Pre-existing hemagglutinin-specific antibody levels are associated with B-cell repertoire maturation and antibody breadth after influenza vaccination

Wei Wang; Yan Sun; Qing Liu; Yidan Xia; Jing Wang; Minhao Hu; Mengyao Kong; Jun He; Rongbao Gao; Yong Gao

doi:10.1016/j.virs.2026.08.007

August 2026

Citation: Wei Wang, Yan Sun, Qing Liu, Yidan Xia, Jing Wang, Minhao Hu, Mengyao Kong, Jun He, Rongbao Gao, Yong Gao. Pre-existing hemagglutinin-specific antibody levels are associated with B-cell repertoire maturation and antibody breadth after influenza vaccination .VIROLOGICA SINICA, 2026, 41(4) : 898-911. http://dx.doi.org/10.1016/j.virs.2026.08.007

Pre-existing hemagglutinin-specific antibody levels are associated with B-cell repertoire maturation and antibody breadth after influenza vaccination

Wei Wang ^a ,
Yan Sun ^a ,
Qing Liu ^a ,
Yidan Xia ^d ,
Jing Wang ^a ,
Minhao Hu ^d ,
Mengyao Kong ^c ,
Jun He ^{c,d
,,} ,
Rongbao Gao ^{b
,,} ,
Yong Gao ^{a,e
,,}

a. The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, Anhui, 230001, China;
b. NHC Key Laboratory of Biosafety, NHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, 102206, China;
c. Microbiological Laboratory, Anhui Provincial Center for Disease Control and Prevention, Hefei, 230601, China;
d. Department of Health Inspection and Quarantine, School of Public Health, Anhui Medical University, Hefei, 230032, China;
e. Hefei Infectious Disease Hospital, Hefei, 230000, China

Corresponding author: Jun He, heliosking@sina.com
Rongbao Gao, rongbaogao@163.com
Yong Gao, ygao387@ustc.edu.cn
Received Date: 25 April 2026
Accepted Date: 07 August 2026
Available online: 10 August 2026

Abstract

Repeated influenza exposures generate complex antibody landscapes, yet how pre-existing antibodies are associated with subsequent vaccine responses remains unclear. We longitudinally tracked 21 adults stratified by pre-vaccination neutralizing titers to examine B-cell dynamics using flow cytometry, B cell receptor (BCR) sequencing, and monoclonal antibody characterization. After vaccination, participants with high baseline titers showed a longitudinal increase in the frequency of HA⁺CD27^-IgD⁺ naive-phenotype B cells. BCRs recovered from hemagglutinin (HA)-binding (HA⁺) B cells in the high-titer group displayed lower somatic hypermutation and shorter CDR3s, whereas those recovered from the lower-titer groups showed higher mutation levels and more extensive class switching. Correspondingly, monoclonal antibodies from the high-titer group generally showed narrow reactivity, while those from lower-titer groups showed broader neutralizing activity against historical strains. Clonal tracing further suggested that mAbs from lower-titer groups were more often linked to pre-existing sequences. Together, these findings suggest that pre-existing antibody levels are associated with differences in B-cell repertoire maturation and antibody breadth after influenza vaccination.
- Influenza vaccination
- , Hemagglutinin (HA)
- , Pre-existing antibodies
- , B-cell receptor repertoire
- , Antibody breadth

Electronic Supplementary Material

10.1016j.virs.2026.08.007-ESM.docx
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