Human noroviruses (HuNoV) are the leading cause of acute gastroenteritis worldwide. The humoral immune response plays an important role in clearing HuNoV infections and elucidating the antigenic landscape of HuNoV during an infection can shed light on antibody targets to inform vaccine design. Here, we utilized Jun-Fos-assisted phage display of a HuNoV genogroup GI.1 genomic library and deep sequencing to simultaneously map the epitopes of serum antibodies of six individuals infected with GI.1 HuNoV. We found both unique and common epitopes that were widely distributed among both nonstructural proteins and the major capsid protein. Recurring epitope profiles suggest immunodominant antibody footprints among these individuals. Analysis of sera collected longitudinally from three individuals showed the presence of existing epitopes in the pre-infection sera, suggesting these individuals had prior HuNoV infections. Nevertheless, newly recognized epitopes surfaced seven days post-infection. These new epitope signals persisted by 180 days post-infection along with the pre-infection epitopes, suggesting a persistent production of antibodies recognizing epitopes from previous and new infections. Lastly, analysis of a GII.4 genotype genomic phage display library with sera of three persons infected with GII.4 virus revealed epitopes that overlapped with those identified in GI.1 affinity selections, suggesting the presence of GI.1/GII.4 cross-reactive antibodies. The results demonstrate that genomic phage display coupled with deep sequencing can characterize HuNoV antigenic landscapes from complex polyclonal human sera to reveal the timing and breadth of the human humoral immune response to infection.

人诺如病毒 （HuNoV） 是全球急性胃肠炎的主要原因。体液免疫反应在清除 HuNoV 感染和阐明感染期间 HuNoV 的抗原景观方面起着重要作用，可以阐明抗体靶点，为疫苗设计提供信息。在这里，我们利用 HuNoV 基因群 GI.1 基因组文库的 Jun-Fos 辅助噬菌体展示和深度测序来同时绘制 6 个感染 GI.1 HuNoV 的个体的血清抗体表位。我们发现了独特和常见的表位，它们广泛分布在非结构蛋白和主要衣壳蛋白中。反复出现的表位谱表明这些个体中存在免疫显性抗体足迹。对从 3 个个体纵向收集的血清的分析显示，感染前血清中存在现有表位，表明这些个体之前曾感染过 HuNoV。尽管如此，新发现的表位在感染后 7 天浮出水面。这些新的表位信号与感染前表位一起在感染后 180 天内持续存在，表明识别来自先前和新感染的表位的抗体持续产生。最后，对 GII.4 基因型基因组噬菌体展示文库和三名 GII.4 病毒感染者的血清进行分析，发现表位与 GI.1 亲和选择中鉴定的表位重叠，表明存在 GI.1/GII.4 交叉反应抗体。结果表明，基因组噬菌体展示与深度测序相结合，可以表征来自复杂多克隆人血清的 HuNoV 抗原景观，以揭示人体液免疫反应对感染的时间和广度。