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. 2022 Jul 13;12:11876. doi: 10.1038/s41598-022-15293-5

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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 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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Number, phenotype and spatial distribution of synovial B cells in Rheumatoid Arthritis. (A) The flow cytometry panels show all CD3-CD14-AnnexinV-CD19+ single B cells from synovial biopsies of 4 ACPA+ and 4 ACPA− RA patients at the time of diagnosis in pseudocolor plots, where red color indicates data point overlap. A5 was taken from a metacarpophalangeal (MCP) 4 joint, A6 and A7 were taken from wrists. Remaining synovial biopsies are arthroscopic biopsies from knees. (B) 5 out of 8 biopsies exceeded a minimum of 382 B cells suitable for phenotypic characterization and scRNAseq analysis. (C) Memory B cells made up the largest population of synovial B cells (24–63%), while naïve cells were a bit fewer (20–45%). Double negative B cells had a variable range (8–29%), while a minority was CD27++ (1–4%) or of the unswitched IgD+ CD27+ memory phenotype (2–4%). (D) H&E-staining shows the tissue architecture of A1–A4. Immunohistochemistry staining with anti-CD19 and anti-CD20 antibody cocktail confirms the presence of B cells. ACPA+ samples are indicated by turquoise color, and ACPA− in orange.