Single-component multilayered self-assembling protein nanoparticles presenting glycan-trimmed uncleaved prefusion optimized envelope trimers as HIV-1 vaccine candidates

Nature Communications, Journal Year: 2023, Volume and Issue: 14(1)

Published: April 8, 2023

Uncleaved prefusion-optimized (UFO) design can stabilize diverse HIV-1 envelope glycoproteins (Envs). Single-component, self-assembling protein nanoparticles (1c-SApNP) display 8 or 20 native-like Env trimers as vaccine candidates. We characterize the biophysical, structural, and antigenic properties of 1c-SApNPs that present BG505 UFO trimer with wildtype modified glycans. For 1c-SApNPs, glycan trimming improves recognition CD4 binding site without affecting broadly neutralizing antibodies (bNAbs) to major epitopes. In mice, rabbits, nonhuman primates, increases frequency responders (FVR) steers antibody responses away from immunodominant holes patches. The mechanism vaccine-induced immunity is examined in mice. Compared trimer, multilayered E2p I3-01v9 show 420 times longer retention lymph node follicles, 20-32 greater presentation on follicular dendritic cell dendrites, up-to-4 stronger germinal center reactions. These findings inform future development.

Language: Английский

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An intranasal influenza virus-vectored vaccine prevents SARS-CoV-2 replication in respiratory tissues of mice and hamsters

Nature Communications, Journal Year: 2023, Volume and Issue: 14(1)

Published: April 12, 2023

Abstract Current available vaccines for COVID-19 are effective in reducing severe diseases and deaths caused by SARS-CoV-2 infection but less optimal preventing infection. Next-generation which able to induce mucosal immunity the upper respiratory prevent or reduce infections highly transmissible variants of urgently needed. We have developed an intranasal vaccine candidate based on a live attenuated influenza virus (LAIV) with deleted NS1 gene that encodes cell surface expression receptor-binding-domain (RBD) spike protein, designated DelNS1-RBD4N-DAF. Immune responses protection against challenge following administration DelNS1-RBD4N-DAF were analyzed mice compared intramuscular injection BioNTech BNT162b2 mRNA hamsters. LAIVs induced high levels neutralizing antibodies various hamsters stimulated robust T mice. Notably, vaccination LAIVs, not mRNA, prevented replication variants, including Delta Omicron BA.2, tissues animals. The LAIV system warrants further evaluation humans control transmission and, more significantly, creating dual function both use annual strategies.

Language: Английский

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Glycosylation shapes the efficacy and safety of diverse protein, gene and cell therapies

Biotechnology Advances, Journal Year: 2023, Volume and Issue: 67, P. 108206 - 108206

Published: June 22, 2023

Over recent decades, therapeutic proteins have had widespread success in treating a myriad of diseases. Glycosylation, near universal feature this class drugs, is critical quality attribute that significantly influences the physical properties, safety profile and biological activity proteins. Optimizing protein glycosylation, therefore, offers an important avenue to developing more efficacious therapies. In review, we discuss specific examples how variations glycan structure glycoengineering impacts stability, safety, clinical efficacy protein-based drugs are already market as well those still preclinical development. We also highlight impact glycosylation on next generation biologics such T cell-based cancer therapy gene therapy.

Language: Английский

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Semisynthesis of homogeneous spike RBD glycoforms from SARS-CoV-2 for profiling the correlations between glycan composition and function

National Science Review, Journal Year: 2024, Volume and Issue: 11(2)

Published: Jan. 9, 2024

ABSTRACT Vaccines have been the primary remedy in global fight against coronavirus disease 2019 (COVID-19). The receptor-binding domain (RBD) of spike protein, a critical viral immunogen, is affected by heterogeneity its glycan structures and relatively low immunogenicity. Here, we describe scalable synthetic platform that enables precise synthesis homogeneously glycosylated RBD, facilitating elucidation carbohydrate structure–function relationships. Five RBDs bearing biantennary glycans were prepared, three which conjugated to T-helper epitope (Tpep) from tetanus toxoid improve their weak immune response. Relative natural HEK293-derived with N-glycan elicited higher level neutralising antibodies SARS-CoV-2 mice. Furthermore, containing Tpep significant responses transgenic mice expressing human angiotensin-converting enzyme 2. Our collective data suggest trimming N-glycans conjugation RBD could potentially serve as an effective strategy for developing subunit vaccines providing efficient protection.

Language: Английский

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Immune Epitopes of SARS-CoV-2 Spike Protein and Considerations for Universal Vaccine Development

ImmunoHorizons, Journal Year: 2024, Volume and Issue: 8(3), P. 214 - 226

Published: March 1, 2024

Abstract Despite the success of global vaccination programs in slowing spread COVID-19, these efforts have been hindered by emergence new SARS-CoV-2 strains capable evading prior immunity. The mutation and evolution created a demand for persistent vaccine development. Spike protein has primary target COVID-19 development, but it is also hotspot mutations directly involved host susceptibility virus immune evasion. Our ability to predict emerging mutants select conserved epitopes critical development broadly neutralizing therapy or universal vaccine. In this article, we review general paradigm responses vaccines, highlighting immunological that are likely associated with eliciting protective immunity resulting from humans. Specifically, analyze structural evolutionary characteristics related activation function via TLRs, B cells, T cells. We aim provide comprehensive analysis protein, thereby contributing strategies broad neutralization vaccination.

