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. 2023 Jan 19;41(9):1229–1238. doi: 10.1038/s41587-022-01637-z

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© The Author(s) 2023, 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. 7 — a) Splenocytes were harvested from select MuSK-immunized mice, and purified B-cells were evaluated using a MuSK-specific and total IgG B-cell ELISpot assay to quantitate the MuSK-specific B-cell frequency. Dots from duplicated wells were summarized after normalization with seeded cells (dots per 100,000 cells). MuSK-specific B-cell frequency is calculated by dividing anti-MuSK B-cells by total IgG⁺ B-cells. b) IgG subclasses of anti-MuSK antibodies were detected in sera from MuSK-immunized mice in reference to sera from negative controls (non-immunized C57BL/6 and Rag2IL2rγ-deficient mice) and median value was plotted. c) Epitope mapping of serum from a mouse immunized and boosted with MuSK Ig1-Ig2, or MuSK Ig1-Ig2 followed by MuSK full length (FL) boost, using Jurkat CAAR T-cells expressing individual MuSK domain CAARs linked to GFP. Representative FACS plots are shown from three independent experiments. d) Schematic of pMSGV1-1D3-CAR (mouse CD8α signal peptide, 1D3 anti-mouse CD19 single chain variable fragment (scFv), mouse CD28 hinge (HD)/transmembrane domain (TMD)/costimulatory domain, and a mouse CD3ζ.1-3mut domain to confer enhanced persistence. Schematic of pMSGV1-MuSK-CAAR (human MuSK extracellular domains (amino acids 24-495), glycine-serine (GS) linker, hCD8α TMD, and human CD137-CD3ζ). e, f) Transduction efficiency of 1D3-CAR and MuSK-CAAR in primary mouse T-cells (Set 1 and Set 2) and g) percentage of CD4⁺/CD8⁺ T-cells in Set 1 was evaluated on day 4 after T-cell activation (one day prior to injection).