Chen Sheng, Xu Chunzuan, Liu Changyun, Li Jiaqi, Ke Shupei, Lu Yingqian, Huang Yali, Chen Jialin, Lin Feifei, Huang Huapin, Zou Zhangyu
Department of Neurology, Fujian Medical University Union Hospital, Fuzhou, China.
Institute of Clinical Neurology, Fujian Medical University, Fuzhou, China.
Ann Med. 2025 Dec;57(1):2540023. doi: 10.1080/07853890.2025.2540023. Epub 2025 Aug 3.
Amyotrophic lateral sclerosis (ALS) urgently requires robust biomarkers for early diagnosis and prognostic stratification. This study aims to investigate the diagnostic and prognostic potential of membrane-bound and soluble immune checkpoint molecules in ALS pathogenesis.
In the present study at Fujian Medical Union Hospital, 72 participants (46 ALS and 26 healthy controls [HC]) underwent flow cytometry analysis of PD-1 expression in CD4 T cells and its subsets. A second cohort ( = 93, 44 ALS, 30 HC and 19 ALS mimics [Mimics]) was evaluated using Luminex technology for 14 serum immune checkpoint molecules. A single-molecule array was used to screen the neurofilament light chain (NFL) in serum.
Flow cytometry revealed elevated PD1 expression in CD4 T cells, particularly in Th9 and Th17 subsets ( < 0.05). ALS patients exhibiting a greater percentage of PD-1 in CD4 T cells showed accelerated functional decline. Serum analyses identified four elevated soluble checkpoints in ALS versus both HCs and Mimics (sPD-1/sBTLA/sCTLA-4/sCD27, < 0.05), with sCD28/TIM-3 showing higher in ALS than in Mimics, and sGITR/sCD137/sIDO/sCD80/sLAG3/sPD-L2 elevating in ALS compared to HCs. Soluble TIM-3 correlated inversely with ALSFRS-R, while sPD-L1 demonstrated dual associations: negative with ALSFRS-R and positive with NFL (all < 0.05).
Our research demonstrated a considerable increase in membrane-bound and soluble PD-1 in ALS patients, correlating with disease progression and worse prognosis. Furthermore, we explored 13 other immune checkpoint molecules. Collectively, these molecules may be implicated in peripheral immune mechanisms underlying ALS pathogenesis. While baseline PD-1 levels show some association with prognosis, their elevation potentially indicates an unfavorable course.
肌萎缩侧索硬化症（ALS）迫切需要强大的生物标志物用于早期诊断和预后分层。本研究旨在探讨膜结合型和可溶性免疫检查点分子在ALS发病机制中的诊断和预后潜力。
在福建医科大学附属协和医院进行的本研究中，72名参与者（46例ALS患者和26名健康对照[HC]）接受了CD4 T细胞及其亚群中PD-1表达的流式细胞术分析。使用Luminex技术对第二个队列（n = 93，44例ALS患者、30名HC和19例ALS模拟患者[模拟患者]）的14种血清免疫检查点分子进行了评估。使用单分子阵列筛选血清中的神经丝轻链（NFL）。
流式细胞术显示CD4 T细胞中PD1表达升高，特别是在Th9和Th17亚群中（P < 0.05）。CD4 T细胞中PD-1百分比更高的ALS患者功能衰退加速。血清分析确定，与HC和模拟患者相比，ALS患者有4种可溶性检查点升高（sPD-1/sBTLA/sCTLA-4/sCD27，P < 0.05），sCD28/TIM-3在ALS患者中高于模拟患者，与HC相比，sGITR/sCD137/sIDO/sCD80/sLAG3/sPD-L2在ALS患者中升高。可溶性TIM-3与ALSFRS-R呈负相关，而sPD-L1表现出双重关联：与ALSFRS-R呈负相关，与NFL呈正相关（均P < 0.05）。
我们的研究表明，ALS患者中膜结合型和可溶性PD-1显著增加，与疾病进展和更差的预后相关。此外，我们还探索了其他13种免疫检查点分子。总体而言，这些分子可能参与了ALS发病机制的外周免疫机制。虽然基线PD-1水平与预后有一定关联，但它们的升高可能预示着病情不利。
