Long-lasting B cell convergence to distinct broadly reactive epitopes following vaccination with chimeric influenza virus hemagglutinins

Jenna J Guthmiller¹, Linda Yu-Ling Lan², Lei Li³, Yanbin Fu⁴, Sean A Nelson⁴, Carole Henry⁵, Christopher T Stamper², Henry A Utset⁵, Alec W Freyn⁶, Julianna Han⁷, Olivia Stovicek⁵, Jiaolong Wang⁵, Nai-Ying Zheng³, Min Huang³, Haley L Dugan², Micah E Tepora⁵, Xueyong Zhu⁷, Yao-Qing Chen⁵, Anna-Karin E Palm⁵, Dustin G Shaw², Madhumathi Loganathan⁶, Benjamin F Francis⁶, Jiayi Sun⁴, Jordan Chervin⁴, Chloe Troxell⁴, Philip Meade⁶, Nancy H L Leung⁸, Sophie A Valkenburg⁹, Sarah Cobey¹⁰, Benjamin J Cowling¹¹, Ian A Wilson¹², Adolfo García-Sastre¹³, Raffael Nachbagauer⁶, Andrew B Ward⁷, Lynda Coughlan¹⁴, Florian Krammer¹⁵, Patrick C Wilson¹⁶

Affiliations

¹ Department of Medicine, Section of Rheumatology, University of Chicago, Chicago, IL 60637, USA; Department on Immunology and Microbiology, University of Colorado Anschutz Medical Campus, Aurora, CO 80045, USA. Electronic address: jenna.guthmiller@cuanschutz.edu.
² Committee on Immunology, University of Chicago, Chicago, IL 60637, USA.
³ Department of Medicine, Section of Rheumatology, University of Chicago, Chicago, IL 60637, USA; Drukier Institute for Children's Health, Department of Pediatrics, Weill Cornell Medicine, New York, NY 10021, USA.
⁴ Drukier Institute for Children's Health, Department of Pediatrics, Weill Cornell Medicine, New York, NY 10021, USA.
⁵ Department of Medicine, Section of Rheumatology, University of Chicago, Chicago, IL 60637, USA.
⁶ Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
⁷ Department of Integrative Structural and Computational Biology, The Scripps Research Institute, La Jolla, CA 92037, USA.
⁸ WHO Collaborating Center for Infectious Disease Epidemiology and Control, School of Public Health, LKS Faculty of Medicine, The University of Hong Kong, Hong Kong, China; Laboratory of Data Discovery for Health Limited, Hong Kong Science and Technology Park, Hong Kong, China.
⁹ Department of Microbiology and Immunology, University of Melbourne, Peter Doherty Institute for Infection and Immunity, Melbourne, VIC 3000, Australia; HKU-Pasteur Research Pole, School of Public Health, The University of Hong Kong, Hong Kong, China.
¹⁰ Department of Ecology and Evolution, University of Chicago, Chicago, IL 60637, USA.
¹¹ WHO Collaborating Center for Infectious Disease Epidemiology and Control, School of Public Health, LKS Faculty of Medicine, The University of Hong Kong, Hong Kong, China.
¹² Department of Integrative Structural and Computational Biology, The Scripps Research Institute, La Jolla, CA 92037, USA; The Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, CA 92037, USA.
¹³ Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA; Department of Medicine, Division of Infectious Diseases, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA; Global Health and Emerging Pathogens Institute, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA; The Tisch Cancer Center, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA; Department of Pathology, Molecular and Cell-Based Medicine, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
¹⁴ Department of Microbiology and Immunology, University of Maryland School of Medicine, Baltimore, MD 21201, USA; Center for Vaccine Development and Global Health (CVD), University of Maryland School of Medicine, Baltimore, MD 21201, USA.
¹⁵ Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA; Department of Pathology, Molecular and Cell-Based Medicine, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA; Center for Vaccine Research and Pandemic Preparedness (C-VaRPP), Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
¹⁶ Department of Medicine, Section of Rheumatology, University of Chicago, Chicago, IL 60637, USA; Committee on Immunology, University of Chicago, Chicago, IL 60637, USA; Drukier Institute for Children's Health, Department of Pediatrics, Weill Cornell Medicine, New York, NY 10021, USA. Electronic address: pcw4001@med.cornell.edu.

PMID: 40132593
PMCID: PMC11981830 (available on 2026-04-08)
DOI: 10.1016/j.immuni.2025.02.025

Long-lasting B cell convergence to distinct broadly reactive epitopes following vaccination with chimeric influenza virus hemagglutinins

Jenna J Guthmiller et al. Immunity. 2025.

. 2025 Apr 8;58(4):980-996.e7.

doi: 10.1016/j.immuni.2025.02.025. Epub 2025 Mar 24.

