第三淋巴结构（TLS）是一群免疫细胞的有组织聚集，与癌症预后和免疫治疗反应有关，但TLS的免疫结构尚不清楚。在这里，我们假设TLS中白细胞的空间结构可以通过自身所含的趋化因子受体来重建，至少是部分地。单细胞RNA测序（scRNA-seq）揭示了头颈鳞状细胞癌（HNSC）中47个亚种白细胞。结合大剂量RNA-seq，我们观察到CXCR3，CCR7，CCR6，CXCR5和CCR1是TLS相关的趋化因子受体。根据空间参照，TLS相关趋化因子受体的细胞图谱在HNSC TLS中得到了复杂的多重免疫组化（mIHC）描绘。随后，我们探索了分布在TLS中的细胞的功能和进化轨迹。我们的研究提出了一个重建TLS免疫结构的方法，可以通过在HNSC中诱导TLS的新生来增强抗肿瘤免疫反应。版权所有©2023 Elsevier B.V. 发布。

Tertiary lymphoid structures (TLSs) are organized aggregates of immune cells associated with favorable prognosis and response to immunotherapy in cancer, but the immune architecture of TLSs remains poorly elucidated. Here, we hypothesize that the spatial architecture of leukocytes in TLSs can be reconstructed de novo, at least partially, by cell-inherent chemokine receptors profiles. Single-cell RNA-sequencing (scRNA-seq) revealed 47 subpopulations of leukocytes in head and neck squamous cell carcinoma (HNSC). Combined with bulk RNA-seq, we observed that CXCR3, CCR7, CCR6, CXCR5, and CCR1 are TLS-associated chemokine receptors. According to the spatial reference, the cellular atlas with TLS-associated chemokine receptors in HNSC TLSs was elaborately portrayed by multiplex immunohistochemistry (mIHC). Subsequently, we explored the functions and evolutionary trajectory of cells distributed in TLSs. Our investigation presents an approach to reconstructing the immune architecture of TLSs, which would help boost the antitumor immune response by inducing neogenesis TLSs in HNSC.Copyright © 2023. Published by Elsevier B.V.