Common variable immunodeficiency (CVID), characterized by recurrent infections, low serum class-switched immunoglobulin isotypes, and poor antigen-specific antibody responses, comprises a heterogeneous patient population in terms of clinical presentation and underlying etiology. The diagnosis is regularly associated with a severe decrease of germinal center (GC)-derived B-cell populations in peripheral blood. However, data from B-cell differentiation within GC is limited. We present a multiplex approach combining histology, flow cytometry, and B-cell receptor repertoire analysis of sorted GC B-cell populations allowing the modeling of distinct disturbances in GCs of three CVID patients. Our results reflect pathophysiological heterogeneity underlying the reduced circulating pool of post-GC memory B cells and plasmablasts in the three patients. In patient 1, quantitative and qualitative B-cell development in GCs is relatively normal. In patient 2, irregularly shaped GCs are associated with reduced somatic hypermutation (SHM), antigen selection, and class-switching, while in patient 3, high SHM, impaired antigen selection, and class-switching with large single clones imply increased re-cycling of cells within the irregularly shaped GCs. In the lymph nodes of patients 2 and 3, only limited numbers of memory B cells and plasma cells are formed. While reduced numbers of circulating post GC B cells are a general phenomenon in CVID, the integrated approach exemplified distinct defects during GC maturation ranging from near normal morphology and function to severe disturbances with different facets of impaired maturation of memory B cells and/or plasma cells. Integrated dissection of disturbed GC B-cell maturation by histology, flow cytometry, and BCR repertoire analysis contributes to unraveling defects in the essential steps during memory formation.
Bibliographical noteFunding Information:
This study was supported by the Bundesministerium für Bildung und Forschung (BMBF), grant number BMBF 01EO1303 and GAIN 01GM1910A to KW, by the Stichting Sophia Kinderziekenhuis Fonds (grant no. S15-07 Genes and Immunity in SCID) (FK and MB.), and by the Dutch Organization for Scientific Research (NWO/ZonMw veni grant 91616058 to PS) and an EFIS-IL-Short-Term Fellowship to PS.
Copyright © 2022 van Schouwenburg, Unger, Payne, Kaiser, Pico-Knijnenburg, Pfeiffer, Hausmann, Friedmann, Erbel, Seidl, van Zessen, Stubbs, van der Burg and Warnatz.