Department of Neurology, Odense University Hospital, Odense, Denmark.
Department of Clinical Research, University of Southern Denmark, Odense, Denmark.
Ann Clin Transl Neurol. 2024 Oct;11(10):2657-2672. doi: 10.1002/acn3.52182. Epub 2024 Sep 15.
Anti-CD20 therapy is a highly effective treatment for multiple sclerosis (MS). In this study, we investigated MS-related changes in peripheral blood mononuclear cell (PBMC) subsets compared to healthy controls and longitudinal changes related to the treatment.
Multicolor spectral flow cytometry analysis was performed on 78 samples to characterize disease- and treatment-related PBMC clusters. Blood samples from MS patients were collected at baseline and up to 8 months post-treatment, with three collection points after treatment initiation. Unsupervised clustering tools and manual gating were applied to identify subclusters of interest and quantify changes.
B cells were depleted from the periphery after anti-CD20 treatment as expected, and we observed an isolated acute, transitory drop in the proportion of natural killer (NK) and NKT cells among the main populations of PBMC (P = 0.03, P = 0.004). Major affected PBMC subpopulations were cytotoxic immune cells (NK, NKT, and CD8 T cells), and we observed a higher proportion of cytotoxic cells with reduced brain-homing ability and a higher regulatory function as a long-term anti-CD20-related effect. Additionally, anti-CD20 therapy altered distributions of memory CD8 T cells and reduced exhaustion markers in both CD4 and CD8 T cells.
The findings of this study elucidate phenotypic clusters of NK and CD8 T cells, which have previously been underexplored in the context of anti-CD20 therapy. Phenotypic modifications towards a more regulatory and controlled phenotype suggest that these subpopulations may play a critical and previously unrecognized role in mediating the therapeutic efficacy of anti-CD20 treatments.
抗 CD20 治疗是治疗多发性硬化症（MS）的一种非常有效的方法。在这项研究中，我们研究了与 MS 相关的外周血单个核细胞（PBMC）亚群的变化，与健康对照组进行了比较，并对与治疗相关的纵向变化进行了研究。
对 78 个样本进行多色光谱流式细胞术分析，以描述疾病和治疗相关的 PBMC 簇。在治疗后，从 MS 患者采集血液样本，直至 8 个月，在治疗开始后的三个采集点。应用无监督聚类工具和手动门控技术来识别感兴趣的亚群并量化变化。
抗 CD20 治疗后，外周 B 细胞被耗尽，我们观察到 NK 和 NKT 细胞在外周血单个核细胞的主要群体中的比例急性、一过性下降（P=0.03，P=0.004）。受影响的主要 PBMC 亚群是细胞毒性免疫细胞（NK、NKT 和 CD8 T 细胞），我们观察到具有降低的脑归巢能力和更高的调节功能的细胞毒性细胞比例更高，这是长期抗 CD20 相关的效应。此外，抗 CD20 治疗改变了记忆 CD8 T 细胞的分布，并减少了 CD4 和 CD8 T 细胞中的耗竭标志物。
这项研究的结果阐明了 NK 和 CD8 T 细胞的表型簇，这些细胞在抗 CD20 治疗背景下以前尚未得到充分探索。表型向更具调节性和控制性的表型的改变表明，这些亚群可能在介导抗 CD20 治疗的疗效方面发挥着关键的、以前未被认识到的作用。
