Rheumatology Discipline, School of Medical Sciences, University of Campinas (UNICAMP), Campinas, Brazil.
Hemocentro UNICAMP, School of Medical Sciences, University of Campinas (UNICAMP), Campinas, Brazil.
J Cell Mol Med. 2022 Oct;26(19):5044-5053. doi: 10.1111/jcmm.17531. Epub 2022 Sep 2.
In patients with ANCA-associated vasculitis, interactions between neutrophils and endothelial cells cause endothelial damage and imbalance. Endothelial colony-forming cells (ECFCs) represent a cellular population of the endothelial lineage with proliferative capacity and vasoreparative properties. This study aimed to evaluate the angiogenic capacity of ECFCs of patients with granulomatosis with polyangiitis (GPA). The ECFCs of 13 patients with PR3-positive GPA and 14 healthy controls were isolated and characterized using fluorescence-activated cell sorting, capillary tube formation measurement, scratching assays and migration assays with and without plasma stimulation. Furthermore, three patients with active disease underwent post-treatment recollection of ECFCs for longitudinal evaluation. The ECFCs from the patients and controls showed similar capillary structure formation. However, the ECFCs from the patients with inactive GPA exhibited early losses of angiogenic capacity. Impairments in the migration capacities of the ECFCs were also observed in patients with GPA and controls (12th h, p = 0.05). Incubation of ECFCs from patients with GPA in remission with plasma from healthy controls significantly decreased migration capacity (p = 0.0001). Longitudinal analysis revealed that treatment significantly lowered ECFC migration rates. This study revealed that ECFCs from the patients with PR3-positive GPA in remission demonstrated early losses of tube formation and reduced migration capacity compared to those of the healthy controls, suggesting impairment of endothelial function.
在抗中性粒细胞胞浆抗体相关性血管炎患者中，中性粒细胞与内皮细胞之间的相互作用导致内皮损伤和失衡。内皮祖细胞（endothelial colony-forming cells，ECFCs）是内皮谱系中具有增殖能力和血管修复特性的细胞群体。本研究旨在评估肉芽肿性多血管炎（granulomatosis with polyangiitis，GPA）患者的 ECFC 血管生成能力。采用荧光激活细胞分选、毛细血管管腔形成测量、划痕实验和迁移实验（有或无血浆刺激），分离并鉴定了 13 名 PR3 阳性 GPA 患者和 14 名健康对照者的 ECFCs。此外，对 3 名处于活动期疾病的患者进行了治疗后 ECFC 采集，进行纵向评估。患者和对照组的 ECFCs 均表现出相似的毛细血管结构形成。然而，处于缓解期的 GPA 患者的 ECFCs 早期表现出血管生成能力丧失。GPA 患者和对照组的 ECFC 迁移能力也出现损伤（第 12 小时，p=0.05）。将处于缓解期的 GPA 患者的 ECFCs 与健康对照者的血浆孵育，显著降低了迁移能力（p=0.0001）。纵向分析显示，治疗显著降低了 ECFC 的迁移率。本研究显示，与健康对照组相比，处于缓解期的 PR3 阳性 GPA 患者的 ECFCs 在管腔形成和迁移能力方面表现出早期丧失，表明内皮功能受损。
