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目的 探究高致病性H5N1禽流感病毒HA基因型更替及其在2024-2025美国奶牛场疫情传播过程中的分子进化轨迹，阐述抗原表位关键氨基酸位点的演化规律及其对抗原谱的影响。方法 基于GISAID数据库高致病性H5N1禽流感病毒HA基因序列，运用系统发育分析、正向选择压力分析及关键氨基酸位点变异频率统计，分析基因进化和抗原位点变化特征，结合线性抗原表位评估技术分析关键位点的结构功能影响。结果 H5N1病毒HA总体上呈现出时间拓扑连续性，分布于侧枝的2.3.4.4b谱系毒株紧邻早期祖先分支，表现出“进化迟滞”，而2.3.4.4h分支虽遗传距离较远，却通过131和139位点的回复突变呈现抗原表型的‘返祖’。2024-2025美国奶牛场H5N1疫情经历了早期主导的B3.13基因型向D1.1基因型显著的更替。B3.13基因型呈现出典型的连续抗原漂移特征，其中位于130环关键抗原区的147位点受到正向选择（P<0.1）；抗原性预测显示该变异改变了局部抗原指数评分，且可能影响表位特征，提示其可能参与免疫逃逸与宿主适应过程。D1.1基因型其131、211及226等关键抗原位点的氨基酸组合趋同于2021年北美早期祖先株HA21-NL，呈现出遗传距离与抗原表型的非线性分离特征，类似于已报道的2.3.4.4h分支的抗原非线性演化模式。结论 H5N1病毒在新型哺乳动物宿主中采用了复杂的双重进化策略，即通过关键位点突变（如V147M）实现适应性漂移，或重现类似2.3.4.4h的机制实现早期抗原表型的回归。这种演化规律重塑了当前的流行毒株抗原谱，为评估现有疫苗匹配度及预警大流行风险提供依据。
Abstract：Objective To investigate hemagglutinin(HA) genotype replacement in highly pathogenic H5N1 avian influenza viruses and to characterize their molecular evolutionary trajectories during the 2024 – 2025 outbreak in U.S. dairy farms. In addition, this study aims to elucidate evolutionary patterns at key amino acid residues within antigenic epitopes and their impact on the antigenic landscape. Methods High-quality HA gene sequences of highly pathogenic avian influenza virus(HPAIV) H5N1 were retrieved from the GISAID database. Phylogenetic analysis, positive selection analysis, and frequency profiling of mutations at key amino acid sites were performed to characterize genetic evolution and antigenic variation. In parallel, linear epitope–based antigenicity assessment was conducted to evaluate the structural and functional implications of these key mutations. Results Overall, the H5N1 HA gene exhibited temporal continuity in phylogenetic topology. Strains within the clade 2.3.4.4b lineage, located on peripheral branches, clustered closely with early ancestral lineages, indicating a pattern of evolutionary stasis. In contrast, the clade 2.3.4.4h lineage, despite greater genetic divergence, displayed antigenic reversion through back mutations at positions 131 and 139. During the 2024– 2025 outbreak in U.S. dairy farms, a marked genotype shift occurred from the initially dominant B3.13 genotype to the D1.1 genotype. The B3.13 genotype exhibited a typical pattern of continuous antigenic drift, with residue 147—located in the critical antigenic 130-loop—under positive selection(P < 0.1). Antigenicity prediction suggested that this substitution alters local antigenic index scores and may affect epitope properties, indicating a potential role in immune escape and host adaptation. In contrast, the amino acid constellation at key antigenic sites(positions 131, 211, and 226) in the D1.1 genotype converged toward that of the early North American ancestral strain HA21-NL(2021). This pattern reflects a nonlinear decoupling between genetic distance and antigenic phenotype, resembling the antigenic evolution previously described for the clade 2.3.4.4h lineage. Conclusion H5N1 viruses appear to adopt a complex dual evolutionary strategy in novel mammalian hosts: either achieving adaptive antigenic drift through key mutations(e. g., V147M) or recapitulating mechanisms similar to those observed in the 2.3.4.4h lineage to revert toward earlier antigenic phenotypes. These evolutionary dynamics reshape the antigenic landscape of circulating strains and provide important insights for vaccine strain selection and pandemic risk assessment.
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基本信息:
DOI：10.13242/j.cnki.bingduxuebao.260084
中图分类号:R373.13
引用信息:
[1]蒲思宇,崔思敏,罗军皓,等.高致病性H5N1禽流感病毒HA抗原谱演化特征和基因型更替分析[J].病毒学报,2026,42(03):912-922.DOI:10.13242/j.cnki.bingduxuebao.260084.
基金信息:
国家重点研发计划（项目号：2023YFC2605602）,题目：高致病性病毒遗传进化特征与溯源关键技术研究;国家重点研发计划（项目号：2022YFC2303402）,题目：病原宿主互作机制研究及广谱新靶点发现; 国家科技重大专项（项目号：2025ZD01903202）,题目：高致病性禽传人流感病毒的人用疫苗技术储备研究; 国家自然科学基金（项目号：81971946）,题目：甲型流感病毒重症感染下C反应蛋白介导的免疫损伤机制研究~~
2026-05-09
2026-05-09
2026-05-09