EB 病毒 (EBV) 与鼻咽癌 (NPC) 密切相关，尤其在中国南方流行。尽管 EBV 的 II 型潜伏期在 NPC 的发展中起着至关重要的作用，但一些裂解基因和间歇性重新激活对于病毒传播和肿瘤进展也至关重要。由于 T 细胞介导的免疫可有效靶向杀死 EBV 阳性细胞，因此在整个 EBV 生命周期中鉴定由高度流行的人类白细胞抗原 I 类 (HLA-I) 分子呈递的 EBV 衍生肽非常重要。在这里，我们构建了 EBV 阳性 NPC 细胞模型，以评估 EBV 裂解期肽在链霉亲和素标记的特定 HLA-I 分子上的呈递。利用基于质谱 (LC-MS/MS) 的免疫肽组学方法，我们表征了 11 种新型 EBV 肽以及两种先前鉴定的肽。此外，我们确定这些肽具有免疫原性，并且可以刺激中国南方鼻咽癌流行人群中 EBV VCA/NA-IgA 阳性供体的 PBMC。总体而言，这项工作表明，可以捕获高度流行的 HLA-I 特异性 EBV 肽并进行功能性呈现，以在体外模型中引发免疫反应，从而深入了解 EBV 裂解周期和再激活期间呈现的表位。它扩大了潜在鼻咽癌早期诊断和治疗的病毒靶标范围。

Epstein-Barr Virus (EBV) is closely linked to nasopharyngeal carcinoma (NPC), notably prevalent in southern China. Although type II latency of EBV plays a crucial role in the development of NPC, some lytic genes and intermittent reactivation are also critical for viral propagation and tumor progression. Since T cell-mediated immunity is effective in targeted killing of EBV-positive cells, it is important to identify EBV-derived peptides presented by highly prevalent human leukocyte antigen class I (HLA-I) molecules throughout the EBV life cycle. Here, we constructed an EBV-positive NPC cell model to evaluate the presentation of EBV lytic phase peptides on streptavidin-tagged specific HLA-I molecules. Utilizing a mass spectrometry (LC-MS/MS)-based immunopeptidomic approach, we characterized eleven novel EBV peptides as well as two previously identified peptides. Furthermore, we determined these peptides were immunogenic and could stimulate PBMCs from EBV VCA/NA-IgA positive donors in an NPC endemic southern Chinese population. Overall, this work demonstrates that highly prevalent HLA-I-specific EBV peptides can be captured and functionally presented to elicit immune responses in an in vitro model, which provides insight into the epitopes presented during EBV lytic cycle and reactivation. It expands the range of viral targets for potential NPC early diagnosis and treatment.