Multiplexed Antibody Sequencing and Profiling of the Human Hemagglutinin-specific Memory B Cell Response following Influenza Vaccination
- PMID: 39441352
- PMCID: PMC11573632
- DOI: 10.4049/jimmunol.2400326
Multiplexed Antibody Sequencing and Profiling of the Human Hemagglutinin-specific Memory B Cell Response following Influenza Vaccination
Abstract
Influenza virus is a highly contagious respiratory pathogen causing between 9.4 and 41 million infections per year in the United States in the last decade. Annual vaccination is recommended by the World Health Organization, with the goal to reduce influenza severity and transmission. Ag-specific single B cell sequencing methodologies have opened up new avenues into the dissection of the Ab response to influenza virus. The improvement of these methodologies is pivotal to reduce the associated costs and optimize the operational workflow and throughput, especially in the context of multiple samples. In this study, PBMCs and serum samples were collected longitudinally from eight influenza vaccinees either vaccinated yearly for four consecutive influenza seasons or once for one season. Following the serological and B cell profiling of their polyclonal Ab response to a panel of historical, recent, and next-generation influenza vaccine hemagglutinin (HA) and virus strains, a single multiplexed Ag-specific single B cell sequencing run allowed to capture HA-specific memory B cells that were analyzed for preferential Ig H chain/L chain pairing, isotype/subclass usage, and the presence of public BCR clonotypes across participants. Binding and functional profiles of representative private and public clonotypes confirmed their HA specificity, and their overall binding and functional activity were consistent with those observed at the polyclonal level. Collectively, this high-resolution and multiplexed Ab repertoire analysis demonstrated the validity of this optimized methodology in capturing Ag-specific BCR clonotypes, even in the context of a rare B cell population, such as in the case of the peripheral Ag-specific memory B cells.
Copyright © 2024 by The American Association of Immunologists, Inc.
Similar articles
-
Molecular features of the serological IgG repertoire elicited by egg-based, cell-based, or recombinant haemagglutinin-based seasonal influenza vaccines: a comparative, prospective, observational cohort study.Lancet Microbe. 2025 Feb;6(2):100935. doi: 10.1016/j.lanmic.2024.06.002. Epub 2024 Dec 9. Lancet Microbe. 2025. PMID: 39667375 Free PMC article.
-
Convergent antibody evolution and clonotype expansion following influenza virus vaccination.PLoS One. 2021 Feb 22;16(2):e0247253. doi: 10.1371/journal.pone.0247253. eCollection 2021. PLoS One. 2021. PMID: 33617543 Free PMC article.
-
Preexisting subtype immunodominance shapes memory B cell recall response to influenza vaccination.JCI Insight. 2020 Jan 16;5(1):e132155. doi: 10.1172/jci.insight.132155. JCI Insight. 2020. PMID: 31794433 Free PMC article.
-
Influenza vaccination strategies targeting the hemagglutinin stem region.Immunol Rev. 2020 Jul;296(1):132-141. doi: 10.1111/imr.12887. Epub 2020 Jun 16. Immunol Rev. 2020. PMID: 32542739 Free PMC article. Review.
-
Next-Generation Influenza HA Immunogens and Adjuvants in Pursuit of a Broadly Protective Vaccine.Viruses. 2021 Mar 24;13(4):546. doi: 10.3390/v13040546. Viruses. 2021. PMID: 33805245 Free PMC article. Review.
Cited by
-
Leveraging large language models to predict antibody biological activity against influenza A hemagglutinin.Comput Struct Biotechnol J. 2025 Mar 24;27:1286-1295. doi: 10.1016/j.csbj.2025.03.038. eCollection 2025. Comput Struct Biotechnol J. 2025. PMID: 40230408 Free PMC article.
References
-
- Vatti A, Monsalve DM, Pacheco Y, Chang C, Anaya JM, and Gershwin ME. 2017. Original antigenic sin: A comprehensive review. J Autoimmun 83: 12–21. - PubMed
-
- Hartley GE, Edwards ESJ, Bosco JJ, Ojaimi S, Stirling RG, Cameron PU, Flanagan K, Plebanski M, Hogarth PM, O’Hehir RE, and van Zelm MC. 2020. Influenza-specific IgG1(+) memory B-cell numbers increase upon booster vaccination in healthy adults but not in patients with predominantly antibody deficiency. Clin Transl Immunology 9: e1199. - PMC - PubMed
MeSH terms
Substances
Grants and funding
LinkOut - more resources
Full Text Sources
Medical