Authors

Affiliations

¹ Department of Medicine, Section of Rheumatology, University of Chicago, Chicago, IL 60637, USA; Department on Immunology and Microbiology, University of Colorado Anschutz Medical Campus, Aurora, CO 80045, USA. Electronic address: jenna.guthmiller@cuanschutz.edu.
² Committee on Immunology, University of Chicago, Chicago, IL 60637, USA.
³ Department of Medicine, Section of Rheumatology, University of Chicago, Chicago, IL 60637, USA; Drukier Institute for Children's Health, Department of Pediatrics, Weill Cornell Medicine, New York, NY 10021, USA.
⁴ Drukier Institute for Children's Health, Department of Pediatrics, Weill Cornell Medicine, New York, NY 10021, USA.
⁵ Department of Medicine, Section of Rheumatology, University of Chicago, Chicago, IL 60637, USA.
⁶ Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
⁷ Department of Integrative Structural and Computational Biology, The Scripps Research Institute, La Jolla, CA 92037, USA.
⁸ WHO Collaborating Center for Infectious Disease Epidemiology and Control, School of Public Health, LKS Faculty of Medicine, The University of Hong Kong, Hong Kong, China; Laboratory of Data Discovery for Health Limited, Hong Kong Science and Technology Park, Hong Kong, China.
⁹ Department of Microbiology and Immunology, University of Melbourne, Peter Doherty Institute for Infection and Immunity, Melbourne, VIC 3000, Australia; HKU-Pasteur Research Pole, School of Public Health, The University of Hong Kong, Hong Kong, China.
¹⁰ Department of Ecology and Evolution, University of Chicago, Chicago, IL 60637, USA.
¹¹ WHO Collaborating Center for Infectious Disease Epidemiology and Control, School of Public Health, LKS Faculty of Medicine, The University of Hong Kong, Hong Kong, China.
¹² Department of Integrative Structural and Computational Biology, The Scripps Research Institute, La Jolla, CA 92037, USA; The Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, CA 92037, USA.
¹³ Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA; Department of Medicine, Division of Infectious Diseases, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA; Global Health and Emerging Pathogens Institute, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA; The Tisch Cancer Center, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA; Department of Pathology, Molecular and Cell-Based Medicine, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
¹⁴ Department of Microbiology and Immunology, University of Maryland School of Medicine, Baltimore, MD 21201, USA; Center for Vaccine Development and Global Health (CVD), University of Maryland School of Medicine, Baltimore, MD 21201, USA.
¹⁵ Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA; Department of Pathology, Molecular and Cell-Based Medicine, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA; Center for Vaccine Research and Pandemic Preparedness (C-VaRPP), Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
¹⁶ Department of Medicine, Section of Rheumatology, University of Chicago, Chicago, IL 60637, USA; Committee on Immunology, University of Chicago, Chicago, IL 60637, USA; Drukier Institute for Children's Health, Department of Pediatrics, Weill Cornell Medicine, New York, NY 10021, USA. Electronic address: pcw4001@med.cornell.edu.

PMID: 40132593
PMCID: PMC11981830 (available on 2026-04-08)
DOI: 10.1016/j.immuni.2025.02.025

Abstract

In a phase 1 clinical trial, a chimeric hemagglutinin (cHA) immunogen induced antibody responses against the conserved hemagglutinin (HA) stalk domain as designed. Here, we determined the specificity, function, and subsets of B cells induced by cHA vaccination by pairing single-cell RNA sequencing and B cell receptor repertoire sequencing. We have shown that the cHA-inactivated vaccine with a squalene-based adjuvant induced a robust activated B cell and memory B cell (MBC) phenotype against two broadly neutralizing epitopes in the stalk domain. The overall specificities of the acute plasmablast (PB) and MBC responses clonally overlapped, suggesting B cell convergence to these broadly protective epitopes. At 1 year post immunization, we identified that cHA vaccination reshaped the HA-specific MBC pool to enrich for stalk-binding B cells. Altogether, these data indicate the cHA vaccine induced robust and durable B cell responses against broadly protective epitopes of the HA stalk domain, in line with serological data.

Keywords: HA anchor epitope; HA stalk epitope; broadly neutralizing epitopes; chimeric hemagglutinin; durable B cell responses; influenza; next-generation influenza vaccine; universal influenza vaccine.

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Conflict of interest statement

Declaration of interests The Icahn School of Medicine at Mount Sinai has filed patent applications regarding influenza virus vaccines and mAbs that list R.N., F.K., and A.G.-S. as inventors, and these authors have received royalty payments for some of this IP. GSK has in the past provided research support for influenza virus vaccine development to the laboratories of F.K. and A.G.-S. Dynavax is currently providing research support to the laboratories of F.K. and A.G.-S. F.K. is currently consulting for Avimex, Pfizer, GSK, and Third Rock Ventures. A.B.W. is currently consulting for Third Rock Ventures. R.N., C.H., A.W.F., and M.E.T. are employees of Moderna Inc., and R.N., C.H., and A.W.F. are shareholders of Moderna Inc. H.L.D. is an employee of Adimab, LLC, and is a shareholder of Adimab, LLC. The A.G.-S. laboratory has received research support from GSK, Pfizer, Senhwa Biosciences, Kenall Manufacturing, Blade Therapeutics, Avimex, Johnson & Johnson, Dynavax, 7Hills Pharma, Pharmamar, ImmunityBio, Accurius, Nanocomposix, Hexamer, N-fold LLC, Model Medicines, Atea Pharma, Applied Biological Laboratories, and Merck. A.G.-S. has consulting agreements for the following companies involving cash and/or stock: Castlevax, Amovir, Vivaldi Biosciences, Contrafect, 7Hills Pharma, Avimex, Vaxalto, Pagoda, Accurius, Esperovax, Farmak, Applied Biological Laboratories, Pharmamar, Paratus, CureLab Oncology, CureLab Veterinary, Synairgen, Paratus, Prosetta, and Pfizer. A.G.-S. has been an invited speaker in meeting events organized by Seqirus, Janssen, Abbott, and Astrazeneca. A.G.-S. is inventor on patents and patent applications on the use of antivirals and vaccines for the treatment and prevention of virus infections and cancer, owned by the Icahn School of Medicine at Mount Sinai, New York.

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Long-lasting B cell convergence to distinct broadly reactive epitopes following vaccination with chimeric influenza virus hemagglutinins

Affiliations

Long-lasting B cell convergence to distinct broadly reactive epitopes following vaccination with chimeric influenza virus hemagglutinins

Authors

Affiliations

Abstract

Conflict of interest statement

References

MeSH terms

Substances

Grants and funding

LinkOut - more resources

Full Text Sources

Medical