背景:三级淋巴结构(TLS)是免疫细胞的有组织聚集体,在非淋巴组织中在出生后发育并与病理状况有关。TLS通常包含含有T细胞区和树突细胞并被T细胞区和树突细胞包围的B细胞滤泡。TLS作为抗肿瘤免疫的潜在介质在肿瘤微环境(TME)中的预后和预测价值已引起人们的兴趣。然而,TLS的定位和成熟与透明细胞肾细胞癌(ccRCC)中存在的临床结果之间的确切关系尚未阐明。
方法:使用免疫组织化学和多光谱荧光来评估TLS异质性以及TME细胞浸润特征。对来自两个独立队列的395例ccRCC患者的TLS异质性的预后意义进行了彻底调查。通过定量免疫细胞浸润来评估TLS异质性与免疫活性之间的关联。
结果:在34.2%的ccRCC样品中发现了浸润的TLS(N=395)。这些TLS被发现是肿瘤近端,肿瘤远端,或者两者都是37.8%,74.1%,和11.9%的TLS阳性病例,分别。在肿瘤远端TLS中发现早期TLS的比例更高(p=0.016),而肿瘤近端TLS主要包含次级滤泡样结构(p=0.004)。在主要研究队列中(复旦大学上海癌症中心,N=290),Kaplan-Meier分析显示,肿瘤近端TLS的存在与无进展生存期的改善之间存在显着相关性(PFS,p<0.001)和总生存期(OS,p=0.002)。相反,肿瘤远端TLS的存在与不良PFS(p=0.02)和OS(p=0.021)相关.这些发现在105例ccRCC患者的外部验证集中得到了进一步验证。值得注意的是,成熟TLS的存在(即次级卵泡样TLS,CD23+生发中心)与ccRCC患者更好的临床结局显着相关。此外,结合肿瘤近端TLS存在的新型列线图显示了ccRCC切除8年结局的显著可预测性(曲线下面积>0.80).此外,富含初级滤泡样TLS的肿瘤远端TLS的ccRCC样品在肿瘤远端区域表现出更高的程序性死亡配体1肿瘤相关巨噬细胞水平和调节性T细胞浸润,指示抑制TME。
结论:本研究首次阐明了TLS定位和成熟异质性对ccRCC不同临床结局的影响。研究结果表明,ccRCC中的大多数TLS位于肿瘤远端区域,并与未成熟相关。免疫抑制特征。此外,我们的研究结果证实了先前的研究,表明肿瘤近端TLS与良好的临床结局相关.
Tertiary lymphoid structures (TLS) are organized aggregates of immune cells that develop postnatally in non-lymphoid tissues and are associated with pathological conditions. TLS typically comprise B-cell follicles containing and are encompassed by T- cell zones and dendritic cells. The prognostic and predictive value of TLS in the tumor microenvironment (TME) as potential mediators of antitumor immunity have gained interest. However, the precise relationship between localization and maturation of TLS and the clinical outcome of their presence in clear cell renal cell carcinoma (ccRCC) is yet to be elucidated.
Immunohistochemistry and multispectral fluorescence were used to evaluate the TLS heterogeneity along with TME cell-infiltrating characterizations. A thorough investigation of the prognostic implications of the TLS heterogeneity in 395 patients with ccRCC from two independent cohorts was conducted. Associations between TLS heterogeneity and immunologic activity were assessed by quantifying the immune cell infiltration.
Infiltrated TLS were identified in 34.2% of the ccRCC samples (N=395). These TLS were found to be tumor-proximal, tumor-distal, or both in 37.8%, 74.1%, and 11.9% of the TLS-positive cases, respectively. A higher proportion of early TLS was found in tumor-distal TLS (p=0.016), while tumor-proximal TLS primarily comprised secondary follicle-like structures (p=0.004). In the main study cohort (Fudan University Shanghai Cancer Center, N=290), Kaplan-Meier analyses revealed a significant correlation between the presence of tumor-proximal TLS and improved progression-free survival (PFS, p<0.001) and overall survival (OS, p=0.002). Conversely, the presence of tumor-distal TLS was associated with poor PFS (p=0.02) and OS (p=0.021). These findings were further validated in an external validation set of 105 patients with ccRCC. Notably, the presence of mature TLS (namely secondary follicle-like TLS, with CD23+ germinal center) was significantly associated with better clinical outcomes in patients with ccRCC. Furthermore, novel nomograms incorporating the presence of tumor-proximal TLS demonstrated remarkable predictability for the 8-year outcomes of resected ccRCC (area under the curve >0.80). Additionally, ccRCC samples with tumor-distal TLS enriched with primary follicle-like TLS exhibited higher programmed death-ligand 1 tumor-associated macrophages levels and regulatory T cells infiltration in the tumor-distal region, indicative of a suppressive TME.
This study for the first time elucidates the impact of TLS localization and maturation heterogeneities on the divergent clinical outcomes of ccRCC. The findings reveal that most TLS in ccRCC are located in the tumor-distal area and are associated with immature, immunosuppressive characterizations. Furthermore, our findings corroborate previous research demonstrating that tumor-proximal TLS were associated with favorable clinical outcomes.