终末淋巴结构（TLSs）与多种癌症的良好预后有关。然而，食管鳞癌（ESCC）的TLSs与预后之间的相关性以及TLSs对肿瘤免疫微环境（TIME）的影响尚不清楚。我们在180个ESCC标本中病理学评估了TLSs的意义，重点关注TLSs的成熟情况，并使用14个ESCC组织进行了单细胞RNA测序（scRNA-seq）以研究免疫细胞根据TLSs的存在的功能差异。TLSs阳性的病例与TLSs阴性的病例相比，复发无病生存（RFS）（p<0.0001）和总生存（OS）（p=0.0016）更好。此外，成熟的TLSs阳性病例的RFS和OS均优于不成熟的TLSs阳性病例（p=0.019和p=0.015）以及TLSs阴性的病例（p<0.0001和p=0.0002）。scRNA-seq显示在TLSs阳性肿瘤中，CD8+T细胞表达细胞毒性标志物水平较高，并且TLSs阳性肿瘤中的树突状细胞（DCs）的抗原递呈增强。免疫组织化学显示TLSs阳性肿瘤中浸润肿瘤的CD8+T细胞和DCs的密度显著高于TLSs阴性肿瘤。这些数据表明了TLSs在ESCC中的预后和功能重要性，并为了解TLSs对TIME提供了新的见解。© 2023. 本文作者，独家许可给 Springer Nature Limited。

Tertiary lymphoid structures (TLSs) are associated with a favorable prognosis in several cancers. However, the correlation between TLSs and outcomes of esophageal squamous cell carcinoma (ESCC) and the impact of TLSs on the tumor immune microenvironment (TIME) remain unknown.We pathologically evaluated the significance of TLSs in ESCC focusing on TLS maturation using 180 ESCC specimens and performed single-cell RNA sequencing (scRNA-seq) using 14 ESCC tissues to investigate functional differences of immune cells according to TLS presence.TLS+ cases had better recurrence-free-survival (RFS) (p < 0.0001) and overall survival (OS) (p = 0.0016) compared with TLS- cases. Additionally, mature TLS+ cases had better RFS and OS compared with immature TLS+ cases (p = 0.019 and p = 0.015) and TLS- cases (p < 0.0001 and p = 0.0002). The scRNA-seq showed that CD8+ T cells in TLS+ tumors expressed high levels of cytotoxic signatures and antigen-presentation of dendritic cells (DCs) was enhanced in TLS+ tumors. Immunohistochemistry showed that the densities of tumor-infiltrating CD8+ T cells and DCs were significantly higher in TLS+ tumors than those in TLS- tumors.These data suggest the prognostic and functional significance of TLSs in ESCC and provides new insights into TLSs on the TIME.© 2023. The Author(s), under exclusive licence to Springer Nature Limited.