高危型人乳头瘤病毒(HPV)是增加口咽部鳞状细胞癌(OPSCCs)数量的主要原因。这些癌症的病毒病因提供了与没有病毒成分的癌症相比范围更小的抗原定向治疗的机会。然而，特定的病毒编码表位及其相应的免疫反应尚未完全定义。为了了解OPSCC的免疫景观，我们对HPV16+和HPV33+原发肿瘤和转移淋巴结进行了全面的单细胞分析。我们使用编码的肽-人白细胞抗原(HLA)四聚体单细胞分析了HPV16+和HPV33+ OPSCC肿瘤，表征了体外细胞对在主要I类和II类HLA等位基因中呈现的HPV衍生抗原的反应。我们确定了针对HPV16蛋白E1和E2的强大细胞毒性T细胞反应，这些反应在多个患者之间共享，特别是在HLA-A*01:01和HLA-B*08:01中。对E2的反应与至少一种肿瘤中E2表达的丧失相关，表明这些E2识别的T细胞的功能能力以及其中许多相互作用已在功能性实验中验证。相反，对E6和E7的细胞反应在数量和细胞毒性方面受到限制，而肿瘤E6和E7表达持续存在。这些数据突显了HPV16 E6和E7以外的抗原性，并提名了抗原定向治疗的候选人。©作者(s)(或其雇主(s))2023年。在CC BY-NC下允许再次使用。不得商业再利用。由BMJ发表。

High-risk human papillomavirus (HPV) is a primary cause of an increasing number of oropharyngeal squamous cell carcinomas (OPSCCs). The viral etiology of these cancers provides the opportunity for antigen-directed therapies that are restricted in scope compared with cancers without viral components. However, specific virally-encoded epitopes and their corresponding immune responses are not fully defined.To understand the OPSCC immune landscape, we conducted a comprehensive single-cell analysis of HPV16+ and HPV33+ primary tumors and metastatic lymph nodes. We used single-cell analysis with encoded peptide-human leukocyte antigen (HLA) tetramers to analyze HPV16+ and HPV33+ OPSCC tumors, characterizing the ex vivo cellular responses to HPV-derived antigens presented in major Class I and Class II HLA alleles.We identified robust cytotoxic T-cell responses to HPV16 proteins E1 and E2 that were shared across multiple patients, particularly in HLA-A*01:01 and HLA-B*08:01. Responses to E2 were associated with loss of E2 expression in at least one tumor, indicating the functional capacity of these E2-recognizing T cells and many of these interactions validated in a functional assay. Conversely, cellular responses to E6 and E7 were limited in quantity and cytotoxic capacity, and tumor E6 and E7 expression persisted.These data highlight antigenicity beyond HPV16 E6 and E7 and nominate candidates for antigen-directed therapies.© Author(s) (or their employer(s)) 2023. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.