Characterization of prefusion-F-specific antibodies elicited by natural infection with human metapneumovirus

Rush, Scott A.; Brar, Gurinder Singh; Hsieh, Ching-Lin; Chautard, Émilie; Rainho-Tomko, Jennifer N; Slade, Chris D; Bricault, Christine A.; Kume, Ana; Kearns, James; Groppo, Rachel; Mundle, Sophia T.; Zhang, Linong; Casimiro, Danilo R.; Fu, Tong-Ming; DiNapoli, Joshua; McLellan, Jason S.

doi:10.1016/j.celrep.2022.111399

Cited by 10 publications

(15 citation statements)

References 82 publications

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“…A consistent bias across all categories towards IGHJ4 was also observed (Supplementary Figure 3). IGHV1-69 has been implicated in several anti-pathogen repertoires [34][35][36][37][38][39][40] and indeed was observed among all specificity categories here. In comparing to previously published repertoires, unselected by any antigen specificity [41,42], several genes were found at a higher frequency in the antigen specificity categories, most notably IGHV1-18 and IGHV3-30.…”

Section: Antigen-specific B Cell Patterns In V Gene Usage Ighv:igl(k)...supporting

confidence: 69%

“…Stabilized ectodomains of hMPV F subtypes A1 and B2 ( [38], as well as RSV strains A2 ( [53,54]and B9320 F (DS-Cav1) [55], were purified over Strep-Tactin Sepharose resin (IBA Lifesciences) in a gravity column. The resin was washed with 4 column volumes of PBS, and purified protein was eluted from the column with 4 column volumes of 100 mM Tris pH 8.0, 150 mM NaCl, 1 mM EDTA, 2.5 mM desthiobiotin.…”

Section: Antigen Expression and Purificationmentioning

confidence: 99%

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Convergent Sequence Features of Antiviral B Cells

Abu-Shmais,

Vukovich,

Wasdin

et al. 2023

Preprint

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Throughout life, humans experience repeated exposure to viral antigens through infection and vaccination, building diverse antigen-specific antibody repertoires. In recent years, these repertoires have become an important source for novel antibody-based antiviral therapeutics, yet there is still limited understanding of the determinants of antibody-antigen specificity. Here, we generated a large dataset mapping antibody sequence to antigen specificity for thousands of B cells, by screening the repertoires of a set of healthy individuals against twenty viral antigens representing diverse pathogens of biomedical significance. Analysis revealed antigen-specific patterns in variable gene usage, gene pairing, and somatic hypermutation, as well as the presence of convergent antiviral signatures across multiple individuals. These results help define the characteristics of human antibody repertoires simultaneously against an unprecedented number and diversity of viral targets. Understanding the fundamental rules of antibody-antigen interactions can lead to transformative new approaches for the development of antibody therapeutics and vaccines against current and emerging viruses.

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Section: Antigen-specific B Cell Patterns In V Gene Usage Ighv:igl(k)...supporting

confidence: 69%

Section: Antigen Expression and Purificationmentioning

confidence: 99%

Convergent Sequence Features of Antiviral B Cells

Abu-Shmais,

Vukovich,

Wasdin

et al. 2023

Preprint

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“…For hMPV, similar design strategies (e.g., disulfide bond, proline, and cavity-filling substitutions) have recently been used to stabilize prefusion F for vaccine development ( 35, 60–62 ). In addition to 101F and MPE8, which are previously identified NAbs targeting both RSV and hMPV ( 63, 64 ), further hMPV NAbs were identified and structurally characterized using prefusion F ( 65–67 ). Although these prefusion hMPV-F constructs showed promising results in animal studies ( 60, 61 ), significant gaps remain between preclinical research and vaccine approval for human use.…”

Section: Introductionmentioning

confidence: 99%

A tale of two fusion proteins: understanding the metastability of human respiratory syncytial virus and metapneumovirus and implications for rational design of uncleaved prefusion-closed trimers

