Structural convergence and water-mediated substrate mimicry enable broad neuraminidase inhibition by human antibodies

Abstract Influenza has been responsible for multiple global pandemics and seasonal epidemics and claimed millions of lives. The imminent threat of a panzootic outbreak of avian influenza H5N1 virus underscores the urgent need for pandemic preparedness and effective countermeasures, including monoclo...

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Main Authors: Julia Lederhofer, Andrew J. Borst, Lam Nguyen, Rebecca A. Gillespie, Connor J. Williams, Emma L. Walker, Julie E. Raab, Christina Yap, Daniel Ellis, Adrian Creanga, Hyon-Xhi Tan, Thi H. T. Do, Michelle Ravichandran, Adrian B. McDermott, Valerie Le Sage, Sarah F. Andrews, Barney S. Graham, Adam K. Wheatley, Douglas S. Reed, Neil P. King, Masaru Kanekiyo
Format: Article
Language:English
Published: Nature Portfolio 2025-08-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-025-62339-z
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author Julia Lederhofer
Andrew J. Borst
Lam Nguyen
Rebecca A. Gillespie
Connor J. Williams
Emma L. Walker
Julie E. Raab
Christina Yap
Daniel Ellis
Adrian Creanga
Hyon-Xhi Tan
Thi H. T. Do
Michelle Ravichandran
Adrian B. McDermott
Valerie Le Sage
Sarah F. Andrews
Barney S. Graham
Adam K. Wheatley
Douglas S. Reed
Neil P. King
Masaru Kanekiyo
author_facet Julia Lederhofer
Andrew J. Borst
Lam Nguyen
Rebecca A. Gillespie
Connor J. Williams
Emma L. Walker
Julie E. Raab
Christina Yap
Daniel Ellis
Adrian Creanga
Hyon-Xhi Tan
Thi H. T. Do
Michelle Ravichandran
Adrian B. McDermott
Valerie Le Sage
Sarah F. Andrews
Barney S. Graham
Adam K. Wheatley
Douglas S. Reed
Neil P. King
Masaru Kanekiyo
author_sort Julia Lederhofer
collection DOAJ
description Abstract Influenza has been responsible for multiple global pandemics and seasonal epidemics and claimed millions of lives. The imminent threat of a panzootic outbreak of avian influenza H5N1 virus underscores the urgent need for pandemic preparedness and effective countermeasures, including monoclonal antibodies (mAbs). Here, we characterize human mAbs that target the highly conserved catalytic site of viral neuraminidase (NA), termed NCS mAbs, and the molecular basis of their broad specificity. Cross-reactive NA-specific B cells were isolated by using stabilized NA probes of non-circulating subtypes. We found that NCS mAbs recognized multiple NAs of influenza A as well as influenza B NAs and conferred prophylactic protections in mice against H1N1, H5N1, and influenza B viruses. Cryo-electron microscopy structures of two NCS mAbs revealed that they rely on structural mimicry of sialic acid, the substrate of NA, by coordinating not only amino acid side chains but also water molecules, enabling inhibition of NA activity across multiple influenza A and B viruses, including avian influenza clade 2.3.4.4b H5N1 viruses. Our results provide a molecular basis for the broad reactivity and inhibitory activity of NCS mAbs targeting the catalytic site of NA through substrate mimicry.
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spelling doaj-art-10602569ec1b463e8e695c3ee4e5716b2025-08-20T03:46:11ZengNature PortfolioNature Communications2041-17232025-08-0116111510.1038/s41467-025-62339-zStructural convergence and water-mediated substrate mimicry enable broad neuraminidase inhibition by human antibodiesJulia Lederhofer0Andrew J. Borst1Lam Nguyen2Rebecca A. Gillespie3Connor J. Williams4Emma L. Walker5Julie E. Raab6Christina Yap7Daniel Ellis8Adrian Creanga9Hyon-Xhi Tan10Thi H. T. Do11Michelle Ravichandran12Adrian B. McDermott13Valerie Le Sage14Sarah F. Andrews15Barney S. Graham16Adam K. Wheatley17Douglas S. Reed18Neil P. King19Masaru Kanekiyo20Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of HealthInstitute for Protein Design, University of WashingtonVaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of HealthVaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of HealthDepartment of Immunology, Center for Vaccine Research, University of PittsburghDepartment of Immunology, Center for Vaccine Research, University of PittsburghVaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of HealthVaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of HealthInstitute for Protein Design, University of WashingtonVaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of HealthDepartment of Microbiology and Immunology, Peter Doherty Institute for Infection and Immunity, University of MelbourneDepartment of Microbiology and Immunology, Peter Doherty Institute for Infection and Immunity, University of MelbourneVaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of HealthVaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of HealthDepartment of Immunology, Center for Vaccine Research, University of PittsburghVaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of HealthVaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of HealthDepartment of Microbiology and Immunology, Peter Doherty Institute for Infection and Immunity, University of MelbourneDepartment of Immunology, Center for Vaccine Research, University of PittsburghInstitute for Protein Design, University of WashingtonVaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of HealthAbstract Influenza has been responsible for multiple global pandemics and seasonal epidemics and claimed millions of lives. The imminent threat of a panzootic outbreak of avian influenza H5N1 virus underscores the urgent need for pandemic preparedness and effective countermeasures, including monoclonal antibodies (mAbs). Here, we characterize human mAbs that target the highly conserved catalytic site of viral neuraminidase (NA), termed NCS mAbs, and the molecular basis of their broad specificity. Cross-reactive NA-specific B cells were isolated by using stabilized NA probes of non-circulating subtypes. We found that NCS mAbs recognized multiple NAs of influenza A as well as influenza B NAs and conferred prophylactic protections in mice against H1N1, H5N1, and influenza B viruses. Cryo-electron microscopy structures of two NCS mAbs revealed that they rely on structural mimicry of sialic acid, the substrate of NA, by coordinating not only amino acid side chains but also water molecules, enabling inhibition of NA activity across multiple influenza A and B viruses, including avian influenza clade 2.3.4.4b H5N1 viruses. Our results provide a molecular basis for the broad reactivity and inhibitory activity of NCS mAbs targeting the catalytic site of NA through substrate mimicry.https://doi.org/10.1038/s41467-025-62339-z
spellingShingle Julia Lederhofer
Andrew J. Borst
Lam Nguyen
Rebecca A. Gillespie
Connor J. Williams
Emma L. Walker
Julie E. Raab
Christina Yap
Daniel Ellis
Adrian Creanga
Hyon-Xhi Tan
Thi H. T. Do
Michelle Ravichandran
Adrian B. McDermott
Valerie Le Sage
Sarah F. Andrews
Barney S. Graham
Adam K. Wheatley
Douglas S. Reed
Neil P. King
Masaru Kanekiyo
Structural convergence and water-mediated substrate mimicry enable broad neuraminidase inhibition by human antibodies
Nature Communications
title Structural convergence and water-mediated substrate mimicry enable broad neuraminidase inhibition by human antibodies
title_full Structural convergence and water-mediated substrate mimicry enable broad neuraminidase inhibition by human antibodies
title_fullStr Structural convergence and water-mediated substrate mimicry enable broad neuraminidase inhibition by human antibodies
title_full_unstemmed Structural convergence and water-mediated substrate mimicry enable broad neuraminidase inhibition by human antibodies
title_short Structural convergence and water-mediated substrate mimicry enable broad neuraminidase inhibition by human antibodies
title_sort structural convergence and water mediated substrate mimicry enable broad neuraminidase inhibition by human antibodies
url https://doi.org/10.1038/s41467-025-62339-z
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