrombocytopenia. Here, we describe preparation well-defined mimetics controlled head-to-tail assembly HS...
Abstract Recombinant soluble trimeric influenza A virus hemagglutinins (HA) and tetrameric neuraminidases (NAs) have proven to be excellent tools decipher biological properties. Receptor binding sialic acid cleavage by recombinant proteins correlate satisfactorily compared whole viruses. Expression of HA NA can achieved in a plethora different laboratory hosts. For immunological receptor interaction studies however, insect mammalian cell expressed are preferred due the presence N‐linked...
Abstract Emerging evidence suggests that host glycans influence infection by SARS-CoV-2. Here, we reveal the receptor-binding domain (RBD) of spike (S)-protein on SARS-CoV-2 recognizes oligosaccharides containing sialic acid (SA), with preference for oligosaccharide monosialylated gangliosides. Gangliosides embedded within an artificial membrane also bind RBD. The monomeric affinities ( K d = 100-200 μM) gangliosides RBD are similar to heparan sulfate, another negatively charged glycan...
Abstract Enveloped viruses carry one or multiple proteins with receptor‐binding functionalities. Functional receptors can be glycans, proteinaceous, both; therefore, recombinant protein approaches are instrumental in attaining new insights regarding viral envelope properties. Visualizing and measuring receptor binding typically entails antibody detection direct labeling, whereas fluorescent fusions attractive tools molecular biology. Here, we report a suite of distinct fusions, both N‐...
Coronaviruses (CoVs) recognize a wide array of protein and glycan receptors by using the S1 subunit spike (S) glycoprotein. The contains two functional domains: N-terminal domain (S1-NTD) C-terminal (S1-CTD). S1-NTD SARS-CoV-2, MERS-CoV, HCoV-HKU1 possesses an evolutionarily conserved binding cleft that facilitates weak interactions with sialic acids on cell surfaces. employs 9-O-acetylated α2-8-linked disialylated structures for initial binding, followed TMPRSS2 receptor virus–cell fusion....
Abstract Emerging evidence suggests that host glycans influence infection by SARS-CoV-2. Here, we reveal the receptor-binding domain (RBD) of spike (S)-protein on SARS-CoV-2 recognizes oligosaccharides containing sialic acid (SA), with preference for oligosaccharide monosialylated gangliosides. Gangliosides embedded within an artificial membrane also bind RBD. The monomeric affinities (Kd = 100-200 μM) gangliosides RBD are similar to heparan sulfate, another negatively charged glycan ligand...
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 necessary epithelial respiratory tract accommodate binding IDV HEF both worldwide clades D/Oklahoma (D/OK) D/Oklahoma/660 (D/660). More recently, seroprevalence studies have identified IDV-seropositive...
Receptor binding studies using recombinant SARS-CoV proteins have been hampered due to challenges in approaches creating spike protein or domains thereof, that recapitulate receptor properties of native viruses. We hypothesized trimeric RBD would be suitable candidates study SARS-CoV-1 and -2. Here we created monomeric fluorescent proteins, derived from adherent HEK293T, as well GnTI mutant cells, analyze the effect complex vs high mannose glycosylation on binding. The results demonstrate...
A novel miniaturized sensor for electrochemical detection that contains graphene- and gold nanoparticles was functionalized with proteins. Using cyclic voltammetry (CV) differential pulse (DPV) it possible to observe quantify interactions of molecules these The protein binders included carbohydrate ligands as small carbohydrates up COVID-19 spike variants engaged in protein-protein interactions. system uses off-the-shelf sensors combined an affordable potentiostat yet is sensitive enough...
Coronaviruses recognize a wide array of protein and glycan receptors using the S1 subunit spike (S) glycoprotein. The contains two functional domains: N-terminal (S1-NTD) C-terminal (S1-CTD). S1-NTD SARS-CoV-2, MERS-CoV, HCoV-HKU1 possess an evolutionarily conserved binding cleft that facilitates weak interactions with sialic acids on cell surfaces. employs 9-
The quest for a universal influenza vaccine holds great promise mitigating the global burden of influenza-related morbidity and mortality. However, challenges persist in identifying conserved epitopes capable inducing protection. In this study, we explore influence glycan evolution on H3 hemagglutinin from 1968 to present day its impacts antigenicity immunogenicity. We observe that appearance potential N-linked glycosylation sites Sing/16 head domain reduces binding broadly neutralizing...
SARS-CoV-2 viruses engage ACE2 as a functional receptor with their spike protein. The S1 domain of the protein contains C-terminal receptor-binding (RBD) and an N-terminal (NTD). NTD other coronaviruses includes glycan-binding cleft. However, for protein-glycan binding was only observed weakly sialic acids highly sensitive methods. Amino acid changes in Variants Concern (VoC) shows antigenic pressure, which can be indication NTD-mediated binding. Trimeric proteins SARS-CoV-2, Alpha, Beta,...
Abstract SARS-CoV-2 attaches to angiotensin-converting enzyme 2 (ACE2) gain entry into cells after which the spike protein is cleaved by transmembrane serine protease (TMPRRS2) facilitate viral-host membrane fusion. ACE2 and TMPRRS2 expression profiles have been analyzed at genomic, transcriptomic, single-cell RNAseq level, however, biologically relevant receptor organization in whole tissues still poorly understood. To describe organ-level architecture of expression, related ability mediate...
Abstract Enveloped viruses carry one or multiple proteins with receptor binding functionalities. Functional receptors can either be glycans, proteinaceous both, recombinant protein approaches are instrumental to gain more insight into these properties. Visualizing and measuring normally entails antibody detection direct labelling, whereas fluorescent fusions attractive tools in molecular biology. Here we report a suite of different fusions, both N- and/or C-terminal, for influenza A virus...