Defects in B-lymphopoiesis and B-cell maturation underlie prolonged B-cell depletion in ANCA-associated vasculitis
Jens Thiel, Franziska 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 Voll, Jan Stuchlý, Marina Bakardjieva, Tomáš Kalina, Cristian R. Smulski, Nils Venhoff, Marta Rizzi
Abstract
<h3>Objectives</h3> 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. <h3>Methods</h3> 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 <i>in vitro</i> modelling and the role of BAFF receptor by quantitative PCR, western blot analysis and <i>in vitro</i> assays. <h3>Results</h3> 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. <i>In vitro</i> 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 <i>in vitro</i>. <h3>Conclusions</h3> 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.