Yang H, Yang L Y, Zhao M Q, Wu L, Yan Y S, Zhao Y M, Xing L G, Sun X R
Department of Graduate, Shandong First Medical University, Jinan 250117, China Department of Nuclear Medicine, Shandong Cancer Hospital and Institute, Shandong First Medical University and Shandong Academy of Medical Sciences, Jinan 250117, China.
Department of Radiation Oncology, Shandong Cancer Hospital and Institute, Shandong First Medical University and Shandong Academy of Medical Sciences, Jinan 250117, China.
Zhonghua Zhong Liu Za Zhi. 2025 Jun 23;47(6):508-516. doi: 10.3760/cma.j.cn112152-20240805-00326.
The impact of bystander CD8 T cells (CD8 Tbys) within the tumor microenvironment on the prognosis of early-stage non-small cell lung cancer (NSCLC) remains unclear, particularly concerning their infiltration differences at the invasive margin (IM) and tumor center (TC). We retrospectively collected postoperative specimens from 173 patients with primary early-stage NSCLC who underwent radical surgery at the Affiliated Tumor Hospital of Shandong First Medical University from 2014 to 2018. Tissue microarrays encompassing IM and TC regions were prepared and subjected to multicolor immunofluorescence staining (CK/CD8/CD103/DAPI). Image processing and phenotype recognition (CD8 T cells, CKCD8; CD8 T, CKCD8CD103) were performed using inFrom software, and automatic quantitative cell density analysis was conducted using R language. Differences in CD8 T and CD8 T densities at IM and TC were analyzed using the Mann-Whitney test. The relationship between CD8 T, CD8 T and clinicopathological features was examined using the Kruskal-Wallis test. The impact of CD8 T and CD8 T on recurrence-free survival (RFS) was evaluated using Kaplan-Meier, log-rank, and Cox proportional hazards models. A total of 173 patients with stage ⅠA-ⅡA NSCLC were included, with a recurrence rate of 26.6% (46/173) and a median RFS of 62.3 months (range: 44.7-71.9 months). CD8 T and CD8 T densities (1/1 000) were significantly higher in the IM region than in the TC region [70(32, 155) vs. 37(18, 88), ＜0.001; 25(11, 46) vs. 18(7, 34), =0.002]. No significant association was found between CD8 T, CD8 T and age, gender, smoking history, histological type, or pathological stage (all ＞0.05). Patients with low-density IM CD8 T cells had lower RFS compared to those with high-density IM CD8 T cells (=0.021; median RFS not reached), Further analysis revealed that patients with low-density IM CD8 T cell had lower RFS compared to those with high-density IM CD8 T (=0.047; median RFS not reached), and low-density IM CD8 T cell was an independent risk factor for postoperative recurrence (=1.836, 95% :1.007-3.347, =0.048). Joint analysis of IM and TC revealed that patients with low IM CD8 T and high TC CD8 T had significantly lower RFS compared to the other three groups (=0.006), and this combination was an independent risk factor for postoperative recurrence in early-stage NSCLC (=2.090, 95% :1.162-3.760, =0.014). The spatial distribution of bystander CD8 T cells within the primary tumor influences the prognosis of patients with early-stage NSCLC. Patients with low-density IM CD8 T and high-density TC CD8 T are more prone to recurrence after radical surgery.
肿瘤微环境中旁观者CD8 T细胞（CD8 Tbys）对早期非小细胞肺癌（NSCLC）预后的影响尚不清楚，尤其是它们在浸润边缘（IM）和肿瘤中心（TC）的浸润差异。我们回顾性收集了2014年至2018年在山东第一医科大学附属肿瘤医院接受根治性手术的173例原发性早期NSCLC患者的术后标本。制备了包含IM和TC区域的组织微阵列，并进行多色免疫荧光染色（CK/CD8/CD103/DAPI）。使用inFrom软件进行图像处理和表型识别（CD8 T细胞，CKCD8；CD8 T，CKCD8CD103），并使用R语言进行自动定量细胞密度分析。使用Mann-Whitney检验分析IM和TC处CD8 T和CD8 T密度的差异。使用Kruskal-Wallis检验检查CD8 T、CD8 T与临床病理特征之间的关系。使用Kaplan-Meier、对数秩和Cox比例风险模型评估CD8 T和CD8 T对无复发生存期（RFS）的影响。共纳入173例ⅠA-ⅡA期NSCLC患者，复发率为26.6%（46/173），中位RFS为62.3个月（范围：44.7-71.9个月）。IM区域的CD8 T和CD8 T密度（每1000个细胞中细胞数）显著高于TC区域[70（32，155）对37（18，88），＜0.001；25（11，46）对18（7，34），=0.002]。未发现CD8 T、CD8 T与年龄、性别、吸烟史、组织学类型或病理分期之间存在显著关联（均＞0.05）。IM CD8 T细胞低密度的患者与IM CD8 T细胞高密度的患者相比，RFS较低（=0.021；中位RFS未达到）。进一步分析显示，IM CD8 T细胞低密度的患者与IM CD8 T高密度的患者相比，RFS较低（=0.047；中位RFS未达到），且IM CD8 T细胞低密度是术后复发 的独立危险因素（=1.836，95%置信区间：1.007-3.347，=0.048）。IM和TC的联合分析显示，IM CD8 T低且TC CD8 T高的患者与其他三组相比，RFS显著较低（=0.006），并且这种组合是早期NSCLC术后复发的独立危险因素（=2.090，95%置信区间：1.162-3.760，=0.014）。原发性肿瘤内旁观者CD8 T细胞的空间分布影响早期NSCLC患者的预后。IM CD8 T低密度且TC CD8 T高密度的患者在根治性手术后更容易复发。
