Department of Clinical and Translational Research Center, Affiliated Hospital of Nantong University & Department of Pathology, Medical School of Nantong University, Jiangsu, China.
Department of Pathology, Affiliated Tumor Hospital of Nantong University, Nantong, Jiangsu Province, China.
Lab Invest. 2024 Nov;104(11):102144. doi: 10.1016/j.labinv.2024.102144. Epub 2024 Sep 27.
An increasing number of studies have revealed a correlation between tertiary lymphoid structures (TLSs) and the outcome of hepatocellular carcinoma (HCC). Nevertheless, the associations between the heterogeneity of cellular composition and the overall survival (OS) in HCC remain unexplored. Here, we evaluated the cancer tissues from 150 HCC individuals using multiplex immunofluorescence to determine the presence and characteristics of TLS and to investigate the relationship between intra-TLS immunologic activity, TLS maturation, and intratumoral immune cell infiltration. Prognostic factors influencing the outcome were identified through both univariate and multivariate analyses. Additionally, the levels of cytotoxic T-lymphocyte antigen-4 (CTLA-4), programmed death 1, programmed death-ligand 1, and lymphocyte activation gene-3 were determined, as well as their relationship with TLS features were determined. TLS was detected in 71 (47.3%) of the 150 HCC cases and was related to higher intratumoral infiltration levels of lymphocytes. Additionally, intra-TLS lymphocyte proliferation correlated with that of intratumoral lymphocytes, and the presence of TLS and a high proportion of mature TLS demonstrated a significant correlation with better prognosis (P = .013 and P = .03, respectively). Among TLS-positive tumors, a high proportion of B cells expressing activation-induced cytidine deaminase and a high proportion of CD8 T cells expressing CD45RO were significantly related to improved OS (P = .01 and P < .001, respectively). Comparatively, a high proportion of CD21CD20 B cells demonstrated a significant correlation with poorer OS (P < .001). A markedly reduced number of CTLA-4 cells in the stromal regions in TLS-negative tumors was observed compared with TLS-positive tumors (P = .01). These findings reveal a correlation between TLS presence and improved OS in HCC patients. However, TLS exhibited significant variation in maturation state, T- and B-cell proliferation, and expression of markers related to B- and T-cell function. Notably, these characteristics were also found to possess prognostic significance, indicating that certain TLS might hinder tumor immunity by inhibiting immune cells, whereas others may foster antigen-driven immune responses, likely influenced by the composition and functional status of intra-TLS lymphocytes.
越来越多的研究揭示了三级淋巴结构（TLS）与肝细胞癌（HCC）结局之间存在相关性。然而，HCC 中细胞组成异质性与总生存期（OS）之间的关联仍未得到探索。在这里，我们使用多重免疫荧光法评估了 150 名 HCC 患者的癌组织，以确定 TLS 的存在和特征，并研究 TLS 内免疫活性、TLS 成熟度与肿瘤内免疫细胞浸润之间的关系。通过单因素和多因素分析确定影响结局的预后因素。此外，还测定了细胞毒性 T 淋巴细胞抗原-4（CTLA-4）、程序性死亡 1（PD-1）、程序性死亡配体 1（PD-L1）和淋巴细胞激活基因-3 的水平，并确定了它们与 TLS 特征的关系。在 150 例 HCC 病例中，有 71 例（47.3%）检测到 TLS，与肿瘤内淋巴细胞浸润水平较高有关。此外，TLS 内淋巴细胞增殖与肿瘤内淋巴细胞增殖相关，TLS 的存在和成熟 TLS 的高比例与预后较好显著相关（P=0.013 和 P=0.03）。在 TLS 阳性肿瘤中，表达激活诱导胞苷脱氨酶的 B 细胞比例较高和表达 CD45RO 的 CD8 T 细胞比例较高与 OS 改善显著相关（P=0.01 和 P<0.001）。相比之下，CD21CD20 B 细胞比例较高与较差的 OS 显著相关（P<0.001）。与 TLS 阳性肿瘤相比，TLS 阴性肿瘤的基质区域 CTLA-4 细胞数量明显减少（P=0.01）。这些发现表明 TLS 的存在与 HCC 患者的 OS 改善相关。然而，TLS 在成熟状态、T 和 B 细胞增殖以及与 B 和 T 细胞功能相关的标志物表达方面存在显著差异。值得注意的是，这些特征也具有预后意义，表明某些 TLS 可能通过抑制免疫细胞来阻碍肿瘤免疫，而其他 TLS 可能通过促进抗原驱动的免疫反应来促进肿瘤免疫，这可能受到 TLS 内淋巴细胞的组成和功能状态的影响。
