A5045696256- HIV Research and Treatment
- SARS-CoV-2 and COVID-19 Research
- Monoclonal and Polyclonal Antibodies Research
- Bacteriophages and microbial interactions
- Glycosylation and Glycoproteins Research
- Animal Virus Infections Studies
- vaccines and immunoinformatics approaches
- HIV/AIDS drug development and treatment
- Photosynthetic Processes and Mechanisms
- Influenza Virus Research Studies
- Immune Cell Function and Interaction
- Protein Structure and Dynamics
- Viral gastroenteritis research and epidemiology
- Bacillus and Francisella bacterial research
- Photoreceptor and optogenetics research
- SARS-CoV-2 detection and testing
- RNA and protein synthesis mechanisms
- Immunotherapy and Immune Responses
- Spectroscopy Techniques in Biomedical and Chemical Research
- T-cell and B-cell Immunology
- Advanced biosensing and bioanalysis techniques
- ATP Synthase and ATPases Research
- thermodynamics and calorimetric analyses
- RNA Interference and Gene Delivery
- Machine Learning in Bioinformatics
Duke University Hospital
2022-2024
Duke Medical Center
2019-2024
International Vaccine Institute
2022-2024
Duke University
2018-2024
Lancaster University
2024
Trinity College Dublin
2020
University of Arkansas at Fayetteville
2012-2016
The coronavirus (CoV) spike (S) protein, involved in viral-host cell fusion, is the primary immunogenic target for virus neutralization and current focus of many vaccine design efforts. highly flexible S-protein, with its mobile domains, presents a moving to immune system. Here, better understand S-protein mobility, we implemented structure-based vector analysis available β-CoV structures. Despite an overall similarity domain organization, found that S-proteins from different β-CoVs display...
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants with multiple spike mutations enable increased transmission and antibody resistance. We combined cryo-electron microscopy (cryo-EM), binding, computational analyses to study variant spikes, including one that was involved in between minks humans, others originated spread human populations. All showed angiotensin-converting enzyme (ACE2) receptor binding propensity for domain (RBD)-up states. While adaptation mink resulted...
The severe acute respiratory coronavirus 2 (SARS-CoV-2) spike (S) protein is the target of vaccine design efforts to end disease 2019 (COVID-19) pandemic. Despite a low mutation rate, isolates with D614G substitution in S appeared early during pandemic and are now dominant form worldwide. Here, we explore conformational changes effects on soluble ectodomain construct. Cryoelectron microscopy (cryo-EM) structures reveal altered receptor binding domain (RBD) disposition; antigenicity...
Engineering better bnAbs A highly effective HIV vaccine has been the goal of vaccinologists for nearly 35 years. successful would need to induce broadly neutralizing antibodies (bnAbs) that are capable multiple strains (see Perspective by Agazio and Torres). Steichen et al. report a strategy in which first shot can lead immune responses generate desired bnAbs. By combining knowledge human antibody repertoires structure guide design, they validated candidate immunogens through functional...
The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) Omicron BA.2 sub-lineage has gained in proportion relative to BA.1. Because spike (S) protein variations may underlie differences their pathobiology, here we determine cryoelectron microscopy (cryo-EM) structures of the S ectodomain and compare these with previously determined BA.1 structures. receptor-binding domain (RBD) mutations induce remodeling RBD structure, resulting tighter packing improved thermostability....
The SARS-CoV-2 spike (S) protein, a primary target for COVID-19 vaccine development, presents its receptor binding domain in two conformations, the receptor-accessible 'up' or receptor-inaccessible 'down' states. Here we report that commonly used stabilized S ectodomain construct '2P' is sensitive to cold temperatures, and this sensitivity abrogated state-stabilized ectodomain. Our findings will impact structural, functional studies use
The CD4 binding site (CD4bs) of the HIV-1 envelope glycoprotein is susceptible to multiple lineages broadly neutralizing antibodies (bnAbs) that are attractive elicit with vaccines. CH235 lineage (VH1-46) CD4bs bnAbs particularly because most mature members neutralize 90% circulating strains, do not possess long HCDR3 regions, and contain insertions deletions may be difficult induce. We used virus neutralization measure interaction unmutated common ancestor (CH235 UCA) functional Env trimers...
