Song Yang, Chen Ji, Zhang Yaqin, Wu Ning, Zhu Yongjun, Chen Gang, Miao Feng, Chen Zhiming, Wang Yiqing
Department of Cardio-Thoracic Surgery, Huashan Hospital, Fudan University, Shanghai, China.
Cell Oncol (Dordr). 2025 Apr 7. doi: 10.1007/s13402-025-01040-1.
Adoptive cell therapy (ACT) mediates durable and complete regression of various cancers. However, its efficacy is limited by the long-term persistence of cytotoxic T lymphocytes, given their irreversible dysfunction within the tumor microenvironment. Herein, we aimed to establish an artificial lung metastasis model to examine T-lymphocyte subsets, in order to identify potential effective cell subsets for ACT.
A metastatic lung melanoma mouse model was established using OVA-expressing melanoma B16 cells. Flow cytometry analysis was conducted to examine the surface markers, transcription factors, and secreted cytokines of tumor-specific CD8 T cells within metastatic tissues. The infiltrated cells were sorted by flow cytometry for in vitro tumor cell killing assays or in vivo cell infusion therapy combined with chemotherapeutic drugs and immune checkpoint blockade antibodies.
Exhausted CD8 T cells (Tex) exhibited high heterogeneity in metastatic tissues. Among Tex cells, the CXCR6 precursor cell showed certain memory characteristics, including phenotype, transcription factors, and maintenance, whereas the CXCR6 subpopulation partially lost these traits. Moreover, CXCR6 precursor cells effectively replenished effector-like Tex cells in metastatic tissues and exerted direct cytotoxicity against tumor cells. Notably, transferring these tumor-specific CXCR6 precursor-exhausted T (Texp) cells into recipients induced a substantial regression of metastasis. In addition, these cells could respond to immune checkpoint blockade, which could better control tumor metastasis.
In our study, a subset of antigen-specific CXCR6-expressing Texp cells was observed within the metastatic tissue. The cells served as a crucial source of effector-like Tex cells and exerted direct cytotoxic effects on tumor cells. Adoptive transfer of CXCR6 Texp cells effectively mitigated lung metastasis in mice. This study helps elucidate the role of Texp cells in metastasis, thereby offering novel insights into enhancing the efficacy and durability of immunotherapy.
过继性细胞疗法（ACT）可介导多种癌症的持久且完全消退。然而，由于细胞毒性T淋巴细胞在肿瘤微环境中发生不可逆功能障碍，其长期存活受限，导致该疗法的疗效受到限制。在此，我们旨在建立一种人工肺转移模型来检测T淋巴细胞亚群，以确定ACT潜在的有效细胞亚群。
利用表达卵清蛋白（OVA）的黑色素瘤B16细胞建立转移性肺黑色素瘤小鼠模型。通过流式细胞术分析检测转移组织中肿瘤特异性CD8 T细胞的表面标志物、转录因子和分泌的细胞因子。通过流式细胞术对浸润细胞进行分选，用于体外肿瘤细胞杀伤试验或体内细胞输注治疗，并联合化疗药物和免疫检查点阻断抗体。
耗竭的CD8 T细胞（Tex）在转移组织中表现出高度异质性。在Tex细胞中，CXCR6前体细胞表现出一定的记忆特征，包括表型、转录因子和维持能力，而CXCR6亚群部分丧失了这些特征。此外，CXCR6前体细胞可有效补充转移组织中效应样Tex细胞，并对肿瘤细胞发挥直接细胞毒性作用。值得注意的是，将这些肿瘤特异性CXCR6前体耗竭T（Texp）细胞转移至受体小鼠可显著减轻转移。此外，这些细胞可对免疫检查点阻断产生反应，从而更好地控制肿瘤转移。
在我们的研究中，在转移组织中观察到了表达抗原特异性CXCR6的Texp细胞亚群。这些细胞是效应样Tex细胞的关键来源，对肿瘤细胞发挥直接细胞毒性作用。过继性转移CXCR6 Texp细胞可有效减轻小鼠的肺转移。本研究有助于阐明Texp细胞在转移中的作用，从而为提高免疫治疗的疗效和持久性提供新的见解。
