神经胶质瘤的特征是有限的淋巴细胞浸润，构成对各种免疫疗法不敏感的“免疫沙漠”肿瘤。本研究旨在探索诱导胶质瘤微环境（GME）内三级淋巴结构（TLS）形成的治疗策略，使其从免疫抵抗状态转变为激活状态。通过颅内注射Toll样药物成功诱导GME中TLS的形成受体 (TLR) 激动剂（OK-432、TLR2/4/9 激动剂）和神经胶质瘤抗原（即 αTLR-mix）。我们采用染色分析、抗体中和、单细胞 RNA 测序 (scRNA-Seq) 和 BCR/TCR 测序来研究 TLS 形成的潜在机制及其在抗神经胶质瘤免疫中的作用。此外，还进行了一项初步的转化临床研究。TLS 的形成与 GME 中淋巴细胞浸润的增加相关，并改善了患有神经胶质瘤的小鼠的预后。在 TLS 诱导机制的研究中，某些巨噬细胞/小胶质细胞和 Th17 分别显示“LTo”和“LTi”细胞标记，通过 LTα/β-LTβR 相互作用促进 TLS 诱导。 TLS 形成后，CD4 和 CD8 T 细胞（而非 CD19 B 细胞）有助于抗神经胶质瘤免疫。对大脑和淋巴结之间 B/T 细胞的比较分析表明，大脑 B/T 细胞从幼稚向成熟转变，一些 B 细胞突出显示 CSR 相关基因的富集，观察到 V 基因的使用和克隆型偏差。在相关的临床研究中，i.c. αTLR-mix治疗表现出耐受性，趋化因子/细胞因子测定提供了支持GME中TLS形成的初步证据。GME中的TLS诱导增强了抗神经胶质瘤免疫力，改善了免疫微环境，并控制了神经胶质瘤的生长，提示了治疗神经胶质瘤的潜在治疗途径future.© 作者 2024。由牛津大学出版社代表神经肿瘤学会出版。版权所有。如需商业重复使用，请联系 reprints@oup.com 获取转载和转载的翻译权。所有其他权限都可以通过我们网站文章页面上的权限链接通过我们的 RightsLink 服务获得 - 如需了解更多信息，请联系journals.permissions@oup.com。

Glioma, characterized by limited lymphocytic infiltration, constitutes an "immune-desert" tumor displaying insensitivity to various immunotherapies. This study aims to explore therapeutic strategies for inducing tertiary lymphoid structure (TLS) formation within the glioma microenvironment (GME) to transition it from an immune-resistant to an activated state.TLS formation in GME was successfully induced by intracranial administration of Toll-like receptor (TLR) agonists (OK-432, TLR2/4/9 agonist) and glioma antigens (i.c. αTLR-mix). We employed staining analysis, antibody neutralization, single-cell RNA sequencing (scRNA-Seq), and BCR/TCR sequencing to investigate the underlying mechanisms of TLS formation and its role in anti-glioma immunity. Additionally, a preliminary translational clinical study was conducted.TLS formation correlated with increased lymphocyte infiltration in GME and led to improved prognosis in glioma-bearing mice. In the study of TLS induction mechanisms, certain macrophages/microglia and Th17 displayed markers of "LTo" and "LTi" cells, respectively, interaction through LTα/β-LTβR promoted TLS induction. Post-TLS formation, CD4+ and CD8+ T cells but not CD19+ B cells contributed to anti-glioma immunity. Comparative analysis of B/T cells between brain and lymph node showed that brain B/T cells unveiled switch from naïve to mature, some B cells highlighted an enrichment of CSR-associated genes, V gene usage and clonotype bias were observed. In related clinical studies, i.c. αTLR-mix treatment exhibited tolerability, and chemokines/cytokines assay provided preliminary evidence supporting TLS formation in GME.TLS induction in GME enhanced anti-glioma immunity, improved the immune microenvironment, and controlled glioma growth, suggesting potential therapeutic avenues for treating glioma in the future.© The Author(s) 2024. Published by Oxford University Press on behalf of the Society for Neuro-Oncology. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.