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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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Extended Data Fig. 10 — (a) Representative multi-color immunofluorescence image of spleen sections. Overlayed markers are shown as CD19 (red), CD4 (green), PD-1 (violet), and DAPI (blue) staining performed simultaneously in a single experiment for spleen sections from a HC and a pRD patient. Red arrow: T cell region, light blue arrow: GC light zone, dark blue arrow: GC dark zone. Magnification: 20x. (b) PD-1 expression. Magnification: 60x. (c) PD-1 expression. Magnification: 60x, yellow arrow depicts Tfh cells (CD4⁺PD-1⁺). (d) Gating for naive, CM, EM TEMRA T_H cell subsets. (e) Enumeration of T cell subsets. Data were obtained from HC-N (n = 1), HC-Ags (n = 10), pRD-N (n = 1) and pRD-Ags (n = 8) subjects. (f) Productive Simpson clonality of cTfh TCR-Vβ repertoires. (g) CDR3 length distribution in cTfh TCR-Vβ repertoires. Means of HCs and pRD patients are depicted as a black and red line, respectively. Individual values are shown for the patients. nt – nucleotides. (h) CDR3 length of cTfh TCR-Vβ repertoires. (i) Differential usage of V genes in cTfh TCR-Vβ repertoires. Relative frequency of TCR-Vβ family usages is shown. (j) Productive Simpson clonality of Treg TCR-Vβ repertoires. (k) CDR3 length distribution in Treg TCR-Vβ repertoires. Means of HCs and pRD patients are depicted as a black and red line, respectively. Individual values are shown for the patients. nt – nucleotides. (l) CDR3 length of Treg TCR-Vβ repertoires. (m) Differential usage of V genes in Treg TCR-Vβ repertoires. Relative frequency of TCR-Vβ family usages is shown. All TCR-Vβ repertoire data were obtained on HC-Ag (n = 7) and pRD-Ag (n = 6) subjects. (n) Gating strategy for sorted CD38^int B and cTfh cells. Gating and sort purity of batch sorted CD38^int B cells and cTfh cells for co-culture assay shown on a representative HC-Ag subject. Four independent experiments were performed with one HC-Ag (n = 4) and one pRD-Ag (n = 4) subjects in each. Data for e-f h-j and l-m are shown as mean±s.e.m. with individual values. Statistical analysis was performed using two-sided unpaired Student’s t-test for panel f-h and j-l, or Mann–Whitney U test with Holm-Šídák multiple comparison for panel e, i and m.

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