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. 2022 Jul 28;23(8):1256–1272. doi: 10.1038/s41590-022-01271-6

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 2 — a, Detection of CD19^hiCD21^lo B cells. b,c, Gating for CD19^hiCD21^lo B cells in a representative HC-Ag and patient with pRD-Ag. Fraction of CD19^hiCD21^lo B cells in total (b) and CD38^int, CD38⁻ and CD27⁺ B cells (c) in HC-Ags (n = 14) and patients with pRD-Ag (n = 6). d, Detection of ABCs. Gating for ABCs in a representative HC-Ag and patient with pRD-Ag. e,f, Fraction of ABCs in total (e) and CD38^int, CD38⁻ and CD27⁺ B cells (f) in HC-Ags (n = 14) and patients with pRD-Ag (n = 6). g, Distribution of ABCs in conventional and CD19^hiCD21^lo B cells. Contour plots show total B cells (gray), gate depicts CD19^highCD21^low B cells and ABCs are overlayed (green) in a representative HC-Ag and patient with pRD-Ag. h, Correlation between CD19^hiCD21^lo B cells and ABCs. Linear regression line is shown with 95% confidence intervals, Pearson r and P values are shown. Data were obtained from HC-Ags (gray, n = 14) and patients with pRD-Ag (red, n = 6). i, T-bet expressions in total B cells are shown as gMFIs. j,k, Detection (j) and fraction (k) of CD21⁺CD11c⁻, CD21^loCD11c⁻ and CD21^loCD11c⁺ B cell populations. Gating is shown in a representative HC-Ag and patient with pRD-Ag. l,m, T-bet expression in CD21⁺CD11c⁻, CD21^loCD11c⁻ and CD21^loCD11c⁺ B cells in a representative HC-Ag and patient with pRD-Ag (l) and shown as gMFIs (m). n–p, Expressions of FcRL4, FcRL5, CD85j and CXCR3 (n), CD95 (o) and BAFF-R (p) in total B cells are shown as gMFIs. Data for i–p were obtained from HC-Ags (n = 7–14) and patients with pRD-Ag (n = 4–5). Data for b,c,e,f,k–o are shown as mean ± s.e.m. with individual values depicted. Statistical analyses were performed using two-sided unpaired Student’s t-test for data on b,c,e,f,I–o or Mann–Whitney U-test with multiple comparison (Holm–Šídák method) for k. q, Overlayed markers (CD19, CD4, T-bet and 4,6-diamidino-2-phenylindole (DAPI)) in a HC-Ags and patients with pRD-Ag. Magnification ×10. Follicular (Fo) and extrafollicular (EF) areas are indicated that were used for higher magnifications. r, T-bet expression by Fo and EF areas. Magnification ×60. s, T-bet expression in B cells by Fo and EF areas. Arrow shows T-bet⁺ B cells. Magnification ×60.

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