Ian A. Wilson

Antibodies to distinct epitopes on MPXV enveloped virion provide additive and complete protection

The continuing evolution of monkeypox virus (MPXV) clades capable of sustained human-to-human transmission poses an escalating global health risk, yet no effective therapy exists for poxvirus infection. Here, we investigate a panel of human monoclonal antibodies (mAbs), derived from an antibody library, which targets A35 and B6 antigens located on the extracellular enveloped virion (EV). Four high-affinity mAbs target the central groove of the A35 dimer, whereas one targets a previously unidentified epitope on the A35 protruding loop. Three other mAbs recognize distinct, previously unmapped epitopes on the multidomain B6 antigen. Those epitopes are conserved across most orthopoxviruses. A combination of an anti-A35 and an anti-B6 antibody provides additive and complete protection from both morbidity and mortality in a lethal VACV mouse model, challenging the previous assumption that targeting both the EV and mature virion (MV) forms is required for full protection. These studies uncover critical information for next-generation therapeutics and vaccines against emerging MPXV as well as other Orthopoxvirus threats.

Affinity Maturation Enables Dual Targeting of Cancer-Associated MUC1-Tn/STn and Drives Potent Tumor Control

A major challenge in solid tumor immunotherapy is identifying tumor-restricted antigens while overcoming intratumoral antigen heterogeneity. The truncated O-glycans Tn (GalNAcα-O-Ser/Thr) and STn (Neu5Acα2-6GalNAcα-O-Ser/Thr) are aberrantly displayed on the transmembrane glycoprotein MUC1 across a broad spectrum of epithelial malignancies, making MUC1-Tn/STn an attractive shared tumor-associated antigen class. However, heterogeneous intratumoral Tn/STn expression renders single-glycoform-specific antibodies vulnerable to antigen-driven escape. To address this challenge, we used structure-guided directed evolution and yeast surface display to affinity-mature the anti-MUC1-Tn antibody 5E5, generating variants with high-affinity dual recognition of MUC1-Tn and MUC1-STn. Co-crystal structures revealed that heavy-chain CDR loops cooperatively engage the GalNAc moiety primarily through hydrogen-bonding networks, in which HCDR3 residue 98 fine-tunes glycoform preference: Tyr98 favors MUC1-Tn recognition, whereas His98 enhances MUC1-STn engagement, while both support high-affinity recognition of both glycoforms. When reformatted as bispecific T-cell engagers, the affinity-matured variants mediated potent and durable cytolytic activity against cancer cells with heterogeneous glycan profiles, suppressed mixed Tn/STn tumors in vivo where the parental antibody failed, and showed enhanced activity in patient-derived tumor-like clusters. Together, these findings establish functional glycoform coverage as a design principle for antibody engineering, whereby a single paratope is optimized to recognize a defined spectrum of tumor-associated glycoforms while preserving antigen specificity, providing a strategy to overcome glycan heterogeneity in solid tumor immunotherapy.

Single-component self-assembling protein nanoparticles displaying stabilized prefusion-closed hemagglutinin trimers for influenza vaccine development

Engineered antibodies preserve structural and functional recognition of Plasmodium falciparum circumsporozoite protein

Substrate and target selectivity of 4′-fluoroadenosine against viral and host polymerases

Developing safe and effective treatments against emerging RNA viruses is an important goal in pandemic preparedness efforts. 4'-fluorouridine (4'-FlU) is a broad-spectrum antiviral that was shown to inhibit viral RNA-dependent RNA polymerases (RdRps). Given its notable range of antiviral activity, this class of nucleoside analogs warrants further investigation. Here, we studied the antiviral activity and underlying mechanism of inhibition of 4'-fluoroadenosine (4'-FlA). Like 4'-FlU, 4'-FlA demonstrates a broad-spectrum of antiviral activity against eight prototypic viruses representing diverse families. Enzyme kinetics show that the triphosphate (4'-FlA-TP) is efficiently incorporated by viral RdRps. A cryo-EM structure of the RdRp of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in complex with double-stranded RNA and the incorporated monophosphate (4'-FlA-MP) characterizes interactions at the active site. The incorporated analog elicits heterogeneous inhibition patterns in primer extension reactions. In contrast, templates with embedded 4'-FlA-MP inhibit incorporation of complementary UTP across the viral RdRps. However, incorporation of 4'-FIA-TP is not limited to viral polymerases and likewise includes human mitochondrial RNA polymerase. These results demonstrate the general potential for 4'-fluorinated nucleotides as antiviral drugs and guide the development of more selective derivatives for medical use in appropriate settings.

CD8 PET imaging with [ ⁸⁹ Zr]Zr-berdoxam-crefmirlimab in patients with solid tumors: the Phase II iCorrelate study

Feasibility of day-case greenlight laser prostatectomy to reduce waiting times: A comparative appraisal with TURP in an NHS setting

VHH antibody loop guides design of a synthetic macrocyclic peptide that potently blocks influenza virus membrane fusion

Miniaturizing biologically complex structural motifs to produce synthetic functional mimetics holds significant promise for development of new therapeutic modalities. Here, we demonstrate a unique approach using the key binding loop of the single variable domain of a heavy chain (V H H) llama antibody as a starting point for peptide design. V H H antibodies of camelids and sharks generally have longer, but more ligand-efficient complementarity determining region 3 (CDR3) loops and are relatively stable structures. We harnessed these attributes as templates for design of a series of synthetic macrocyclic peptides. The designed peptides exhibit nanomolar binding to influenza hemagglutinin (HA) and heterosubtypic in vitro neutralization breadth against influenza A viruses by inhibiting the low pH mediated HA conformational changes that lead to membrane fusion. X-ray structures of peptide-HA complexes reveal high structural mimicry with the parent V H H antibody. One such macrocycle peptide candidate is promising for further development of broad protection against influenza A group 1 viruses.