Language: Английский

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Water–glycan interactions drive the SARS-CoV-2 spike dynamics: insights into glycan-gate control and camouflage mechanisms

Chemical Science, Journal Year: 2024, Volume and Issue: 15(35), P. 14177 - 14187

Published: Jan. 1, 2024

To develop therapeutic strategies against COVID-19, we introduce a high-resolution all-atom polarizable model capturing many-body effects of protein, glycan, solvent, and membrane components in SARS-CoV-2 spike protein open closed states. Employing μs-long molecular dynamics simulations powered by high-performance cloud-computing unsupervised density-driven adaptive sampling, investigated the differences bulk-solvent-glycan protein-solvent-glycan interfaces between these We unraveled sophisticated solvent-glycan polarization interaction network involving N165/N343 glycan-gate patterns that provide structural support for state identified key water molecules could potentially be targeted to destabilize this configuration. In state, reduced solvent diminishes overall dipoles, yet internal interactions reorganized sugar coat stabilize state. Despite variations, our glycan-solvent accessibility analysis reveals glycan shield capability conserve constant with effectively camouflaging virus from immune detection both The presented insights advance comprehension viral pathogenesis at an atomic level, offering potential combat COVID-19.

Language: Английский

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Conserved role of spike S2 domain N-glycosylation across betacoronaviruses

npj Viruses, Journal Year: 2025, Volume and Issue: 3(1)

Published: Jan. 25, 2025

Abstract Besides acting as an immunological shield, the N-glycans of SARS-CoV-2 are also critical for viral life cycle. As S2 subunit spike is highly conserved across betacoronaviruses, we determined functional significance five ‘stem N-glycans’ located in between N1098-N1194. Studies were performed with 31 Asn-to-Gln mutants, betacoronavirus virus-like particles and single-cycle replicons. Deletions stem enhanced S1 shedding from trimeric spike, reduced ACE2 binding abolished syncytia formation. When three or more deleted, expression on cell surface incorporation into virions was both reduced. Viral entry function progressively lost upon deleting N1098 glycan combination additional glycosite modifications. In addition to SARS-CoV-2, SARS-CoV MERS-CoV prevented target cells. These data suggest multiple roles N-glycans, evolutionarily properties these complex carbohydrates human betacoronaviruses.

Language: Английский

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Cell-based glycoengineering for production of homogeneous and specific glycoform-enriched antibodies with improved effector functions

Proceedings of the National Academy of Sciences, Journal Year: 2025, Volume and Issue: 122(8)

Published: Feb. 19, 2025

Glycosylation of humanized antibody at Fc-Asn297 significantly affects the Fc-mediated killing target cells through effector functions, especially antibody-dependent cellular cytotoxicity (ADCC), cell-mediated phagocytosis (ADCP), and vaccinal effect (ADVE). Previous studies showed that therapeutic immunoglobulin G (IgG) antibodies with α2,6-sialyl complex type (SCT) glycan attached to exhibited optimal binding Fc receptors on associated ADCC, ADCP, ADVE. However, production homogeneous Fc-SCT requires multiple in vitro enzymatic purification steps. In this study, we report two cell-based methods produce Fc-GlcNAc Fc-SCT-enriched improved functions. First, expressed endoglycosidase S2 Expi293F GnT1- trim all N-glycans for transglycosylation generate well-defined glycan. Second, engineered glycosylation pathway HEK293T knock-out undesired glycosyltransferases knock-in desired evaluated their receptors, found is like or better than

Language: Английский

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The diversity of the glycan shield of sarbecoviruses related to SARS-CoV-2

Cell Reports, Journal Year: 2023, Volume and Issue: 42(4), P. 112307 - 112307

Published: March 15, 2023

Animal reservoirs of sarbecoviruses represent a significant risk emergent pandemics, as evidenced by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic. Vaccines remain successful at limiting disease and death, but potential for further zoonosis motivates search pan-coronavirus vaccines. This necessitates better understanding glycan shields coronaviruses, which can occlude antibody epitopes on spike glycoproteins. Here, we compare structure 12 sarbecovirus shields. Of 22 N-linked attachment sites present SARS-CoV-2, 15 are shared all sarbecoviruses. However, there differences in processing state N-terminal domain, such N165. Conversely, glycosylation S2 domain highly conserved contain low abundance oligomannose-type glycans, suggesting shield density. The may therefore provide more attractive target immunogen design efforts aiming to generate response.

Language: Английский

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Role for N -glycans and calnexin-calreticulin chaperones in SARS-CoV-2 Spike maturation and viral infectivity

Science Advances, Journal Year: 2022, Volume and Issue: 8(38)

Published: Sept. 23, 2022

Functional and epidemiological data suggest that N-linked glycans on the SARS-CoV-2 Spike protein may contribute to viral infectivity. To investigate this, we created a panel of N-to-Q mutations at N-glycosylation sites proximal S1-S2 (N61, N603, N657, N616) S2' (N603 N801) proteolysis sites. Some these mutations, particularly N61Q N801Q, reduced incorporation into Spike-pseudotyped lentivirus authentic virus-like particles (VLPs). These also pseudovirus VLP entry ACE2-expressing cells by 80 90%. In contrast, glycan had relatively minor effect cell surface expression Spike, ACE2 binding, syncytia formation. A similar dichotomy in function was observed when virus produced host lacking ER chaperones, calnexin calreticulin. Here, while both chaperones regulated function, only VLPs KOs were less infectious. Overall, N-glycans are likely critical for could serve as drug targets COVID-19.

Language: Английский

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