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. 2024 May 30;25(6):1073–1082. doi: 10.1038/s41590-024-01833-w

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© The Author(s) 2024, corrected publication 2024

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Extended Data Fig. 8 — a, Representative gating strategy of splenic B cells sorted for sequencing of BCRs from hD3-3/J_H6 mice six weeks after immunization with 10E8-GT12 24mer. b, Percentage of 10E8-GT-binding cells (10E8-GT9-KO⁺⁺, 10E8-GT10.1⁺⁺ or 10E8-GT12⁺⁺⁻) among IgM⁻IgD⁻ B cells, 42 days after immunization of hD3-3/J_H6 mice with 10E8-GT9-KO 12mer (control, n = 3), 10E8-GT12 12mer (n = 12), 10E8-GT12 12mer (n = 5) or 10E8-GT12 24mer (n = 12) delivered as protein in SMNP, respectively, or 10E8-GT12 24mer delivered by mRNA (n = 11). Each symbol indicates an animal, lines indicate median values. c, Percentage of 10E8-class HCDR3s among all IgM⁻IgD⁻ B cells after immunization as in b. d, Enrichment ratio for HCDR3 amino acid (aa) length distribution for epitope-specific (10E8-GT10.1⁺⁺10E8-GT10.1-KO⁻ or 10E8-GT12⁺⁺10E8-GT12-KO⁻) IgG⁺ B cells from animals immunized with the indicated 10E8-GT immunogens, relative to HCDR3 amino acid length distribution for epitope-specific (10E8-GT9⁻KO⁺⁺10E8-GT9⁻) IgG⁺ B cells from animals immunized with 10E8-GT9-KO 12mer as in b. HCDR3 lengths longer than 22 were only found in the 10E8-GT-immunized groups, precluding calculation of enrichment scores for longer HCDR3s. e, Frequency of long (>=20 aa) HCDR3s among epitope-specific IgG⁺ B cells as in d. f, Percentage of HCDR3s containing the YxFW motif among epitope-specific IgG⁺ B cells as in c. *p < 0.05, Kruskal-Wallis test with Dunn’s multiple comparison correction. P value (control vs. GT10 12mer): 0.02, p value (control vs. GT12 24mer): 0.01. g, Fold-change in SPR-measured K_D for 10E8-GT immunogens binding to 10E8-iGL3 upon addition of the indicated HCDR3 mutations. h, Percentage of epitope-specific (10E8-GT9-KO⁺⁺10E8-GT9⁻,10E8-GT10.1⁺⁺10E8-GT10.1-KO⁻, or 10E8-GT12⁺⁺10E8-GT12-KO⁻) with 10E8⁻class or LN01-class HCDR3s among IgG⁺ BCRs from day 42 after immunization of hD3⁻3/J_H6 as in b. Symbols represent individual animals; bars indicate median values.