Li Enhao, Li Zekun, Wang Jinzeng, Wu Haoyang, Xue Yilun, Lou Can, Chen Zhenping, Liu Feng, Wu Wenman, Ding Qiulan, Wu Runhui, Wang Xuefeng, Dai Jing
Department of Laboratory Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
College of Health Science and Technology, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
Blood Adv. 2025 Aug 12;9(15):3967-3978. doi: 10.1182/bloodadvances.2025015799.
Inhibitors are the most severe complication of replacement therapy in patients with hemophilia. Previous studies, along with our clinical observations, have identified distinct incidence rates and clinical manifestations of factor VIII (FVIII) and FIX inhibitors in patients with severe hemophilia A (HA) and HB. To explore different immune responses to FVIII and FIX in patients with HA and HB and elucidate the mechanisms underlying the varying clinical manifestations of these patients, we performed single-cell sequencing on peripheral blood mononuclear cells (PBMCs) collected from 5 patients with HA and 5 with HB with inhibitors. After quality control, a total of 75 051 cells were clustered into 19 subsets. Transcriptome analysis revealed differences in the composition of lymphocyte subsets and the functional status of immune cells between the HA and HB groups. Additionally, immune repertoire analysis indicated variations in the diversity of B- and T-cell clones between the 2 groups. HA group exhibited a relatively higher proportion of B cells and more active B cells, whereas HB group demonstrated a higher proportion of T cells, with more active CD4+ T helper cells. Our study provides insights into the distinct biological processes underlying the distinct immune responses to therapeutic FVIII and FIX in patients with HA and HB, as revealed through single-cell sequencing of PBMCs from patients with hemophilia with inhibitors. The data generated will serve as a valuable resource for future research on how the immune system recognizes and initiates responses to antigens with varying molecular characteristics.
抑制剂是血友病患者替代治疗最严重的并发症。既往研究以及我们的临床观察已经明确了重度甲型血友病（HA）和乙型血友病（HB）患者中因子VIII（FVIII）和FIX抑制剂的不同发病率及临床表现。为探究HA和HB患者对FVIII和FIX的不同免疫反应，并阐明这些患者临床表现各异的潜在机制，我们对5例患有抑制剂的HA患者和5例患有抑制剂的HB患者采集的外周血单个核细胞（PBMC）进行了单细胞测序。经过质量控制，共75051个细胞被聚类为19个亚群。转录组分析揭示了HA组和HB组之间淋巴细胞亚群组成及免疫细胞功能状态的差异。此外，免疫组库分析表明两组之间B细胞和T细胞克隆的多样性存在差异。HA组B细胞比例相对较高且B细胞更活跃，而HB组T细胞比例较高，CD4+辅助性T细胞更活跃。我们的研究通过对患有抑制剂的血友病患者的PBMC进行单细胞测序，深入了解了HA和HB患者对治疗性FVIII和FIX产生不同免疫反应背后的独特生物学过程。所产生的数据将为未来关于免疫系统如何识别并启动对具有不同分子特征抗原的反应的研究提供宝贵资源。
