Introduction: The analysis of exhaustion marker expression, like immune checkpoint molecules (ICMs) on tumor-infiltrating lymphocytes as well as peripheral immune cells of patients with head and neck squamous cell carcinoma (HNSCC), is of high interest since it reveals information about the patient’s immune status and the effectiveness of immune modulation. However, data regarding age-dependent expression are lacking. Here, we demonstrate an increase in most ICMs associated with age and disease, suggesting immune checkpoint modulation as an evidence-based option for the treatment of aged HNSCC patients. Material and Methods: Peripheral blood mononuclear cells (PBMCs) (healthy: n = 30 with an age range from 21 to 84 years/HNSCC: n = 37 with an age range from 37 to 94 years) were analyzed via flow cytometry following (density gradient separation) standard protocol. Flow cytometry was performed using CD4 and CD8 as backbone markers as well as co-stimulatory molecules like CD137, OX40, GITR, and CD27 or ICM like PD-1, CTLA4, BTLA, LAG3, or TIM3 using a Gallios flow cytometer (Beckman Coulter, Brea, CA, USA). Results: We found a statistically significant decreased ICM expression on the PBMCs of healthy donors with increasing age. However, the expression of ICMs in HNSCC was significantly increased (PD1 on CD4+T cells:p= 0.001), while we found a decreased co-stimulatory molecule expression (e.g., CD27 on CD4+T cells and CD39+T cells:p= 0.003 andp= 0.009, respectively). Immune cells of HPVnegHNSCC have a significant age-dependent decrease in CD27 expression on CD8+, CD4+, and CD39+T cells (p= 0.0426,p= 0.0078, andp= 0.0078, respectively). Conclusions: This study sheds light on the changing co-stimulatory molecule/ICM expression regarding the patient’s age and reveals an increase in most immune checkpoint molecules for HNSCC patients according to their age. This new evidence is valuable in ensuring individualized therapeutic approaches in the increasingly relevant field of checkpoint inhibition, even in old age.
引言：分析头颈部鳞状细胞癌（HNSCC）患者肿瘤浸润淋巴细胞及外周免疫细胞中免疫耗竭标志物（如免疫检查点分子）的表达水平具有重要意义，因其能揭示患者的免疫状态及免疫调节治疗效果。然而，目前尚缺乏关于年龄依赖性表达的数据。本研究证实大多数免疫检查点分子随年龄增长和疾病进展而表达上调，提示免疫检查点调节可作为老年HNSCC患者循证治疗的选择方案。 材料与方法：通过密度梯度分离标准流程获取外周血单个核细胞（健康供体30例，年龄21-84岁；HNSCC患者37例，年龄37-94岁），采用Gallios流式细胞仪（贝克曼库尔特，美国）进行检测。以CD4和CD8作为基础标记物，同时检测共刺激分子（CD137、OX40、GITR、CD27）及免疫检查点分子（PD-1、CTLA4、BTLA、LAG3、TIM3）。 结果：健康供体PBMCs中免疫检查点分子表达随年龄增长呈统计学显著下降。而HNSCC患者免疫检查点分子表达显著升高（CD4+T细胞PD1：p=0.001），共刺激分子表达则降低（如CD4+T细胞CD27及CD39+T细胞：p=0.003和p=0.009）。HPV阴性HNSCC患者的免疫细胞中，CD8+、CD4+及CD39+T细胞的CD27表达呈现显著的年龄依赖性下降（p值分别为0.0426、0.0078和0.0078）。 结论：本研究揭示了共刺激分子/免疫检查点分子表达随患者年龄变化的规律，证实HNSCC患者多数免疫检查点分子表达随年龄增长而升高。这一新证据对于在日益重要的检查点抑制治疗领域（包括老年患者）实现个体化治疗方案具有重要价值。
肿瘤(癌症)患者之家