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. 2024 Feb 21;627(8003):407–415. doi: 10.1038/s41586-024-07079-8

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© The Author(s) 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 — Wild-type, B cell-deficient (Mb1-Cre^KI/KI), Aqp4^–/–, and Aqp4^ΔB mice were immunized with full-length AQP4 or human recombinant full-length MOG protein and tested for germinal centre (GC) responses on d12 after immunization. (a) Representative immunostainings of Bcl-6 in spleens of full-length AQP4-immunized mice (n = 2 independent experiments, scale bar 100 µm). (b) Representative cytograms and gating strategy of splenic Tfh cells (live CD19^–CD3⁺CD4⁺Bcl-6⁺PD-1⁺) and GC B cells (live CD19⁺CD3^–B220⁺Bcl-6⁺CD95⁺). (c) Flow cytometric quantification of splenic Tfh cell and GC B cell frequencies in spleens of AQP4-immunized wild-type, B cell-deficient (Mb1-Cre^KI/KI), Aqp4^–/–, and Aqp4^ΔB mice. (d, e) The AQP4-specific serum response was tested with serial dilutions in a cell-based assay with sera isolated prior to (d-1) and on d21 after immunization with full-length AQP4. (d) Representative histograms of the anti-AQP4-serum response tested in LN18^AQP4 cells at d-1 (grey histograms) and d21 (blue histograms). (e) Quantification of the anti-AQP4-serum response for Aqp4^–/– (n = 2, upper panel) and Aqp4^ΔB (n = 3, lower panel) alongside wild-type (n = 4) and Mb1-Cre^KI/KI (n = 2, depicted in both panels for reference), tested in serial dilutions as indicated on the x-axis. (f) Quantification of Tfh cell and GC B cell frequencies in secondary lymphoid tissues of MOG protein-immunized wild-type (n = 3), B cell-deficient Mb1-Cre^KI/KI (n = 2), Aqp4^–/– (n = 3), and Aqp4^ΔB (n = 3) mice. (g) The MOG-specific serum response was tested with serial dilutions in a cell-based assay with sera isolated on d-1 and d21 after immunization with full-length MOG protein. Quantification of the anti-MOG-serum response tested in serial dilutions as indicated on the x-axis. Data in (c, f) are shown as mean ± SD and tested by one-way ANOVA with Tukey’s post-test. Data in (e, g) are shown as delta mean fluorescence intensity (ΔMFI = MFI_d21 – MFI_d-1) ± SD and tested by two-way ANOVA with Sidak’s post-test. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. ns = not significant. Symbols indicate biological replicates.

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