Elicitation of VRC01-class broadly neutralizing antibodies (bnAbs) is an appealing approach for a preventative HIV-1 vaccine. Despite extensive investigations, strategies to induce bnAbs and overcome the barrier posed by envelope N276 glycan have not been successful. Here, we inferred high-probability unmutated common ancestor (UCA) VRC01 lineage reconstructed stages maturation. Env immunogens designed on reverted bound UCA with affinity sufficient activate naive B cells. Early mutations...
The glycan shield of the beta-coronavirus (β-CoV) Spike (S) glycoprotein provides protection from host immune responses, acting as a steric block to potentially neutralizing antibody responses. conformationally dynamic S-protein is primary immunogenic target vaccine design owing its role in host-cell fusion, displaying multiple receptor binding domain (RBD) 'up' and 'down' state configurations. Here, we investigated potential for RBD adjacent, N-terminal (NTD) glycans influence...
Summary New SARS-CoV-2 variants that have accumulated multiple mutations in the spike (S) glycoprotein enable increased transmission and resistance to neutralizing antibodies. Here, we study antigenic structural impacts of S protein from four variants, one was involved between minks humans, three rapidly spread human populations originated United Kingdom, Brazil or South Africa. All either retained improved binding ACE2 receptor. The B.1.1.7 (UK) B.1.1.28 (Brazil) showed reduced NTD RBD...
Aided by extensive spike protein mutation, the SARS-CoV-2 Omicron variant overtook previously dominant Delta variant. Spike conformation plays an essential role in evolution via changes receptor binding domain (RBD) and neutralizing antibody epitope presentation affecting virus transmissibility immune evasion. Here, we determine cryo-EM structures of spikes to understand conformational impacts mutations each. The structure revealed unusually tightly packed RBD organization with long range...
The HIV-1 Envelope (Env) glycoprotein facilitates host cell fusion through a complex series of receptor-induced structural changes. Although remarkable progress has been made in understanding the structures various Env conformations, microsecond timescale dynamics have not studied experimentally. Here, we used time-resolved, temperature-jump small-angle x-ray scattering to monitor rearrangements an SOSIP ectodomain construct with precision. In two distinct variants, detected transition that...
The trimeric HIV-1 Envelope protein (Env) mediates viral-host cell fusion via a network of conformational transitions, with allosteric elements in each protomer orchestrating host receptor-induced exposure the co-receptor binding site and elements. To understand molecular details this allostery, here, we introduce Env mutations aimed to prevent CD4-induced rearrangements BG505 trimer. Binding analysis single-molecule Förster Resonance Energy Transfer confirm that these transitions Env....
Abstract The coronavirus (CoV) viral host cell fusion spike (S) protein is the primary immunogenic target for virus neutralization and current focus of many vaccine design efforts. highly flexible S-protein, with its mobile domains, presents a moving to immune system. Here, better understand S-protein mobility, we implemented structure-based vector analysis available β-CoV structures. We found that despite overall similarity in domain organization, different strains display distinct...
Abstract Somatic mutations within antibody variable and framework regions (FWR) can alter thermostability structural flexibility, but their impact on functional potency is unclear. Here we study use molecular dynamics (MD) simulations to assess the role of FWR during maturation HIV-1 broadly neutralizing antibodies (bnAbs). The tested bnAbs show lower than unmutated ancestor antibodies. in Fab elbow region are frequently observed MD that such interdomain flexibility two bnAbs. In a...
Abstract Antibody affinity maturation enables adaptive immune responses to a wide range of pathogens. In some individuals broadly neutralizing antibodies develop recognize rapidly mutating pathogens with extensive sequence diversity. Vaccine design for such as HIV-1 and influenza has therefore focused on recapitulating the natural process. Here, we determine structures in complex Envelope all observed members ancestral states V3-glycan targeting DH270 antibody clonal B cell lineage. These...
Summary The COVID-19 pandemic caused by SARS-CoV-2 has escalated into a global crisis. spike (S) protein that mediates cell entry and membrane fusion is the current focus of vaccine therapeutic antibody development efforts. S protein, like many other viral proteins such as HIV-1 envelope (Env) influenza hemagglutinin, glycosylated with both complex high mannose glycans. Here we demonstrate binding to category Fab-dimerized glycan-reactive (FDG) HIV-1-induced broadly neutralizing antibodies...