Jens Thiel, Franziska M Schmidt, Raquel Lorenzetti, Arianna Troilo, Iga Janowska, Lena Nießen, Sophie Pfeiffer, Julian Staniek, Bruno Benassini, Marei-Theresa Bott, Jakov Korzhenevich, Lukas Konstantinidis, Frank Burgbacher, Ann-Katrin Dufner, Natalie Frede, Reinhard E Voll, Jan Stuchly, Marina Bakardjieva, Tomas Kalina, Cristian Roberto Smulski, Nils Venhoff, Marta Rizzi
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引用次数: 0
Abstract
Objectives: B-cell depletion time after rituximab (RTX) treatment is prolonged in antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV) compared with other autoimmune diseases. We investigated central and peripheral B-cell development to identify the causes for the defect in B-cell reconstitution after RTX therapy.
Methods: We recruited 91 patients with AAV and performed deep phenotyping of the peripheral and bone marrow B-cell compartment by spectral flow and mass cytometry. B-cell development was studied by in vitro modelling and the role of BAFF receptor by quantitative PCR, western blot analysis and in vitro assays.
Results: Treatment-naïve patients with AAV showed low transitional B-cell numbers, suggesting impaired B-lymphopoiesis. We analysed bone marrow of treatment-naïve and RTX-treated patients with AAV and found reduced B-lymphoid precursors. In vitro modelling of B-lymphopoiesis from AAV haematopoietic stem cells showed intact, but slower and reduced immature B-cell development. In a subgroup of patients, after RTX treatment, the presence of transitional B cells did not translate in replenishment of naïve B cells, suggesting an impairment in peripheral B-cell maturation. We found low BAFF-receptor expression on B cells of RTX-treated patients with AAV, resulting in reduced survival in response to BAFF in vitro.
Conclusions: Prolonged depletion of B cells in patients with AAV after RTX therapy indicates a B-cell defect that is unmasked by RTX treatment. Our data indicate that impaired bone marrow B-lymphopoiesis results in a delayed recovery of peripheral B cells that may be further aggravated by a survival defect of B cells. Our findings contribute to the understanding of AAV pathogenesis and may have clinical implications regarding RTX retreatment schedules and immunomonitoring after RTX therapy.
研究目的与其他自身免疫性疾病相比,抗中性粒细胞胞浆抗体(ANCA)相关性血管炎(AAV)患者接受利妥昔单抗(RTX)治疗后的B细胞耗竭时间延长。我们研究了中枢和外周 B 细胞的发育情况,以找出 RTX 治疗后 B 细胞重建缺陷的原因:我们招募了 91 名 AAV 患者,通过光谱流式细胞仪和质谱细胞仪对外周和骨髓 B 细胞区进行了深入的表型分析。通过体外建模研究了B细胞的发育,并通过定量PCR、Western印迹分析和体外试验研究了BAFF受体的作用:结果:未经治疗的 AAV 患者显示过渡性 B 细胞数量较少,表明 B 淋巴细胞生成受损。我们分析了未经治疗和经 RTX 治疗的 AAV 患者的骨髓,发现 B 淋巴细胞前体减少。用 AAV 造血干细胞制作的 B 淋巴细胞生成体外模型显示,未成熟 B 细胞的发育完好无损,但速度较慢且数量减少。在一个亚组患者中,经过RTX治疗后,过渡性B细胞的存在并没有转化为幼稚B细胞的补充,这表明外周B细胞成熟受到了损害。我们发现,经AAV治疗的RTX患者的B细胞上BAFF受体表达较低,导致体外BAFF反应存活率降低:结论:RTX 治疗后,AAV 患者 B 细胞的长期耗竭表明,RTX 治疗可消除 B 细胞缺陷。我们的数据表明,骨髓B淋巴细胞生成受损导致外周B细胞恢复延迟,而B细胞的存活缺陷可能会进一步加剧这种延迟。我们的研究结果有助于人们了解 AAV 的发病机制,并可能对 RTX 治疗后的 RTX 再治疗计划和免疫监测产生临床影响。
期刊介绍:
Annals of the Rheumatic Diseases (ARD) is an international peer-reviewed journal covering all aspects of rheumatology, which includes the full spectrum of musculoskeletal conditions, arthritic disease, and connective tissue disorders. ARD publishes basic, clinical, and translational scientific research, including the most important recommendations for the management of various conditions.