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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. 7 — a, In vitro CD38^int B cell differentiation. Sorted CD38^int B cells were activated with CpG, CD40L and IL-21. Differentiated cells (CD27⁺CD38^lo and CD27⁺CD38^hi) were quantified on day 7. Experimental design is shown and contour plots are representative of an individual with HC-Ag and a patient with pRD-Ag from six independent experiment. b, Enumeration of effector cells. The proportion of CD27⁺CD38^lo and CD27⁺CD38^hi from HC-Ags (n = 6), pRD-N (P1) (n = 1) and patients with pRD-Ag (n = 5) are shown as percentage of total live B cells in the culture. c, Experimental design for testing B cell activation by plasma. Sorted CD38^int HC B cells were cultured in vitro in the presence of IL-21 and plasma from either a patient with pRD-Ag (n = 5) or independent HC-Ag (n = 5) and analyzed on day 2 for CD25 and CD86 expression. d, Activation of sorted CD38^int B cells with plasma. Surface expression of CD25 and CD86 are shown as overlaid histogram plots depicting representative samples of HC-Ag plasma-stimulated, patient with pRD-Ag plasma-stimulated and unstimulated HC. Dashed line indicated marker expression level on day 0. Expression levels are indicated as gMFIs. e, Enumeration of B cell activation. Graphs show expression of CD25 and CD86 at day 0 and 2 days of stimulation with HC-Ags (n = 5) or patients with pRD-Ag (n = 6) plasma at the indicated plasma concentration in culture medium. Data for d,e were obtained from a single experiment performed on all plasma samples simultaneously. f, Plasma endotoxin levels. HC-Ags (n = 13), pRD-N (P1) (n = 1) and pRD-Ags (n = 10). g,h, Plasma virus levels. Epstein–Barr nuclear antigen and CMV titers were detected in the plasma of HC-Ags (n = 15), pRD-N (P1) (n = 1) and pRD-Ags (n = 12). i, Plasma soluble IL-2R levels. HC-Ags (n = 11), pRD-N (P1) (n = 1) and pRD-Ags (n = 10). j, Plasma BAFF levels. HC-Ags (n = 20), pRD-N (P1) (n = 1) and pRD-Ags (n = 14). k,l, Plasma IFN-γ and IL-10. HC-Ags (n = 11), pRD-N (P1) (n = 1) and pRD-Ags (n = 14). Data are shown as mean ± s.e.m. with individual values depicted on each graph. For statistical analysis Mann–Whitney U-test was performed for b; and two-sided unpaired Student’s t-test was used for e–l to compare patients with pRD-Ag to individuals with HC-Ag.

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