Lee,

Han,

Zhang

et al. 2024

Preprint

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Respiratory syncytial virus (RSV) and human metapneumovirus (hMPV) cause human respiratory diseases and are major targets for vaccine development. In this study, we designed uncleaved prefusion-closed (UFC) trimers for the fusion (F) proteins of both viruses by examining mutations critical to F metastability. For RSV, we assessed four previous prefusion F designs, including the first and second generations of DS-Cav1, SC-TM, and 847A. We then identified key mutations that can maintain prefusion F in a native-like, closed trimeric form (up to 76%) without introducing any interprotomer disulfide bond. For hMPV, we developed a stable UFC trimer with a truncated F2-F1 linkage and an interprotomer disulfide bond. Tens of UFC constructs were characterized by negative-stain electron microscopy (nsEM), x-ray crystallography (11 RSV-F and one hMPV-F structures), and antigenic profiling. Using an optimized RSV-F UFC trimer as bait, we identified three potent RSV neutralizing antibodies (NAbs) from a phage-displayed human antibody library, with a public NAb lineage targeting sites Ø and V and two cross-pneumovirus NAbs recognizing site III. In mouse immunization, rationally designed RSV-F and hMPV-F UFC trimers induced robust antibody responses with high neutralizing titers. Our study provides a foundation for future prefusion F-based RSV and hMPV vaccine development.

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“…26–28 Silva et al 26 showed that accessibility of the full-length SARS-CoV-2 spike’s hinge epitope necessitates both RBD transitioning to the ‘up’ conformation and S2 subunits ‘breathing.’ This aligns with HDX-MS experiments showing the existence of an open conformation of the full-length spike where the S2 apex is solvent-exposed. 28 However, S2 apex epitopes elicit non-neutralizing antibodies, 26,27 which are dominant in the B cell population 27,37 and can interfere with binding of neutralizing antibodies. 38,39 This is also the case for other trimer interface epitopes in influenza HA 34 and human metapneumovirus (hMPV) F. 37 Hence, preventing the S2 trimer’s apex opening may offer the foundations for the design of improved S2-based immunogens that may also afford binding and elicitation of quaternary epitope-specific antibodies.…”

Section: Introductionmentioning

confidence: 99%

“…28 However, S2 apex epitopes elicit non-neutralizing antibodies, 26,27 which are dominant in the B cell population 27,37 and can interfere with binding of neutralizing antibodies. 38,39 This is also the case for other trimer interface epitopes in influenza HA 34 and human metapneumovirus (hMPV) F. 37 Hence, preventing the S2 trimer’s apex opening may offer the foundations for the design of improved S2-based immunogens that may also afford binding and elicitation of quaternary epitope-specific antibodies.…”

Section: Introductionmentioning

confidence: 99%

Simulation-Driven Design of Stabilized SARS-CoV-2 Spike S2 Immunogens

Nuqui,

Casalino,

Zhou

et al. 2023

Preprint

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The full-length prefusion-stabilized SARS-CoV-2 spike (S) is the principal antigen of COVID-19 vaccines. Vaccine efficacy has been impacted by emerging variants of concern that accumulate most of the sequence modifications in the immunodominant S1 subunit. S2, in contrast, is the most evolutionarily conserved region of the spike and can elicit broadly neutralizing and protective antibodies. Yet, the usage of S2 as an alternative vaccine strategy is hampered by its general instability. Here, we use a simulation-driven approach to design highly stable S2-only antigens retaining a closed prefusion conformation. Weighted ensemble simulations provide mechanistic characterization of the S2 trimer opening, informing the design of tryptophan substitutions that impart kinetic and thermodynamic stabilization. Alchemical free energy perturbation calculations and a corroborating set of experiments confirm that V991W and T998W in the central helices of S2 stabilize the trimer in the closed prefusion conformation, producing an antigen with increased protein expression, superior thermostability, and preserved immunogenicity against sarbecoviruses.

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Characterization of prefusion-F-specific antibodies elicited by natural infection with human metapneumovirus

Cited by 10 publications

References 82 publications

Convergent Sequence Features of Antiviral B Cells

Convergent Sequence Features of Antiviral B Cells

A tale of two fusion proteins: understanding the metastability of human respiratory syncytial virus and metapneumovirus and implications for rational design of uncleaved prefusion-closed trimers

Simulation-Driven Design of Stabilized SARS-CoV-2 Spike S2 Immunogens

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