文献类型：期刊文献

英文题名：Structure-Guided Glycosylation of Hemagglutinin Enhances Stability and Modulates Immunogenicity of Influenza Vaccines

作者：Zhang, Zheng[1,2];Xiao, Zhiying[1,2];Zhang, Xu[1,2];Ye, Qian[1,2];Zhang, Xin[1,3];Tan, Wen-Song[1,2,4]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[2]Shanghai Collaborat Innovat Ctr Biomfg Technol SCI, Shanghai 200237, Peoples R China;[3]Shanghai Inst Biol Prod, Dept Virol & Vaccine, Shanghai 200052, Peoples R China;[4]Shanghai BioEngine Sci Tech Co Ltd, Shanghai 201203, Peoples R China

年份：2026

卷号：14

期号：5

外文期刊名：VACCINES

收录：;WOS:【SCI-EXPANDED(收录号:WOS:001775514000001)】；

语种：英文

外文关键词：hemagglutinin; glycosylation; influenza virus; vaccines

摘要：Background: Antigenic drift limits the protective efficacy of influenza vaccine. Glycosylation of hemagglutinin (HA) represents a promising immunofocusing strategy that enhances neutralizing antibody responses by masking immunodominant non-neutralizing epitopes. Methods: B-cell epitopes of influenza viruses were retrieved from the Immune Epitope Database and were mapped onto the HA structure of A/Puerto Rico/8/1934 (H1N1). Structure-guided analysis identified residues 136 and 137 as candidate sites for N-linked glycosylation (NLG). Single-site mutants (136NLG and 137NLG) were generated using reverse genetics and evaluated for stability, receptor binding, viral replication, and immunogenicity in a murine model with inactivated whole-virus vaccines. Results: Both mutants exhibited increased thermostability at 42 degrees C. Glycosylation reduced the HA-sialic acid affinity, resulting in decreased viral adsorption and internalization efficiency in MDCK cells, and delayed viral replication at low multiplicity of infection (MOI). In vivo, all vaccine groups provided complete protection against lethal challenge; notably, the 136NLG group exhibited reduced weight loss, indicating improved protective efficacy compared with wild-type (WT). Conclusions: Targeted glycosylation at residue 136 in the HA head domain effectively enhances the viral stability and elicits a 1.78-fold increase in hemagglutination inhibition titer (GMT) relative to the WT, thereby improving vaccine performance. These findings establish a rational and structure-based design strategy for developing more stable and effective influenza vaccines.