在长期的慢性炎症过程中形成了第三级淋巴结构（TLSs），促进局部淋巴细胞的招募、抗原呈递和免疫反应调节，与癌症患者预后良好相关。虽然已经有研究探索了加速TLSs建立的方法，但在头颈鳞状细胞癌（HNSCC）中相关研究仍然缺乏。在本研究中，我们分析了The Cancer Genome Atlas（TCGA）的数据，并对188个患者样本进行了免疫组化染色分析。结果显示，TLSs促进了免疫细胞的浸润。TLSs中CD8 +细胞浸润高的患者预后最好。由于在TLSs组织中Lymphotoxinα（LTα）明显增加，我们在SCC7细胞中过表达了LTα（一种小鼠源性HNSCC细胞系）并建立了舌肿瘤携带模型。组织切片的多色观察显示，在LTα细胞组簇增加了T细胞集聚，第12天接种细胞时形成了1级TLS。此外，与对照组相比，LTα组的肿瘤体积显着减少，而浸润的CD8 + T细胞数量和比例均增加。去除小鼠周围的CD8 +细胞未发现肿瘤大小或TLS形成的差异。值得注意的是，TLS通过招募HNSCC中的CD8 + T细胞增强了抗肿瘤效应，显示出CD8 + T细胞介导的抗肿瘤效应。此外，肿瘤中LTα的过表达促进了TLSs的形成。本文受版权保护。保留所有权利。

Tertiary lymphoid structures (TLSs) are formed in long-term chronic inflammation, promoting the local recruitment of lymphocytes, antigen presentation, and regulation of immune response, correlated with a better prognosis for cancer patients. Although studies have been conducted to explore methods that accelerate the establishment of TLSs, related research in head and neck squamous cell carcinoma (HNSCC) is still lacking. In the current study, we analyzed data from The Cancer Genome Atlas (TCGA) and performed immunohistochemical (IHC) staining analyses of 188 patient samples. The results showed that TLSs promoted the infiltration of immune cells. Patients with TLSs with high infiltration of CD8+ cells showed the best prognosis. Since lymphotoxin α (LTα) was significantly increased in tissues with TLSs, we overexpressed LTα in SCC7 cells (a mouse-derived HNSCC cell line) and established tongue-tumour-bearing models. The polychromatic observation of tissue sections showed that T-cell aggregation increased in the LTα cell group, and a grade 1 TLS was formed on the 12th day after inoculating the cells. Moreover, the tumour volume in the LTα group was significantly less compared to the control group, whereas both the number and proportion of infiltrated CD8+ T cells were increased. The peripheral CD8+ cells in mice were removed and no difference was observed in tumour size or TLS formation. Remarkably, we found that TLS led to an increase in the anti-tumour effect by recruiting CD8+ T cells in HNSCC, showing a CD8+ T-cell-dependent anti-tumour effect. Moreover, LTα overexpression in the tumour promoted the formation of TLSs.This article is protected by copyright. All rights reserved.