Rational design of next-generation filovirus vaccines combining glycoprotein stabilization and nanoparticle display with glycan modification

Filoviruses pose a significant threat to human health due to frequent outbreaks and high mortality. Although two vector-based vaccines are available for Ebola virus, a broadly protective filovirus vaccine remains elusive. Here, we evaluate a general strategy for stabilizing glycoproteins (GP) from Ebola, Sudan, and Bundibugyo orthoebolaviruses, as well as Ravn orthomarburgvirus. A 3.2 Å crystal structure provides atomic-level details of the redesigned Ebola virus GP, while cryo-electron microscopy reveals how a pan-orthoebolavirus neutralizing antibody targets a conserved site on the stabilized Sudan virus GP (3.13 Å resolution), along with a low-resolution model of antibody-bound Ravn virus GP. A self-assembling protein nanoparticle (SApNP), I3-01v9, is redesigned at the N terminus to enable optimal surface display of filovirus GP trimers. Following detailed in vitro characterization, we examine the lymph node dynamics of Sudan virus GP and GP-presenting SApNPs in mice. Compared with the soluble trimer, SApNPs exhibit ~112-fold longer retention in lymph node follicles, up to 28-fold greater presentation on follicular dendritic cell dendrites, and up to 3-fold stronger germinal center reactions. Functional antibody responses induced by filovirus GP trimers and SApNPs bearing wild-type or modified glycans are assessed in mice. This study provides a foundation for next-generation filovirus vaccine development. There are limited vaccines available for Ebola virus and none for broad protection from filoviruses. Here, the authors rationally design vaccines using nanoparticles and stabilized Ebola virus and other filovirus glycoproteins, characterize antibody epitopes and profile lymph node and antibody responses in mice.

Structural hijacking of FcRn by human astrovirus spikes reveals conserved epitopes for broad-spectrum antivirals

Human astroviruses (HAstVs) are a leading cause of pediatric gastroenteritis and emerging systemic infections; however, no targeted therapies exist. A critical barrier to intervention has been the lack of molecular insights into viral entry, particularly the interaction between the HAstV capsid spike and its receptor, the neonatal Fc receptor (FcRn). Here, we report crystal structures of the HAstV spike from classical serotypes 2 and 6 in complex with human FcRn at 3 Å resolution, defining a conserved receptor-binding interface at atomic resolution. These structures reveal serotype-specific variations that dictate receptor affinity and demonstrate that reported neutralizing antibodies can inhibit infection by sterically blocking the receptor-binding site. Mapping conserved epitopes across classical HAstV serotypes provides a blueprint for designing broad-spectrum antivirals that disrupt viral entry. Notably, our structural data rationalize the potential repurposing of clinical FcRn inhibitors, such as nipocalimab, to block HAstV infection, bridging critical gaps in astrovirus biology and antiviral development.

A highly divergent cryptic SARS-CoV-2 lineage exhibits strong receptor binding and immune evasion

SARS-CoV-2 genomic surveillance in wastewater enables the identification of cryptic lineages, often with numerous mutations not observed in circulating variants. Here, we discovered a unique cryptic SARS-CoV-2 lineage (designated NJ) from New Jersey wastewater that carries many rare mutations and exhibits the highest binding affinity to the hACE2 receptor among all tested SARS-CoV-2 variant RBDs, including the latest Omicron lineages. Moreover, NJ shows extensive evasion against all five major classes of neutralizing antibodies to SARS-CoV-2. Deep mutational scanning further demonstrates the involvement of epistasis in the evolution of the NJ-RBD. Crystal structures of NJ-RBD in complex with hACE2 and with a broadly neutralizing antibody CC25.4 reveal key amino acid substitutions that affect receptor binding, antibody evasion, and explain its unique molecular characteristics. Overall, these findings not only elucidate the structural and functional features of a cryptic SARS-CoV-2 lineage, but also provide insights into potential future evolution of SARS-CoV-2.

Human coronavirus HKU1 neutralization by glycan receptor mimicry

Common cold embecovirus imprinting primes broadly neutralizing antibody responses to SARS-CoV-2 S2

Structure and function of a cross-neutralizing influenza neuraminidase antibody that accommodates recent N2 NA Asn245 glycosylation

Childhood immunological imprinting of cross-subtype antibodies targeting the hemagglutinin head domain of influenza viruses

Chemoenzymatic synthesis of N-linked glycan receptors of H1N1 influenza virus on human airway epithelial cells

HIV Env trimers elicit NHP apex cross-neutralizing antibodies mimicking human bNAbs

Structural Hijacking of FcRn by Human Astrovirus Spikes Reveals Conserved Epitopes for Broad-Spectrum Antivirals

Inactivation of peptidoglycan remodeling promotes antibiotic susceptibility in vancomycin-resistant Enterococcus faecium

In Vivo Evolution of Monoclonal Antibody CR3022 to Achieve Cross-Neutralization of SARS-CoV-2 and Implications for Vaccine Strategies Against SARS-Related Viruses